# Patent

## Title
Stress-regulated genes of plants, transgenic plants containing same, and methods of use

## Abstract

Clusters of plant genes that are regulated in response to one or more stress conditions are provided, as are isolated plant stress-regulated genes, including portions thereof comprising a coding sequence or a regulatory element, and to consensus sequences comprising a plant stress-regulated regulatory element. In addition, a recombinant polynucleotide, which includes a plant stress-regulated gene, or functional portion thereof, operatively linked to a heterologous nucleotide sequence, is provided, as are transgenic plants, which contain a plant stress-regulated gene or functional portion thereof that was introduced into a progenitor cell of the plant. Also provided are methods of using a plant stress-regulated gene to confer upon a plant a selective advantage to a stress condition, methods of identifying an agent that modulates the activity of a plant stress-regulated regulatory element, and methods of determining whether a plant has been exposed to a stress.

## Background
This application claims the benefit under 35 U.S.C. 119(e) of U.S. Ser. No. 60/227,866, filed Aug. 24, 2000; U.S. Ser. No. 60/264,647, filed Jan. 26, 2001; and U.S. Ser. No. 60/300,111, filed Jun. 22, 2001, each of which is incorporated herein by reference in its entirety.


Three CD-R compact discs, labeled “Copy 1”, “Copy 2”, and “CRF” and having the files listed below, are submitted herewith and are incorporated herein by reference. Copy 1 and Copy 2 each contain two text documents: 1) a file named SCRIP1300-3_SEQUENCE_LISTING, which contains the Sequence Listing, was created on Aug. 20, 2001 (and recorded on the CD-R on Aug. 21, 2001), and is 9,972 KB in size; and 2) a file named SCRIP1300-3_Table—32, which contains Table 32, was created on Aug. 20, 2001 (and recorded on the CD-R on Aug. 21, 2001), and is 1,251 KB in size. The CRF contains a single file named SCRIP1300-3_SEQUENCE_LISTING, which contains the Sequence Listing, was created on Aug. 20, 2001 (and recorded on the CD-R on Aug. 21, 2001), is 9,972 KB in size, and is identical to the files having the same name on Copy 1 and Copy 2.
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates generally to plant genes, the expression of which are regulated in response to stress, and more specifically to the gene regulatory elements involved in a stress-induced response in plants, to uses of the coding sequences and regulatory elements of such plant stress-regulated genes, and to transgenic plants genetically modified to express such a coding sequence or to express a heterologous polynucleotide from such a regulatory element.
2. Background Information
Microarray technology is a powerful tool that can be used to identify the presence and level of expression of a large number of polynucleotides in a single assay. A microarray is formed by linking a large number of discrete polynucleotide sequences, for example, a population of polynucleotides representative of a genome of an organism, to a solid support such as a microchip, glass slide, or the like, in a defined pattern. By contacting the microarray with a nucleic acid sample obtained from a cell of interest, and detecting those polynucleotides expressed in the cell can hybridize specifically to complementary sequences on the chip, the pattern formed by the hybridizing polynucleotides allows the identification of clusters of genes that are expressed in the cell. Furthermore, where each polynucleotide linked to the solid support is known, the identity of the hybridizing sequences from the nucleic acid sample can be identified.
A strength of microarray technology is that it allows the identification of differential gene expression simply by comparing patterns of hybridization. For example, by comparing the hybridization pattern of nucleic acid molecules obtained from cells of an individual suffering from a disease with the nucleic acids obtained from the corresponding cells of a healthy individual, genes that are differentially expressed can be identified. The identification of such differentially expressed genes provides a means to identify new genes, and can provide insight as to the etiology of a disease.
Microarray technology has been widely used to identify patterns of gene expression associated with particular stages of development or of disease conditions in animal model systems, and is being applied to the identification of specific patterns of gene expression in humans. The recent availability of information for the genomes of plants provides a means to adapt microarray technology to the study of plant gene expression.
Plants and plant products provide the primary sustenance, either directly or indirectly, for all animal life, including humans. For the majority of the world's human population and for many animals, plants and plant products provide the sole source of nutrition. As the world population increases, the best hope to prevent widespread famine is to increase the quantity and improve the quality of food crops, and to make the crops available to the regions of the world most in need of food.
Throughout history, a continual effort has been made to increase the yield and nutritious value of food crops. For centuries, plants having desirable characteristics such as greater resistance to drought conditions or increased size of fruit were crossbred and progeny plants exhibiting the desired characteristics were selected and used to produce seed or cuttings for propagation. Using such classical genetic methods, plants having, for example, greater disease resistance, increased yield, and better flavor have been obtained. The identification of plant genes involved in conferring a selective advantage on the plant to an environmental challenge would facilitate the generation and yield of plants, thereby increasing the available food supply to an increasing world population. The involvement of these genes in a single organism to responses to multiple stress conditions, however, remains unknown. Thus, a need exists to identify plant genes and polynucleotides that are involved in modulating the response of a plant to changing environmental conditions. The present invention satisfies this need and provides additional advantages.
SUMMARY OF THE INVENTION
The present invention relates to clusters of genes that are regulated in response to a stress condition in plants. Such clusters include, for example, plant polynucleotides whose expression is altered in response to two or more different stress conditions; and plant polynucleotides the expression of which are altered in response to one stress condition, but not to others. The identification of such clusters, using microarray technology, has allowed the identification of plant stress-regulated genes in Arabidopsis thaliana (see Tables 1 and 2); and homologs and orthologs thereof in other plant species (see Table 32). Thus, the invention provides isolated polynucleotide portions of Arabidopsis plant stress-regulated genes, and homologs and orthologs thereof; variants of such sequences, and polynucleotides encoding substantially similar plant stress-regulated polypeptides expressed therefrom. Such sequences include, for example, sequences encoding transcription factors; enzymes, including kinases; and structural proteins, including channel proteins (see Tables 29–31). Accordingly, the present invention also relates to an isolated polynucleotide comprising all or a portion of a plant stress-regulated gene, and to polynucleotide portions thereof, including a coding region (open reading frame), which encodes all or a portion of a stress-regulated polypeptide, for example, as set forth in SEQ ID NOS:1–2703; and a regulatory element involved in regulating the response of the plant to a stress condition such exposure to an abnormal level of salt, osmotic pressure, temperature or any combination thereof, for example, as set forth in SEQ ID NOS:2704–5379.
The present invention also relates to a recombinant polynucleotide, which contains a nucleotide sequence of a plant stress-regulated gene or functional portion thereof operatively linked to a heterologous nucleotide sequence. In one embodiment, the recombinant polynucleotide comprises a plant stress-regulated gene regulatory element operatively linked to a heterologous nucleotide sequence, which is not regulated by the regulatory element in a naturally occurring plant. The heterologous nucleotide sequence, when expressed from the regulatory element, can confer a desirable phenotype to a plant cell containing the recombinant polynucleotide. In another embodiment, the recombinant polynucleotide comprises a coding region, or portion thereof, of a plant stress-regulated gene operatively linked to a heterologous promoter. The heterologous promoter provides a means to express an encoded stress-regulated polypeptide constitutively, or in a tissue-specific or phase-specific manner.
Accordingly, in one aspect, the present invention provides an isolated polynucleotide comprising a nucleotide sequence of a plant gene that hybridizes under stringent conditions, preferably high stringency conditions, to any one of SEQ ID NOS:1–5379 (see Tables 1 and 2), including to a coding region (SEQ ID NOS:1–2703) or a regulatory region, which can alter transcription of an operatively linked nucleic acid sequence in response to an abiotic stress (SEQ ID NOS:2704–5379; see Table 2), or to a complement thereof. Additional aspects provide sequences that hybridize under stringent conditions, preferably high stringency conditions, to the complements of SEQ ID NO 1–1261 (cold responsive genes; Tables 3–6), SEQ ID NOS:2227–2427 (saline responsive genes; Tables 7–10), SEQ ID NOS:2428–2585 (osmotic responsive genes; Tables 11–14), SEQ ID NOS:1699–1969 (cold and osmotic responsive genes; Tables 15–17), SEQ ID NOS:1970–2226 (cold and saline responsive genes; Tables 18–20), SEQ ID NOS:2586–2703 (osmotic and saline responsive genes; Tables 21–23), and SEQ ID NOS:1262–1698 (cold, osmotic and saline responsive genes; Tables 24–26), and which can comprise regulatory regions that can alter transcription in response to cold stress, osmotic stress, saline stress, or combinations thereof (SEQ ID NOS:2704–5379; see Table 2). Also provided are nucleotide sequences complementary thereto, and expression cassettes, plants and seeds comprising any of the above isolated sequences.
In another aspect, the present invention provides an isolated polynucleotide comprising a plant nucleotide sequence that hybridizes under stringent conditions, preferably high stringency conditions, to the complement of any one of SEQ ID NOS:1–2703 (Table 1), including to a coding region thereof (SEQ ID NOS:2704–5379), wherein expression of said coding region is altered in response to an abiotic stress. Additional aspects provide sequences that hybridize under high stringency conditions to the complements of SEQ ID NO 1–1261 (cold responsive genes; Tables 3–6), SEQ ID NOS:2227–2427 (saline responsive genes; Tables 7–10), SEQ ID NOS:2428–2585 (osmotic responsive genes; Tables 11–14), SEQ ID NOS:1699–1969 (cold and osmotic responsive genes; Tables 15–17), SEQ ID NOS:1970–2226 (cold and saline responsive genes; Tables 18–20), SEQ ID NOS:2586–2703 (osmotic and saline responsive genes; Tables 21–23), and SEQ ID NOS:1262–1698 (cold, osmotic and saline responsive genes; Tables 24–26), and which can comprise a coding region whose transcription is altered in response to cold stress, osmotic stress, saline stress, or a combination thereof. Also provided are nucleotide sequences complementary thereto, and expression cassettes, plants and seeds comprising any of the above sequences.
The invention further relates to a method of producing a transgenic plant, which comprises at least one plant cell that exhibits altered responsiveness to a stress condition. In one embodiment, the method can be performed by introducing a polynucleotide portion of plant stress-regulated gene into a plant cell genome, whereby the polynucleotide portion of the plant stress-regulated gene modulates a response of the plant cell to a stress condition.
The polynucleotide portion of the plant stress-regulated gene can encode a stress-regulated polypeptide or functional peptide portion thereof (see SEQ ID NOS:1–2703), wherein expression of the stress-regulated polypeptide or functional peptide portion thereof either increases the stress tolerance of the transgenic plant, or decreases the stress tolerance of the transgenic plant. The polynucleotide portion of the plant stress-regulated gene encoding the stress-regulated polypeptide or functional peptide portion thereof can be operatively linked to a heterologous promoter. The polynucleotide portion of the plant stress-regulated gene also can comprise a stress-regulated gene regulatory element (see SEQ ID NOS:2704–5379). The stress-regulated gene regulatory element can integrate into the plant cell genome in a site-specific manner, whereupon it can be operatively linked to a heterologous nucleotide sequence, which can be expressed in response to a stress condition specific for the regulatory element; or can be a mutant regulatory element, which is not responsive to the stress condition, whereby upon integrating into the plant cell genome, the mutant regulatory element disrupts an endogenous stress-regulated regulatory element of a plant stress-regulated gene, thereby altering the responsiveness of the plant stress-regulated gene to the stress condition.
In one aspect, the invention provides a method for producing a transgenic plant by introducing into at least one plant cell a recombinant nucleic acid construct comprising i) all or a portion of any one of SEQ ID NOS:1–5379; ii) a polynucleotide comprising a coding region that hybridizes under conditions of high stringency to all or a portion of the complement of any one of SEQ ID NOS:1–2703; iii) a polynucleotide comprising a sequence that alters transcription of an operatively linked coding region in response to abiotic stress, and that hybridizes under conditions of high stringency to the complement of any one of SEQ ID NOS:2704–5379; iv) a polynucleotide having at least 90% sequence identity with any one of SEQ ID NO:1–5379; v) a fragment of any one of the sequences of iv), wherein the fragment comprises a coding region; or vi) a fragment of any one of the sequences of iv), wherein the fragment comprises a nucleotide sequence that alters transcription of an operatively linked coding region in response to abiotic stress; and regenerating a plant from the at least one plant cell.
Another aspect provides a method for producing a transgenic plant comprising introducing into at least one plant cell a recombinant nucleic acid construct comprising i) any one of SEQ ID NOS:1–1261 or 2704–3955; ii) a polynucleotide comprising a coding region that hybridizes under conditions of high stringency to the complement of any one of SEQ ID NOS:1–1261; iii) a polynucleotide comprising a sequence that alters transcription of an operatively linked coding region in response to cold stress that hybridizes under conditions of high stringency to the complement of any one of SEQ ID NOS:2704–3955; iv) a polynucleotide that has at least 90% sequence identity with any one of SEQ ID NOS:1–1261 or 2704–3955; v) a fragment of any one of the sequences of iv), wherein the fragment comprises a coding region; or vi) a fragment of any one of the sequences of iv) wherein the fragment comprises a sequence or region that alters transcription of an operatively linked coding region in response to cold stress; and regenerating a plant from the at least one plant cell.
In another aspect, the invention provides a method for producing a transgenic plant by introducing into at least one plant cell a recombinant nucleic acid construct comprising i) any one of SEQ ID NOS:2428–2585 or 5108–5263; ii) a polynucleotide comprising a coding region that hybridizes under conditions of high stringency to the complement of any one of SEQ ID NOS:2428–2585; iii) a polynucleotide comprising a sequence that alters transcription of an operatively linked coding region in response to osmotic stress that hybridizes under conditions of high stringency to the complement of any one of SEQ ID NOS:5108–5263; iv) a polynucleotide that has at least 90% sequence identity with any one of SEQ ID NOS:2428–2585 or 5108–5263; v) a fragment of any one of the sequences of iv), wherein the fragment comprises a coding region; or vi) a fragment of any one of the sequences of iv), wherein the fragment comprises a sequence or region that alters transcription of an operatively linked coding region in response to osmotic stress; and regenerating a plant from the at least one plant cell.
Still another aspect provides a method for producing a transgenic plant comprising introducing into at least one plant cell a recombinant nucleic acid construct comprising i) any one of SEQ ID NOS:2227–2427 or 4910–5107; ii) a polynucleotide comprising a coding region that hybridizes under conditions of high stringency to the complement of any one of SEQ ID NOS:2227–2427; iii) a polynucleotide comprising a sequence that alters transcription of an operatively linked coding region in response to saline stress that hybridizes under conditions of high stringency to the complement of any one of SEQ ID NOS:2227–2427; iv) a polynucleotide that has at least 90% sequence identity with any one of SEQ ID NOS:4910–5107; v) a fragment of any one of the sequences of iv), wherein the fragment comprises a coding region; or vi) a fragment of any one of the sequences of iv) wherein the fragment comprises a sequence or region that alters transcription of an operatively linked coding region in response to saline stress; and regenerating a plant from the at least one plant cell.
Yet another aspect provides a method for producing a transgenic plant comprising introducing into at least one plant cell a recombinant nucleic acid construct comprising i) any one of SEQ ID NOS:1699–1969 or 4389–4654; ii) a polynucleotide comprising a coding region that hybridizes under conditions of high stringency to the complement of any one of SEQ ID NOS:1699–1969; iii) a polynucleotide comprising a sequence that alters transcription of an operatively linked coding region in response to a combination of cold and osmotic stress that hybridizes under conditions of high stringency to the complement of any one of SEQ ID NOS:4389–4654; iv) a polynucleotide that has at least 90% sequence identity with any one of SEQ ID NOS:1699–1969 or 4389–4654; v) a fragment of any one of the sequences of iv), wherein the fragment comprises a coding region; or vi) a fragment of any one of the sequences of iv), wherein the fragment comprises a sequence or region that alters transcription of an operatively linked coding region in response to a combination of cold and osmotic stress; and regenerating a plant from the at least one plant cell.
Yet another aspect provides a method for producing a transgenic plant comprising introducing into at least one plant cell a recombinant nucleic acid construct comprising i) any one of SEQ ID NOS:1970–2226 or 4655–4909; ii) a polynucleotide comprising a coding region that hybridizes under conditions of high stringency to the complement of any one of SEQ ID NOS:1970–2226; iii) a polynucleotide comprising a sequence that alters transcription of an operatively linked coding region in response to a combination of cold and saline stress that hybridizes under conditions of high stringency to the complement of any one of SEQ ID NOS:4655–4909; iv) a polynucleotide that has at least 90% sequence identity with any one of SEQ ID NOS:1970–2226 or 4655–4909; v) a fragment of any one of the sequences of iv), wherein the fragment comprises a coding region; or vi) a fragment of any one of the sequences of iv), wherein the fragment comprises a sequence or region that alters transcription of an operatively linked coding region in response to a combination of cold and saline stress; and regenerating a plant from the at least one plant cell.
A further aspect provides a method for producing a transgenic plant comprising introducing into at least one plant cell a recombinant nucleic acid construct comprising i) any one of SEQ ID NOS:2586–2703 or 5264–5379; ii) a polynucleotide comprising a coding region that hybridizes under conditions of high stringency to the complement of any one of SEQ ID NOS:2586–2703; iii) a polynucleotide comprising a sequence that alters transcription of an operatively linked coding region in response to a combination of osmotic and saline stress that hybridizes under conditions of high stringency to the complement of any one of SEQ ID NOS: 5264–5379; iv) a polynucleotide that has at least 90% sequence identity with any one of SEQ ID NOS:2586–2703 or 5264–5379; v) a fragment of any one of the sequences of iv), wherein the fragment comprises a coding region; or vi) a fragment of any one of the sequences of iv), wherein the fragment comprises a sequence or region that alters transcription of an operatively linked coding region in response to a combination of osmotic and saline stress; and regenerating a plant from the at least one plant cell.
Another aspect provides a method for producing a transgenic plant comprising introducing into at least one plant cell a recombinant nucleic acid construct comprising i) any one of SEQ ID NOS:1262–1698 or 3956–4388; ii) a polynucleotide comprising a coding region that hybridizes under conditions of high stringency to the complement of any one of SEQ ID NOS:1262–1698; iii) a polynucleotide comprising a sequence that alters transcription of an operatively linked coding region in response to a combination of cold, osmotic and saline stress that hybridizes under conditions of high stringency to the complement of any one of SEQ ID NOS:3956–4388; iv) a polynucleotide that has at least 90% sequence identity with any one of SEQ ID NOS:1262–1698 or 3956–4388; v) a fragment of any one of the sequences of iv), wherein the fragment comprises a coding region; or vi) a fragment of any one of the sequences of iv) wherein the fragment comprises a sequence or region that alters transcription of an operatively linked coding region in response to a combination of cold, osmotic and saline stress; and regenerating a plant from the at least one plant cell. Further aspects include plants and uniform populations of plants made by the above methods as well as seeds and progeny from such plants.
In another embodiment, a transgene introduced into a plant cell according to a method of the invention can encode a polypeptide that regulates expression from an endogenous plant stress-regulated gene. Such a polypeptide can be, for example, a recombinantly produced polypeptide comprising a zinc finger domain, which is specific for the regulatory element, and an effector domain, which can be a repressor domain or an activator domain. The polynucleotide encoding the recombinant polypeptide can be operatively linked to and expressed from a constitutively active, inducible or tissue specific or phase specific regulatory element. Expression of the recombinant polypeptide from a plant stress-regulated promoter as disclosed herein can be particularly advantageous in that the polypeptide can be coordinately expressed with the endogenous plant stress-regulated genes upon exposure to a stress condition. The invention also provides transgenic plants produced by a method as disclosed, as well as to a plant cell obtained from such transgenic plant, wherein said plant cell exhibits altered responsiveness to the stress condition; a seed produced by the transgenic plant; and a cDNA or genomic DNA library prepared from the transgenic plant, or from a plant cell from said transgenic plant, wherein said plant cell exhibits altered responsiveness to the stress condition.
In one aspect, the invention provides an isolated nucleic acid molecule comprising a nucleotide sequence substantially similar to a sequence of any one of SEQ ID NOS:2704–5379, which can alter transcription of an operatively linked polynucleotide in a plant cell in response to an abiotic stress. Additional aspects of the invention provide isolated polynucleotides, including, for example, sequences substantially similar to any of SEQ ID NOS:2704–3955, which can alter transcription of an operatively linked polynucleotide in response to a cold stress; isolated polynucleotides substantially similar to a sequence of any of SEQ ID NOS:5108–5263, which can alter transcription of an operatively linked polynucleotide in response to an osmotic stress; isolated polynucleotides substantially similar to a sequence of any of SEQ ID NOS:4910–5107, which can alter transcription of an operatively linked polynucleotide in response to a saline stress; isolated polynucleotides substantially similar to a sequence of any of SEQ ID NOS:4389–4654, which can alter transcription of an operatively linked polynucleotide in response to a combination of cold and osmotic stresses; isolated polynucleotides substantially similar to a sequence of any of SEQ ID NOS:4655–4909, which can alter transcription of an operatively linked polynucleotide in response to a combination of cold and saline stresses; isolated polynucleotides substantially similar to a sequence of any of SEQ ID NOS:5264–5379, which can alter transcription of an operatively linked polynucleotide in response to a combination of osmotic and saline stresses; and isolated polynucleotides substantially similar to a sequence of any of SEQ ID NOS:3956–4388, which can alter transcription of an operatively linked polynucleotide in response to a combination of cold, osmotic and saline stresses.
Related aspects of the invention provide an isolated nucleotide sequences that can alter transcription of an operatively linked polynucleotide in response to an abiotic stress, and that hybridize under stringent conditions, preferably highly stringent conditions, to the complement of any one of SEQ ID NOS:2704–5379. Additional aspects provide an isolated nucleotide sequence that can alter transcription of an operatively linked polynucleotide in response to cold stress, and that hybridizes under stringent conditions, preferably highly stringent conditions, to the complement of any one of SEQ ID NOS:2704–3955; a nucleotide sequence that alters transcription of an operatively linked polynucleotide in response to osmotic stress, and that hybridizes under stringent conditions, preferably highly stringent conditions, to the complement of any one of SEQ ID NOS:5108–5263; a nucleotide sequence that alters transcription of an operatively linked polynucleotide in response to saline stress, and that hybridizes under stringent conditions, preferably highly stringent conditions, to the complement of any one of SEQ ID NOS:4910–5107; a nucleotide sequence that alters transcription of an operatively linked polynucleotide in response to a combination of cold and osmotic stress, and that hybridizes under stringent conditions, preferably highly stringent conditions, to the complement of any one of SEQ ID NOS:4389–4654; a nucleotide sequence that alters transcription of an operatively linked polynucleotide in response to a combination of cold and saline stress, and that hybridizes under stringent conditions, preferably highly stringent conditions, to the complement of any one of SEQ ID NOS:4655–4909; a nucleotide sequence that alters transcription of an operatively linked polynucleotide in response to an combination of osmotic and saline stress, and that hybridizes under stringent conditions, preferably highly stringent conditions, to the complement of any one of SEQ ID NOS:5264–5379; and a nucleotide sequence that alters transcription of an operatively linked polynucleotide in response to a combination of cold, osmotic and saline stress, and that hybridizes under stringent conditions, preferably highly stringent conditions, to the complement of any one of SEQ ID NOS:3956–4388.
Further aspects provide an expression cassette comprising as operatively linked components any of the above isolated nucleic acid sequences that alter transcription, a coding region, and a termination sequence. Also provided are host cells and seeds comprising such expression cassettes, plants containing such host cells and seeds and progeny of plants containing said host cells. In related aspects, the coding region of the expression cassettes comprise sequences encoding marker proteins and sequences involved in gene silencing such as antisense sequences, double stranded RNAi sequences, a triplexing agent, and sequences comprising dominant negative mutations. In additional related aspects, the coding regions comprise sequences encoding polypeptides that alter the response of a plant to an abiotic stress.
The present invention also relates to a method of modulating the responsiveness of a plant cell to a stress condition. Such a method can be performed, for example, by introducing a polynucleotide portion of a plant stress-regulated genes described herein into the plant cell, thereby modulating the responsiveness of the plant cell to a stress condition. Such a method can result in the responsiveness of the plant cell being increased upon exposure to the stress condition, which, in turn, can result in increased or decreased tolerance of the plant cell to a stress condition; or can result in the responsiveness of the plant cell to the stress condition being decreased, which, in turn, can result in increased or decreased tolerance of the plant cell to a stress condition. In one embodiment, the polynucleotide portion of the plant stress-regulated gene can integrate into the genome of the plant cell, thereby modulating the responsiveness of the plant cell to the stress condition. In another embodiment, the polynucleotide portion of the plant stress-regulated gene encodes a stress-regulated polypeptide or functional peptide portion thereof, and can be operatively linked to a heterologous promoter. The polynucleotide portion of the plant stress-regulated gene also can contain a mutation, whereby upon integrating into the plant cell genome, the polynucleotide disrupts (knocks-out) an endogenous plant stress-regulated sequence, thereby modulating the responsiveness of the plant cell to the stress condition. Depending on whether the knocked-out gene encodes an adaptive or a maladaptive stress-regulated polypeptide, the responsiveness of the plant will be modulated accordingly.
The present invention further relates to a method of modulating the activity of a biological pathway in a plant cell, wherein the pathway involves a stress-regulated polypeptide or a non-protein regulatory molecule. Such a method can be performed by introducing a polynucleotide portion of a plant stress-regulated gene, or a polynucleotide derived therefrom, for example a ribozyme derived from a nucleotide sequence as set forth in any of SEQ ID NOS:1–2703, into the plant cell, thereby modulating the activity of the biological pathway. The method can be performed with respect to a pathway involving any of the stress-regulated polypeptides as disclosed herein or encoded by the polynucleotides disclosed herein, as well as using homologs or orthologs thereof. In one embodiment, the method is performed by introducing a polynucleotide portion of a plant stress-regulated gene into the plant cell, wherein the plant stress-regulated gene comprises a nucleotide sequence as set forth in any of SEQ ID NOS:1–155, 157–228, 230–232, 234–557, 559–572, 574–605, 607–634, 636–786, 788–812, 814–1262, 1264–1386, 1387–1390, 1392–1404, 1406–1444, 1446–1483, 1485–1588, 1590–1608, 1610–1633, 1634–1725, 1727–1865, 1867–1917, 1919–1927, 1929–2855, 2857–2928, 2930–2932, 2934–3256, 3258–3271, 3273–3304, 3306–3323, 3325–3333, 3335–3485, 3487–3511, 3313–3956, 3958–4078, 4080–4097, 4099–4136, 4138–4175, 4177–4279, 4281–4299, 4301–4324, 4326–4414, 4416–4552, 4554–4602, and 4604–5379, thereby modulating the activity of the biological pathway.
The present invention also relates to a method of identifying a polynucleotide that modulates a stress response in a plant cell. In one embodiment the method comprises determining gene expression in a plant exposed to at least one stress to produce an expression profile and identifying sequences whose expression is altered at least two fold compared to plants not exposed to the stress. Such an expression profile can be obtained, for example, by contacting an array of probes representative of a plant cell genome with nucleic acid molecules expressed in a plant cell exposed to the stress; and detecting one or more nucleic acid molecules expressed at a level different from a level of expression in the absence of the stress. The method can further comprise introducing the differentially expressed nucleic acid molecule into a plant cell; and detecting a modulated response of the genetically modified plant cell to a stress, thereby identifying a polynucleotide that modulates a stress response in a plant cell. The stress can be any stress, for example, an abiotic stress such as exposure to an abnormal level of cold, osmotic pressure, and salinity. The contacting is under conditions that allow for selective hybridization of a nucleic acid molecule with probe having sufficient complementarity, for example, under stringent hybridization conditions. Expression of the nucleic acid molecule can increase or decrease the tolerance of the plant cell to the stress, and the nucleic acid molecule can be expressed at a level that is less than or greater than the level of expression in the absence of the stress.
In still another embodiment, the polynucleotide portion of the plant stress-regulated gene can comprise a stress-regulated regulatory element, which can be operatively linked to a heterologous nucleotide sequence, the expression of which can modulate the responsiveness of the plant cell to a stress condition. Such a heterologous nucleotide sequence can encode, for example, a stress-inducible transcription factor such as DREB1A. The heterologous nucleotide sequence also can encode a polynucleotide that is specific for a plant stress-regulated gene, for example, an antisense molecule, an RNAi molecule, a ribozyme, and a triplexing agent, any of which, upon expression in the plant cell, reduces or inhibits expression of a stress-regulated polypeptide encoded by the gene, thereby modulating the responsiveness of the plant cell to a stress condition, for example, an abnormal level of cold, osmotic pressure, and salinity. In another aspect, the method can include introducing a polynucleotide portion of a plant stress-regulated gene into the plant cell, wherein the plant stress-regulated gene includes a nucleotide sequence of a polynucleotide as set forth in any of SEQ ID NOS:1–155, 157–228, 230–232, 234–557, 559–572, 574–605, 607–634, 636–786, 788–812, 814–1262, 1264–1386, 1387–1390, 1392–1404, 1406–1444, 1446–1483, 1485–1588, 1590–1608, 1610–1633, 1634–1725, 1727–1865, 1867–1917, 1919–1927, 1929–2855, 2857–2928, 2930–2932, 2934–3256, 3258–3271, 3273–3304, 3306–3323, 3325–3333, 3335–3485, 3487–3511, 3313–3956, 3958–4078, 4080–4097, 4099–4136, 4138–4175, 4177–4279, 4281–4299, 4301–4324, 4326–4414, 4416–4552, 4554–4602, and 4604–5379, thereby modulating the responsiveness of the plant cell to a stress condition. The invention also relates to a plant cell obtained by any of the methods of modulating the responsiveness of a plant to a stress condition or combination of stress conditions, and to a plant comprising such a plant cell.
The present invention further relates to a method of selecting a plant having an altered resistance to an abiotic stress condition or a combination of abiotic stress conditions, such a method being useful for marker-assisted breeding. Such a method can be performed, for example, by contacting nucleic acid molecules representative of expressed polynucleotides in a plant cell of a plant to be examined for having an altered resistance to an abiotic stress with a nucleic acid probes that selectively hybridizes under stringent conditions to a plant stress-regulated gene comprising a nucleotide sequence as set forth in any of SEQ ID NO:1–5379; detecting a level of selective hybridization of the nucleic acid probes to a nucleic acid molecule representative of an expressed polynucleotide in the plant cell, wherein the level of selective hybridization corresponds to the level of the expressed polynucleotide in the plant cell, which is indicative of resistance of the plant to an abiotic stress; and selecting a plant having a level of expression of a polynucleotide indicative of altered resistance to an abiotic stress condition. For example, the abiotic stress condition can be cold stress, and the nucleic acid probe can include at least about 15 nucleotides of a nucleotide sequence as set forth in any of SEQ ID NOS:1–1261 and 2704–3955, for example, at least about 15 nucleotides of a nucleotide sequence as set forth in any of SEQ ID NOS:1–155, 157–228, 230–232, 234–557, 559–572, 574–605, 607–634, 636–786, 788–812, 814–1261, 2704–2855, 2857–2928, 2930–2932, 2934–3256, 3258–3271, 3273–3304, 3306–3323, 3325–3333, 3335–3485, 3487–3511, and 3313–3955; or the abiotic stress condition can be saline stress, and the nucleic acid probe can include at least about 15 nucleotides of a nucleotide sequence as set forth in any of SEQ ID NOS:2226–2427 and 4910–5107; or the abiotic stress condition can be osmotic stress, and the nucleic acid probe can include at least about 15 nucleotides of a nucleotide sequence as set forth in any of SEQ ID NOS:2428–2585 and 5108–5263. In addition, a combination of abiotic stress conditions can be a combination of cold stress and osmotic stress, and the nucleic acid probe can include at least about 15 nucleotides of a nucleotide sequence as set forth in any of SEQ ID NOS:1669–1969 and 4389–4654, for example, at least about 15 nucleotides of a nucleotide sequence as set forth in any of SEQ ID NOS:1699–1725, 1727–1865, 1867–1917, 1919–1927, 1929–1969, 4389–4414, 4416–4552, 4554–4602, 4604–4612, and 4613–4654; or the combination of abiotic stress conditions can be a combination of cold stress and saline stress, and the nucleic acid probe can include at least about 15 nucleotides of a nucleotide sequence as set forth in any of SEQ ID NOS: 1970–2226 and 4655–4909; or the combination of abiotic stress conditions can be a combination of osmotic stress and saline stress, and the nucleic acid probe can include at least about 15 nucleotides of a nucleotide sequence as set forth in any of SEQ ID NOS:2586–2703 and 5264–5379; or the combination of abiotic stress conditions can be a combination of cold stress, osmotic stress and saline stress, and the nucleic acid probe can include at least about 15 nucleotides of a nucleotide sequence as set forth in any of SEQ ID NOS:1262–1698 and 3956–4388, for example, at least about 15 nucleotides of a nucleotide sequence as set forth in any of SEQ ID NOS:1262, 1264–1386, 1387–1390, 1392–1404, 1406–1444, 1446–1483, 1485–1588, 1590–1608, 1610–1633, 1634–1698, 3956, 3958–4078, 4080–4097, 4099–4136, 4138–4175, 4177–4279, 4281–4299, 4301–4324, and 4326–4388.
The present invention also relates to a method of expressing a heterologous nucleotide sequence in a plant cell. Such a method can be performed, for example, by introducing into the plant cell a plant stress-regulated regulatory element operatively linked to the heterologous nucleotide sequence, whereby, upon exposure of the plant cell to a stress condition, the heterologous nucleotide sequence is expressed in the plant cell. In one embodiment, the stress-regulated gene regulatory element is any of the sequences described herein that are capable of altering transcription of an operatively linked sequence in response to an abiotic stress, for example, SEQ ID NOS:2704–5379. In another embodiment, stress-regulated gene regulatory element comprises a nucleotide sequence as set forth in any of SEQ ID NOS:2704–2855, 2857–2928, 2930–2932, 2934–3256, 3258–3271, 3273–3304, 3306–3323, 3325–3333, 3335–3485, 3487–3511, 3313–3956, 3958–4078, 4080–4097, 4099–4136, 4138–4175, 4177–4279, 4281–4299, 4301–4324, 4326–4414, 4416–4552, 4554–4602, and 4604–5379, whereby, upon exposure of the plant cell to stress condition, the heterologous nucleotide sequence is expressed in the plant cell. The heterologous nucleotide sequence can encode a selectable marker, a diagnostic marker, or a polypeptide that confers a desirable trait upon the plant cell, for example, a polypeptide that improves the nutritional value, digestibility or ornamental value of the plant cell, or a plant comprising the plant cell.
The present invention additionally relates to a method of identifying a stress condition to which a plant cell was exposed by comparing an expression profile from a test plant suspected of having been exposed to at least one stress condition to an expression profile obtained from a reference plant, preferably of the same species, which has been exposed to the suspected stress condition. Such a method can be performed, for example, by contacting nucleic acid molecules representative of expressed polynucleotides in cells of the test plant with at least one nucleic acid probe under conditions suitable for selective hybridization to a complementary nucleotide sequence, wherein the probe comprises at least 15 nucleotides of a plant stress-regulated gene, wherein the stress-regulated gene does not have a nucleotide sequence of a polynucleotide as set forth in any of SEQ ID NOS:156, 229, 233, 558, 573, 606, 635, 787, 813, 1263, 1386, 1391, 1405, 1445, 1484, 1589, 1609, 1634, 1726, 1866, 1918 or 1928, or a nucleotide sequence complementary thereto, whereby detecting selective hybridization of at least one nucleic acid probe, or detecting a change in a level of selective hybridization as compared to a level of selective hybridization obtained using nucleic acid molecules representative of expressed polynucleotides in cells of a plant known not have been exposed to an abiotic stress, indicates that the test plant has been exposed to an abiotic stress, and whereby an absence of selective hybridization of at least one nucleic acid probe indicates that the test plant has not been exposed to an abiotic stress. For example, the abiotic stress is cold stress, and the probe can include at least 15 nucleotides of a nucleotide sequence as set forth in any of SEQ ID NOS:1–155, 157–228, 230–232, 234–557, 559–572, 574–605, 607–634, 636–786, 788–812, 814–1261 or a nucleotide sequence complementary thereto; or the abiotic stress can be a saline stress, and the probe can include at least 15 nucleotides of a nucleotide sequence as set forth in any of SEQ ID NOS:2226–2427 or a nucleotide sequence complementary thereto; or the abiotic stress can be an osmotic stress, and the probe can include at least 15 nucleotides of a nucleotide sequence as set forth in two or more of SEQ ID NOS:2428–2585 or a nucleotide sequence complementary thereto.
A method of identifying a stress condition to which a plant cell was exposed also can be performed, for example, by contacting nucleic acid molecules expressed in the test plant cell with an array of probes representative of the plant cell genome; detecting a profile of expressed nucleic acid molecules characteristic of a stress response, and comparing the expression pattern in the test plant to the expression pattern obtained from a reference plant thereby identifying the stress condition to which the plant cell was exposed. The contacting is under conditions that allow for selective hybridization of a nucleic acid molecule with probes having sufficient complementarity, for example, under stringent hybridization conditions. The profile can be characteristic of exposure to a single stress condition, for example, an abnormal level of cold, osmotic pressure, or salinity, or can be characteristic of exposure to more than one stress condition, for example, cold, increased osmotic pressure and increased salinity. In one embodiment, the nucleotide sequence of a gene whose expression is detected is selected from a polynucleotide comprising any of SEQ ID NOS:1–2703. In further embodiments, the nucleotide sequence of a gene that is expressed in response a particular stress or combination of stresses can comprise a polynucleotide expressed in response to cold stress (SEQ ID NOS:1–1261), osmotic stress (SEQ ID NOS:2428–2585), saline (salt) stress (SEQ ID NOS:2227–2427), a combination of cold and osmotic stress (SEQ ID NOS:1699–1969), a combination of saline and osmotic stress (SEQ ID NOS:1970–2226), a combination of osmotic and saline stress (SEQ ID NOS:2586–2703), or a combination of cold, osmotic and saline stress (SEQ ID NOS:1262–1698).
In another embodiment, the method can be used for determining whether a test plant has been exposed to a combination of abiotic stress conditions. Such a method can be performed, for example, by contacting nucleic acid molecules representative of expressed polynucleotides in cells of the test plant with at least one nucleic acid probe under conditions suitable for selective hybridization to a complementary nucleotide sequence, whereby detecting selective hybridization of at least one nucleic acid probe, or detecting a change in a level of selective hybridization as compared to a level of selective hybridization obtained using nucleic acid molecules representative of expressed polynucleotides in cells of a plant known not have been exposed to a combination of stress conditions, indicates that the test plant has been exposed to a combination of abiotic stress conditions, and whereby an absence of selective hybridization of at least one nucleic acid probe indicates that the test plant has not been exposed to a combination of abiotic stress conditions. For example, the combination of abiotic stress conditions can be a combination of a cold stress and an osmotic stress, and the probe can include at least 15 nucleotides of a nucleotide sequence as set forth in any of SEQ ID NOS:1699–1969, or a nucleotide sequence complementary thereto; or the combination of abiotic stress conditions can be a combination of a cold stress and a saline stress, and the probe can include at least 15 nucleotides of a nucleotide sequence as set forth in any of SEQ ID NOS:1970–2226, or a nucleotide sequence complementary thereto; or the combination of abiotic stress conditions can be a combination of an osmotic stress and a saline stress, and the probe can included at least 15 nucleotides of a nucleotide sequence as set forth in any of SEQ ID NOS:2586–2703, or a nucleotide sequence complementary thereto; or the combination of abiotic stress conditions can be a combination of a cold stress, a saline stress and an osmotic stress, and the probe can include at least 15 nucleotides of a nucleotide sequence as set forth in any of SEQ ID NOS:1262–1698, or a nucleotide sequence complementary thereto.
The present invention also relates to a method for monitoring a population of plants for exposure to a stress condition or combination of stress conditions. Such a method can be performed, for example, by introducing into the population of a plants a sentinel plant, wherein said sentinel plant is a transgenic plant, which contains plant cells containing a stress-regulated regulatory element operatively linked to a polynucleotide encoding a detectable marker; and examining the sentinel plant for expression of the detectable marker, which is indicative of exposure of the population of plants to a stress condition or combination of stress conditions. The stress condition or combination of stress conditions can be any such condition or conditions, particularly an abiotic stress condition or combination of abiotic stress conditions. The detectable marker can be any reporter molecule that is readily or conveniently detectable, particularly a marker that is visibly detectable, for example, a luminescent detectable marker such as luciferin, or a fluorescent detectable marker such as a green fluorescent protein, a yellow fluorescent protein, a cyan fluorescent protein, a red fluorescent protein, or an enhanced or modified form thereof.
The present invention further relates to a transgenic plant, which contains a nucleic acid construct comprising a polynucleotide portion of plant stress-regulated polynucleotide. In one embodiment, the transgenic plant exhibits altered responsiveness to a stress condition as compared to a corresponding reference plant not containing the construct. Such a transgenic plant can contain, for example, a construct that disrupts an endogenous stress-regulated gene in the plant, thereby reducing or inhibiting expression of the gene in response to a stress condition. Such a knock-out can increase or decrease tolerance of the plant to a stress condition. The transgene also can comprise a coding sequence of a plant stress-regulated gene, which can be operatively linked to a heterologous regulatory element such as a constitutively active regulatory element, an regulated regulatory element, a tissues specific or phase specific regulatory element, or the like. In another embodiment, the transgenic plant contains a nucleic acid construct comprising a plant stress-regulated regulatory element, which can be operatively linked to a heterologous nucleotide sequence that can encode a polypeptide. Expression of the heterologous polypeptide can confer a desirable characteristic on the plant, for example, can improve the nutritional or ornamental value of the transgenic plant. In still another embodiment, the transgenic plant contains multiple nucleic acid constructs, which can be multiple copies of the same construct, or can be two or more different constructs.
The present invention also relates to a plant stress-regulated regulatory element, which is obtained from a plant stress-regulated polynucleotide disclosed herein for example any of SEQ ID NOS:2704–5379; a homolog or ortholog thereof The invention also provides a method of identifying an agent, for example a transcription factor, that specifically binds to or activates a plant stress-regulated regulatory element. Such a method can be performed, for example, by contacting the regulatory element with a plant cell extract, and identifying polypeptides that specifically bind to the regulatory element. Confirmation that the specifically binding polypeptide is a transcription factor can be demonstrated using, for example, the stress-regulated regulatory element operably linked to a reporter gene, and detecting expression of the reporter gene. Control constructs comprising a regulatory element, other than a plant stress-regulated regulatory element, operatively linked to a reporter molecule can be used to confirm that the transcription factor is specific for the plant stress-regulated regulatory element. A polynucleotide encoding such a transcription factor also can be obtained.
The present invention also relates to a method of using a polynucleotide portion of a plant stress-regulated gene to confer a selective advantage on a plant cell. In one embodiment, such a method is performed by introducing a plant stress-regulated regulatory element into a plant cell such as those described herein, wherein, upon exposure of the plant cell to a stress condition to which the regulatory element is responsive, a nucleotide sequence operatively linked to the regulatory element is expressed, thereby conferring a selective advantage to plant cell. The operatively linked nucleotide sequence can be, for example, a transcription factor, the expression of which induces the further expression of polynucleotides involved in a stress response, thereby enhancing the response of a plant to the stress condition. In another embodiment, a coding sequence of a plant stress-regulated gene as disclosed herein is introduced into the cell, thereby providing the plant with a selective advantage in response to a stress condition. In still another embodiment, the method results in the knock-out of a plant stress-regulated gene as disclosed herein in a first population of plants, thereby providing a selective advantage to a stress condition in a second population of plants.
The invention further relates to a method of identifying an agent that modulates the activity of a stress-regulated regulatory element of a plant. In a particular embodiment, is provided a method for identifying an agent that alters the activity of an abiotic stress responsive regulatory element comprising contacting the agent or a composition containing an agent to be tested with at least one abiotic stress responsive regulatory element, preferably selected from the group consisting of SEQ ID NOS:2704–5379 (see Table 2), and determining the effect of the agent on the ability of the regulatory sequence to regulate transcription. In further embodiments, the regulatory elements are associated with particular stresses or combination of stresses such as cold stress (SEQ ID NOS:2704–3955), osmotic stress (SEQ ID NOS:5108–5263), saline stress (SEQ ID NOS:4910–5107), a combination of cold and osmotic stress (SEQ ID NOS:4389–4654), a combination of cold and saline stress (SEQ ID NOS:4655–4909), a combination of osmotic and saline stress (SEQ ID NOS:5264–5379), or a combination of cold, osmotic and saline stress (SEQ ID NOS:3956–4388). In one embodiment, the regulatory element can be operatively linked to a heterologous polynucleotide encoding a reporter molecule, and an agent that modulates the activity of the stress-regulated regulatory element can be identified by detecting a change in expression of the reporter molecule due to contacting the regulatory element with the agent. Such a method can be performed in vitro in a plant cell-free system, or in a plant cell in culture or in a plant in situ. In another embodiment, the agent is contacted with a transgenic plant containing an introduced plant stress-regulated regulatory element, and an agent that modulates the activity of the regulatory element is identified by detecting a phenotypic change in the transgenic plant. The methods of the invention can be performed in the presence or absence of the stress condition to which the particularly regulatory element is responsive.
Another aspect provides a method for identifying an agent that alters abiotic stress responsive polynucleotide expression in a plant or plant cell comprising contacting a plant or plant cell with a test agent; subjecting the plant cell or plant cell to an abiotic stress or combination of stresses before, during or after contact with the agent to be tested; obtaining an expression profile of the plant or plant cell and comparing the expression profile of the plant or plant cell to an expression profile from a plant or plant cell not exposed to the abiotic stress or combination of stresses. In one embodiment, the expression profile comprises expression data for at least one nucleotide sequence comprising any of SEQ ID NOS:1–5379 (see Tables 1 and 2). In additional embodiments, the expression profile comprises expression data for at least one, and preferably two or more sequences associated with a particular abiotic stress or combination of stresses such as cold stress (SEQ ID NOS:1–1261 and 2704–3955), osmotic stress (SEQ ID NOS:2428–2585 and 5108–5263), saline stress (SEQ ID NOS:2227–2427 and 4910–5107), a combination of cold and osmotic stress (SEQ ID NOS:1699–1969 and 4389–4654), a combination of cold and saline stress (SEQ ID NOS:1970–2226 and 4655–4909), a combination of osmotic and saline stress (SEQ ID NOS:2586–2703 and 5264–5379), or a combination of cold, osmotic and saline stress (SEQ ID NOS:1262–1698 and 3956–4388).
Still another aspect provides nucleotide probes useful for detecting an abiotic stress response in plants, the probes comprising a nucleotide sequence of at least 15, 25, 50 or 100 nucleotides that hybridizes under stringent, preferably highly stringent, conditions to at least one sequence comprising any of SEQ ID NOS:1–2703. Also provided are nucleotide probes comprising at least 15, 25, 50 or 100 nucleotides in length that hybridize under stringent, preferably highly stringent conditions, to at least one gene associated with a particular stress or combination of stresses, for example cold stress, (SEQ ID NOS:1–1261), osmotic stress (SEQ ID NOS:2428–2585), saline stress (SEQ ID NOS:2227–2427), a combination of cold and osmotic stress (SEQ ID NOS:1699–1969), a combination of cold and saline stress (SEQ ID NOS: 1970–2226), a combination of osmotic and saline stress (SEQ ID NOS:2586–2703), or a combination of cold, osmotic, and saline stress (SEQ ID NOS:1262–1698).
An additional aspect provides a method for marker-assisted breeding to select plants having an altered resistance to abiotic stress comprising obtaining nucleic acid molecules from the plants to be selected; contacting the nucleic acid molecules with one or more probes that selectively hybridize under stringent, preferably highly stringent, conditions to a nucleic acid sequence selected from the group consisting of SEQ ID NOS:1–2703; detecting the hybridization of the one or more probes to the nucleic acid sequences wherein the presence of the hybridization indicates the presence of a gene associated with altered resistance to abiotic stress; and selecting plants on the basis of the presence or absence of such hybridization. Marker-assisted selection can also be accomplished using one or more probes which selectively hybridize under stringent, preferably highly stringent conditions, to a nucleotide sequence comprising a polynucleotide expressed in response associated with a particular stress, for example, a nucleotide sequence comprising any of SEQ ID NOS:1–1261 (cold stress), SEQ ID NOS:2428–2585 (osmotic stress), SEQ ID NOS:2227–2427 (saline stress), SEQ ID NOS:1699–1969 (cold and osmotic stress), SEQ ID NOS:1970–2226 (cold and saline stress), SEQ ID NOS:2586–2703 (osmotic and saline stress), or SEQ ID NOS:1262–1698 (cold, osmotic and saline stress). In each case marker-assisted selection can be accomplished using a probe or probes to a single sequence or multiple sequences. If multiple sequences are used they can be used simultaneously or sequentially.
A further aspect provides a method for monitoring a population of plants comprising providing at least one sentinel plant containing a recombinant polynucleotide comprising a stress responsive regulatory sequence selected from the group consisting of SEQ ID NOS:2704–5379 which is operatively linked to a nucleotide sequence encoding a detectable marker, for example a fluorescent protein. Additional aspects provide the use of various regulatory sequences including those associated with cold stress (SEQ ID NOS:2704–3955), osmotic stress (SEQ ID NOS:5108–5263), saline stress (SEQ ID NOS:4910–5107), cold and osmotic stress (SEQ ID NOS:4389–4654), cold and saline stress (SEQ ID NOS:4655–4909), osmotic and saline stress (SEQ ID NOS:5264–5379), and cold, osmotic and saline stress (SEQ ID NOS:3956–4388), or fragments thereof wherein such fragments can alter transcription of an operatively linked nucleotide sequence in response to an abiotic stress.
A further aspect provides a computer readable medium having stored thereon computer executable instructions for performing a method comprising receiving data on gene expression in a test plant of at least one nucleic acid molecule having at least 70%, preferably at least 80%, more preferably at least 90%, and most preferably at least 95% nucleotide sequence identity to one or more polynucleotide sequences as set forth in any of SEQ ID NOS:1–2703; and comparing expression data from the test plant to expression data for the same polynucleotide sequence or sequences in a plant that has been exposed to at least one abiotic stress.
Yet a further aspect provides a computer readable medium having stored thereon a data structure comprising, sequence data for at least one, and preferably a plurality of nucleic acid molecules having at least 70%, preferably at least 80%, more preferably at least 90%, and most preferably at least 95% nucleotide sequence identity to a polynucleotide comprising any of SEQ ID NOS:1–2703, or the complement thereof; and a module receiving the nucleic acid molecule sequence data which compares the nucleic acid molecule sequence data to at least one other nucleic acid sequence.


DETAILED DESCRIPTION OF THE INVENTION
The present invention relates to clusters of genes that are induced in response to one or a combination of abiotic stress conditions. Abiotic stress conditions, such as a shortage or excess of solar energy, water and nutrients, and salinity, high and low temperature, or pollution (e.g., heavy metals), can have a major impact on plant growth and can significantly reduce the yield, for example, of cultivars. Under conditions of abiotic stress, the growth of plant cells is inhibited by arresting the cell cycle in late G1, before DNA synthesis, or at the G2/M boundary (see Dudits, Plant Cell Division, Portland Press Research, Monograph; Francis, Dudits, and Inze, eds., 1997; chap. 2, page 21; Bergounioux, Protoplasma 142:127–136, 1988). The identification of stress-regulated gene clusters, using microarray technology, provides a means to identify plant stress-regulated genes.
As used herein, the term “cluster,” when used in reference to stress-regulated genes, refers to nucleotide sequences of genes that have been selected by drawing Venn diagrams, and selecting those genes that are regulated only by a selected stress condition. In general, a cluster of stress-regulated genes includes at least 5, 10, 15, or 20 genes, including polynucleotide portions thereof, each of which is responsive to the same selected stress condition or conditions. The selected stress condition can be a single stress condition, for example, cold, osmotic stress or salinity stress (see Tables 3–14), or can be a selected combination of stress conditions, for example, cold, osmotic stress and salinity stress (see Tables 15–26). In addition, a cluster can be selected based on specifying that all of the genes are coordinately regulated, for example, they all start at a low level and are induced to a higher level. However, a cluster of saline stress-regulated genes, for example, that was selected for coordinate regulation from low to high, also can be decreased in response to cold or mannitol. By varying the parameters used for selecting a cluster of gene nucleotide sequences, those genes that are expressed in a specific manner following a stress can be identified.
As used herein in reference to a polynucleotide or polynucleotide portion of a gene or nucleic acid molecule, the term “isolated” means a polynucleotide, polynucleotide portion of a gene, or nucleic acid molecule that is free of one or both of the nucleotide sequences that normally flank the polynucleotide in a genome of a naturally-occurring organism from which the polynucleotide is derived. The term includes, for example, a polynucleotide or fragment thereof that is incorporated into a vector or expression cassette; into an autonomously replicating plasmid or virus; into the genomic DNA of a prokaryote or eukaryote; or that exists as a separate molecule independent of other polynucleotides. It also includes a recombinant polynucleotide that is part of a hybrid polynucleotide, for example, one encoding a polypeptide sequence.
The terms “polynucleotide,” “oligonucleotide,” and “nucleic acid sequence” are used interchangeably herein to refer to a polymeric (2 or more monomers) form of nucleotides of any length, either ribonucleotides or deoxyribonucleotides. Although nucleotides are usually joined by phosphodiester linkages, the term also includes polymers containing neutral amide backbone linkages composed of aminoethyl glycine units. The terms are used only to refer to the primary structure of the molecule. Thus, the term includes double stranded and single stranded DNA molecules, including a sense strand or an antisense strand, and RNA molecules as well as genomic DNA, cDNA, mRNA and the like. It will be recognized that such polynucleotides can be modified, for example, by including a label such as a radioactive, fluorescent or other tag, by methylation, by the inclusion of a cap structure, by containing a substitution of one or more of the naturally occurring nucleotides with a nucleotide analog, by containing an internucleotide modification such as having uncharged linkages (e.g., methyl phosphonates, phosphotriesters, phosphoramidates, carbamates, or the like), by containing a pendant moiety such as a protein (e.g., a nuclease, toxin, antibody, signal peptide, poly-L-lysine, or the like), by containing an intercalator such as acridine or psoralen, by containing a chelator, which can be a metal such as boron, an oxidative metal, or a radioactive metal, by containing an alkylator, or by having a modified linkage (e.g., an alpha anomeric nucleic acid).
The term “recombinant nucleic acid molecule” refers to a polynucleotide produced by human intervention. A recombinant nucleic acid molecule can contain two or more nucleotide sequences that are linked in a manner such that the product is not found in a cell in nature. In particular, the two or more nucleotide sequences can be operatively linked and, for example, can encode a fusion polypeptide, or can comprise a nucleotide sequence and a regulatory element. A recombinant nucleic acid molecule also can be based on, but different, from a naturally occurring polynucleotide, for example, a polynucleotide having one or more nucleotide changes such that a first codon, which normally is found in the polynucleotide, is replaced with a degenerate codon that encodes the same or a conservative amino acid, or such that a sequence of interest is introduced into the polynucleotide, for example, a restriction endonuclease recognition site or a splice site, a promoter, a DNA replication initiation site, or the like.
As used herein, the term “abiotic stress” or “abiotic stress condition” refers to the exposure of a plant, plant cell, or the like, to a non-living (“abiotic”) physical or chemical agent or condition that has an adverse effect on metabolism, growth, development, propagation and/or survival of the plant (collectively “growth”). An abiotic stress can be imposed on a plant due, for example, to an environmental factor such as water (e.g., flooding, drought, dehydration), anaerobic conditions (e.g., a low level of oxygen), abnormal osmotic conditions, salinity or temperature (e.g., hot/heat, cold, freezing, frost), a deficiency of nutrients or exposure to pollutants, or by a hormone, second messenger or other molecule. Anaerobic stress, for example, is due to a reduction in oxygen levels (hypoxia or anoxia) sufficient to produce a stress response. A flooding stress can be due to prolonged or transient immersion of a plant, plant part, tissue or isolated cell in a liquid medium such as occurs during monsoon, wet season, flash flooding or excessive irrigation of plants, or the like. A cold stress or heat stress can occur due to a decrease or increase, respectively, in the temperature from the optimum range of growth temperatures for a particular plant species. Such optimum growth temperature ranges are readily determined or known to those skilled in the art. Dehydration stress can be induced by the loss of water, reduced turgor, or reduced water content of a cell, tissue, organ or whole plant. Drought stress can be induced by or associated with the deprivation of water or reduced supply of water to a cell, tissue, organ or organism. Saline stress (salt stress) can be associated with or induced by a perturbation in the osmotic potential of the intracellular or extracellular environment of a cell. Osmotic stress also can be associated with or induced by a change, for example, in the concentration of molecules in the intracellular or extracellular environment of a plant cell, particularly where the molecules cannot be partitioned across the plant cell membrane.
As disclosed herein, clusters of plant stress-regulated genes (Example 1; see, also, Tables 1–31) and homologs and orthologs thereof (Table 32) have been identified. Remarkably, several of the stress-regulated genes previously were known to encode polypeptides having defined cellular functions, including roles as transcription factors, enzymes such as kinases, and structural proteins such as channel proteins (see Tables 29–31). The identification of Arabidopsis stress-regulated genes provides a means to identify homologous and orthologous genes and gene sequences in other plant species using well known procedures and algorithms based on identity (or homology) to the disclosed sequences. Thus, the invention provides polynucleotide sequences comprising plant stress-regulated genes that are homologs or orthologs, variants, or otherwise substantially similar to the polynucleotides disclosed herein, and having an E value≦1×10−8, which can be identified, for example, by a BLASTN search using the Arabidopsis polynucleotides of Tables 1 and 2 (SEQ ID NOS:1–5379) as query sequences (see Table 32, on CD).
A polynucleotide sequence of a stress-regulated gene as disclosed herein can be particularly useful for performing the methods of the invention on a variety of plants, including but not limited to, corn (Zea mays), Brassica sp. (e.g., B. napus, B. rapa, B. juncea), particularly those Brassica species useful as sources of seed oil, alfalfa (Medicago sativa), rice (Oryza sativa), rye (Secale cereale), sorghum (Sorghum bicolor, Sorghum vulgare), millet (e.g., pearl millet (Pennisetum glaucum), proso millet (Panicum miliaceum), foxtail millet (Setaria italica), finger millet (Eleusine coracana)), sunflower (Helianthus annuus), safflower (Carthamus tinctorius), wheat (Triticum aestivum), soybean (Glycine max), tobacco (Nicotiana tabacum), potato (Solanum tuberosum), peanuts (Arachis hypogaea), cotton (Gossypium barbadense, Gossypium hirsutum), sweet potato (Ipomoea batatus), cassava (Manihot esculenta), coffee (Cofea spp.), coconut (Cocos nucifera), pineapple (Ananas comosus), citrus trees (Citrus spp.), cocoa (Theobroma cacao), tea (Camellia sinensis), banana (Musa spp.), avocado (Persea ultilane), fig (Ficus casica), guava (Psidium guajava), mango (Mangifera indica), olive (Olea europaea), papaya (Carica papaya), cashew (Anacardium occidentale), macadamia (Macadamia integrifolia), almond (Prunus amygdalus), sugar beets (Beta vulgaris), sugarcane (Saccharum spp.), oats, duckweed (Lemna), barley, tomatoes (Lycopersicon esculentum), lettuce (e.g., Lactuca sativa), green beans (Phaseolus vulgaris), lima beans (Phaseolus limensis), peas (Lathyrus spp.), and members of the genus Cucumis such as cucumber (C. sativus), cantaloupe (C. cantalupensis), and musk melon (C. melo). Ornamentals such as azalea (Rhododendron spp.), hydrangea (Macrophylla hydrangea), hibiscus (Hibiscus rosasanensis), roses (Rosa spp.), tulips (Tulipa spp.), daffodils (Narcissus spp.), petunias (Petunia hybrida), carnation (Dianthus caryophyllus), poinsettia (Euphorbia pulcherrima), and chrysanthemum are also included. Additional ornamentals within the scope of the invention include impatiens, Begonia, Pelargonium, Viola, Cyclamen, Verbena, Vinca, Tagetes, Primula, Saint Paulia, Agertum, Amaranthus, Antihirrhinum, Aquilegia, Cineraria, Clover, Cosmo, Cowpea, Dahlia, Datura, Delphinium, Gerbera, Gladiolus, Gloxinia, Hippeastrum, Mesembryanthemum, Salpiglossos, and Zinnia. Conifers that may be employed in practicing the present invention include, for example, pines such as loblolly pine (Pinus taeda), slash pine (Pinus elliotii), ponderosa pine (Pinus ponderosa), lodgepole pine (Pinus contorta), and Monterey pine (Pinus radiata), Douglas-fir (Pseudotsuga menziesii); Western hemlock (Tsuga ultilane); Sitka spruce (Picea glauca); redwood (Sequoia sempervirens); true firs such as silver fir (Abies amabilis) and balsam fir (Abies balsamea); and cedars such as Western red cedar (Thuja plicata) and Alaska yellow-cedar (Chamaecyparis nootkatensis).
Leguminous plants which may be used in the practice of the present invention include beans and peas. Beans include guar, locust bean, fenugreek, soybean, garden beans, cowpea, mung bean, lima bean, fava bean, lentils, chickpea, etc. Legumes include, but are not limited to, Arachis, e.g., peanuts, Vicia, e.g., crown vetch, hairy vetch, adzuki bean, mung bean, and chickpea, Lupinus, e.g., lupine, trifolium, Phaseolus, e.g., common bean and lima bean, Pisum, e.g., field bean, Melilotus, e.g., clover, Medicago, e.g., alfalfa, Lotus, e.g., trefoil, lens, e.g., lentil, and false indigo. Preferred forage and turf grass for use in the methods of the invention include alfalfa, orchard grass, tall fescue, perennial ryegrass, creeping bent grass, and redtop.
Other plants within the scope of the invention include Acacia, aneth, artichoke, arugula, blackberry, canola, cilantro, clementines, escarole, eucalyptus, fennel, grapefruit, honey dew, jicama, kiwifruit, lemon, lime, mushroom, nut, okra, orange, parsley, persimmon, plantain, pomegranate, poplar, radiata pine, radicchio, Southern pine, sweetgum, tangerine, triticale, vine, yams, apple, pear, quince, cherry, apricot, melon, hemp, buckwheat, grape, raspberry, chenopodium, blueberry, nectarine, peach, plum, strawberry, watermelon, eggplant, pepper, cauliflower, Brassica, e.g., broccoli, cabbage, ultilan sprouts, onion, carrot, leek, beet, broad bean, celery, radish, pumpkin, endive, gourd, garlic, snapbean, spinach, squash, turnip, ultilane, chicory, groundnut and zucchini.
As used herein, the term “substantially similar”, when used herein with respect to a nucleotide sequence, means a nucleotide sequence corresponding to a reference nucleotide sequence, wherein the corresponding sequence encodes a polypeptide or comprises a regulatory element having substantially the same structure and function as the polypeptide encoded by the reference nucleotide sequence, for example, where only changes in amino acids not affecting the polypeptide function occur. For purposes of the present invention, a reference (or query) sequence is a polynucleotide sequence as set forth in any of SEQ ID NOS:1–2703 or a polypeptide encoded thereby. Desirably, a substantially similar nucleotide sequence encodes the polypeptide encoded by the reference nucleotide sequence. The percentage of identity between the substantially similar nucleotide sequence and the reference nucleotide sequence desirably is at least 60%, more desirably at least 75%, preferably at least 90%, more preferably at least 95%, still more preferably at least 99% and including 100%. A nucleotide sequence is “substantially similar” to reference nucleotide sequence hybridizes to the reference nucleotide sequence in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO4, 1 mM EDTA at 50° C. with washing in 2×SSC, 0.1% SDS at 50° C., more desirably in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO4, 1 mM EDTA at 50° C. with washing in 1×SSC, 0.1% SDS at 50° C. (stringent conditions), more desirably still in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO4, 1 mM EDTA at 50° C. with washing in 0.5×SSC, 0.1% SDS at 50° C. (high stringency), preferably in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO4, 1 mM EDTA at 50° C. with washing in 0.1×SSC, 0.1% SDS at 50° C. (very high stringency), more preferably in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO4, 1 mM EDTA at 50° C. with washing in 0.1×SSC, 0.1% SDS at 65° C. (extremely high stringency).
In addition, the term “substantially similar,” when used in reference to a polypeptide sequence, means that an amino acid sequence relative to a reference (query) sequence shares at least about 65% amino acid sequence identity, particularly at least about 75% amino acid sequence identity, and preferably at least about 85%, more preferably at least about 90%, and most preferably at least about 95% or greater amino acid sequence identity. Generally, sequences having an E≦10−8 are considered to be substantially similar to a query sequence. Such sequence identity can take into account conservative amino acid changes that do not substantially affect the function of a polypeptide. As such, homologs or orthologs of the Arabidopsis stress-regulated nucleotide sequences disclosed herein, variants thereof, and polypeptides substantially similar to the polynucleotide sequence of Arabidopsis stress-regulated genes set forth in SEQ ID NOS:1–5379 are encompassed within the present invention and, therefore, useful for practicing the methods of the invention (see, for example, Table 32, which is on the CD-R filed herewith, and incorporated herein by reference).
Homology or identity is often measured using sequence analysis software such as the Sequence Analysis Software Package of the Genetics Computer Group (University of Wisconsin Biotechnology Center, 1710 University Avenue, Madison, Wis. 53705). Such software matches similar sequences by assigning degrees of homology to various deletions, substitutions and other modifications. The terms “homology” and “identity,” when used herein in the context of two or more nucleic acids or polypeptide sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or of nucleotides that are the same when compared and aligned for maximum correspondence over a comparison window or designated region as measured using any number of sequence comparison algorithms or by manual alignment and visual inspection.
For sequence comparison, typically one sequence acts as a reference sequence, to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. Default program parameters can be used, or alternative parameters can be designated. The sequence comparison algorithm then calculates the percent sequence identities for the test sequences relative to the reference sequence, based on the program parameters.
The term “comparison window” is used broadly herein to include reference to a segment of any one of the number of contiguous positions, for example, about 20 to 600 positions, for example, amino acid or nucleotide position, usually about 50 to about 200 positions, more usually about 100 to about 150 positions, in which a sequence may be compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned. Methods of alignment of sequence for comparison are well-known in the art. Optimal alignment of sequences for comparison can be conducted, for example, by the local homology algorithm of Smith and Waterman (Adv. Appl. Math. 2:482, 1981), by the homology alignment algorithm of Needleman and Wunsch (J. Mol. Biol. 48:443, 1970), by the search for similarity method of Person and Lipman (Proc. Natl. Acad. Sci., USA 85:2444, 1988), each of which is incorporated herein by reference; by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.); or by manual alignment and visual inspection. Other algorithms for determining homology or identity include, for example, in addition to a BLAST program (Basic Local Alignment Search Tool at the National Center for Biological Information), ALIGN, AMAS (Analysis of Multiply Aligned Sequences), AMPS (Protein Multiple Sequence Alignment), ASSET (Aligned Segment Statistical Evaluation Tool), BANDS, BESTSCOR, BIOSCAN (Biological Sequence Comparative Analysis Node), BLIMPS (BLocks IMProved Searcher), FASTA, Intervals & Points, BMB, CLUSTAL V, CLUSTAL W, CONSENSUS, LCONSENSUS, WCONSENSUS, Smith-Waterman algorithm, DARWIN, Las Vegas algorithm, FNAT (Forced Nucleotide Alignment Tool), Framealign, Framesearch, DYNAMIC, FILTER, FSAP (Fristensky Sequence Analysis Package), GAP (Global Alignment Program), GENAL, GIBBS, GenQuest, ISSC (Sensitive Sequence Comparison), LALIGN (Local Sequence Alignment), LCP (Local Content Program), MACAW (Multiple Alignment Construction & Analysis Workbench), MAP (Multiple Alignment Program), MBLKP, MBLKN, PIMA (Pattern-Induced Multi-sequence Alignment), SAGA (Sequence Alignment by Genetic Algorithm) and WHAT-IF. Such alignment programs can also be used to screen genome databases to identify polynucleotide sequences having substantially identical sequences.
A number of genome databases are available for comparison. Several databases containing genomic information annotated with some functional information are maintained by different organizations, and are accessible via the internet, for example, at world wide web addresses (url's) “wwwtigr.org/tdb”; “genetics.wisc.edu”; “genome-www.stanford.edu/ ̃ball”; “hiv-web.lanl.gov”; “ncbi.nlm.nih.gov”; “ebi.ac.uk”; “Pasteur.fr/other/biology”; and “genome.wi.mit.edu”.
In particular, the BLAST and BLAST 2.0 algorithms using default parameters are particularly useful for identifying polynucleotide and polypeptides encompassed within the present invention (Altschul et al. (Nucleic Acids Res. 25:3389–3402, 1977; J. Mol. Biol. 215:403–410, 1990, each of which is incorporated herein by reference). Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov). This algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul et al., supra, 1977, 1990). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always >0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a wordlength (W) of 11, an expectation (E) of 10, M=5, N=4 and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a wordlength of 3, and expectations (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff and Henikoff, Proc. Natl. Acad. Sci., USA 89:10915, 1989) alignments (B) of 50, expectation (E) of 10, M=5, N=4, and a comparison of both strands.
The BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, for example, Karlin and Altschul, Proc. Natl. Acad. Sci. USA 90:5873, 1993, which is incorporated herein by reference). One measure of similarity provided by BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability by which a match between two nucleotide or amino acid sequences would occur by chance. For example, a nucleic acid is considered similar to a references sequence if the smallest sum probability in a comparison of the test nucleic acid to the reference nucleic acid is less than about 0.2, more preferably less than about 0.01, and most preferably less than about 0.001. Significantly, upon identifying polynucleotides that are substantially similar to those of SEQ ID NOS:1–5379, the identified polynucleotides can be used as query sequences in a BLAST search to identify polynucleotides and polypeptides substantially similar thereto.
It should be noted that the nucleotide sequences set forth as SEQ ID NOS:1–2703 comprise coding sequences, whereas the nucleotide sequences set forth as SEQ ID NOS:2704–5379 comprise regulatory sequences. In addition, the coding sequences and regulatory sequences are related in that, for example, SEQ ID NO:1 is the coding sequence of a plant cold regulated gene having a 5′ upstream (regulatory) sequence set forth as SEQ ID NO:2704 (see Table 2). Similarly, SEQ ID NO:2705 comprises a regulatory region of SEQ ID NO:2, SEQ ID NO:2706 comprises a regulatory region of SEQ ID NO:3, and so forth as shown in Table 2. As such, reference herein, for example, to a “polynucleotide comprising SEQ ID NO:1” can, unless indicated otherwise, include at least SEQ ID NO:2704. In some cases, the entire coding region of a plant stress regulated gene or the 5′ upstream sequence has not yet been determined (see, for example, SEQ ID NO:43 in Table 3, where “none” indicates that 5′ upstream regulatory sequences have not yet been determined). However, the determination of a complete coding sequence where only a portion is known or of regulatory sequences where a portion of the coding sequence is known can be made using methods as disclosed herein or otherwise known in the art.
In one embodiment, protein and nucleic acid sequence homologies are evaluated using the Basic Local Alignment Search Tool (“BLAST”). In particular, five specific BLAST programs are used to perform the following task:

    
    
        (1) BLASTP and BLAST3 compare an amino acid query sequence against a protein sequence database;
        (2) BLASTN compares a nucleotide query sequence against a nucleotide sequence database;
        (3) BLASTX compares the six-frame conceptual translation products of a query nucleotide sequence (both strands) against a protein sequence database;
        (4) TBLASTN compares a query protein sequence against a nucleotide sequence database translated in all six reading frames (both strands); and
        (5) TBLASTX compares the six-frame translations of a nucleotide query sequence against the six-frame translations of a nucleotide sequence database.
    
    


The BLAST programs identify homologous sequences by identifying similar segments, which are referred to herein as “high-scoring segment pairs,” between a query amino or nucleic acid sequence and a test sequence which is preferably obtained from a protein or nucleic acid sequence database. High-scoring segment pairs are preferably identified (i.e., aligned) by means of a scoring matrix, many of which are known in the art. Preferably, the scoring matrix used is the BLOSUM62 matrix (Gonnet et al., Science 256:1443–1445, 1992; Henikoff and Henikoff, Proteins 17:49–61, 1993, each of which is incorporated herein by reference). Less preferably, the PAM or PAM250 matrices may also be used (Schwartz and Dayhoff, eds., “Matrices for Detecting Distance Relationships: Atlas of Protein Sequence and Structure” (Washington, National Biomedical Research Foundation 1978)). BLAST programs are accessible through the U.S. National Library of Medicine, for example, on the world wide web at address (url) “ncbi.nlm.nih.gov”.
The parameters used with the above algorithms may be adapted depending on the sequence length and degree of homology studied. In some embodiments, the parameters may be the default parameters used by the algorithms in the absence of instructions from the user.
The term “substantially similar” also is used in reference to a comparison of expression profiles of nucleotide sequences, wherein a determination that an expression profile characteristic of a stress response is substantially similar to the profile of nucleic acid molecules expressed in a plant cell being examined (“test plant”) is indicative of exposure of the test plant cell to one or a combination of abiotic stress conditions. When used in reference to such a comparison of expression profiles, the term “substantially similar” means that the individual nucleotide sequences in the test plant cell profile are altered in the same manner as the corresponding nucleotide sequences in the expression profile characteristic of the stress response.
By way of example, where exposure to saline results in an increased expression of nucleotide sequences A, B and C, and a decreased expression of nucleotide sequences D and E, as indicated by the expression profile characteristic of a saline stress response, a determination that corresponding nucleotide sequences A, B and C in the test plant cell are increased and that nucleotides sequences D and E are decreased is indicative of exposure of the test plant cell to a saline stress condition. It should be recognized that, where, for example, only nucleotide sequences A, B, D and E are examined in the test plant cell, an increase in A and B and a decrease in D and E expression of the test plant cells is considered to be substantially similar to the expression profile characteristic of a saline stress condition and, therefore, is indicative of exposure of the plant cell to a saline stress condition. Similarly, where the levels of expression of the nucleotide sequences examined in a test plant are altered in the same manner, i.e., are increased or are decreased, as that observed in an expression profile characteristic of a particular stress response, the absolute levels of expression may vary, for example, two-fold, five-fold, ten-fold, or the like. Nevertheless, the expression profile of the test plant cell is considered to be substantially similar to the expression profile characteristic of the particular stress response and, therefore, indicative of exposure of the plant cell to the stress condition.
As disclosed herein, clusters of stress-regulated genes (and their products), some of which also have been described as having cellular functions such as enzymatic activity or roles as transcription factors, are involved in the response of plant cells to various abiotic stresses (see Tables 29–31; see, also, Tables 1 and 32). As such, the polynucleotide sequences comprising the genes in a cluster likely share common stress-regulated regulatory elements, including, for example, cold-regulated regulatory elements (SEQ ID NOS:2704–3955), salinity-regulated regulatory elements (SEQ ID NOS:4910–5107, and osmotic pressure-regulated regulatory elements (SEQ ID NO:5108–5263), as well as regulatory elements that are responsive to a combination of stress conditions, but not to any of the individual stress conditions, alone (SEQ ID NOS:3956–4909 and 5263–5379). The identification of such clusters of genes thus provides a means to identify the stress-regulated regulatory elements that control the level of expression of these genes.
As used herein, the term “plant stress-regulated gene” means a polynucleotide sequence of a plant, the transcription of which is altered in response to exposure to a stress condition, and the regulatory elements linked to such a polynucleotide sequence and involved in the stress response, which can be induction or repression. In general, plant stress gene regulatory elements are contained within a sequence including approximately two kilobases upstream (5′) of the transcription or translation start site and two kilobases downstream (3′) of the transcription or translation termination site. In the absence of an abiotic stress condition, the stress-regulated gene can normally be unexpressed in the cells, can be expressed at a basal level, which is induced to a higher level in response to the stress condition, or can be expressed at a level that is reduced (decreased) in response to the stress condition. The coding region of a plant stress-regulated gene encodes a stress-regulated polypeptide, and also can be the basis for expression of a functional RNA molecule such as an antisense molecule or ribozyme. A stress-regulated polypeptide can have an adaptive effect on a plant, thereby allowing the plant to better tolerate stress conditions; or can have a maladaptive effect, thereby decreasing the ability of the plant to tolerate the stress conditions.
The present invention provides an isolated plant stress-regulated regulatory element, which regulates expression of an operatively linked nucleotide sequence in a plant in response a stress condition. As disclosed herein, a plant stress-regulated regulatory element can be isolated from a polynucleotide sequence of a plant stress-regulated gene comprising a nucleotide sequence as set forth in SEQ ID NOS:1–2703, for example any of SEQ ID NOS:2704–5379 (see Table 2). It is recognized that certain of the polynucleotides set forth as SEQ ID NOS:1–5379 previously have been described as being involved in a stress-regulated response in plants, including SEQ ID NOS:156, 229, 233, 558, 573, 606, 625, 635, 787, 813, 1263, 1386, 1391, 1405, 1445, 1484, 1589, 1609, 1634, 1726, 1866, 1918, and 1928 and, therefore, are not encompassed, in whole or in part, within the compositions of the invention, and are encompassed within only certain particular methods of the invention, for example, methods of making a transgenic plant that is resistant to two or more stress conditions, since, even where such a gene was known to be expressed in response to a single stress condition such as cold or saline (e.g., SEQ ID NO:1263), it was not known prior to the present disclosure that any of these genes was responsive to a combination of stress conditions (for example, a combination of cold and osmotic stress for SEQ ID NOS:1726, 1866, 1918, and 1928; or a combination of cold, osmotic and saline stress for SEQ ID NOS:1263, 1386, 1391, 1405, 1445, 1484, 1589, 1609, and 1634).
Methods for identifying and isolating the stress-regulated regulatory element from the disclosed polynucleotides, or genomic DNA clones corresponding thereto, are well known in the art. For example, methods of making deletion constructs or linker-scanner constructs can be used to identify nucleotide sequences that are responsive to a stress condition. Generally, such constructs include a reporter gene operatively linked to the sequence to be examined for regulatory activity. By performing such assays, a plant stress-regulated regulatory element can be defined within a sequence of about 500 nucleotides or fewer, generally at least about 200 nucleotides or fewer, particularly about 50 to 100 nucleotides, and more particularly at least about 20 nucleotides or fewer. Preferably the minimal (core) sequence required for regulating a stress response of a plant is identified.
The nucleotide sequences of the genes of a cluster also can be examined using a homology search engine such as described herein to identify sequences of conserved identity, particularly in the nucleotide sequence upstream of the transcription start site. Since all of the genes in a cluster as disclosed are induced in response to a particular stress condition or a particular combination of stress conditions, some or all of the nucleotide sequences can share conserved stress-regulated regulatory elements. By performing such a homology search, putative stress-regulated regulatory elements can be identified. The ability of such identified sequences to function as a plant stress-regulated regulatory element can be confirmed, for example, by operatively linking the sequence to a reporter gene and assaying the construct for responsiveness to a stress condition.
As used herein, the term “regulatory element” means a nucleotide sequence that, when operatively linked to a coding region of a gene, effects transcription of the coding region such that a ribonucleic acid (RNA) molecule is transcribed from the coding region. A regulatory element generally can increase or decrease the amount of transcription of a nucleotide sequence, for example, a coding sequence, operatively linked to the element with respect to the level at which the nucleotide sequence would be transcribed absent the regulatory element. Regulatory elements are well known in the art and include promoters, enhancers, silencers, inactivated silencer intron sequences, 3′-untranslated or 5′-untranslated sequences of transcribed sequence, for example, a poly-A signal sequence, or other protein or RNA stabilizing elements, or other gene expression control elements known to regulate gene expression or the amount of expression of a gene product. A regulatory element can be isolated from a naturally occurring genomic DNA sequence or can be synthetic, for example, a synthetic promoter.
Regulatory elements can be constitutively expressed regulatory element, which maintain gene expression at a relative level of activity (basal level), or can be regulated regulatory elements. Constitutively expressed regulatory elements can be expressed in any cell type, or can be tissue specific, which are expressed only in particular cell types, phase specific, which are expressed only during particular developmental or growth stages of a plant cell, or the like. A regulatory element such as a tissue specific or phase specific regulatory element or an inducible regulatory element useful in constructing a recombinant polynucleotide or in a practicing a method of the invention can be a regulatory element that generally, in nature, is found in a plant genome. However, the regulatory element also can be from an organism other than a plant, including, for example, from a plant virus, an animal virus, or a cell from an animal or other multicellular organism.
A regulatory element useful for practicing method of the present is a promoter element. Useful promoters include, but are not limited to, constitutive, inducible, temporally regulated, developmentally regulated, spatially-regulated, chemically regulated, stress-responsive, tissue-specific, viral and synthetic promoters. Promoter sequences are known to be strong or weak. A strong promoter provides for a high level of gene expression, whereas a weak promoter provides for a very low level of gene expression. An inducible promoter is a promoter that provides for the turning on and off of gene expression in response to an exogenously added agent, or to an environmental or developmental stimulus. A bacterial promoter such as the Ptac promoter can be induced to varying levels of gene expression depending on the level of isothiopropylgalactoside added to the transformed bacterial cells. An isolated promoter sequence that is a strong promoter for heterologous nucleic acid is advantageous because it provides for a sufficient level of gene expression to allow for easy detection and selection of transformed cells and provides for a high level of gene expression when desired.
Within a plant promoter region there are several domains that are necessary for full function of the promoter. The first of these domains lies immediately upstream of the structural gene and forms the “core promoter region” containing consensus sequences, normally 70 base pairs immediately upstream of the gene. The core promoter region contains the characteristic CAAT and TATA boxes plus surrounding sequences, and represents a transcription initiation sequence that defines the transcription start point for the structural gene.
The presence of the core promoter region defines a sequence as being a promoter: if the region is absent, the promoter is non-functional. The core promoter region, however, is insufficient to provide full promoter activity. A series of regulatory sequences upstream of the core constitute the remainder of the promoter. These regulatory sequences determine expression level, the spatial and temporal pattern of expression and, for an important subset of promoters, expression under inductive conditions (regulation by external factors such as light, temperature, chemicals, hormones).
To define a minimal promoter region, a DNA segment representing the promoter region is removed from the 5′ region of the gene of interest and operably linked to the coding sequence of a marker (reporter) gene by recombinant DNA techniques well known to the art. The reporter gene is operably linked downstream of the promoter, so that transcripts initiating at the promoter proceed through the reporter gene. Reporter genes generally encode proteins which are easily measured, including, but not limited to, chloramphenicol acetyl transferase (CAT), beta-glucuronidase (GUS), green fluorescent protein (GFP), β-galactosidase (β-GAL), and luciferase.
The construct containing the reporter gene under the control of the promoter is then introduced into an appropriate cell type by transfection techniques well known to the art. To assay for the reporter protein, cell lysates are prepared and appropriate assays, which are well known in the art, for the reporter protein are performed. For example, if CAT were the reporter gene of choice, the lysates from cells transfected with constructs containing CAT under the control of a promoter under study are mixed with isotopically labeled chloramphenicol and acetyl-coenzyme A (acetyl-CoA). The CAT enzyme transfers the acetyl group from acetyl-CoA to the 2-position or 3-position of chloramphenicol. The reaction is monitored by thin layer chromatography, which separates acetylated chloramphenicol from unreacted material. The reaction products are then visualized by autoradiography.
The level of enzyme activity corresponds to the amount of enzyme that was made, which in turn reveals the level of expression from the promoter of interest. This level of expression can be compared to other promoters to determine the relative strength of the promoter under study. In order to be sure that the level of expression is determined by the promoter, rather than by the stability of the mRNA, the level of the reporter mRNA can be measured directly, for example, by northern blot analysis.
Once activity is detected, mutational and/or deletional analyses may be employed to determine the minimal region and/or sequences required to initiate transcription. Thus, sequences can be deleted at the 5′ end of the promoter region and/or at the 3′ end of the promoter region, and nucleotide substitutions introduced. These constructs are then introduced to cells and their activity determined.
The choice of promoter will vary depending on the temporal and spatial requirements for expression, and also depending on the target species. In some cases, expression in multiple tissues is desirable. While in others, tissue-specific, e.g., leaf-specific, seed-specific, petal-specific, anther-specific, or pith-specific, expression is desirable. Although many promoters from dicotyledons have been shown to be operational in monocotyledons and vice versa, ideally dicotyledonous promoters are selected for expression in dicotyledons, and monocotyledonous promoters for expression in monocotyledons. There is, however, no restriction to the origin or source of a selected promoter. It is sufficient that the promoters are operational in driving the expression of a desired nucleotide sequence in the particular cell.
A range of naturally-occurring promoters are known to be operative in plants and have been used to drive the expression of heterologous (both foreign and endogenous) genes and nucleotide sequences in plants: for example, the constitutive 35S cauliflower mosaic virus (CaMV) promoter, the ripening-enhanced tomato polygalacturonase promoter (Bird et al., 1988), the E8 promoter (Diekman and Fischer, 1988) and the fruit specific 2A1 promoter (Pear et al., 1989). Many other promoters, e.g., U2 and U5 snRNA promoters from maize, the promoter from alcohol dehydrogenase, the Z4 promoter from a gene encoding the Z4 22 kD zein protein, the Z10 promoter from a gene encoding a 10 kD zein protein, a Z27 promoter from a gene encoding a 27 kD zein protein, the A20 promoter from the gene encoding a 19 kD zein protein, inducible promoters, such as the light inducible promoter derived from the pea rbcS gene and the actin promoter from rice, e.g., the actin 2 promoter (WO 00/70067); seed specific promoters, such as the phaseolin promoter from beans, may also be used. The nucleotide sequences of the stress-regulated genes of this invention can also be expressed under the regulation of promoters that are chemically regulated. This enables the nucleic acid sequence or encoded polypeptide to be synthesized only when the crop plants are treated with the inducing chemicals. Chemical induction of gene expression is detailed in EP 0 332 104 and U.S. Pat. No. 5,614,395.
In some instances it may be desirable to link a constitutive promoter to a polynucleotide comprising a stress regulated gene of the invention. Examples of some constitutive promoters include the rice actin 1 (Wang et al., 1992; U.S. Pat. No. 5,641,876), CaMV 35S (Odell et al., 1985), CaMV 19S (Lawton et al., 1987), nos, Adh, sucrose synthase; and the ubiquitin promoters.
In other situations it may be desirable to limit expression of stress-related sequences to specific tissues or stages of development. As used herein, the term “tissue specific or phase specific regulatory element” means a nucleotide sequence that effects transcription in only one or a few cell types, or only during one or a few stages of the life cycle of a plant, for example, only for a period of time during a particular stage of growth, development or differentiation. The terms “tissue specific” and “phase specific” are used together herein in referring to a regulatory element because a single regulatory element can have characteristics of both types of regulatory elements. For example, a regulatory element active only during a particular stage of plant development also can be expressed only in one or a few types of cells in the plant during the particular stage of development. As such, any attempt to classify such regulatory elements as tissue specific or as phase specific can be difficult. Accordingly, unless indicated otherwise, all regulatory elements having the characteristic of a tissue specific regulatory element, or a phase specific regulatory element, or both are considered together for purposes of the present invention.
Examples of tissue specific promoters which have been described include the lectin (Vodkin, 1983; Lindstrom et al., 1990) corn alcohol dehydrogenase 1 (Vogel et al., 1989; Dennis et al., 1984), corn light harvesting complex (Simpson, 1986; Bansal et al., 1992), corn heat shock protein (Odell et al., 1985), pea small subunit RuBP carboxylase (Poulsen et al., 1986), Ti plasmid mannopine synthase and Ti plasmid nopaline synthase (Langridge et al., 1989), petunia chalcone isomerase (vanTunen et al., 1988), bean glycine rich protein 1 (Keller et al., 1989), truncated CaMV 35s (Odell et al., 1985), potato patatin (Wenzler et al., 1989), root cell (Yamamoto et al., 1990), maize zein (Reina et al., 1990; Kriz et al., 1987; Wandelt et al., 1989; Langridge et al., 1983; Reina et al., 1990), globulin-1 (Belanger et al., 1991), α-tubulin, cab (Sullivan et al., 1989), PEPCase (Hudspeth & Grula, 1989), R gene complex-associated promoters (Chandler et al., 1989), histone, and chalcone synthase promoters (Franken et al., 1991). Tissue specific enhancers are described by Fromm et al. (1989).
Several other tissue-specific regulated genes and/or promoters have been reported in plants, including genes encoding seed storage proteins such as napin, cruciferin, beta-conglycinin, and phaseolin, zein or oil body proteins such as oleosin, genes involved in fatty acid biosynthesis, including acyl carrier protein, stearoyl-ACP desaturase, fatty acid desaturases (fad 2-1), and other genes expressed during embryonic development such as Bce4 (see, for example, EP 255378 and Kridl et al., 1991). Particularly useful for seed-specific expression is the pea vicilin promoter (Czako et al., 1992). (See also U.S. Pat. No. 5,625,136, which is incorporated herein by reference.) Other useful promoters for expression in mature leaves are those that are switched on at the onset of senescence, such as the SAG promoter from Arabidopsis (Gan et al., 1995).
A class of fruit-specific promoters expressed at or during antithesis through fruit development, at least until the beginning of ripening, is discussed in U.S. Pat. No. 4,943,674. cDNA clones that are preferentially expressed in cotton fiber have been isolated (John et al., 1992). cDNA clones from tomato displaying differential expression during fruit development have been isolated and characterized (Mansson et al., 1985, Slater et al., 1985). The promoter for polygalacturonase gene is active in fruit ripening. The polygalacturonase gene is described in U.S. Pat. Nos. 4,535,060, 4,769,061, 4,801,590, and 5,107,065, each of which is incorporated herein by reference.
Other examples of tissue-specific promoters include those that direct expression in leaf cells following damage to the leaf (for example, from chewing insects), in tubers (for example, patatin gene promoter), and in fiber cells (an example of a developmentally-regulated fiber cell protein is E6 (John et al., 1992). The E6 gene is most active in fiber, although low levels of transcripts are found in leaf, ovule and flower.
Additional tissue specific or phase specific regulatory elements include, for example, the AGL8/FRUITFULL regulatory element, which is activated upon floral induction (Hempel et al., Development 124:3845–3853, 1997, which is incorporated herein by reference); root specific regulatory elements such as the regulatory elements from the RCP1 gene and the LRP1 gene (Tsugeki and Fedoroff, Proc. Natl. Acad., USA 96:12941–12946, 1999; Smith and Fedoroff, Plant Cell 7:735–745, 1995, each of which is incorporated herein by reference); flower specific regulatory elements such as the regulatory elements from the LEAFY gene and the APETELA1 gene (Blazquez et al., Development 124:3835–3844, 1997, which is incorporated herein by reference; Hempel et al., supra, 1997); seed specific regulatory elements such as the regulatory element from the oleosin gene (Plant et al., Plant Mol. Biol. 25:193–205, 1994, which is incorporated herein by reference), and dehiscence zone specific regulatory element. Additional tissue specific or phase specific regulatory elements include the Zn13 promoter, which is a pollen specific promoter (Hamilton et al., Plant Mol. Biol. 18:211–218, 1992, which is incorporated herein by reference); the UNUSUAL FLORAL ORGANS (UFO) promoter, which is active in apical shoot meristem; the promoter active in shoot meristems (Atanassova et al., Plant J. 2:291, 1992, which is incorporated herein by reference), the cdc2a promoter and cyc07 promoter (see, for example, Ito et al., Plant Mol. Biol. 24:863, 1994; Martinez et al., Proc. Natl. Acad. Sci., USA 89:7360, 1992; Medford et al., Plant Cell 3:359, 1991; Terada et al., Plant J. 3:241, 1993; Wissenbach et al., Plant J. 4:411, 1993, each of which is incorporated herein by reference); the promoter of the APETELA3 gene, which is active in floral meristems (Jack et al., Cell 76:703, 1994, which is incorporated herein by reference; Hempel et al., supra, 1997); a promoter of an agamous-like (AGL) family member, for example, AGL8, which is active in shoot meristem upon the transition to flowering (Hempel et al., supra, 1997); floral abscission zone promoters; L1-specific promoters; and the like.
The tissue-specificity of some “tissue-specific” promoters may not be absolute and may be tested by one skilled in the art using the diphtheria toxin sequence. One can also achieve tissue-specific expression with “leaky” expression by a combination of different tissue-specific promoters (Beals et al., 1997). Other tissue-specific promoters can be isolated by one skilled in the art (see U.S. Pat. No. 5,589,379). Several inducible promoters (“gene switches”) have been reported, many of which are described in the review by Gatz (1996) and Gatz (1997). These include tetracycline repressor system, Lac repressor system, copper inducible systems, salicylate inducible systems (such as the PR1a system), glucocorticoid (Aoyama et al., 1997) and ecdysone inducible systems. Also included are the benzene sulphonamide (U.S. Pat. No. 5,364,780) and alcohol (WO 97/06269 and WO 97/06268) inducible systems and glutathione S-transferase promoters.
In some instances it might be desirable to inhibit expression of a native DNA sequence within a plant's tissues to achieve a desired phenotype. In this case, such inhibition might be accomplished with transformation of the plant to comprise a constitutive, tissue-independent promoter operably linked to an antisense nucleotide sequence, such that constitutive expression of the antisense sequence produces an RNA transcript that interferes with translation of the mRNA of the native DNA sequence.
Inducible regulatory elements also are useful for purposes of the present invention. As used herein, the term “inducible regulatory element” means a regulatory element that, when exposed to an inducing agent, effects an increased level of transcription of a nucleotide sequence to which it is operatively linked as compared to the level of transcription, if any, in the absence of an inducing agent. Inducible regulatory elements can be those that have no basal or constitutive activity and only effect transcription upon exposure to an inducing agent, or those that effect a basal or constitutive level of transcription, which is increased upon exposure to an inducing agent. Inducible regulatory elements that effect a basal or constitutive level of expression generally are useful in a method or composition of the invention where the induced level of transcription is substantially greater than the basal or constitutive level of expression, for example, at least about two-fold greater, or at least about five-fold greater. Particularly useful inducible regulatory elements do not have a basal or constitutive activity, or increase the level of transcription at least about ten-fold greater than a basal or constitutive level of transcription associated with the regulatory element.
Inducible promoters that have been described include the ABA- and turgor-inducible promoters, the promoter of the auxin-binding protein gene (Schwob et al., 1993), the UDP glucose flavonoid glycosyl-transferase gene promoter (Ralston et al., 1988), the MPI proteinase inhibitor promoter (Cordero et al., 1994), and the glyceraldehyde-3-phosphate dehydrogenase gene promoter (Kohler et al., 1995; Quigley et al., 1989; Martinez et al., 1989).
The term “inducing agent” is used to refer to a chemical, biological or physical agent or environmental condition that effects transcription from an inducible regulatory element. In response to exposure to an inducing agent, transcription from the inducible regulatory element generally is initiated de novo or is increased above a basal or constitutive level of expression. Such induction can be identified using the methods disclosed herein, including detecting an increased level of RNA transcribed from a nucleotide sequence operatively linked to the regulatory element, increased expression of a polypeptide encoded by the nucleotide sequence, or a phenotype conferred by expression of the encoded polypeptide.
An inducing agent useful in a method of the invention is selected based on the particular inducible regulatory element. For example, the inducible regulatory element can be a metallothionein regulatory element, a copper inducible regulatory element or a tetracycline inducible regulatory element, the transcription from which can be effected in response to metal ions, copper or tetracycline, respectively (Furst et al., Cell 55:705–717, 1988; Mett et al., Proc. Natl. Acad. Sci. USA 90:4567–4571, 1993; Gatz et al., Plant J. 2:397–404, 1992; Roder et al., Mol. Gen. Genet. 243:32–38, 1994, each of which is incorporated herein by reference). The inducible regulatory element also can be an ecdysone regulatory element or a glucocorticoid regulatory element, the transcription from which can be effected in response to ecdysone or other steroid (Christopherson et al., Proc. Natl. Acad. Sci., USA 89:6314–6318, 1992; Schena et al., Proc. Natl. Acad. Sci., USA 88:10421–10425, 1991, each of which is incorporated herein by reference). In addition, the regulatory element can be a cold responsive regulatory element or a heat shock regulatory element, the transcription of which can be effected in response to exposure to cold or heat, respectively (Takahashi et al., Plant Physiol. 99:383–390, 1992, which is incorporated herein by reference). Additional regulatory elements useful in the methods or compositions of the invention include, for example, the spinach nitrite reductase gene regulatory element (Back et al., Plant Mol. Biol. 17:9, 1991, which is incorporated herein by reference); a light inducible regulatory element (Feinbaum et al., Mol. Gen. Genet. 226:449, 1991; Lam and Chua, Science 248:471, 1990, each of which is incorporated herein by reference), a plant hormone inducible regulatory element (Yamaguchi-Shinozaki et al., Plant Mol. Biol. 15:905, 1990; Kares et al., Plant Mol. Biol. 15:225, 1990, each of which is incorporated herein by reference), and the like.
An inducible regulatory element also can be a plant stress-regulated regulatory element of the invention. In addition to the known stress conditions that specifically induce or repress expression from such elements, the present invention provides methods of identifying agents that mimic a stress condition. Accordingly, such stress mimics are considered inducing or repressing agents with respect to a plant stress-regulated regulatory element. In addition, a recombinant polypeptide comprising a zinc finger domain, which is specific for the regulatory element, and an effector domain, particularly an activator, can be useful as an inducing agent for a plant stress-regulated regulatory element. Furthermore, such a recombinant polypeptide provides the advantage that the effector domain can be a repressor domain, thereby providing a repressing agent, which decreases expression from the regulatory element. In addition, use of such a method of modulating expression of an endogenous plant stress-regulated gene provides the advantage that the polynucleotide encoding the recombinant polypeptide can be introduced into cells of the plant, thus providing a transgenic plant that can be regulated coordinately with the endogenous plant stress-regulated gene upon exposure to a stress condition. A polynucleotide encoding such a recombinant polypeptide can be operatively linked to and expressed from a constitutively active, inducible or tissue specific or phase specific regulatory element.
In one embodiment, the promoter may be a gamma zein promoter, an oleosin ole16 promoter, a globulin I promoter, an actin I promoter, an actin c1 promoter, a sucrose synthetase promoter, an INOPS promoter, an EXM5 promoter, a globulin2 promoter, a b-32, ADPG-pyrophosphorylase promoter, an LtpI promoter, an Ltp2 promoter, an oleosin ole17 promoter, an oleosin ole18 promoter, an actin 2 promoter, a pollen-specific protein promoter, a pollen-specific pectate lyase promoter, an anther-specific protein promoter (Huffman), an anther-specific gene RTS2 promoter, a pollen-specific gene promoter, a tapeturn-specific gene promoter, tapeturn-specific gene RAB24 promoter, a anthranilate synthase alpha subunit promoter, an alpha zein promoter, an anthranilate synthase beta subunit promoter, a dihydrodipicolinate synthase promoter, a Thi 1 promoter, an alcohol dehydrogenase promoter, a cab binding protein promoter, an H3C4 promoter, a RUBISCO SS starch branching enzyme promoter, an ACCase promoter, an actin3 promoter, an actin7 promoter, a regulatory protein GF14-12 promoter, a ribosomal protein L9 promoter, a cellulose biosynthetic enzyme promoter, an S-adenosyl-L-homocysteine hydrolase promoter, a superoxide dismutase promoter, a C-kinase receptor promoter, a phosphoglycerate mutase promoter, a root-specific RCc3 mRNA promoter, a glucose-6 phosphate isomerase promoter, a pyrophosphate-fructose 6-phosphate-1-phosphotransferase promoter, an ubiquitin promoter, a beta-ketoacyl-ACP synthase promoter, a 33 kDa photosystem 11 promoter, an oxygen evolving protein promoter, a 69 kDa vacuolar ATPase subunit promoter, a metallothionein-like protein promoter, a glyceraldehyde-3-phosphate dehydrogenase promoter, an ABA- and ripening-inducible-like protein promoter, a phenylalanine ammonia lyase promoter, an adenosine triphosphatase S-adenosyl-L-homocysteine hydrolase promoter, an a-tubulin promoter, a cab promoter, a PEPCase promoter, an R gene promoter, a lectin promoter, a light harvesting complex promoter, a heat shock protein promoter, a chalcone synthase promoter, a zein promoter, a globulin-1 promoter, an ABA promoter, an auxin-binding protein promoter, a UDP glucose flavonoid glycosyl-transferase gene promoter, an NTI promoter, an actin promoter, an opaque 2 promoter, a b70 promoter, an oleosin promoter, a CaMV 35S promoter, a CaMV 19S promoter, a histone promoter, a turgor-inducible promoter, a pea small subunit RuBP carboxylase promoter, a Ti plasmid mannopine synthase promoter, Ti plasmid nopaline synthase promoter, a petunia chalcone isomerase promoter, a bean glycine rich protein I promoter, a CaMV 35S transcript promoter, a potato patatin promoter, or a S-E9 small subunit RuBP carboxylase promoter.
In addition to promoters, a variety of 5′ and 3′ transcriptional regulatory sequences are also available for use in the present invention. Transcriptional terminators are responsible for the termination of transcription and correct mRNA polyadenylation. The 3′-untranslated regulatory DNA sequence preferably includes from about 50 to about 1,000, more preferably about 100 to about 1,000, nucleotide base pairs and contains plant transcriptional and translational termination sequences. Appropriate transcriptional terminators and those which are known to function in plants include the CaMV 35S terminator, the tml terminator, the nopaline synthase terminator, the pea rbcS E9 terminator, the terminator for the T7 transcript from the octopine synthase gene of Agrobacterium tumefaciens, and the 3′ end of the protease inhibitor I or II genes from potato or tomato, although other 3′ elements known to those of skill in the art can also be employed. Alternatively, one also could use a gamma coixin, oleosin 3 or other terminator from the genus Coix. Preferred 3′ elements include those from the nopaline synthase gene of Agrobacterium tumefaciens (Bevan et al., 1983), the terminator for the T7 transcript from the octopine synthase gene of Agrobacterium tumefaciens, and the 3′ end of the protease inhibitor I or II genes from potato or tomato.
As the DNA sequence between the transcription initiation site and the start of the coding sequence, i.e., the untranslated leader sequence, can influence gene expression, one may also wish to employ a particular leader sequence. Preferred leader sequences are contemplated to include those that include sequences predicted to direct optimum expression of the attached sequence, i.e., to include a preferred consensus leader sequence that may increase or maintain mRNA stability and prevent inappropriate initiation of translation. The choice of such sequences will be known to those of skill in the art in light of the present disclosure. Sequences that are derived from genes that are highly expressed in plants will be most preferred.
Other sequences that have been found to enhance gene expression in transgenic plants include intron sequences (e.g., from Adh1, bronze1, actin1, actin2 (WO 00/760067), or the sucrose synthase intron) and viral leader sequences (e.g., from TMV, MCMV and AMV). For example, a number of non-translated leader sequences derived from viruses are known to enhance expression. Specifically, leader sequences from tobacco mosaic virus (TMV), maize chlorotic mottle virus (MCMV), and alfalfa mosaic virus (AMV) have been shown to be effective in enhancing expression (e.g., Gallie et al., 1987; Skuzeski et al., 1990). Other leaders known in the art include but are not limited to picornavirus leaders, for example, EMCV leader (encephalomyocarditis virus 5′ non-coding region; Elroy-Stein et al., 1989); potyvirus leaders, for example, TEV leader (tobacco etch virus); MDMV leader (maize dwarf mosaic virus); human immunoglobulin heavy chain binding protein (BiP) leader, (Macejak et al., 1991); untranslated leader from the coat protein mRNA of AMV (AMV RNA 4; Jobling et al., 1987), TMV (Gallie et al., 1989), and MCMV (Lommel et al., 1991; see also, della Cioppa et al., 1987).
Regulatory elements such as Adh intron 1 (Callis et al., 1987), sucrose synthase intron (Vasil et al., 1989) or TMV omega element (Gallie, et al., 1989), may further be included where desired. Examples of enhancers include elements from the CaMV 35S promoter, octopine synthase genes (Ellis et al., 1987), the rice actin I gene, the maize alcohol dehydrogenase gene (Callis et al., 1987), the maize shrunken I gene (Vasil et al., 1989), TMV Omega element (Gallie et al., 1989) and promoters from non-plant eukaryotes (e.g. yeast; Ma et al., 1988).
Vectors for use in accordance with the present invention may be constructed to include the ocs enhancer element, which was first identified as a 16 bp palindromic enhancer from the octopine synthase (ocs) gene of ultilane (Ellis et al., 1987), and is present in at least 10 other promoters (Bouchez et al., 1989). The use of an enhancer element, such as the ocs element and particularly multiple copies of the element, will act to increase the level of transcription from adjacent promoters when applied in the context of monocot transformation.
The methods of the invention provide genetically modified plant cells, which can contain, for example, a coding region, or peptide portion thereof, of a plant stress-regulated gene operatively linked to a heterologous inducible regulatory element; or a plant stress-regulated regulatory element operatively linked to a heterologous nucleotide sequence encoding a polypeptide of interest. In such a plant, the expression from the inducible regulatory element can be effected by exposing the plant cells to an inducing agent in any of numerous ways depending, for example, on the inducible regulatory element and the inducing agent. For example, where the inducible regulatory element is a cold responsive regulatory element present in the cells of a transgenic plant, the plant can be exposed to cold conditions, which can be produced artificially, for example, by placing the plant in a thermostatically controlled room, or naturally, for example, by planting the plant in an environment characterized, at least in part, by attaining temperatures sufficient to induce transcription from the promoter but not so cold as to kill the plants. By examining the phenotype of such transgenic plants, those plants that ectopically express a gene product that confers increased resistance of the plant to cold can be identified. Similarly, a transgenic plant containing a metallothionein promoter can be exposed to metal ions such as cadmium or copper by watering the plants with a solution containing the inducing metal ions, or can be planted in soil that is contaminated with a level of such metal ions that is toxic to most plants. The phenotype of surviving plants can be observed, those expressing desirable traits can be selected.
As used herein, the term “phenotype” refers to a physically detectable characteristic. A phenotype can be identified visually by inspecting the physical appearance of a plant following exposure, for example, to increased osmotic conditions; can be identified using an assay to detecting a product produced due to expression of reporter gene, for example, an RNA molecule, a polypeptide such as an enzyme, or other detectable signal such as disclosed herein; or by using any appropriate tool useful for identifying a phenotype of a plant, for example, a microscope, a fluorescence activated cell sorter, or the like.
A transgenic plant containing an inducible regulatory element such as a steroid inducible regulatory element can be exposed to a steroid by watering the plants with a solution containing the steroid. The use of an inducible regulatory element that is induced upon exposure to a chemical or biological inducing agent that can be placed in solution or suspension in an aqueous medium can be particularly useful because the inducing agent can be applied conveniently to a relatively large crop of transgenic plants containing the inducible regulatory element, for example, through a watering system or by spraying the inducing agent over the field. As such, inducible regulatory elements that are responsive to an environmental inducing agent, for example, cold; heat; metal ions or other potentially toxic agents such as a pesticides, which can contaminate a soil; or the like; or inducible regulatory elements that are regulated by inducing agents that conveniently can be applied to plants, can be particularly useful in a method or composition of the invention, and allow the identification and selection of plants that express desirable traits and survive and grow in environments that otherwise would not support growth of the plants.
As disclosed herein, the present invention provides plant stress-regulated regulatory elements, which are identified based on the expression of clusters of plant genes in response to stress. As used herein, the term “stress-regulated regulatory element of a plant” or “plant stress-regulated regulatory element” means a nucleotide sequence of a plant genome that can respond to a stress such that expression of a gene product encoded by a gene comprising the regulatory element (a stress-inducible gene) is increased above or decreased below the level of expression of the gene product in the absence of the stress condition. The regulatory element can be any gene regulatory element, including, for example, a promoter, an enhancer, a silencer, or the like. In one embodiment, the plant stress-regulated regulatory element is a plant stress-regulated promoter.
For purposes of modulating the responsiveness of a plant to a stress condition, it can be useful to introduce a modified plant stress-regulated regulatory element into a plant. Such a modified regulatory element can have any desirable characteristic, for example, it can be inducible to a greater level than the corresponding wild-type promoter, or it can be inactivated such that, upon exposure to a stress, there is little or no induction of expression of a nucleotide sequence operatively linked to the mutant element. A plant stress-regulated regulatory element can be modified by incorporating random mutations using, for example, in vitro recombination or DNA shuffling (Stemmer et al., Nature 370: 389–391, 1994; U.S. Pat. No. 5,605,793, each of which is incorporated herein by reference). Using such a method, millions of mutant copies of the polynucleotide, for example, stress-regulated regulatory element, can be produced based on the original nucleotide sequence, and variants with improved properties, such as increased inducibility can be recovered.
A mutation method such as DNA shuffling encompasses forming a mutagenized double-stranded polynucleotide from a template double-stranded polynucleotide, wherein the template double-stranded polynucleotide has been cleaved into double stranded random fragments of a desired size, and comprises the steps of adding to the resultant population of double-stranded random fragments one or more single or double stranded oligonucleotides, wherein the oligonucleotides comprise an area of identity and an area of heterology to the double stranded template polynucleotide; denaturing the resultant mixture of double stranded random fragments and oligonucleotides into single stranded fragments; incubating the resultant population of single stranded fragments with a polymerase under conditions that result in the annealing of the single stranded fragments at the areas of identity to form pairs of annealed fragments, the areas of identity being sufficient for one member of a pair to prime replication of the other, thereby forming a mutagenized double-stranded polynucleotide; and repeating the second and third steps for at least two further cycles, wherein the resultant mixture in the second step of a further cycle includes the mutagenized double-stranded polynucleotide from the third step of the previous cycle, and the further cycle forms a further mutagenized double-stranded polynucleotide. Preferably, the concentration of a single species of double stranded random fragment in the population of double stranded random fragments is less than 1% by weight of the total DNA. In addition, the template double stranded polynucleotide can comprise at least about 100 species of polynucleotides. The size of the double stranded random fragments can be from about 5 base pairs to 5 kilobase pairs. In a further embodiment, the fourth step of the method comprises repeating the second and the third steps for at least 10 cycles.
A plant stress-regulated regulatory element of the invention is useful for expressing a nucleotide sequence operatively linked to the element in a cell, particularly a plant cell. As used herein, the term “expression” refers to the transcription and/or translation of an endogenous gene or a transgene in plants. In the case of an antisense molecule, for example, the term “expression” refers to the transcription of the polynucleotide encoding the antisense molecule.
As used herein, the term “operatively linked,” when used in reference to a plant stress-regulated regulatory element, means that the regulatory element is positioned with respect to a second nucleotide sequence such that the regulatory element effects transcription or transcription and translation of the nucleotide sequence in substantially the same manner, but not necessarily to the same extent, as it does when the regulatory element is present in its natural position in a genome. Transcriptional promoters, for example, generally act in a position and orientation dependent manner and usually are positioned at or within about five nucleotides to about fifty nucleotides 5′ (upstream) of the start site of transcription of a gene in nature. In comparison, enhancers and silencers can act in a relatively position or orientation independent manner and, therefore, can be positioned several hundred or thousand nucleotides upstream or downstream from a transcription start site, or in an intron within the coding region of a gene, yet still be operatively linked to a coding region so as to effect transcription.
The second nucleotide sequence, i.e., the sequence operatively linked to the plant stress-regulated regulatory element, can be any nucleotide sequence, including, for example, a coding region of a gene or cDNA; a sequence encoding an antisense molecule, an RNAi molecule, ribozyme, triplexing agent (see, for example, Frank-Kamenetskii and Mirkin, Ann. Rev. Biochem. 64:65–95, 1995), or the like; or a sequence that, when transcribed, can be detected in the cell using, for example, by hybridization or amplification, or when translated produces a detectable signal. The term “coding region” is used broadly herein to include a nucleotide sequence of a genomic DNA or a cDNA molecule comprising all or part of a coding region of the coding strand. A coding region can be transcribed from an operatively linked regulatory element, and can be translated into a full length polypeptide or a peptide portion of a polypeptide. It should be recognized that, in a nucleotide sequence comprising a coding region, not all of the nucleotides in the sequence need necessarily encode the polypeptide and, particularly, that a gene transcript can contain one or more introns, which do not encode an amino acid sequence of a polypeptide but, nevertheless, are part of the coding region, particularly the coding strand, of the gene.
The present invention also relates to a recombinant polynucleotide, which contains a polynucleotide portion of a plant stress-regulated gene operatively linked to a heterologous nucleotide sequence. As used herein, the term “polynucleotide portion of plant stress-regulated sequence” means a contiguous nucleotide sequence of the plant stress-regulated gene that provides a function. The portion can be any portion of the sequence, particularly a coding sequence, or a sequence encoding a peptide portion of the stress-regulated polypeptide; the stress-regulated regulatory element; a sequence useful as an antisense molecule or triplexing agent; or a sequence useful for disrupting (knocking-out) an endogenous plant stress-regulated gene.
A heterologous nucleotide sequence is a nucleotide sequence that is not normally part of the plant stress-regulated gene from which the polynucleotide portion of the plant stress-regulated gene-component of the recombinant polynucleotide is obtained; or, if it is a part of the plant stress-regulated gene from which the polynucleotide portion is obtained, it is an orientation other than it would normally be in, for example, is an antisense sequence, or comprises at least partially discontinuous as compared to the genomic structure, for example, a single exon operatively linked to the regulatory element. In general, where the polynucleotide portion of the plant stress-regulated gene comprises the coding sequence in a recombinant polynucleotide of the invention, the heterologous nucleotide sequence will function as a regulatory element. The regulatory element can be any heterologous regulatory element, including, for example, a constitutively active regulatory element, an inducible regulatory element, or a tissue specific or phase specific regulatory element, as disclosed above. Conversely, where the polynucleotide portion of the plant stress-regulated polynucleotide comprises the stress-regulated regulatory element of a recombinant polynucleotide of the invention, the heterologous nucleotide sequence generally will be a nucleotide sequence that can be transcribed and, if desired, translated. Where the heterologous nucleotide sequence is expressed from a plant stress-regulated regulatory element, it generally confers a desirable phenotype to a plant cell containing the recombinant polynucleotide, or provides a means to identify a plant cell containing the recombinant polynucleotide. It should be recognized that a “desirable” phenotype can be one that decreases the ability of a plant cell to compete where the plant cell, or a plant containing the cell, is an undesired plant cell. Thus, a heterologous nucleotide sequence can allow a plant to grow, for example, under conditions in which it would not normally be able to grow.
A heterologous nucleotide sequence can be, or encode, a selectable marker. As used herein, the term “selectable marker” is used herein to refer to a molecule that, when present or expressed in a plant cell, provides a means to identify a plant cell containing the marker. As such, a selectable marker can provide a means for screening a population of plants, or plant cells, to identify those having the marker. A selectable marker also can confer a selective advantage to the plant cell, or a plant containing the cell. The selective advantage can be, for example, the ability to grow in the presence of a negative selective agent such as an antibiotic or herbicide, compared to the growth of plant cells that do not contain the selectable marker. The selective advantage also can be due, for example, to an enhanced or novel capacity to utilize an added compound as a nutrient, growth factor or energy source. A selectable advantage can be conferred, for example, by a single polynucleotide, or its expression product, or to a combination of polynucleotides whose expression in a plant cell gives the cell with a positive selective advantage, a negative selective advantage, or both.
Examples of selectable markers include those that confer antimetabolite resistance, for example, dihydrofolate reductase, which confers resistance to methotrexate (Reiss, Plant Physiol. (Life Sci. Adv.) 13:143–149, 1994); neomycin phosphotransferase, which confers resistance to the aminoglycosides neomycin, kanamycin and paromycin (Herrera-Estrella, EMBO J. 2:987–995, 1983) and hygro, which confers resistance to hygromycin (Marsh, Gene 32:481–485, 1984), trpB, which allows cells to utilize indole in place of tryptophan; hisD, which allows cells to utilize histinol in place of histidine (Hartman, Proc. Natl. Acad. Sci., USA 85:8047, 1988); mannose-6-phosphate isomerase which allows cells to utilize mannose (WO 94/20627); ornithine decarboxylase, which confers resistance to the ornithine decarboxylase inhibitor, 2-(difluoromethyl)-DL-ornithine (DFMO; McConlogue, 1987, In: Current Communications in Molecular Biology, Cold Spring Harbor Laboratory ed.); and deaminase from Aspergillus terreus, which confers resistance to Blasticidin S (Tamura, Biosci. Biotechnol. Biochem. 59:2336–2338, 1995). Additional selectable markers include those that confer herbicide resistance, for example, phosphinothricin acetyltransferase gene, which confers resistance to phosphinothricin (White et al., Nucl. Acids Res. 18:1062, 1990; Spencer et al., Theor. Appl. Genet. 79:625–631, 1990), a mutant EPSPV-synthase, which confers glyphosate resistance (Hinchee et al., Bio/Technology 91:915–922, 1998), a mutant acetolactate synthase, which confers imidazolione or sulfonylurea resistance (Lee et al., EMBO J. 7:1241–1248, 1988), a mutant psbA, which confers resistance to atrazine (Smeda et al., Plant Physiol. 103:911–917, 1993), or a mutant protoporphyrinogen oxidase (see U.S. Pat. No. 5,767,373), or other markers conferring resistance to an herbicide such as glufosinate. In addition, markers that facilitate identification of a plant cell containing the polynucleotide encoding the marker include, for example, luciferase (Giacomin, Plant Sci. 116:59–72, 1996; Scikantha, J. Bacteriol. 178:121, 1996), green fluorescent protein (Gerdes, FEBS Lett. 389:44–47, 1996) or fl-glucuronidase (Jefferson, EMBO J. 6:3901–3907, 1997), and numerous others as disclosed herein or otherwise known in the art. Such markers also can be used as reporter molecules.
A heterologous nucleotide sequence can encode an antisense molecule, particularly an antisense molecule specific for a nucleotide sequence of a plant stress-regulated gene, for example, the gene from which the regulatory component of the recombinant polynucleotide is derived. Such a recombinant polynucleotide can be useful for reducing the expression of a plant stress-regulated polypeptide in response to a stress condition because the antisense molecule, like the polypeptide, only will be induced upon exposure to the stress. A heterologous nucleotide sequence also can be, or can encode, a ribozyme or a triplexing agent. In addition to being useful as heterologous nucleotide sequences, such molecules also can be used directly in a method of the invention, for example, to modulate the responsiveness of a plant cell to a stress condition. Thus, an antisense molecule, ribozyme, or triplexing agent can be contacted directly with a target cell and, upon uptake by the cell, can effect their antisense, ribozyme or triplexing activity; or can be encoded by a heterologous nucleotide sequence that is expressed in a plant cell from a plant stress-regulated regulatory element, whereupon it can effect its activity.
An antisense polynucleotide, ribozyme or triplexing agent is complementary to a target sequence, which can be a DNA or RNA sequence, for example, messenger RNA, and can be a coding sequence, a nucleotide sequence comprising an intron-exon junction, a regulatory sequence such as a Shine-Delgarno-like sequence, or the like. The degree of complementarity is such that the polynucleotide, for example, an antisense polynucleotide, can interact specifically with the target sequence in a cell. Depending on the total length of the antisense or other polynucleotide, one or a few mismatches with respect to the target sequence can be tolerated without losing the specificity of the polynucleotide for its target sequence. Thus, few if any mismatches would be tolerated in an antisense molecule consisting, for example, of twenty nucleotides, whereas several mismatches will not affect the hybridization efficiency of an antisense molecule that is complementary, for example, to the full length of a target mRNA encoding a cellular polypeptide. The number of mismatches that can be tolerated can be estimated, for example, using well known formulas for determining hybridization kinetics (see Sambrook et al., “Molecular Cloning; A Laboratory Manual” 2nd Edition (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.; 1989)) or can be determined empirically using methods as disclosed herein or otherwise known in the art, particularly by determining that the presence of the antisense polynucleotide, ribozyme, or triplexing agent in a cell decreases the level of the target sequence or the expression of a polypeptide encoded by the target sequence in the cell.
A nucleotide sequence useful as an antisense molecule, a ribozyme or a triplexing agent can inhibit translation or cleave a polynucleotide encoded by plant stress-regulated gene, thereby modulating the responsiveness of a plant cell to a stress condition. An antisense molecule, for example, can bind to an mRNA to form a double stranded molecule that cannot be translated in a cell. Antisense oligonucleotides of at least about 15 to 25 nucleotides are preferred since they are easily synthesized and can hybridize specifically with a target sequence, although longer antisense molecules can be expressed from a recombinant polynucleotide introduced into the target cell. Specific nucleotide sequences useful as antisense molecules can be identified using well known methods, for example, gene walking methods (see, for example, Seimiya et al., J. Biol. Chem. 272:4631–4636 (1997), which is incorporated herein by reference). Where the antisense molecule is contacted directly with a target cell, it can be operatively associated with a chemically reactive group such as iron-linked EDTA, which cleaves a target RNA at the site of hybridization. A triplexing agent, in comparison, can stall transcription (Maher et al., Antisense Res. Devel. 1:227 (1991); Helene, Anticancer Drug Design 6:569 (1991)).
A plant stress-regulated regulatory element can be included in an expression cassette. As used herein, the term “expression cassette” refers to a nucleotide sequence that can direct expression of an operatively linked polynucleotide. Thus, a plant stress-regulated regulatory element can constitute an expression cassette, or component thereof. An expression cassette is particularly useful for directing expression of a nucleotide sequence, which can be an endogenous nucleotide sequence or a heterologous nucleotide sequence, in a cell, particularly a plant cell. If desired, an expression cassette also can contain additional regulatory elements, for example, nucleotide sequences required for proper translation of a polynucleotide sequence into a polypeptide. In general, an expression cassette can be introduced into a plant cell such that the plant cell, a plant resulting from the plant cell, seeds obtained from such a plant, or plants produced from such seeds are resistant to a stress condition.
Additional regulatory sequences as disclosed above or other desirable sequences such as selectable markers or the like can be incorporated into an expression cassette containing a plant stress-regulated regulatory element (see, for example, WO 99/47552). Examples of suitable markers include dihydrofolate reductase (DHFR) or neomycin resistance for eukaryotic cells and tetracycline or ampicillin resistance for E. coli. Selection markers in plants include bleomycin, gentamycin, glyphosate, hygromycin, kanamycin, methotrexate, phleomycin, phosphinotricin, spectinomycin, streptomycin, sulfonamide and sulfonylureas resistance (see, for example, Maliga et al., Methods in Plant Molecular Biology, Cold Spring Harbor Laboratory Press, 1995, page 39). The selection marker can have its own promoter or its expression can be driven by the promoter operably linked to the sequence of interest. Additional sequences such as intron sequences (e.g. from Adh1 or bronze1) or viral leader sequences (e.g. from TMV, MCMV and AIVIV), all of which can enhance expression, can be included in the cassette. In addition, where it is desirable to target expression of a nucleotide sequence operatively linked to the stress-regulated regulatory element, a sequence encoding a cellular localization motif can be included in the cassette, for example, such that an encoded transcript or translation product is translocated to and localizes in the cytosol, nucleus, a chloroplast, or another subcellular organelle. Examples of useful transit peptides and transit peptide sequences can be found in Von Heijne et al., Plant Mol. Biol. Rep. 9: 104, 1991; Clark et al., J. Biol. Chem. 264:17544, 1989; della Cioppa et al., Plant Physiol. 84:965, 1987; Romer et al., Biochem. Biophys. Res. Comm. 196:1414, 1993; Shah et al., Science 233:478, 1986; Archer et al., J. Bioenerg Biomemb. 22:789, 1990; Scandalios, Prog. Clin. Biol. Res. 344:515, 1990; Weisbeek et al., J. Cell Sci. Suppl. 11: 199, 1989; Bruce, Trends Cell Biol. 10:440, 2000. The present invention can utilize native or heterologous transit peptides. The encoding sequence for a transit peptide can include all or a portion of the encoding sequence for a particular transit peptide, and may also contain portions of the mature protein encoding sequence associated with a particular transit peptide.
A polynucleotide portion of a plant stress-regulated plant gene, or an expression cassette, can be introduced into a cell as a naked DNA molecule, can be incorporated in a matrix such as a liposome or a particle such as a viral particle, or can be incorporated into a vector. Such vectors can be cloning or expression vectors, but other uses are within the scope of the present invention. A cloning vector is a self-replicating DNA molecule that serves to transfer a DNA segment into a host cell. The three most common types of cloning vectors are bacterial plasmids, phages, and other viruses. An expression vector is a cloning vector designed so that a coding sequence inserted at a particular site will be transcribed and translated into a protein. Incorporation of the polynucleotide into a vector can facilitate manipulation of the polynucleotide, or introduction of the polynucleotide into a plant cell. A vector can be derived from a plasmid or a viral vector such as a T-DNA vector (Horsch et al., Science 227:1229–1231, 1985, which is incorporated herein by reference). If desired, the vector can comprise components of a plant transposable element, for example, a Ds transposon (Bancroft and Dean, Genetics 134:1221–1229, 1993, which is incorporated herein by reference) or an Spm transposon (Aarts et al., Mol. Gen. Genet. 247:555–564, 1995, which is incorporated herein by reference).
In addition to containing the polynucleotide portion of a plant stress-regulated gene, a vector can contain various nucleotide sequences that facilitate, for example, rescue of the vector from a transformed plant cell; passage of the vector in a host cell, which can be a plant, animal, bacterial, or insect host cell; or expression of an encoding nucleotide sequence in the vector, including all or a portion of a rescued coding region. As such, the vector can contain any of a number of additional transcription and translation elements, including constitutive and inducible promoters, enhancers, and the like (see, for example, Bitter et al., Meth. Enzymol. 153:516–544, 1987). For example, a vector can contain elements useful for passage, growth or expression in a bacterial system, including a bacterial origin of replication; a promoter, which can be an inducible promoter; and the like. In comparison, a vector that can be passaged in a mammalian host cell system can have a promoter such as a metallothionein promoter, which has characteristics of both a constitutive promoter and an inducible promoter, or a viral promoter such as a retrovirus long terminal repeat, an adenovirus late promoter, or the like. A vector also can contain one or more restriction endonuclease recognition and cleavage sites, including, for example, a polylinker sequence, to facilitate rescue of a nucleotide sequence operably linked to the polynucleotide portion.
The present invention also relates to a method of using a polynucleotide portion of a plant stress-regulated gene to confer a selective advantage on a plant cell. Such a method can be performed by introducing, for example, a plant stress-regulated regulatory element into a plant cell, wherein, upon exposure of the plant cell to a stress condition to which the regulatory element is responsive, a nucleotide sequence operatively linked to the regulatory element is expressed, thereby conferring a selective advantage to plant cell. The operatively linked nucleotide sequence can be a heterologous nucleotide sequence, which can be operatively linked to the regulatory element prior to introduction of the regulatory sequence into the plant cell; or can be an endogenous nucleotide sequence into which the regulatory element was targeted by a method such as homologous recombination. The selective advantage conferred by the operatively linked nucleotide sequence can be such that the plant is better able to tolerate the stress condition; or can be any other selective advantage.
As used herein, the term “selective advantage” refers to the ability of a particular organism to better propagate, develop, grow, survive, or otherwise tolerate a condition as compared to a corresponding reference organism that does not contain a plant-stress regulated polynucleotide portion of the present invention. In one embodiment, a selective advantage is exemplified by the ability of a desired plant, plant cell, or the like, that contains an introduced plant stress-regulated regulatory element, to grow better than an undesired plant, plant cell, or the like, that does not contain the introduced regulatory element. For example, a recombinant polynucleotide comprising a plant stress-regulated regulatory element operatively linked to a heterologous nucleotide sequence encoding an enzyme that inactivates an herbicide can be introduced in a desired plant. Upon exposure of a mixed population of plants comprising the desired plants, which contain the recombinant polynucleotide, and one or more other populations of undesired plants, which lack the recombinant polynucleotide, to a stress condition that induces expression of the regulatory element and to the herbicide, the desired plants will have a greater likelihood of surviving exposure to the toxin and, therefore, a selective advantage over the undesired plants.
In another embodiment, a selective advantage is exemplified by the ability of a desired plant, plant cell, or the like, to better propagate, develop, grow, survive, or otherwise tolerate a condition as compared to an undesired plant, plant cell, or the like, that contains an introduced plant stress-regulated regulatory element. For example, a recombinant polynucleotide comprising a plant stress-regulated regulatory element operatively linked to a plant cell toxin can be introduced into cells of an undesirable plant present in a mixed population of desired and undesired plants, for example, food crops and weeds, respectively, then the plants can be exposed to stress conditions that induce expression from the plant stress-regulated regulatory element, whereby expression of the plant cell toxin results in inhibition of growth or death of the undesired plants, thereby providing a selective advantage to the desired plants, which no longer have to compete with the undesired plants for nutrients, light, or the like. In another example, a plant stress-regulated regulatory element operatively linked to a plant cell toxin can be introduced into cells of plants used as a nurse crop. Nurse crops, also called cover or companion crops, are planted in combination with plants of interest to provide, among other things, shade and soil stability during establishment of the desired plants. Once the desired plants have become established, the presence of the nurse crop may no longer be desirable. Exposure to conditions inducing expression of the gene linked to the plant stress-regulated regulatory element allows elimination of the nurse crop. Alternatively nurse crops can be made less tolerate to abiotic stress by the inhibition of any of the stress-regulated sequences disclosed herein. Inhibition can be accomplished by any of the method described herein. Upon exposure of the nurse crop to the stress, the decreased ability of the nurse crop to respond to the stress will result in elimination of the nurse crop, leaving only the desired plants.
The invention also provides a means of producing a transgenic plant, which comprises plant cells that exhibit altered responsiveness to a stress condition. As such, the present invention further provides a transgenic plant, or plant cells or tissues derived therefrom, which are genetically modified to respond to stress differently than a corresponding wild-type plant or plant not containing constructs of the present invention would respond. As used herein, the term “responsiveness to a stress condition” refers to the ability of a plant to express a plant stress-regulated gene upon exposure to the stress condition. A transgenic plant cell contains a polypeptide portion of a plant stress-regulated gene, or a mutant form thereof, for example, a knock-out mutant. A knock-out mutant form of a plant stress-regulated gene can contain, for example, a mutation such that a STOP codon is introduced into the reading frame of the translated portion of the gene such that expression of a functional stress-regulated polypeptide is prevented; or a mutation in the stress-regulated regulatory element such that inducibility of the element in response to a stress condition is inhibited. Such transgenic plants of the invention can display any of various idiotypic modifications is response to an abiotic stress, including altered tolerance to the stress condition, as well as increased or decreased plant growth, root growth, yield, or the like, as compared to the corresponding wild-type plant.
The term “plant” is used broadly herein to include any plant at any stage of development, or to part of a plant, including a plant cutting, a plant cell, a plant cell culture, a plant organ, a plant seed, and a plantlet. A plant cell is the structural and physiological unit of the plant, comprising a protoplast and a cell wall. A plant cell can be in the form of an isolated single cell or a cultured cell, or can be part of higher organized unit, for example, a plant tissue, plant organ, or plant. Thus, a plant cell can be a protoplast, a gamete producing cell, or a cell or collection of cells that can regenerate into a whole plant. As such, a seed, which comprises multiple plant cells and is capable of regenerating into a whole plant, is considered plant cell for purposes of this disclosure. A plant tissue or plant organ can be a seed, protoplast, callus, or any other groups of plant cells that is organized into a structural or functional unit. Particularly useful parts of a plant include harvestable parts and parts useful for propagation of progeny plants. A harvestable part of a plant can be any useful part of a plant, for example, flowers, pollen, seedlings, tubers, leaves, stems, fruit, seeds, roots, and the like. A part of a plant useful for propagation includes, for example, seeds, fruits, cuttings, seedlings, tubers, rootstocks, and the like.
A transgenic plant can be regenerated from a transformed plant cell. As used herein, the term “regenerate” means growing a whole plant from a plant cell; a group of plant cells; a protoplast; a seed; or a piece of a plant such as a callus or tissue. Regeneration from protoplasts varies from species to species of plants. For example, a suspension of protoplasts can be made and, in certain species, embryo formation can be induced from the protoplast suspension, to the stage of ripening and germination. The culture media generally contains various components necessary for growth and regeneration, including, for example, hormones such as auxins and cytokinins; and amino acids such as glutamic acid and proline, depending on the particular plant species. Efficient regeneration will depend, in part, on the medium, the genotype, and the history of the culture. If these variables are controlled, however, regeneration is reproducible.
Regeneration can occur from plant callus, explants, organs or plant parts. Transformation can be performed in the context of organ or plant part regeneration. (see Meth. Enzymol. Vol. 118; Klee et al. Ann. Rev. Plant Physiol. 38:467, 1987, which is incorporated herein by reference). Utilizing the leaf disk-transformation-regeneration method, for example, disks are cultured on selective media, followed by shoot formation in about two to four weeks (see Horsch et al., supra, 1985). Shoots that develop are excised from calli and transplanted to appropriate root-inducing selective medium. Rooted plantlets are transplanted to soil as soon as possible after roots appear. The plantlets can be repotted as required, until reaching maturity.
In vegetatively propagated crops, the mature transgenic plants are propagated utilizing cuttings or tissue culture techniques to produce multiple identical plants. Selection of desirable transgenotes is made and new varieties are obtained and propagated vegetatively for commercial use. In seed propagated crops, the mature transgenic plants can be self crossed to produce a homozygous inbred plant. The resulting inbred plant produces seeds that contain the introduced plant stress-induced regulatory element, and can be grown to produce plants that express a polynucleotide or polypeptide in response to a stress condition that induces expression from the regulatory element. As such, the invention further provides seeds produced by a transgenic plant obtained by a method of the invention.
In addition, transgenic plants comprising different recombinant sequences can be crossbred, thereby providing a means to obtain transgenic plants containing two or more different transgenes, each of which contributes a desirable characteristic to the plant. Methods for breeding plants and selecting for crossbred plants having desirable characteristics or other characteristics of interest are well known in the art.
A method of the invention can be performed by introducing a polynucleotide portion of a plant stress-regulated gene into the plant. As used herein, the term “introducing” means transferring a polynucleotide into a plant cell. A polynucleotide can be introduced into a cell by a variety of methods well known to those of ordinary skill in the art. For example, the polynucleotide can be introduced into a plant cell using a direct gene transfer method such as electroporation or microprojectile mediated transformation, or using Agrobacterium mediated transformation. Non-limiting examples of methods for the introduction of polynucleotides into plants are provided in greater detail herein. As used herein, the term “transformed” refers to a plant cell containing an exogenously introduced polynucleotide portion of a plant stress-regulated gene that is or can be rendered active in a plant cell, or to a plant comprising a plant cell containing such a polynucleotide.
It should be recognized that one or more polynucleotides, which are the same or different can be introduced into a plant, thereby providing a means to obtain a genetically modified plant containing multiple copies of a single transgenic sequence, or containing two or more different transgenic sequences, either or both of which can be present in multiple copies. Such transgenic plants can be produced, for example, by simply selecting plants having multiple copies of a single type of transgenic sequence; by cotransfecting plant cells with two or more populations of different transgenic sequences and identifying those containing the two or more different transgenic sequences; or by crossbreeding transgenic plants, each of which contains one or more desired transgenic sequences, and identifying those progeny having the desired sequences.
Methods for introducing a polynucleotide into a plant cell to obtain a transformed plant also include direct gene transfer (see European Patent A 164 575), injection, electroporation, biolistic methods such as particle bombardment, pollen-mediated transformation, plant RNA virus-mediated transformation, liposome-mediated transformation, transformation using wounded or enzyme-degraded immature embryos, or wounded or enzyme-degraded embryogenic callus, and the like. Transformation methods using Agrobacterium tumefaciens tumor inducing (Ti) plasmids or root-inducing (Ri) plasmids, or plant virus vectors are well known in the art (see, for example, WO 99/47552; Weissbach & Weissbach, “Methods for Plant Molecular Biology” (Academic Press, NY 1988), section VIII, pages 421–463; Grierson and Corey, “Plant Molecular Bioloy” 2d Ed. (Blackie, London 1988), Chapters 7–9, each of which is incorporated herein by reference; Horsch et al., supra, 1985). The wild-type form of Agrobacterium, for example, contains a Ti plasmid, which directs production of tumorigenic crown gall growth on host plants. Transfer of the tumor inducing T-DNA region of the Ti plasmid to a plant genome requires the Ti plasmid-encoded virulence genes as well as T-DNA borders, which are a set of direct DNA repeats that delineate the region to be transferred. An Agrobacterium based vector is a modified form of a Ti plasmid, in which the tumor inducing functions are replaced by a nucleotide sequence of interest that is to be introduced into the plant host.
Methods of using Agrobacterium mediated transformation include cocultivation of Agrobacterium with cultured isolated protoplasts; transformation of plant cells or tissues with Agrobacterium; and transformation of seeds, apices or meristems with Agrobacterium. In addition, in planta transformation by Agrobacterium can be performed using vacuum infiltration of a suspension of Agrobacterium cells (Bechtold et al., C. R. Acad. Sci. Paris 316:1194, 1993, which is incorporated herein by reference).
Agrobacterium mediated transformation can employ cointegrate vectors or binary vector systems, in which the components of the Ti plasmid are divided between a helper vector, which resides permanently in the Agrobacterium host and carries the virulence genes, and a shuttle vector, which contains the gene of interest bounded by T-DNA sequences. Binary vectors are well known in the art (see, for example, De Framond, BioTechnology 1:262, 1983; Hoekema et al., Nature 303:179, 1983, each of which is incorporated herein by reference) and are commercially available (Clontech; Palo Alto Calif.). For transformation, Agrobacterium can be cocultured, for example, with plant cells or wounded tissue such as leaf tissue, root explants, hypocotyledons, stem pieces or tubers (see, for example, Glick and Thompson, “Methods in Plant Molecular Biology and Biotechnology” (Boca Raton Fla., CRC Press 1993), which is incorporated herein by reference). Wounded cells within the plant tissue that have been infected by Agrobacterium can develop organs de novo when cultured under the appropriate conditions; the resulting transgenic shoots eventually give rise to transgenic plants, which contain an exogenous polynucleotide portion of a plant stress-regulated gene.
Agrobacterium mediated transformation has been used to produce a variety of transgenic plants, including, for example, transgenic cruciferous plants such as Arabidopsis, mustard, rapeseed and flax; transgenic leguminous plants such as alfalfa, pea, soybean, trefoil and white clover; and transgenic solanaceous plants such as eggplant, petunia, potato, tobacco and tomato (see, for example, Wang et al., “Transformation of Plants and Soil Microorganisms” (Cambridge, University Press 1995), which is incorporated herein by reference). In addition, Agrobacterium mediated transformation can be used to introduce an exogenous polynucleotide sequence, for example, a plant stress-regulated regulatory element into apple, aspen, belladonna, black currant, carrot, celery, cotton, cucumber, grape, horseradish, lettuce, morning glory, muskmelon, neem, poplar, strawberry, sugar beet, sunflower, walnut, asparagus, rice and other plants (see, for example, Glick and Thompson, supra, 1993; Hiei et al., Plant J. 6:271–282, 1994; Shimamoto, Science 270:1772–1773, 1995).
Suitable strains of Agrobacterium tumefaciens and vectors as well as transformation of Agrobacteria and appropriate growth and selection media are well known in the art (GV3101, pMK90RK), Koncz, Mol. Gen. Genet. 204:383–396, 1986; (C58C1, pGV3850kan), Deblaere, Nucl. Acid Res. 13:4777, 1985; Bevan, Nucl. Acid Res. 12:8711, 1984; Koncz, Proc. Natl. Acad. Sci. USA 86:8467–8471, 1986; Koncz, Plant Mol. Biol. 20:963–976, 1992; Koncz, Specialized vectors for gene tagging and expression studies. In: Plant Molecular Biology Manual Vol. 2, Gelvin and Schilperoort (Eds.), Dordrecht, The Netherlands: Kluwer Academic Publ. (1994), 1–22; European Patent A-1 20 516; Hoekema: The Binary Plant Vector System, Offsetdrukkerij Kanters B. V., Alblasserdam (1985), Chapter V; Fraley, Crit. Rev. Plant. Sci., 4:1–46; An, EMBO J. 4:277–287, 1985).
Where a polynucleotide portion of a plant stress-regulated gene is contained in vector, the vector can contain functional elements, for example “left border” and “right border” sequences of the T-DNA of Agrobacterium, which allow for stable integration into a plant genome. Furthermore, methods and vectors that permit the generation of marker-free transgenic plants, for example, where a selectable marker gene is lost at a certain stage of plant development or plant breeding, are known, and include, for example, methods of co-transformation (Lyznik, Plant Mol. Biol. 13:151–161, 1989; Peng, Plant Mol. Biol. 27:91–104, 1995), or methods that utilize enzymes capable of promoting homologous recombination in plants (see, e.g., WO97/08331; Bayley, Plant Mol. Biol. 18:353–361, 1992; Lloyd, Mol. Gen. Genet. 242:653–657, 1994; Maeser, Mol. Gen. Genet. 230:170–176, 1991; Onouchi, Nucl. Acids Res. 19:6373–6378, 1991; see, also, Sambrook et al., supra, 1989).
A direct gene transfer method such as electroporation also can be used to introduce a polynucleotide portion of a plant stress-regulated gene into a cell such as a plant cell. For example, plant protoplasts can be electroporated in the presence of the regulatory element, which can be in a vector (Fromm et al., Proc. Natl. Acad. Sci., USA 82:5824, 1985, which is incorporated herein by reference). Electrical impulses of high field strength reversibly permeabilize membranes allowing the introduction of the nucleic acid. Electroporated plant protoplasts reform the cell wall, divide and form a plant callus. Microinjection can be performed as described in Potrykus and Spangenberg (eds.), Gene Transfer To Plants (Springer Verlag, Berlin, N.Y. 1995). A transformed plant cell containing the introduced polynucleotide can be identified by detecting a phenotype due to the introduced polynucleotide, for example, increased or decreased tolerance to a stress condition.
Microprojectile mediated transformation also can be used to introduce a polynucleotide into a plant cell (Klein et al., Nature 327:70–73, 1987, which is incorporated herein by reference). This method utilizes microprojectiles such as gold or tungsten, which are coated with the desired nucleic acid molecule by precipitation with calcium chloride, spermidine or polyethylene glycol. The microprojectile particles are accelerated at high speed into a plant tissue using a device such as the BIOLISTIC PD-1000 (BioRad; Hercules Calif.).
Microprojectile mediated delivery (“particle bombardment”) is especially useful to transform plant cells that are difficult to transform or regenerate using other methods. Methods for the transformation using biolistic methods are well known (Wan, Plant Physiol. 104:37–48, 1984; Vasil, Bio/Technology 11:1553–1558, 1993; Christou, Trends in Plant Science 1:423–431, 1996). Microprojectile mediated transformation has been used, for example, to generate a variety of transgenic plant species, including cotton, tobacco, corn, hybrid poplar and papaya (see Glick and Thompson, supra, 1993). Important cereal crops such as wheat, oat, barley, sorghum and rice also have been transformed using microprojectile mediated delivery (Duan et al., Nature Biotech. 14:494–498, 1996; Shimamoto, Curr. Opin. Biotech. 5:158–162, 1994). A rapid transformation regeneration system for the production of transgenic plants such as a system that produces transgenic wheat in two to three months (see European Patent No. EP 0709462A2, which is incorporated herein by reference) also can be useful for producing a transgenic plant using a method of the invention, thus allowing more rapid identification of gene functions. The transformation of most dicotyledonous plants is possible with the methods described above. Transformation of monocotyledonous plants also can be transformed using, for example, biolistic methods as described above, protoplast transformation, electroporation of partially permeabilized cells, introduction of DNA using glass fibers, Agrobacterium mediated transformation, and the like.
Plastid transformation also can be used to introduce a polynucleotide portion of a plant stress-regulated gene into a plant cell (U.S. Pat. Nos. 5,451,513, 5,545,817, and 5,545,818; WO 95/16783; McBride et al., Proc. Natl. Acad. Sci., USA 91:7301–7305, 1994). Chloroplast transformation involves introducing regions of cloned plastid DNA flanking a desired nucleotide sequence, for example, a selectable marker together with polynucleotide of interest into a suitable target tissue, using, for example, a biolistic or protoplast transformation method (e.g., calcium chloride or PEG mediated transformation). One to 1.5 kb flanking regions (“targeting sequences”) facilitate homologous recombination with the plastid genome, and allow the replacement or modification of specific regions of the plastome. Using this method, point mutations in the chloroplast 16S rRNA and rps12 genes, which confer resistance to spectinomycin and streptomycin, can be utilized as selectable markers for transformation (Svab et al., Proc. Natl. Acad. Sci., USA 87:8526–8530, 1990; Staub and Maliga, Plant Cell 4:39–45, 1992), resulted in stable homopiasmic transformants; at a frequency of approximately one per 100 bombardments of target leaves. The presence of cloning sites between these markers allowed creation of a plastid targeting vector for introduction of foreign genes (Staub and Maliga, EMBO J. 12:601–606, 1993). Substantial increases in transformation frequency are obtained by replacement of the recessive rRNA or r-protein antibiotic resistance genes with a dominant selectable marker, the bacterial aadA gene encoding the spectinomycin-detoxifying enzyme aminoglycoside-3′-adenyltransferase (Svab and Maliga, Proc. Natl. Acad. Sci., USA 90:913–917, 1993). Approximately 15 to 20 cell division cycles following transformation are generally required to reach a homoplastidic state. Plastid expression, in which genes are inserted by homologous recombination into all of the several thousand copies of the circular plastid genome present in each plant cell, takes advantage of the enormous copy number advantage over nuclear-expressed genes to permit expression levels that can readily exceed 10% of the total soluble plant protein.
Plants suitable to treatment according to a method of the invention can be monocots or dicots and include, but are not limited to, corn (Zea mays), Brassica sp. (e.g., B. napus, B. rapa, B. juncea), particularly those Brassica species useful as sources of seed oil, alfalfa (Medicago sativa), rice (Oryza sativa), rye (Secale cereale), sorghum (Sorghum bicolor, Sorghum vulgare), millet (e.g., pearl millet (Pennisetum glaucum), proso millet (Panicum miliaceum), foxtail millet (Setaria italica), finger millet (Eleusine coracana)), sunflower (Helianthus annuus), safflower (Carthamus tinctorius), wheat (Triticum aestivum), soybean (Glycine max), tobacco (Nicotiana tabacum), potato (Solanum tuberosum), peanuts (Arachis hypogaea), cotton (Gossypium barbadense, Gossypium hirsutum), sweet potato (Ipomoea batatus), cassava (Manihot esculenta), coffee (Cofea spp.), coconut (Cocos nucifera), pineapple (Ananas comosus), citrus trees (Citrus spp.), cocoa (Theobroma cacao), tea (Camellia sinensis), banana (Musa spp.), avocado (Persea ultilane), fig (Ficus casica), guava (Psidium guajava), mango (Mangifera indica), olive (Olea europaea), papaya (Carica papaya), cashew (Anacardium occidentale), macadamia (Macadamia integrifolia), almond (Prunus amygdalus), sugar beets (Beta vulgaris), sugarcane (Saccharum spp.), oats, duckweed (Lemna), barley, tomatoes (Lycopersicon esculentum), lettuce (e.g., Lactuca sativa), green beans (Phaseolus vulgaris), lima beans (Phaseolus limensis), peas (Lathyrus spp.), and members of the genus Cucumis such as cucumber (C. sativus), cantaloupe (C. cantalupensis), and musk melon (C. melo).
Ornamentals such as azalea (Rhododendron spp.), hydrangea (Macrophylla hydrangea), hibiscus (Hibiscus rosasanensis), roses (Rosa spp.), tulips (Tulipa spp.), daffodils (Narcissus spp.), petunias (Petunia hybrida), carnation (Dianthus caryophyllus), poinsettia (Euphorbia pulcherrima), and chrysanthemum are also included. Additional ornamentals within the scope of the invention include impatiens, Begonia, Pelargonium, Viola, Cyclamen, Verbena, Vinca, Tagetes, Primula, Saint Paulia, Agertum, Amaranthus, Antihirrhinum, Aquilegia, Cineraria, Clover, Cosmo, Cowpea, Dahlia, Datura, Delphinium, Gerbera, Gladiolus, Gloxinia, Hippeastrum, Mesembryanthemum, Salpiglossos, and Zinnia.
Conifers that may be employed in practicing the present invention include, for example, pines such as loblolly pine (Pinus taeda), slash pine (Pinus elliotii), ponderosa pine (Pinus ponderosa), lodgepole pine (Pinus contorta), and Monterey pine (Pinus radiata), Douglas-fir (Pseudotsuga menziesii); Western hemlock (Tsuga ultilane); Sitka spruce (Picea glauca); redwood (Sequoia sempervirens); true firs such as silver fir (Abies amabilis) and balsam fir (Abies balsamea); and cedars such as Western red cedar (Thuja plicata) and Alaska yellow-cedar (Chamaecyparis nootkatensis).
Leguminous plants which may be used in the practice of the present invention include beans and peas. Beans include guar, locust bean, fenugreek, soybean, garden beans, cowpea, mungbean, lima bean, fava bean, lentils, chickpea, etc. Legumes include, but are not limited to, Arachis, e.g., peanuts, Vicia, e.g., crown vetch, hairy vetch, adzuki bean, mung bean, and chickpea, Lupinus, e.g., lupine, trifolium, Phaseolus, e.g., common bean and lima bean, Pisum, e.g., field bean, Melilotus, e.g., clover, Medicago, e.g., alfalfa, Lotus, e.g., trefoil, lens, e.g., lentil, and false indigo. Preferred forage and turf grass for use in the methods of the invention include alfalfa, orchard grass, tall fescue, perennial ryegrass, creeping bent grass, and redtop.
Other plants within the scope of the invention include Acacia, aneth, artichoke, arugula, blackberry, canola, cilantro, clementines, escarole, eucalyptus, fennel, grapefruit, honey dew, jicama, kiwifruit, lemon, lime, mushroom, nut, okra, orange, parsley, persimmon, plantain, pomegranate, poplar, radiata pine, radicchio, Southern pine, sweetgum, tangerine, triticale, vine, yams, apple, pear, quince, cherry, apricot, melon, hemp, buckwheat, grape, raspberry, chenopodium, blueberry, nectarine, peach, plum, strawberry, watermelon, eggplant, pepper, cauliflower, Brassica, e.g., broccoli, cabbage, ultilan sprouts, onion, carrot, leek, beet, broad bean, celery, radish, pumpkin, endive, gourd, garlic, snapbean, spinach, squash, turnip, ultilane, chicory, groundnut and zucchini.
Angiosperms are divided into two broad classes based on the number of cotyledons, which are seed leaves that generally store or absorb food; a monocotyledonous angiosperm has a single cotyledon, and a dicotyledonous angiosperm has two cotyledons. Angiosperms produce a variety of useful products including materials such as lumber, rubber, and paper; fibers such as cotton and linen; herbs and medicines such as quinine and vinblastine; ornamental flowers such as roses and orchids; and foodstuffs such as grains, oils, fruits and vegetables.
Angiosperms encompass a variety of flowering plants, including, for example, cereal plants, leguminous plants, oilseed plants, hardwood trees, fruit-bearing plants and ornamental flowers, which general classes are not necessarily exclusive. Cereal plants, which produce an edible grain cereal, include, for example, corn, rice, wheat, barley, oat, rye, orchardgrass, guinea grass, sorghum and turfgrass. Leguminous plants include members of the pea family (Fabaceae) and produce a characteristic fruit known as a legume. Examples of leguminous plants include, for example, soybean, pea, chickpea, moth bean, broad bean, kidney bean, lima bean, lentil, cowpea, dry bean, and peanut, as well as alfalfa, birdsfoot trefoil, clover and sainfoin. Oilseed plants, which have seeds that are useful as a source of oil, include soybean, sunflower, rapeseed (canola) and cottonseed.
Angiosperms also include hardwood trees, which are perennial woody plants that generally have a single stem (trunk). Examples of such trees include alder, ash, aspen, basswood (linden), beech, birch, cherry, cottonwood, elm, eucalyptus, hickory, locust, maple, oak, persimmon, poplar, sycamore, walnut, sequoia, and willow. Trees are useful, for example, as a source of pulp, paper, structural material and fuel.
Angiosperms are fruit-bearing plants that produce a mature, ripened ovary, which generally contains seeds. A fruit can be suitable for human or animal consumption or for collection of seeds to propagate the species. For example, hops are a member of the mulberry family that are prized for their flavoring in malt liquor. Fruit-bearing angiosperms also include grape, orange, lemon, grapefruit, avocado, date, peach, cherry, olive, plum, coconut, apple and pear trees and blackberry, blueberry, raspberry, strawberry, pineapple, tomato, cucumber and eggplant plants. An ornamental flower is an angiosperm cultivated for its decorative flower. Examples of commercially important ornamental flowers include rose, orchid, lily, tulip and chrysanthemum, snapdragon, camellia, carnation and petunia plants. The skilled artisan will recognize that the methods of the invention can be practiced using these or other angiosperms, as desired, as well as gymnosperms, which do not produce seeds in a fruit.
A method of producing a transgenic plant can be performed by introducing a polynucleotide portion of plant stress-regulated gene into a plant cell genome, whereby the polynucleotide portion of the plant stress-regulated gene modulates a response of the plant cell to a stress condition, thereby producing a transgenic plant, which comprises plant cells that exhibit altered responsiveness to the stress condition. In one embodiment, the polynucleotide portion of the plant stress-regulated gene encodes a stress-regulated polypeptide or functional peptide portion thereof, wherein expression of the stress-regulated polypeptide or functional peptide portion thereof either increases the stress tolerance of the transgenic plant, or decreases the stress tolerance of the transgenic plant. The polynucleotide portion of the plant stress-regulated gene encoding the stress-regulated polypeptide or functional peptide portion thereof can be operatively linked to a heterologous promoter.
In another embodiment, the polynucleotide portion of the plant stress-regulated gene comprises a stress-regulated regulatory element. The stress-regulated regulatory element can integrate into the plant cell genome in a site-specific manner, whereupon it can be operatively linked to an endogenous nucleotide sequence, which can be expressed in response to a stress condition specific for the regulatory element; or can be a mutant regulatory element, which is not responsive to the stress condition, whereby upon integrating into the plant cell genome, the mutant regulatory element disrupts an endogenous stress-regulated regulatory element of a plant stress-regulated gene, thereby altering the responsiveness of the plant stress-regulated gene to the stress condition. Accordingly, the invention also provides genetically modified plants, including transgenic plants, produced by such a method, and a plant cell obtained from such genetically modified plant, wherein said plant cell exhibits altered responsiveness to the stress condition; a seed produced by a transgenic plant; and a cDNA library prepared from a transgenic plant.
Also provided is a method of modulating the responsiveness of a plant cell to a stress condition. Such a method can be performed, for example, by introducing a polynucleotide portion of a plant stress-regulated gene into the plant cell, thereby modulating the responsiveness of the plant cell to a stress condition. As disclosed herein, the responsiveness of the plant cell can be increased or decreased upon exposure to the stress condition, and the altered responsiveness can result in increased or decreased tolerance of the plant cell to a stress condition. The polynucleotide portion of the plant stress-regulated gene can, but need not, be integrated into the genome of the plant cell, thereby modulating the responsiveness of the plant cell to the stress condition. Accordingly, the invention also provide a genetically modified plant, including a transgenic plant, which contains an introduced polynucleotide portion of a plant stress-regulated gene, as well as plant cells, tissues, and the like, which exhibit modulated responsiveness to a stress condition.
The polynucleotide portion of the plant stress-regulated gene can encode a stress-regulated polypeptide or functional peptide portion thereof, which can be operatively linked to a heterologous promoter. As used herein, reference to a “functional peptide portion of a plant stress-regulated polypeptide” means a contiguous amino acid sequence of the polypeptide that has an activity of the full length polypeptide, or that has an antagonist activity with respect to the fall length polypeptide, or that presents an epitope unique to the polypeptide. Thus, by expressing a functional peptide portion of a plant stress-regulated polypeptide in a plant cell, the peptide can act as an agonist or an antagonist of the polypeptide, thereby modulating the responsiveness of the plant cell to a stress condition.
A polynucleotide portion of the plant stress-regulated nucleotide sequence also can contain a mutation, whereby upon integrating into the plant cell genome, the polynucleotide disrupts (knocks-out) an endogenous plant stress-regulated nucleotide sequence, thereby modulating the responsiveness of said plant cell to the stress condition. Depending on whether the knocked-out gene encodes an adaptive or a maladaptive stress-regulated polypeptide, the responsiveness of the plant will be modulated accordingly. Thus, a method of the invention provides a means of producing a transgenic plant having a knock-out phenotype of a plant stress-regulated nucleotide sequence.
Alternatively, the responsiveness of a plant or plant cell to a stress condition can be modulated by use of a suppressor construct containing dominant negative mutation for any of the stress-regulated sequences described herein. Expression of a suppressor construct containing a dominant mutant mutation generates a mutant transcript that, when coexpressed with the wild-type transcript inhibits the action of the wild-type transcript. Methods for the design and use of dominant negative constructs are well known (see, for example, in Herskowitz, Nature 329:219–222, 1987; Lagna and Hemmati-Brivanlou, Curr. Topics Devel. Biol. 36:75–98, 1998).
The polynucleotide portion of the plant stress-regulated gene also can comprise a stress-regulated regulatory element, which can be operatively linked to a heterologous nucleotide sequence, which, upon expression from the regulatory element in response to a stress condition, modulates the responsiveness of the plant cell to the stress condition. Such a heterologous nucleotide sequence can encode, for example, a stress-inducible transcription factor such as DREB1A, which, upon exposure to the stress condition, is expressed such that it can amplify the stress response (see Kasuga et al., supra, 1999). The heterologous nucleotide sequence also can encode a polynucleotide that is specific for a plant stress-regulated gene, for example, an antisense molecule, a ribozyme, and a triplexing agent, either of which, upon expression in the plant cell, reduces or inhibits expression of a stress-regulated polypeptide encoded by the gene, thereby modulating the responsiveness of the plant cell to a stress condition, for example, an abnormal level of cold, osmotic pressure, and salinity. As used herein, the term “abnormal,” when used in reference to a condition such as temperature, osmotic pressure, salinity, or any other condition that can be a stress condition, means that the condition varies sufficiently from a range generally considered optimum for growth of a plant that the condition results in an induction of a stress response in a plant. Methods of determining whether a stress response has been induced in a plant are disclosed herein or otherwise known in the art.
A plant stress-regulated regulatory element can be operatively linked to a heterologous polynucleotide sequence, such that the regulatory element can be introduced into a plant genome in a site-specific matter by homologous recombination. For example, a mutant plant stress-regulated regulatory element for a maladaptive stress-induced polypeptide can be transformed into a plant genome in a site specific manner by in vivo mutagenesis, using a hybrid RNA-DNA oligonucleotide (“chimeroplast” (TIBTECH 15:441–447, 1997; WO 95/15972; Kren, Hepatology 25:1462–1468, 1997; Cole-Strauss, Science 273:1386–1389, 1996, each of which is incorporated herein by reference). Part of the DNA component of the RNA-DNA oligonucleotide is homologous to a nucleotide sequence comprising the regulatory element of the maladaptive gene, but includes a mutation or contains a heterologous region which is surrounded by the homologous regions. By means of base pairing of the homologous regions of the RNA-DNA oligonucleotide and of the endogenous nucleic acid molecule, followed by a homologous recombination the mutation contained in the DNA component of the RNA-DNA oligonucleotide or the heterologous region can be transferred to the plant genome, resulting in a “mutant” gene that, for example, is not induced in response to a stress and, therefore, does not confer the maladaptive phenotype. Such a method similarly can be used to knock-out the activity of a stress-regulated gene, for example, in an undesirable plant. Such a method can provide the advantage that a desirable wild-type plant need not compete with the undesirable plant, for example, for light, nutrients, or the like.
A method of modulating the responsiveness of a plant cell to a stress condition also can be performed by introducing a mutation in the chromosomal copy of a plant stress-regulated gene, for example, in the stress-regulated regulatory element, by transforming a cell with a chimeric oligonucleotide composed of a contiguous stretch of RNA and DNA residues in a duplex conformation with double hairpin caps on the ends. An additional feature of the oligonucleotide is the presence of 2′-0-methylation at the RNA residues. The RNA/DNA sequence is designed to align with the sequence of a chromosomal copy of the target regulatory element and to contain the desired nucleotide change (see U.S. Pat. No. 5,501,967, which is incorporated herein by reference).
A plant stress-regulated regulatory element also can be operatively linked to a heterologous polynucleotide such that, upon expression from the regulatory element in the plant cell, confers a desirable phenotype on the plant cell. For example, the heterologous polynucleotide can encode an aptamer, which can bind to a stress-induced polypeptide. Aptamers are nucleic acid molecules that are selected based on their ability to bind to and inhibit the activity of a protein or metabolite. Aptamers can be obtained by the SELEX (Systematic Evolution of Ligands by Exponential Enrichment) method (see U.S. Pat. No. 5,270,163), wherein a candidate mixture of single stranded nucleic acids having regions of randomized sequence is contacted with a target, and those nucleic acids having a specific affinity to the target are partitioned from the remainder of the candidate mixture, and amplified to yield a ligand enriched mixture. After several iterations a nucleic acid molecule (aptamer) having optimal affinity for the target is obtained. For example, such a nucleic acid molecule can be operatively linked to a plant stress-regulated regulatory element and introduced into a plant. Where the aptamer is selected for binding to a polypeptide that normally is expressed from the regulatory element and is involved in an adaptive response of the plant to a stress, the recombinant molecule comprising the aptamer can be useful for inhibiting the activity of the stress-regulated polypeptide, thereby decreasing the tolerance of the plant to the stress condition.
The invention provides a genetically modified plant, which can be a transgenic plant, that is tolerant or resistant to a stress condition. As used herein, the term “tolerant” or “resistant,” when used in reference to a stress condition of a plant, means that the particular plant, when exposed to a stress condition, shows less of an effect, or no effect, in response to the condition as compared to a corresponding reference plant (naturally occurring wild-type plant or a plant not containing a construct of the present invention). As a consequence, a plant encompassed within the present invention grows better under more widely varying conditions, has higher yields and/or produces more seeds. Thus, a transgenic plant produced according to a method of the invention can demonstrate protection (as compared to a corresponding reference plant) from a delay to complete inhibition of alteration in cellular metabolism, or reduced cell growth or cell death caused by the stress. Preferably, the transgenic plant is capable of substantially normal growth under environmental conditions where the corresponding reference plant shows reduced growth, metabolism or viability, or increased male or female sterility.
The determination that a plant modified according to a method of the invention has increased resistance to a stress-inducing condition can be made by comparing the treated plant with a control (reference) plant using well known methods. For example, a plant having increased tolerance to saline stress can be identified by growing the plant on a medium such as soil, which contains a higher content of salt in the order of at least about 10% compared to a medium the corresponding reference plant is capable of growing on. Advantageously, a plant treated according to a method of the invention can grow on a medium or soil containing at least about 50%, or more than about 75%, particularly at least about more than 100%, and preferably more than about 200% salt than the medium or soil on which a corresponding reference plant can grow. In particular, such a treated plant can grow on medium or soil containing at least 40 mM, generally at least 100 mM, particularly at least 200 mM, and preferably at least 300 mM salt, including, for example, a water soluble inorganic salt such as sodium sulfate, magnesium sulfate, calcium sulfate, sodium chloride, magnesium chloride, calcium chloride, potassium chloride, or the like; salts of agricultural fertilizers, and salts associated with alkaline or acid soil conditions; particularly NaCl.
In another embodiment, the invention provides a plant that is less tolerant or less resistant to a stress condition as compared to a corresponding reference plant. As used herein, the term “less tolerant” or “less resistant,” when used in reference to a stress condition of a plant, means that the particular plant, when exposed to a stress condition, shows an alteration in response to the condition as compared to a corresponding reference plant. As a consequence, such a plant, which generally is an undesirable plant species, is less likely to grow when exposed to a stress condition than an untreated plant.
The present invention also relates to a method of expressing a heterologous nucleotide sequence in a plant cell. Such a method can be performed, for example, by introducing into the plant cell a plant stress-regulated regulatory element operatively linked to the heterologous nucleotide sequence, whereby, upon exposure of the plant cell to stress condition, the heterologous nucleotide sequence is expressed in the plant cell. The heterologous nucleotide sequence can encode a selectable marker, or preferably, a polypeptide that confers a desirable trait upon the plant cell, for example, a polypeptide that improves the nutritional value, digestibility or ornamental value of the plant cell, or a plant comprising the plant cell. Accordingly, the invention provides a transgenic plant that, in response to a stress condition, can produce a heterologous polypeptide from a plant stress-regulated regulatory element. Such transgenic plants can provide the advantage that, when grown in a cold environment for example, expression of the heterologous polypeptide from a plant cold-regulated regulatory element can result in increased nutritional value of the plant.
The present invention further relates to a method of modulating the activity of a biological pathway in a plant cell, wherein the pathway involves a stress-regulated polypeptide. As used herein, reference to a pathway that “involves” a stress-regulated polypeptide means that the polypeptide is required for normal function of the pathway. For example, plant stress-regulated polypeptides as disclosed herein include those acting as kinases or as transcription factors, which are well known to be involved in signal transduction pathways. As such, a method of the invention provides a means to modulate biological pathways involving plant stress-regulated polypeptides, for example, by altering the expression of the polypeptides in response to a stress condition. Thus, a method of the invention can be performed, for example, by introducing a polynucleotide portion of a plant stress-regulated gene into the plant cell, thereby modulating the activity of the biological pathway.
A method of the invention can be performed with respect to a pathway involving any of the stress-regulated polypeptides as encoded by a polynucleotide of SEQ ID NOS:1–2703, including for example, a stress-regulated transcription factor, an enzyme, including a kinase, a channel protein (see, for example, Tables 29–31; see, also, Table 1). Pathways in which the disclosed stress-regulated stress factors are involved can be identified, for example, by searching the Munich Information Center for Protein Sequences (MIPS) Arabidopsis thaliana database (MATDB), which is at http://www.mips.biochem.mpg.de/proj/thal/.
The present invention also relates to a method of identifying a polynucleotide that modulates a stress response in a plant cell. Such a method can be performed, for example, by contacting an array of probes representative of a plant cell genome and nucleic acid molecules expressed in plant cell exposed to the stress; detecting a nucleic acid molecule that is expressed at a level different from a level of expression in the absence of the stress; introducing the nucleic acid molecule that is expressed differently into a plant cell; and detecting a modulated response of the plant cell containing the introduced nucleic acid molecule to a stress, thereby identifying a polynucleotide that modulates a stress response in a plant cell. The contacting is under conditions that allow for selective hybridization of a nucleic acid molecule with probe having sufficient complementarity, for example, under stringent hybridization conditions.
As used herein, the term “array of probes representative of a plant cell genome” means an organized group of oligonucleotide probes that are linked to a solid support, for example, a microchip or a glass slide, wherein the probes can hybridize specifically and selectively to nucleic acid molecules expressed in a plant cell. Such an array is exemplified herein by a GeneChip® Arabidopsis Genome Array (Affymetrix; see Example 1). In general, an array of probes that is “representative” of a plant genome will identify at least about 30% or the expressed nucleic acid molecules in a plant cell, generally at least about 50% or 70%, particularly at least about 80% or 90%, and preferably will identify all of the expressed nucleic acid molecules. It should be recognized that the greater the representation, the more likely all nucleotide sequences of cluster of stress-regulated genes will be identified.
A method of the invention is exemplified in Example 1, wherein clusters of Arabidopsis genes induced to cold, to increased salinity, to increased osmotic pressure, and to a combination of the above three stress conditions were identified. Based on the present disclosure, the artisan readily can obtain nucleic acid samples for Arabidopsis plants exposed to other stress conditions, or combinations of stress conditions, and identify clusters of genes induced in response to the stress conditions. Similarly, the method is readily adaptable to identifying clusters of stress-regulated genes expressed in other plant species, particularly commercially valuable plant species, where a substantial amount of information is known regarding the genome.
The clusters of genes identified herein include those clusters of genes that are induced or repressed in response to a combination of stress conditions, but not to any of the stress conditions alone; and clusters of genes that are induced or repressed in response to a selected stress condition, but not to other stress conditions tested. Furthermore, clusters of genes that respond to a stress condition in a temporally regulated manner are also included, such as gene clusters that are induced early (for example, within about 3 hours), late (for example, after about 8 to 24 hours), or continuously in a stress response. In addition, the genes within a cluster are represented by a variety of cellular proteins, including transcription factors, enzymes such as kinases, channel proteins, and the like (see Tables 1 and 29–31). Thus, the present invention further characterizes nucleotide sequences that previously were known to encode cellular peptides by classifying them within clusters of stress-regulated genes.
The present invention additionally relates to a method of identifying a stress condition to which a plant cell was exposed. Such a method can be performed, for example, by contacting nucleic acid molecules expressed in the plant cell and an array of probes representative of the plant cell genome; and detecting a profile of expressed nucleic acid molecules characteristic of a stress response, thereby identifying the stress condition to which the plant cell was exposed. The contacting generally is under conditions that allow for selective hybridization of a nucleic acid molecule with probe having sufficient complementarity, for example, under stringent hybridization conditions. The profile can be characteristic of exposure to a single stress condition, for example, an abnormal level of cold, osmotic pressure, or salinity (Tables 3–14), or can be characteristic of exposure to more than one stress condition (Tables 15–26, for example, cold, increased osmotic pressure and increased salinity (see Tables 24–26).
The method can be practiced using at least one nucleic acid probe and can identify one or combination of stress conditions by detecting altered expression of one or a plurality of polynucleotides representative of plant stress-regulated genes. As used herein, the term “at least one” includes one, two, three or more, for example, five, ten, twenty, fifty or more polynucleotides, nucleic acid probes, and the like. The term “plurality” is used herein to mean two or more, for example, three, four, five or more, including ten, twenty, fifty or more polynucleotides, nucleic acid probes, and the like.
In a method of the invention, nucleic acid samples from the plant cells to be collected can be contacted with an array, then the profile can be compared with known expression profiles prepared from nucleic acid samples of plants exposed to a known stress condition or combination of stress conditions. By creating a panel of such profiles, representative of various stress conditions, an unknown stress condition to which a plant was exposed can be identified simply by comparing the unknown profile with the known profiles and determining which known profile that matches the unknown profile. Preferably, the comparison is automated. Such a method can be useful, for example, to identify a cause of damage to a crop, where the condition causing the stress is not known or gradually increases over time. For example, accumulation in soils over time of salts from irrigation water can result in gradually decreasing crop yields. Because the accumulation is gradual, the cause of the decreased yield may not be readily apparent. Using the present methods, it is possible to evaluate the stress to which the plants are exposed, thus revealing the cause of the decreased yields.
The present invention, therefore includes a computer readable medium containing executable instructions form receiving expression data for sequences substantially similar to any of those disclosed herein and comparing expression data from a test plant to a reference plant that has been exposed to an abiotic stress. Also provided is a computer-readable medium containing sequence data for sequences substantially similar to any of the sequences described herein, or the complements thereof, and a module for comparing such sequences to other nucleic acid sequences.
Also provided are plants and plant cells comprising plant stress-regulatory elements of the present invention operably linked to a nucleotide sequence encoding a detectable signal. Such plants can be used as diagnostic or “sentinel” plants to provide early warning that nearby plants are being stressed so that appropriate actions can be taken. In one embodiment, the signal is one that alters the appearance of the plant. For example, an osmotic stress regulatory element of the present invention can be operably linked to a nucleotide sequence encoding a fluorescent protein such as green fluorescent protein. When subjected to osmotic stress, the expression of the green fluorescent protein in the sentinel plant provides a visible signal so that appropriate actions can be taken to remove or alleviate the stress. The use of fluorescent proteins in plants is well known (see, for example, in Leffel et al., BioTechniques 23:912, 1997).
The invention further relates to a method of identifying an agent that modulates the activity of a stress-regulated regulatory element of a plant. As used herein, the term “modulate the activity,” when used in reference to a plant stress-regulated regulatory element, means that expression of a polynucleotide from the regulatory element is increased or decreased. In particular, expression can be increased or decreased with respect to the basal activity of the promoter, i.e., the level of expression, if any, in the absence of a stress condition that normally induces expression from the regulatory element; or can be increased or decreased with respect to the level of expression in the presence of the inducing stress condition. As such, an agent can act as a mimic of a stress condition, or can act to modulate the response to a stress condition.
Such a method can be performed, for example, by contacting the regulatory element with an agent suspected of having the ability to modulate the activity of the regulatory element, and detecting a change in the activity of the regulatory element. In one embodiment, the regulatory element can be operatively linked to a heterologous polynucleotide encoding a reporter molecule, and an agent that modulates the activity of the stress-regulated regulatory element can be identified by detecting a change in expression of the reporter molecule due to contacting the regulatory element with the agent. Such a method can be performed in vitro in a plant cell-free system, or in a plant cell in culture or in a plant in situ.
A method of the invention also can be performed by contacting the agent is contacted with a genetically modified cell or a transgenic plant containing an introduced plant stress-regulated regulatory element, and an agent that modulates the activity of the regulatory element is identified by detecting a phenotypic change in the modified cell or transgenic plant.
A method of the invention can be performed in the presence or absence of the stress condition to which the particularly regulatory element is responsive. As such, the method can identify an agent that modulates the activity of plant stress-regulated promoter in response to the stress, for example, an agent that can enhance the stress response or can reduce the stress response. In particular, a method of the invention can identify an agent that selectively activates the stress-regulated regulatory elements of a cluster of plant stress-regulated genes, but does not affect the activity of other stress-regulated regulatory genes. As such, the method provides a means to identify an agent that acts as a stress mimic. Such agents can be particularly useful to prepare a plant to an expected stress condition. For example, a agent that acts as a cold mimic can be applied to a field of plants prior to the arrival of an expected cold front. Thus, the cold stress response can be induced prior to the actual cold weather, thereby providing the plants with the protection of the stress response, without the plants suffering from any initial damage due to the cold. Similarly, an osmotic pressure mimic can be applied to a crop of plants prior a field being flooded by a rising river.
In one embodiment, the present invention provides a method for marker-assisted selection. Marker-assisted selection involves the selection of plants having desirable phenotypes based on the presence of particular nucleotide sequences (“markers”). The use of markers allows plants to be selected early in development, often before the phenotype would normally be manifest. Because it allows for early selection, marker-assisted selection decreases the amount of time need for selection and thus allows more rapid genetic progress.
Briefly, marker-assisted selection involves obtaining nucleic acid from a plant to be selected. The nucleic acid obtained is then probed with probes that selectively hybridize under stringent, preferably highly stringent, conditions to a nucleotide sequence or sequences associated with the desired phenotype. In one embodiment, the probes hybridize to any of the stress-responsive genes or regulatory regions disclosed herein, for example, any one of SEQ ID NOS:1–2703. The presence of any hybridization products formed is detected and plants are then selected on the presence or absence of the hybridization products.
The following examples are intended to illustrate but not limit the invention.
EXAMPLE 1
Profiling of Plant Stress-Regulated Genes
This example demonstrates that clusters of stress-regulated genes can be identified in plant cells exposed to various stress conditions, either alone or in combination.
A GeneChip® Arabidopsis Genome Array (Affymetrix, Santa Clara, Calif.) was used to identify clusters of genes that were coordinately induced in response to various stress conditions. The GeneChip® Arabidopsis Genome Array contains probes synthesized in situ and is designed to measure temporal and spatial gene expression of approximately 8700 genes in greater than 100 EST clusters. The sequences used to develop the array were obtained from GenBank (http://www.ncbi.nlm.nih.gov/) in collaboration with Torrey Mesa Research Institute (San Diego, Calif.), formerly known as Novartis Agriculture Discovery Institute. Eighty percent of the nucleotide sequences represented on the array are predicted coding sequences from genomic BAC entries; twenty percent are high quality cDNA sequences. The array also contains over 100 EST clusters that share homology with the predicted coding sequences from BAC clones (see, for example, world wide web at address (url) “affymetrix.com/products/Arabidopsis_content.html”.
The Affymetrix GeneChip® array was used to define nucleotide sequences/pathways affected by various abiotic stresses and to define which are uniquely regulated by one stress and those that respond to multiple stress, and to identify candidate nucleotide sequences for screening for insertional mutants. Of the approximately 8,700 nucleotide sequences represented on the Affymetrix GeneChip® array, 2862 nucleotide sequences showed at least a 2-fold change in expression in at least one sample, relative to no-treatment controls. Of those 2,862 nucleotide sequences 1,335 were regulated only by cold stress, 166 were regulated only mannitol stress and 209 were regulated only by saline stress. Furthermore, of the 2,862 nucleotide sequences 123 nucleotide sequences were regulated by salt and mannitol stress, 293 were regulated by mannitol and cold stress, 274 were regulated by cold and saline stress and 462 were regulated by cold, mannitol and salt. Of the 2,862 nucleotide sequences, 771 passed the higher stringency of showing at least a 2-fold change in expression in at least 2 samples, relative to control. And, 508 of the 771 nucleotide sequences were found in an in-house collection of insertion mutants.
The following describes in more detail how the experiments were done. Transcriptional profiling was performed by hybridizing fluorescence labeled cRNA with the oligonucleotides probes on the chip, washing, and scanning. Each gene is represented on the chip by about sixteen oligonucleotides (25-mers). Expression level is related to fluorescence intensity. Starting material contained 1 to 10 μg total RNA; detection specificity was about 1:106; approximately a 2-fold change was detectable, with less than 2% false positive; the dynamic range was approximately 500×. Nucleotide sequences having up to 70% to 80% identity could be discriminated using this system.
Seven day old axenic Arabidopsis seedlings were transferred to Magenta boxes with rafts floating on MS medium. Three weeks later (28 day old seedlings), stresses were applied as follows: Control−no treatment; Cold−Magenta box placed in ice; Mannitol−medium+200 mM mannitol; Salt−medium+100 mM NaCl. Tissue samples were collected at 3 hours and 27 hours into the stress, roots and aerial portions were harvested, RNA was purified, and the samples were analyzed using the GeneChip® Arabidopsis Genome Array (Affymetrix, Santa Clara, Calif.) following the manufacturer's protocol.
Raw fluorescence values as generated by Affymetrix software were processed as follows: the values were brought into Microsoft Excel and values of 25 or less were set to 25 (an empirically determined baseline, Zhu and Wang, Plant Physiol. 124:1472–1476; 2000). The values from the stressed samples were then converted to fold change relative to control by dividing the values from the stressed samples by the values from the no-treatment control samples. Expression patterns that were altered at least 2-fold with respect to the control were selected. This method gave very robust results and resulted in a larger number of nucleotide sequences called as stress-regulated than previous methods had permitted.
Based on the profiles obtained following hybridization of nucleic acid molecules obtained from plant cells exposed to various stress conditions to the probes in the microarray, clusters of nucleotide sequences that were altered in response to the stress conditions were identified (see Tables 3–6, cold responsive; Tables 7–10, salt (saline) responsive; Tables 11 to 14, mannitol (osmotic) responsive; Tables 15–17, cold and mannitol responsive; Tables 18–20, 6 salt and cold responsive; Tables 21–23, salt and mannitol responsive; Tables 24–26, cold, salt and mannitol responsive. Examples of plant gene sequences that varied in expression at least two-fold in response to a combination of cold, saline and osmotic stress in root cells and leaf cells are shown in Tables 27 and 28, respectively. In addition, examples of plant gene sequences that encode transcription factors (Table 29), phosphatases (Table 30), and kinases (Table 31) and that varied at least two-fold in response to a combination of cold, saline and osmotic stress are provided.
Affymetrix ID numbers and corresponding SEQ ID NOS: for the respective Arabidopsis nucleotide sequences are provided Tables 3–26, and can be used to determine SEQ ID NOS: for the sequences shown by Affymetrix ID number in Tables 27–31. The Affymetrix ID number refers to a particular nucleotide sequence on the GeneChip® Arabidopsis Genome Array. In some cases, a particular plant stress-regulated gene sequence hybridized to more than one nucleotide sequence on the GeneChip® Arabidopsis Genome Array (see, for example, Table 3, where SEQ ID NO:36 is shown to have hybridized to the 12187_AT and 15920_I_AT nucleotide sequences on the GeneChip®). In addition, it should be recognized that the disclosed sequences are not limited to coding sequences but, in some cases, include 5′ untranslated sequences (see Table 2) or a longest coding region. As such, while the sequences set forth as SEQ ID NOS:1–2073 generally start with an ATG codon, in most cases each comprises a longer nucleotide sequence, including a regulatory region (see Table 2).
The results disclosed herein demonstrate that several polynucleotides, some of which were known to function as transcription factors, enzymes, and structural proteins, also are involved in the response of a plant cell to stress. The identification of the clusters of stress-regulated genes as disclosed herein provides a means to identify stress-regulated regulatory elements present in Arabidopsis thaliana nucleotide sequences, including consensus regulatory elements. It should be recognized, however that the regulatory elements of the plant genes comprising a sequence as set forth in SEQ ID NOS:156, 229, 233, 558, 573, 606, 625, 635, 787, and 813, which previously have been described as cold regulated genes, are not encompassed within the stress-regulated gene regulatory element of the invention, and the regulatory elements of the plant genes comprising the nucleotide sequences set forthas SEQ ID NOS:1263, 1386, 1391, 1405, 1445, 1484, 1589, 1609, 1634, 1726, 1866, 1918, and 1928, which previously have been identified as genes that are responsive to a single stress condition such as cold or saline stress, are not encompassed within the plant stress-regulated gene regulatory elements of the invention to the extent that they confer stress-regulated expression only with respect to the known single stress. Furthermore, the identification of the Arabidopsis stress-regulated genes provides a means to identify the corresponding homologs and orthologs in other plants, including commercially valuable food crops such as wheat, rice, soy, and barley, and ornamental plants. BLASTN and BLASTP searches to identify such sequences revealed the polynucleotide sequences set forth in Table 32, which is on the CD-R compact disc submitted herewith.
Although the invention has been described with reference to the above example, it will be understood that modifications and variations are encompassed within the spirit and scope of the invention. Accordingly, the invention is limited only by the claims, which follow Tables 1 to 31.







TABLE 1







SEQUENCE DESCRIPTIONS










Seq

Seq



ID
Description
ID
Description













1
unknown protein
1352
bZIP transcription factor-like





protein


2
unknown protein
1353
Medicago nodulin N21-like protein


3
unknown protein
1354
putative endo-1,4-beta glucanase


4
putative auxin-induced
1355
1-aminocyclopropane-1-



protein

carboxylate oxidase


5
unknown protein
1356
putative anion exchange protein


6
hypothetical protein
1357
SRG1-like protein


7
putative protein
1358
putative protein


8
unknown protein
1359
putative phi-1-like phosphate-





induced protein


9
unknown protein
1360
putative protein


10
unknown protein
1361
putative embryo-abundant protein


11
putative protein
1362
putative hydrolase


12
Thioredoxin-like protein
1363
unknown protein


13
putative RNA helicase
1364
unknown protein


14
putative protein
1365
hexose transporter-like protein


15
putative protein
1366
unknown protein


16
RING zinc finger protein,
1367
unknown protein



putative


17
putative cyclin
1368
peptide transport-like protein


18
putative protein
1369
unknown protein


19
putative protein
1370
putative peptide transporter


20
unknown protein
1371
disease resistance protein, putative


21
putative protein
1372
cysteine protease component of





protease-inhibitor complex


22
putative protein
1373
putative cytochrome P450


23
hypothetical protein
1374
putative protein


24
unknown protein
1375
hypothetical protein


25
hypothetical protein
1376
unknown protein


26
unknown protein
1377
putative





phosphoribosylaminoimidazolecar





boxamide formyltransferase


27
unknown protein
1378
putative protein


28
unknown protein
1379
HSP like protein


29
unknown protein
1380
unknown protein


30
putative protein
1381
unknown protein


31
putative protein
1382
putative cytochrome P450


32
putative protein
1383
similar to pectinesterase


33
unknown protein
1384
putative glucosyltransferase


34
putative ribonuclease III
1385
thaumatin-like protein


35
unknown protein
1386
drought-inducible cysteine





proteinase RD19A precursor


36
unknown protein
1387
vegetative storage protein Vsp2


37
unknown protein
1388
unknown protein


38
unknown protein
1389
unknown protein


39
unknown protein
1390
anthranilate N-





benzoyltransferase-like





protein


40
putative histidine kinase
1391
delta-1-pyrroline 5-





carboxylase synthetase





(P5C1)


41
scarecrow-like 7 (SCL7)
1392
glutathione S-conjugate





transporting ATPase





(AtMRP1)


42
putative protein
1393
hypothetical protein


43
No function assigned by TIGR
1394
hypothetical protein


44
unknown protein
1395
unknown protein


45
unknown protein
1396
putative protein


46
succinyl-CoA-ligase alpha subunit
1397
putative protein


47
putative protein
1398
No function assigned by





TIGR


48
CLV1 receptor kinase like protein
1399
unknown protein


49
putative receptor-like protein
1400
putative protein kinase



kinase


50
putative squalene synthase
1401
unknown protein


51
putative receptor protein kinase
1402
hypothetical protein


52
somatic embryogenesis receptor-
1403
unknown protein



like kinase, putative


53
putative protein
1404
putative calcium-binding





EF-hand protein


54
putative beta-glucosidase
1405
cinnamyl-alcohol





dehydrogenase ELI3-1


55
multi-drug resistance protein
1406
putative protein


56
receptor protein kinase (TMK1),
1407
unknown protein



putative


57
putative receptor-like protein
1408
senescence-associated



kinase

protein sen1


58
putative pectate lyase
1409
hypothetical protein


59
putative protein kinase
1410
putative cytochrome P450


60
putative peroxidase
1411
proline oxidase,





mitochondrial precursor





(osmotic stress-induced





proline dehydrogenase)


61
cytochrome P450-like protein
1412
putative response regulator





3


62
putative beta-amylase
1413
hypothetical protein


63
monosaccharide transporter STP3
1414
glutamine-dependent





asparagine synthetase


64
Lycopersicon esculentum
1415
lysine-ketoglutarate



proteinase TMP, Pir2:T07617

reductase/saccharopine


65
putative receptor-like protein
1416
En/Spm-like transposon



kinase

protein


66
G-box-binding factor 1
1417
G-box binding bZIP transcription





factor


67
amino acid carrier, putative
1418
putative protein


68
myb-related protein
1419
putative protein


69
No function assigned by TIGR
1420
putative protein


70
SNF1 like protein kinase
1421
ATFP4-like


71
Cu/Zn superoxide dismutase-like
1422
unknown protein



protein


72
putative protein kinase
1423
unknown protein


73
small nuclear ribonucleoprotein
1424
putative protein



U1A


74
ras-like GIF-binding
1425
invertase inhibitor homolog



protein

(emb|CAA73335.1)


75
oleoyl-[acyl-carrier-protein]
1426
unknown protein



hydrolase-like protein


76
putative heat shock
1427
unknown protein



transcription factor


77
putative protein
1428
putative cytochrome b5


78
membrane-bound small
1429
putative protein



GTP-binding-like protein


79
putative protein (fragment)
1430
putative protein


80
indole-3-acetate beta-
1431
putative protein



glucosyltransferase like



protein


81
HD-zip transcription factor
1432
No function assigned by TIGR



(athb-8)


82
putative cAMP-dependent
1433
putative copper/zinc superoxide



protein kinase

dismutase


83
glucuronosyl transferase-
1434
protein phosphatase ABI1



like protein


84
putative leucine-rich repeat
1435
glutamate dehydrogenase 2



disease resistance protein


85
98b like protein
1436
No function assigned by TIGR


86
putative receptor-like
1437
low-temperature-induced protein



protein kinase

78 (sp|Q06738)


87
IAA-Ala hydrolase (IAR3)
1438
putative myo-inositol 1-phosphate





synthase


88
putative AP2 domain
1439
phosphate transporter



transcription factor

(gb|AAB17265.1)


89
putative expansin
1440
4-hydroxyphenylpyruvate





dioxygenase (HPD)


90
putative Ap2 domain
1441
histone H1



protein


91
expansin (At-EXP1)
1442
hypothetical protein


92
cytochrome P450-like
1443
No function assigned by TIGR



protein


93
putative ATP-dependent
1444
neoxanthin cleavage enzyme-like



RNA helicase A

protein


94
unknown protein
1445
dehydration-induced protein RD22


95
predicted protein
1446
zinc finger protein ZAT7


96
putative glucosyltransferase
1447
unknown protein


97
unknown protein
1448
unknown protein


98
putative xyloglucan-
1449
unknown protein



specific glucanase


99
cysteine synthase
1450
unknown protein


100
clathrin assembly protein
1451
putative protein



AP19 homolog


101
dynein light chain like protein
1452
putative protein


102
chaperonin CPN10
1453
RNA helicase, putative


103
putative bHLH transcription factor
1454
putative glycine-rich





protein


104
putative glyoxysomal malate
1455
hypothetical protein



dehydrogenase precursor


105
ATP-dependent RNA helicase,
1456
putative protein



putative


106
chlorophyll synthetase
1457
peroxidase


107
similar to epoxide hydrolases
1458
peroxidase ATP3a





(emb|CAA67340.1)


108
putative protein
1459
metallothionein-like protein


109
unknown protein
1460
endomembrane-associated





protein


110
hypothetical protein
1461
ferritin 1 precursor


111
putative membrane transporter
1462
dehydrin RAB18-like





protein (sp|P30185)


112
putative tyrosyl-tRNA synthetase
1463
HSR201 like protein


113
ARGININE/SERINE-RICH
1464
light regulated protein,



SPLICING FACTOR RSP31

putative


114
putative oxidoreductase
1465
Dr4 (protease inhibitor)


115
unknown protein
1466
mitogen activated protein





kinase kinase (nMAPKK)


116
linker histone protein, putative
1467
glutathione S-transferase


117
hypothetical protein
1468
transcriptional activator





CBF1/CRT/CRE binding





factor 1


118
putative protein
1469
homeobox-leucine zipper





protein ATHB-12


119
putative mitochondrial carrier
1470
amino acid permease I



protein


120
putative transcription factor
1471
MAP kinase (ATMPK7)


121
MYB-related protein
1472
potassium channel protein





AKT3


122
myb-related transcription factor,
1473
cytochrome P450



putative

monooxygenase





(CYP91A2)


123
unknown protein
1474
putative transport protein


124
unknown protein
1475
putative protein


125
putative glycine-rich protein
1476
hypothetical protein


126
No function assigned by TIGR
1477
putative protein


127
unknown protein
1478
hypothetical protein


128
unknown protein
1479
receptor protein kinase-like





protein


129
unknown protein
1480
serine/threonine protein





kinase-like protein


130
unknown protein
1481
putative auxin-regulated





protein


131
putative membrane channel protein
1482
amino acid transport protein





AAP2


132
putative protein
1483
unknown protein


133
unknown protein
1484
cold and ABA inducible protein





kin1


134
gamma glutamyl hydrolase,
1485
gamma-VPE (vacuolar processing



putative

enzyme)


135
40S ribosomal protein S5
1486
putative protein 1 photosystem II





oxygen-evolving complex


136
DnaJ-like protein
1487
myrosinase-associated protein,





putative


137
40S ribosomal protein S26
1488
transcription factor ATMYB4


138
putative WRKY-type DNA binding
1489
H-protein promoter binding factor-



protein

2a


139
putative protein
1490
ammonium transporter, puitative


140
hypothetical protein
1491
putative zeta-carotene desaturase





precursor


141
putative ubiquitin-
1492
high-affinity nitrate transporter



conjugating enzyme

NRT2


142
peptidylprolyl isomerase
1493
light induced protein like



ROC1


143
glyceraldehyde-3-
1494
putative AT-hook DNA-binding



phosphate dehydrogenase C

protein



subunit (GapC)


144
No function assigned by
1495
putative glycogenin



TIGR


145
putative protein
1496
putative light repressible receptor





protein kinase


146
putative thioredoxin
1497
serine/threonine kinase-like





protein


147
thioredoxin h, putative
1498
putative peroxidase


148
thioredoxin-like
1499
cytochrome P450 monooxygenase





(CYP83A1)


149
allene oxide synthase
1500
MYB-related transcription factor



(emb|CAA73184.1)

(CCA1)


150
anthranilate synthase
1501
Terminal flower1 (TFL1)



component I-1 precursor



(sp|P32068)


151
CELL DIVISION
1502
sulfate transporter ATST1



CONTROL PROTEIN 2



HOMOLOG A


152
protein kinase cdc2
1503
RING-H2 finger protein RHA3b



homolog B


153
ethylene responsive
1504
lipoxygenase, putative



element binding factor 1



(frameshift !)


154
ethylene responsive
1505
serine O-acetyltransferase (EC



element binding factor 2

2.3.1.30) Sat-52 (pir||S71207)



(ATERF2) (sp|O80338)


155
ethylene responsive
1506
ferulate-5-hydroxylase (FAH1)



element binding factor 5



(ATERF5) (sp|O80341)


156
glucose-6-phosphate
1507
En/Spm-like transposon protein,



dehydrogenase

putative


157
photomorphogenesis
1508
calmodulin-binding-like protein



represser (COP1)


158
unknown protein
1509
hypothetical protein


159
DNA (cytosine-5)- methyltransferase (DNA
1510
somatic embryogenesis receptor-



methyltransferase) (DNA

like kinase-like protein



metase) (sp|P34881)


160
PROLIFERA
1511
putative giberellin beta-





hydroxylase


161
putative photomorphogenesis
1512
putative pectinesterase



represser protein


162
SNF1-like protein kinase (AKin11)
1513
putative protein


163
thioredoxin h
1514
unknown protein


164
thioredoxin
1515
ribosomal protein


165
Ca2+-dependent lipid-binding
1516
low-temperature-induced



protein, putative

65 kD protein (sp|Q04980)


166
putative auxin-induced protein
1517
putative glucosyltransferase


167
putative bZIP transcription factor
1518
peroxidase





(emb|CAA67551.1)


168
hypothetical protein
1519
ankyrin-like protein


169
putative AVR9 elicitor response
1520
ribosomal protein S11-like



protein


170
putative serine/threonine protein
1521
hypothetical protein



kinase


171
bZIP transcription factor ATB2
1522
glycoprotein (EP1), putative


172
putative spliceosome associated
1523
calnexin-like protein



protein


173
3-hydroxyisobutyryl-coenzyme A
1524
SRG1-like protein



hydrolase-like protein


174
putative protein
1525
ethylene response factor 1





(ERF1)


175
putative Mutator-like transposase
1526
transcriptional activator





CBF1-like protein


176
putative protein
1527
xyloglucan endo-1,4-beta-





D-glucanase (XTR-6)


177
unknown protein
1528
putative cinnamyl alcohol





dehydrogenase


178
putative protein
1529
gibberellin 3 beta-





hydroxylase, putative


179
putative protein
1530
auxin response transcription





factor 3 (ETTIN/ARF3)


180
putative galactinol synthase
1531
No function assigned by





TIGR


181
putative transcriptional regulator
1532
putative protein


182
nuclear matrix constituent protein 1
1533
similar to avrRpt2-induced



(NMCP1)-like

protein 1


183
putative DNA-binding protein
1534
unknown protein



RAV2


184
No function assigned by TIGR
1535
hypothetical protein


185
basic blue protein, 5′ partial
1536
putative protein kinase


186
unknown protein
1537
respiratory burst oxidase-





like protein


187
putative calcium-binding protein,
1538
glucose-6-



calreticulin

phosphate/phosphate-





translocator precursor,





putative


188
putative pyrophosphate-fructose-6-
1539
class 1 non-symbiotic



phosphate 1-phosphotransferase

hemoglobin (AHB1)


189
ribosomal protein L11, cytosolic
1540
endochitinase isolog


190
putative dTDP-glucose 4-6-
1541
putative cytochrome P450



dehydratase


191
40S ribosomal protein S20-like
1542
60S acidic ribosomal protein P0



protein


192
60S ribosomal protein L24
1543
putative protein


193
coatomer-like protein,
1544
auxin-induced protein, putative



epsilon subunit


194
glycoprotein (EP1), putative
1545
unknown protein


195
putative SPL1-related
1546
hypothetical protein



protein


196
unknown protein
1547
protein phosphatase 2C ABI2





(PP2C) (sp|O04719)


197
putative transport protein
1548
peroxidase, prxr2



SEC61 beta-subunit


198
unknown protein
1549
putative peroxidase ATP12a


199
putative cytochrome P450
1550
putative beta-amylase


200
UTP-glucose
1551
putative acetone-cyanohydrin lyase



glucosyltransferase-like



protein


201
60S ribosomal protein L23
1552
fatty acid elongase 3-ketoacyl-CoA





synthase 1


202
40S ribosomal protein S17
1553
putative citrate synthase


203
40S ribosomal protein S26
1554
pEARLI 1-like protein


204
protein translation factor
1555
putative MYB family transcription



Sui1 homolog, putative

factor


205
unknown protein
1556
putative transcription factor





MYB28


206
gamma glutamyl hydrolase,
1557
RNA helicase-like protein



putative


207
dTDP-glucose 4,6-
1558
snoRNA



dehydratase, putative


208
extensin-like protein
1559
putative protein kinase


209
unknown protein
1560
growth regulator like protein


210
protein phosphatase 2C-
1561
putative potassium transporter



like protein


211
ubiquitin-like protein
1562
putative protein


212
protein phosphatase 2C-like
1563
60S ribosomal protein L14



protein


213
unknown protein
1564
unknown protein


214
putative RING zinc finger
1565
putative RING-H2 zinc finger



ankyrin protein

protein


215
unknown protein
1566
putative pollen surface protein


216
putative rubisco subunit
1567
unknown protein



binding-protein alpha



subunit


217
putative acetone-
1568
unknown protein



cyanohydrin lyase


218
putative isoamylase
1569
unknown protein


219
putative protein
1570
putative Ca2+-ATPase


220
HSP associated protein like
1571
1-aminocyclopropane-1-





carboxylate synthase-like protein


221
60S ribosomal protein L39
1572
putative beta-glucosidase


222
unknown protein
1573
transcription factor ZAP1


223
putative SF16 protein {Helianthus
1574
oligopeptide transporter, putative



annuus}


224
unknown protein
1575
putative protein


225
thioredoxin
1576
putative glucosyltransferase


226
trehalose-6-phosphate phosphatase
1577
putative serine/threonine kinase



(AtTPPB)


227
chlorophyll a/b-binding protein
1578
squalene epoxidase-like protein


228
class IV chitinase (CHIV)
1579
similar to 14 KD proline-rich





protein DC2.15 precursor





(sp|P14009); similar to





ESTs emb|Z17709 and





emb|Z47685


229
chalcone synthase (naringenin-
1580
unknown protein



chalcone synthase) (testa 4 protein)



(sp|P13114)


230
unknown protein
1581
unknown protein


231
cinnamyl-alcohol dehydrogenase
1582
hypothetical protein



ELI3-2


232
farnesyl-pyrophosphate synthetase
1583
60S ribosomal protein L38



FPS2


233
phospholipid hydroperoxide
1584
flavin-containing



glutathione peroxidase

monooxygenase, putative


234
heat shock transcription factor
1585
remorin



HSF4


235
heat shock protein 101
1586
unknown protein


236
17.6 kDa heat shock protein (AA
1587
putative protein



1-156)


237
heat shock protein 17.6A
1588
lipoxygenase


238
heat-shock protein
1589
cold-regulated protein





COR6.6 (KIN2)


239
HY5
1590
Myb transcription factor





homolog (ATR1)


240
putative auxin-induced protein,
1591
putative protein



IAA12


241
early auxin-induced protein,
1592
unknown protein



IAA19


242
auxin-inducible gene (IAA2)
1593
unknown protein


243
putative protein
1594
Ca2+-transporting ATPase-





like protein


244
putative choline kinase
1595
protein phosphatase 2C





(AtP2C-HA)


245
thymidylate kinase-like protein
1596
peroxidase ATP24a


246
CTP synthase like protein
1597
branched-chain alpha keto-





acid dehydrogenase,





putative


247
putative protein
1598
putative beta-ketoacyl-CoA





synthase


248
putative amidase
1599
putative protein


249
4-alpha-glucanotransferase
1600
putative beta-galactosidase


250
hypothetical protein
1601
putative protein


251
similar to auxin-induced protein
1602
60S ribosomal protein L27


252
putative protein
1603
putative annexin


253
putative protein
1604
NAC domain protein,





putative


254
putative protein
1605
unknown protein


255
hyuC-like protein
1606
late embryogenesis





abundant protein LEA like


256
putative tetracycline
1607
unknown protein



transporter protein


257
similar to early nodulins
1608
putative protein


258
putative protein
1609
dehydrin Xero2


259
putative peptidyl-prolyl cis-
1610
putative zinc finger protein



trans isomerase


260
unknown protein
1611
unknown protein


261
unknown protein
1612
DnaJ-like protein


262
putative endochitinase
1613
putative inositol polyphosphate-5-





phosphatase


263
putative ABC transporter
1614
putative cytochrome P450


264
No function assigned by
1615
putative protein



TIGR


265
CONSTANS-like B-box
1616
unknown protein



zinc finger protein


266
unknown protein
1617
putative protein


267
unknown protein
1618
hypothetical protein


268
putative mitochondrial
1619
putative protein



processing peptidase alpha



subunit


269
putative pre-mRNA
1620
sucrose-UDP glucosyltransferase



splicing factor


270
putative phosphatidylserine
1621
glucose-6-phosphate 1-



decarboxylase

dehydrogenase


271
unknown protein
1622
unknown protein


272
unknown protein
1623
mitochondrial chaperonin (HSP60)


273
unknown protein
1624
sucrose transport protein SUC1


274
putative casein kinase I
1625
putative protein disulfide isomerase


275
unknown protein
1626
putative pollen-specific protein


276
60S ribosomal protein
1627
integral membrane protein,



L23A

putative


277
putative mitochondrial
1628
rubredoxin, putative



dicarboxylate carrier



protein


278
enoyl-ACP reductase (enr-
1629
putative protein



A)


279
putative isoamylase
1630
disease resistance protein RPS4,





putative


280
formamidase-like protein
1631
putative peptide/amino acid





transporter


281
reticuline oxidase-like
1632
peroxidase, putative



protein


282
unknown protein
1633
ethylene receptor, putative (ETR2)


283
putative transketolase
1634
protein phosphatase 2C (PP2C)



precursor


284
putative protein
1635
putative glutathione S-transferase


285
unknown protein
1636
homeodomain transcription factor





(ATHB-7)


286
unknown protein
1637
putative nitrate transporter


287
unknown protein
1638
putative ribosomal protein L9,





cytosolic


288
putative esterase D
1639
putative DNA-binding protein


289
predicted protein of unknown
1640
beta-1,3-glucanase-like protein



function


290
unknown protein
1641
putative zinc transporter


291
putative indole-3-glycerol
1642
transcription factor TINY



phosphate synthase


292
isopentenyl
1643
putative aspartate kinase-



pyrophosphate:dimethyllallyl

homoserine dehydrogenase



pyrophosphate isomerase


293
kinase associated protein
1644
ethylene reponse factor-like AP2



phosphatase

domain transcription factor


294
putative K+ channel, beta subunit
1645
peptide transporter-like protein


295
KNAT1 homeobox-like protein
1646
trehalose-6-phosphate synthase like





protein


296
PSI type II chlorophyll a/b-binding
1647
putative ribonuclease



protein, putative


297
transcription factor
1648
hypothetical protein


298
putative WD-40 repeat protein,
1649
putative DNA-binding



MSI2

protein


299
WD-40 repeat protein (MSI3)
1650
nodulin-like protein


300
putative WD-40 repeat protein,
1651
trehalose-6-phosphate



MSI4

phosphatase-like protein


301
unknown protein
1652
succinate dehydrogenase





flavoprotein alpha subunit





(emb|CAA05025.1)


302
hypothetical protein
1653
unknown protein


303
putative protein
1654
stress related protein,





putative


304
No function assigned by TIGR
1655
putative chloroplast





initiation factor 3


305
polyphosphoinositide binding
1656
putative protein



protein, putative


306
hypothetical protein
1657
hypothetical protein


307
unknown protein
1658
putative CCCH-type zinc





finger protein


308
chloroplast ribosomal L1-like
1659
similar to harpin-induced



protein

protein hin1 from tobacco


309
cold-regulated protein cor15b
1660
unknown protein



precursor


310
cyanohydrin lyase like protein
1661
unknown protein


311
putative replication protein A1
1662
hypothetical protein


312
putative protein
1663
No function assigned by





TIGR


313
possible apospory-associated like
1664
putative protein



protein


314
DNA binding protein GT-1,
1665
putative glutathione S-



putative

transferase TSI-1


315
AT-hook DNA-binding protein
1666
putative protein



(AHP1)


316
putative phospholipase
1667
putative PTR2 family





peptide transporter


317
chloroplast FtsH protease, putative
1668
receptor kinase-like protein


318
enoyl-CoA hydratase like
1669
putative sugar transport



protein

protein, ERD6


319
berberine bridge enzyme-
1670
putative protein



like protein


320
putative sugar transporter
1671
nodulin-like protein


321
unknown protein
1672
unknown protein


322
No function assigned by
1673
putative receptor-like



TIGR

protein kinase


323
hypothetical protein
1674
glutathione-conjugate





transporter AtMRP4


324
putative acidic ribosomal
1675
ascorbate oxidase-like



protein

protein


325
putative protein
1676
pathogenesis-related protein 1





precursor, 19.3K


326
unknown protein
1677
R2R3-MYB transcription factor


327
hypothetical protein
1678
hypothetical protein


328
putative protein
1679
putative chitinase


329
dihydroxypolypreny
1680
Mlo protein, putative



lbenzoate methyltransferase


330
unknown protein
1681
putative WRKY-type DNA binding





protein


331
myb-related protein
1682
putative acyl-CoA synthetase


332
No function assigned by
1683
putative pathogenesis-related



TIGR

protein


333
putative protein
1684
putative chitinase


334
putative disease resistance
1685
germin precursor oxalate oxidase



response protein


335
hypothetical protein
1686
endoxyloglucan transferase,





putative


336
No function assigned by
1687
putative protein



TIGR


337
starch branching enzyme II
1688
putative cytochrome P450


338
No function assigned by
1689
similar to Mlo proteins from H.



TIGR

vulgare


339
putative enolase (2-
1690
putative tropinone reductase



phospho-D-glycerate



hydroylase)


340
putative protein kinase
1691
extensin-like protein


341
HD-Zip protein, putative
1692
putative sarcosine oxidase


342
putative protein kinase
1693
putative protein


343
phenylalanyl-trna
1694
hypothetical protein



synthetase-like protein


344
putative aconitase
1695
late embryogenesis-abundant





protein, putative


345
NAM (no apical meristem)
1696
beta-carotene hydroxylase



protein, putative


346
unknown protein
1697
putative calcium binding protein


347
putative
1698
unknown protein



phosphomannomutase


348
putative farnesylated protein
1699
unknown protein


349
unknown protein
1700
predicted glycosyl transferase


350
water stress-induced protein,
1701
hypothetical protein



putative


351
unknown protein
1702
hypothetical protein


352
unknown protein
1703
hypothetical protein


353
PEROXISOMAL MEMBRANE
1704
putative protein



PROTEIN PMP22


354
putative peroxisomal membrane
1705
unknown protein



carrier protein


355
putative protein
1706
putative protein


356
unknown protein
1707
putative protein


357
putative protein
1708
serine/threonine kinase-like





protein


358
putative protein
1709
No function assigned by TIGR


359
argininosuccinate synthase-like
1710
putative pectinesterase



protein


360
1-phosphatidylinositol-4,5-
1711
peroxidase like protein



bisphosphate phosphodiesterase


361
putative JUN kinase activator
1712
No function assigned by TIGR



protein


362
putative 60S ribosomal protein L35
1713
phenylalanine ammonia





lyase (PALI)


363
nucleoid DNA-binding protein
1714
peroxidase



cnd41-like protein

(emb|CAA68212.1)


364
SigA binding protein
1715
putative AMP deaminase


365
hypothetical protein
1716
putative MYB family





transcription factor


366
putative protein kinase
1717
DNA-directed RNA





polymerase II, third largest subunit


367
unknown protein
1718
nucleotide pyrophosphatase-





like protein


368
regulatory protein NPR1-like;
1719
putative peroxidase



transcription factor inhibitor I



kappa B-like


369
putative protein
1720
calcium sensor homolog





(gb|AAC26110.1)


370
hypothetical protein
1721
putative GDSL-motif





lipase/hydrolase


371
phosphoribosylanthranilate
1722
putative nonspecific lipid-



isomerase

transfer protein


372
phosphoribosylanthranilate
1723
acyl-carrier protein (ACP),



isomerase

putative


373
sterol glucosyltransferase, putative
1724
putative glycine





dehydrogenase


374
putative gigantea protein
1725
AIG1


375
putative MYB family transcription
1726
ACC synthase (AtACS-6)



factor


376
hypothetical protein
1727
cyclin delta-3


377
hypothetical protein
1728
putative RING zinc finger





protein


378
predicted protein
1729
aldose 1-epimerase-like





protein


379
cytochrome P450, putative
1730
putative phospholipase


380
putative Na+ dependent
1731
phosphoenolpyruvate



ileal bile acid transporter

carboxylase


381
unknown protein
1732
putative galactinol synthase


382
RING-H2 finger protein
1733
unknown protein



RHF1a


383
putative protein
1734
putative protein


384
unknown protein
1735
1-aminocyclopropane-1-





carboxylate oxidase


385
putative protein
1736
thioredoxin (clone GIF1)





(pir||S58118)


386
putative auxin-regulated
1737
trehalose-6-phosphate



protein

phosphatase


387
hypothetical protein
1738
beta-1,3-glucanase 2 (BG2)





(PR-2)


388
unknown protein
1739
putative S-adenosyl-L-





methionine:trans-caffeoyl-





Coenzyme A 3-O-





methyltransferase


389
unknown protein
1740
disease resistance protein EDS1


390
putative protein
1741
putative protein kinase


391
putative protein
1742
Gluthatione reductase, chloroplast





precursor


392
unknown protein
1743
putative heat shock protein


393
histone H1
1744
aspartate kinase


394
Argonaute (AGO1)-like
1745
putative major intrinsic (channel)



protein

protein


395
unknown protein
1746
matrix metalloproteinase, putative


396
putative protein with C-
1747
putative GDSL-motif



terminal RING finger

lipase/hydrolase


397
unknown protein
1748
putative protein


398
unknown protein
1749
DAG-like protein


399
unknown protein
1750
serine/threonine kinase-like





protein


400
unknown protein
1751
formamidase-like protein


401
unknown protein
1752
CER2


402
putative copper amine
1753
26S proteasome subunit 4



oxidase


403
unknown protein
1754
pectinesterase like protein


404
unknown protein
1755
putative disease resistance protein


405
unknown protein
1756
putative RNA methyltransferase


406
putative protein
1757
unknown protein


407
putative protein
1758
HOMEOBOX PROTEIN





KNOTTED-1 LIKE 4 (KNAT4)


408
unknown protein
1759
glycine-rich RNA-binding protein





AtGRP2-like


409
unknown protein
1760
putative acetylornithine





transaminase


410
putative protein
1761
putative Sec24-like COPII protein


411
putative protein
1762
putative berberine bridge enzyme


412
unknown protein
1763
putative GH3-like protein


413
serine/threonine kinase-
1764
putative ABC transporter



like protein


414
alcohol dehydrogenase,
1765
putative reticuline oxidase-like



putative

protein


415
anthranilate
1766
pectate lyase-like protein



phosphoribosyltransferase,



chloroplast precursor (sp|Q02166)


416
phytochrome C (sp|P14714)
1767
protein disulfide-isomerase-like





protein


417
putative phytochrome-associated
1768
putative protein



protein 3


418
receptor serine/threonine kinase
1769
putative membrane transporter



PR5K


419
Ran-binding protein (atranbp1a)
1770
unknown protein


420
small Ras-like GTP-binding
1771
unknown protein



protein (gb|AAB58478.1)


421
sterol-C5-desaturase
1772
putative RING-H2 zinc finger





protein


422
tryptophan synthase beta chain 1
1773
unknown protein



precursor (sp|P14671)


423
thioredoxin f2 (gb|AAD35004.1)
1774
unknown protein


424
No function assigned by TIGR
1775
unknown protein


425
putative WRKY DNA-binding
1776
MADS-box protein



protein

(AGL20)


426
putative protein
1777
amidophosphoribosyltransfer-





ase 2 precursor


427
unknown protein
1778
putative dihydrodipicolinate





synthase


428
unknown protein
1779
hypothetical protein


429
14-3-3 protein homolog RCI1
1780
ABA-responsive protein-



(pir||S47969)

like


430
unknown protein
1781
putative protein


431
putative CCCH-type zinc finger
1782
hypothetical protein



protein


432
PINHEAD (gb|AAD40098.1);
1783
DNA-binding protein-like



translation initiation factor


433
plasma membrane proton ATPase
1784
No function assigned by



(PMA)

TIGR


434
CHLOROPHYLL A-B BINDING
1785
transcription factor,



PROTEIN 4 PRECURSOR

putative



homolog


435
membrane related protein CP5,
1786
nitrate reductase, putative



putative


436
ABC transporter (AtMRP2)
1787
putative protein


437
putative embryo-abundant protein
1788
putative protein


438
putative anthocyanidin-3-glucoside
1789
putative protein



rhamnosyltransferase


439
putative lipid transfer protein
1790
putative protein


440
unknown protein
1791
unknown protein


441
unknown protein
1792
unknown protein


442
galactinol synthase, putative
1793
tryptophan synthase beta-





subunit (TSB2)


443
putative protein
1794
hypothetical protein


444
putative protein
1795
putative protein


445
SCARECROW-like protein
1796
putative DNA-binding





protein


446
unknown protein
1797
putative 40S ribosomal





protein S10


447
unknown protein
1798
putative protein


448
unknown protein
1799
putative cytochrome P450


449
unknown protein
1800
putative protein


450
asparagine--tRNA ligase
1801
putative protein


451
putative protein
1802
putative glucosyltransferase


452
glutamate-1-semialdehyde
1803
No function assigned by



2,1-aminomutase 1

TIGR



precursor (GSA 1)



(glutamate-1-semialdehyde



aminotransferase 1) (GSA-



AT 1) (sp|P42799)


453
hypothetical protein
1804
putative protein


454
putative serine protease-like
1805
putative protein



protein


455
No function assigned by
1806
unknown protein



TIGR


456
unknown protein
1807
glycine-rich RNA binding protein





7


457
unknown protein
1808
dehydrin, putative


458
gamma-adaptin, putative
1809
putative endoxyloglucan





glycosyltransferase


459
UDP rhamnose--
1810
glutamate decarboxylase 1 (GAD



anmocyanidin-3-glucoside

1) (sp|Q42521)



rhamnosyltransferase-like



protein


460
carbonate dehydratase-like
1811
delta 9 desaturase



protein


461
putative microtubule-
1812
UDP-glucose glucosyltransferase



associated protein


462
putative ribophorin I
1813
CARBONIC ANHYDRASE 2


463
putative zinc finger protein
1814
response reactor 2 (ATRR2)


464
chloroplast FtsH protease,
1815
S-adenosyl-methionine-sterol-C-



putative

methyltransferase, putative


465
putative protein
1816
putative DNA-binding protein





(RAV2-like)


466
unknown protein
1817
gamma glutamyl hydrolase,





putative


467
putative LEA protein
1818
protein phosphatase-like


468
putative protein
1819
unknown protein


469
putative protein
1820
unknown protein


470
unknown protein
1821
unknown protein


471
putative purple acid
1822
copper transport protein-like



phosphatase

protein


472
unknown protein
1823
hypothetical protein


473
putative protein
1824
unknown protein


474
unknown protein
1825
putative peptide methionine





sulfoxide reductase


475
chlorophyll binding protein,
1826
putative obtusifoliol 14-alpha



putative

demethylase


476
phosphoenolpyruvate carboxylase
1827
glutamate dehydrogenase (EC



(PPC)

1.4.1.-) 1 (pir||S71217)


477
chlorophyll a/b-binding protein-
1828
unknown protein



like


478
AtAGP4
1829
xyloglucan endo-1,4-beta-D-





glucanase precursor


479
putative cryptochrome 2 apoprotein
1830
unknown protein


480
type 2 peroxiredoxin, putative
1831
SNF1 related protein kinase





(ATSRPK1)


481
Atpm24.1 glutathione S transferase
1832
putative protein


482
delta tonoplast integral protein
1833
putative chloroplast nucleoid DNA



(delta-TIP)

binding protein


483
20S proteasome subunit (PAA2)
1834
hypothetical protein


484
dormancy-associated protein,
1835
putative protein



putative


485
putative cytidine deaminase
1836
putative thiamin biosynthesis





protein


486
No function assigned by TIGR
1837
unknown protein


487
putative phospholipase D-gamma
1838
unknown protein


488
cell elongation protein, Dwarf1
1839
putative RNA helicase


489
germin-like protein
1840
putative SF21 protein





{Helianthus annuus}


490
hevein-like protein precursor (PR-
1841
unknown protein



4)


491
rac-like GTP binding protein
1842
NBS/LRR disease



(ARAC5)

resistance protein, putative


492
phosphoprotein phosphatase, type
1843
hypothetical protein



1 catalytic subunit


493
ubiquitin-protein ligase UBC9
1844
unknown protein


494
xyloglucan endotransglycosylase-
1845
No function assigned by



related protein XTR-7

TIGR


495
cysteine synthase
1846
glycine-rich protein





(AtGRP2)


496
putative villin 2
1847
No function assigned by





TIGR


497
glutathione S-transferase
1848
putative protein


498
5-adenylylsulfate reductase
1849
putative glucosyltransferase


499
arginine decarboxylase
1850
hypothetical protein


500
ATHP2, putative
1851
hypothetical protein


501
ornithine carbamoyltransferase
1852
putative protein



precursor


502
puative protein
1853
putative disease resistance





protein


503
putative protein
1854
thaumatin, putative


504
unknown protein
1855
putative proline-rich protein


505
putative protein
1856
sterol-C-methyltransferase


506
putative protein
1857
superoxidase dismutase


507
unknown protein
1858
TINY-like protein


508
unknown protein
1859
calcium-dependent protein





kinase, putative


509
unknown protein
1860
hypothetical protein


510
unknown protein
1861
putative protein kinase


511
hypothetical protein
1862
DNA-directed RNA





polymerase (mitochondrial)


512
putative protein
1863
putaive DNA-binding





protein


513
putative DnaJ protein
1864
late embryogenesis





abundant M17 protein


514
plastocyanin
1865
putative protein


515
unknown protein
1866
delta-1-pyrroline-5-





carboxylate synthetase


516
unknown protein
1867
putative 60s ribosomal





protein L10


517
unknown protein
1868
cytochrome P450





CYP86A1


518
unknown protein
1869
putative tyrosine aminotransferase


519
unknown protein
1870
thionin


520
unknown protein
1871
No function assigned by TIGR


521
putative ATP-dependent
1872
APETALA2 protein



RNA helicase


522
non-race specific disease
1873
MADS-box protein (AGL3)



resistance protein (NDR1)


523
hypothetical protein
1874
putative monooxygenase


524
putative protein
1875
ZFP3 zinc finger protein


525
putative protein
1876
cell division protein FtsZ





chloroplast homolog precursor





(sp|Q42545)


526
putative protein
1877
calreticulin, putative


527
copper transport protein
1878
phosphoserine aminotransferase


528
putative protein
1879
12-oxophytodienoate-10,11-





reductase


529
unknown protein
1880
putative bHLH transcription factor


530
unknown protein
1881
pectin methylesterase (PMEU1),





putative


531
unknown protein
1882
DNA-binding protein


532
putative protein kinase
1883
carnitine racemase like protein


533
unknown protein
1884
putative protein


534
putative protein
1885
endoxyloglucan transferase





(dbj|BAA81669.1)


535
putative protein
1886
RMA1 RING zinc finger protein


536
hypothetical protein
1887
ammonium transporter


537
putative protein
1888
apyrase (gb|AAF00612.1)


538
putative AP2 domain
1889
potassium uptake transporter-like



transcription factor

protein


539
putative nitrilase
1890
putative ABC transporter


540
putative protein
1891
potassium transporter-like protein


541
putative tetrahydrofolate
1892
integral membrane protein,



synthase

putative


542
heat-shock protein
1893
putative protein


543
unknown protein
1894
pyruvate decarboxylase-1 (Pdc1)


544
unknown protein
1895
putative malate oxidoreductase


545
histone H4
1896
putative histone H2B


546
hypothetical protein
1897
snoRNA


547
unknown protein
1898
symbiosis-related like protein


548
putative protein
1899
unknown protein


549
predicted protein
1900
unknown protein


550
putative dihydrolipoamide
1901
hypothetical protein



succinyltransferase


551
actin 3
1902
putative protein


552
putative CCCH-type zinc finger
1903
copper-binding protein-like



protein


553
MAP kinase kinase 2
1904
putative protein


554
ethylene-insensitive3-like1 (EIL1)
1905
unknown protein


555
histidine transport protein (PTR2-
1906
putative glyoxalase II



B)


556
putative auxin-induced protein
1907
No function assigned by



AUX2-11

TIGR


557
hydroxyacylglutathione hydrolase
1908
hypothetical protein



cytoplasmic (glyoxalase II) (GLX



II)


558
delta-8 sphingolipid desaturase
1909
flavanone 3-hydroxylase





(FH3)


559
cellulose synthase catalytic subunit
1910
putative laccase



(Ath-A)


560
nitrate transporter (NTL1)
1911
putative protein kinase


561
DNA-binding homeotic protein
1912
myb-related protein, 33.3K



Athb-2

(pir||S71284)


562
hypothetical protein
1913
unknown protein


563
aspartate aminotransferase
1914
endo-xyloglucan transferase-





like protein


564
4-coumarate:CoA ligase 1
1915
TMV resistance protein N-





like


565
pyruvate dehydrogenase E1 beta
1916
putative xyloglucan



subunit, putative

endotransglycosylase


566
nucleotide diphosphate kinase Ia
1917
unknown protein



(emb|CAB58230.1)


567
chloroplast Cpn21 protein
1918
proline transporter 2


568
ATP dependent copper transporter
1919
resistance protein, putative


569
very-long-chain fatty acid
1920
actin, putative



condensing enzyme (CUT1)


570
putative purine-rich single-stranded
1921
putative related to microbial



DNA-binding protein

divalent cation tolerance





proteins


571
serine/threonine protein
1922
unknown protein



phosphatase (type 2A)


572
isopentenyl
1923
putative glycosyl



diphosphate:dimethylallyl

transferase



diphosphate isomerase (IPP2)


573
putative c2h2 zinc finger
1924
unknown protein



transcription factor


574
putative 20S proteasome beta
1925
putative protein



subunit PBC2

phosphatase 2C


575
nucleoside diphosphate kinase 3
1926
unknown protein



(ndpk3)


576
ras-related small GTP-binding
1927
serpin, putative



protein


577
putative 4-coumarate:CoA ligase 2
1928
cinnamyl-alcohol





dehydrogenase CAD1


578
transcription factor HBP-1b
1929
putative protein import



homolog (sp|P43273)

receptor


579
biotin synthase (Bio B)
1930
unknown protein


580
homeobox protein HAT22
1931
unknown protein


581
putative preprotein
1932
putative protein



translocase SECY protein


582
carbamoylphosphate
1933
putative CDP-



synthetase, putative

diacylglycerol--glycerol-3-





phosphate 3-





phosphatidyltransferase


583
putative protein kinase,
1934
unknown protein



ADK1


584
putative nuclear DNA-
1935
putative LRR receptor-like



binding protein G2p

protein kinase


585
hypothetical protein
1936
serine/threonine protein kinase,





putative


586
hypothetical protein
1937
potassium transporter-like protein


587
unknown protein
1938
lactate dehydrogenase (LDH1)


588
unknown protein
1939
hypothetical protein


589
molybdopterin synthase
1940
unknown protein



(CNX2)


590
putative ribosomal protein
1941
putative thaumatin



L6


591
unknown protein
1942
putative reticuline oxidase-like





protein


592
En/Spm-like transposon
1943
uracil phosphoribosyltransferase,



protein

putative


593
putative protein
1944
transcription factor, putative


594
putative protein
1945
unknown protein


595
unknown protein
1946
unknown protein


596
hypothetical protein
1947
GATA transcription factor 4


597
unknown protein
1948
unknown protein


598
unknown protein
1949
unknown protein


599
putative lysosomal acid
1950
senescence-associated protein-like



lipase


600
unknown protein
1951
putative pollen allergen


601
unknown protein
1952
unknown protein


602
NifS-like aminotranfserase
1953
putative protein


603
actin 8
1954
glycine-rich protein


604
hypothetical protein
1955
putative protein


605
putative protein
1956
3-methyladenine DNA glycosylase,





putative


606
heat-shock protein (At-
1957
endoplasmic reticulum-type



hsc70-3)

calcium-transporting ATPase 4


607
putative protein disulfide
1958
putative pectinesterase



isomerase precursor


608
adenosine nucleotide
1959
cytochrome P450-like protein



translocator


609
photosystem II oxygen-evolving
1960
RNA-binding protein (cp33)



complex protein 3-like


610
sedoheptulose-bisphosphatase
1961
CONSTANS-like 1



precursor


611
glutathione S-transferase (GST6)
1962
putative small heat shock protein


612
geranylgeranyl reductase
1963
hypothetical protein


613
hypothetical protein
1964
unknown protein


614
hypothetical protein
1965
cytochrome P450-like protein


615
phosphoribulokinase precursor
1966
cysteine proteinase inhibitor like





protein


616
high mobility group protein
1967
nicotianamine synthase



(HMG1), putative

(dbj|BAA74589.1)


617
protease inhibitor II
1968
copper amine oxidase like protein





(fragment2)


618
protease inhibitor II
1969
putative SCARECROW gene





regulator


619
cytochrome P450 90A1
1970
unknown protein



(sp|Q42569)


620
unknown protein
1971
unknown protein


621
heat shock protein 90
1972
putative alanine acetyl





transferase


622
tubulin beta-9 chain
1973
unknown protein


623
putative ubiquitin carboxyl
1974
unknown protein



terminal hydrolase


624
protein kinase
1975
unknown protein


625
DRE/CRT-binding protein
1976
putative extensin



DREB1C


626
histidyl-tRNA synthetase
1977
putative protein kinase


627
splicing factor, putative
1978
putative protein kinase


628
glutamyl-tRNA synthetase
1979
NADPH-dependent





codeinone reductase,





putative


629
putative RING zinc finger protein
1980
peroxidase


630
phytochelatin synthase
1981
putative cytochrome P450



(gb|AAD41794.1)


631
putative C2H2-type zinc finger
1982
No function assigned by



protein

TIGR


632
putative ligand-gated ion channel
1983
putative zinc-finger protein



protein

(B-box zinc finger domain)


633
putative ribosomal-protein S6
1984
putative tyrosine



kinase (ATPK6)

aminotransferase


634
MOLYBDOPTERIN
1985
hypothetical protein



BIOSYNTHESIS CNX1



PROTEIN


635
temperature-sensitive omega-3
1986
DNA binding protein



fatty acid desaturase, chloroplast



precursor (sp|P48622)


636
adenylosuccinate synthetase
1987
putative fatty acid elongase


637
putative 14-3-3 protein
1988
bZIP transcription factor-





like protein


638
putative cytochrome P450
1989
xyloglucan





fucosyltransferase, putative


639
putative two-component
1990
unknown protein



response regulator 3 protein


640
putative RING-H2 zinc
1991
unknown protein



finger protein ATL6


641
No function assigned by
1992
putative protein



TIGR


642
small zinc finger-like
1993
myb factor, putative



protein


643
hypothetical protein
1994
Myb-family transcription





factor, putative


644
MAP kinase (ATMPK6)
1995
putative fructose





bisphosphate aldolase


645
vacuolar ATP synthase,
1996
myrosinase-associated



putative

protein, putative


646
kinesin-like protein
1997
cytochrome P450 like





protein


647
serine/threonine-specific
1998
similar to SOR1 from the



protein kinase NAK

fungus Cercospora





nicotianae


648
No function assigned by
1999
similar to embryo-abundant



TIGR

protein GB:L47672 GI:1350530





[Picea glauca]


649
ACTIN 2/7 (sp|P53492)
2000
alcohol dehydrogenase


650
phosphoglycerate kinase,
2001
auxin response factor 1



putative


651
homeotic protein BEL1
2002
pathogenesis-related protein 1



homolog

precursor, 18.9K


652
proline iminopeptidase
2003
hypothetical protein


653
pasticcino 1
2004
unknown protein


654
serine/threonine protein
2005
zinc finger protein Zat12



kinase


655
cytochrome P450
2006
unknown protein



monooxygenase



(CYP71B4)


656
No function assigned by
2007
unknown protein



TIGR


657
putative GDSL-motif
2008
cyclin, putative



lipase/hydrolase


658
putative protein
2009
2-dehydro-3-





deoxyphosphoheptonate aldolase


659
unknown protein
2010
glutathione synthetase gsh2


660
hypothetical protein
2011
heat shock protein 17


661
putative glycosylation
2012
putative Na+-dependent inorganic



enzyme

phosphate cotransporter


662
No function assigned by
2013
No function assigned by TIGR



TIGR


663
No function assigned by
2014
unknown protein



TIGR


664
unknown protein
2015
putative protein


665
putative ABC transporter
2016
similar to RING-H2 finger protein





RHC1a GB:AAC69854





GI:3790583 from [Arabidopsis





thaliana]


666
nifU-like protein
2017
calcium-binding protein-like


667
putative receptor-like protein
2018
putative protein



kinase


668
putative disease resistance protein
2019
putative aldehyde dehydrogenase


669
receptor-like protein kinase-like
2020
auxin-responsive GH3-like





protein


670
ubiquitin activating enzyme 2
2021
putative protein



(gb|AAB37569.1)


671
No function assigned by TIGR
2022
Phosphoglycerate dehydrogenase-





like protein


672
putative receptor-like protein
2023
unknown protein



kinase


673
K+ transporter, AKT1
2024
unknown protein


674
shaggy-like kinase beta
2025
PSI type III chlorophyll a/b-





binding protein, putative


675
heat shock protein 70
2026
putative protein


676
plasma membrane intrinsic protein
2027
putative protein



1a


677
HSP90-like protein
2028
glutaredoxin, putative


678
histone H1, putative
2029
hypothetical protein


679
unknown protein
2030
No function assigned by TIGR


680
dnaK-type molecular chaperone
2031
putative protein



hsc70.1-like


681
gamma-glutamylcysteine
2032
jasmonate inducible protein,



synthetase

putative


682
peroxidase (ATP22a)
2033
putative polygalacuronase





isoenzyme 1 beta subunit


683
putative serine carboxypeptidase
2034
putative small heat shock protein



precursor


684
putative dioxygenase
2035
unknown protein


685
glucose transporter
2036
putative disease resistance





protein


686
NOI protein, nitrate-induced
2037
putative protein


687
putative protein
2038
ethylene-responsive





element binding factor,





putative


688
putative protein
2039
putative protein


689
unknown protein
2040
Pollen-specific protein





precursor like


690
putative photosystem I reaction
2041
putative protein



center subunit II precursor


691
putative protein
2042
unknown protein


692
unknown protein
2043
EF-Hand containing protein-





like


693
cobalamin biosynthesis protein
2044
unknown protein


694
adenine nucleotide translocase
2045
puative calcium-





transporting ATPase


695
glutathione transferase, putative
2046
antifungal protein-like





(PDF1.2)


696
putative 60S ribosomal protein L21
2047
pathogenesis-related PR-1-





like protein


697
cytochrome P450 like protein
2048
similar to Mlo proteins





from H. vulgare


698
cytochrome b245 beta chain
2049
putative steroid



homolog RbohAp108, putative

sulfotransferase


699
RNA helicase, DRH1
2050
trehalase-like protein


700
putative aldolase
2051
thioredoxin f1


701
farnesyltransferase subunit A
2052
unknown protein



(FTA)


702
No function assigned by
2053
alanine-glyoxylate



TIGR

aminotransferase


703
putative putative sister-
2054
integral membrane protein,



chromatide cohesion

putative



protein


704
calcium-dependent protein
2055
hypothetical protein



kinase


705
serine/threonine protein
2056
unknown protein



phosphatase type 2A,



putative


706
40S ribosomal protein S28
2057
hypothetical protein



(sp|P34789)


707
RNA polymerase subunit
2058
unknown protein


708
DNA-damage-
2059
unknown protein



repair/toleration protein



DRT102


709
putative C2H2-type zinc
2060
unknown protein



finger protein


710
putative adenosine
2061
drought-induced-19-like 1



phosphosulfate kinase


711
lipase
2062
unknown protein


712
putative violaxanthin de-
2063
putative protein



epoxidase precursor



(U44133)


713
aromatic rich glycoprotein,
2064
putative protein



putative


714
putative fumarase
2065
AIG2-like protein


715
flavonol synthase (FLS)
2066
Lhca2 protein



(sp|Q96330)


716
response regulator 5,
2067
phytocyanin



putative


717
sulfate transporter
2068
putative chlorophyll A-B binding





protein


718
putative floral homeotic
2069
Lhcb3 chlorophyll a/b binding



protein, AGL9

protein (gb|AAD28773.1)


719
putative ethylene-inducible
2070
luminal binding protein



protein

(dbj|BAA13948.1)


720
C-8,7 sterol isomerase
2071
hydroxypyruvate reductase (HPR)


721
TCH4 protein
2072
epoxide hydrolase (ATsEH)



(gb|AAA92363.1)


722
hypothetical protein
2073
putative protein (fragment)


723
putative urease accessory
2074
unknown protein



protein


724
molybdopterin synthase
2075
hypothetical protein



sulphurylase



(gb|AAD18050.1)


725
putative protein
2076
putative glucosyl transferase


726
NBD-like protein
2077
putative glucosyl transferase



(gb|AAD20643.1)


727
AtHVA22c
2078
putative 3-methylcrotonyl-CoA





carboxylase


728
unknown protein
2079
putative peroxidase


729
phytoene synthase
2080
acyl-CoA oxidase



(gb|AAB65697.1)

(gb|AAC13497.1)


730
protein kinase (AME2/AFC1)
2081
alternative oxidase 1a precursor


731
hypothetical protein
2082
putative transcription factor





(MYB4)


732
cyclin-dependent protein kinase-
2083
serine acetyltransferase



like protein


733
photosystem II stability/assembly
2084
ATP-sulfurylase



factor HCF136 (sp|O82660)


734
hypothetical protein
2085
calreticulin (crt1)


735
DNA binding-like protein
2086
putative prohibitin 2


736
putative protein
2087
putative monodehydroascorbate





reductase


737
chorismate mutase
2088
branched-chain alpha-keto acid





decarboxylase E1 beta subunit


738
putative LRR receptor protein
2089
cytokinin oxidase-like protein



kinase


739
putative chalcone synthase
2090
putative receptor-like protein





kinase


740
putative protein kinase
2091
unknown protein


741
replicase, putative
2092
hypothetical protein


742
putative cysteine proteinase
2093
No function assigned by TIGR


743
60S ribosomal protein L36
2094
putative APG protein


744
unknown protein
2095
glutathione S-transferase, putative


745
CLC-b chloride channel protein
2096
phytochrome-associated protein 1





(PAP1)


746
putative ribosomal protein S14
2097
amidophosphoribosyltransferase


747
histone H2B like protein
2098
nonphototropic hypocotyl 1



(emb|CAA69025.1)


748
60S ribosomal protein L2
2099
3-keto-acyl-CoA thiolase 2





(gb|AAC17877.1)


749
60S ribosomal protein L15
2100
pEARLI 1



homolog


750
ribosomal protein S27
2101
glutathione reductase, cytosolic


751
ribosomal protein
2102
putative protein


752
60S ribosomal protein L12
2103
putative protein


753
60s ribosomal protein L34
2104
putative aldehyde oxidase


754
putative ribosomal protein S10
2105
probable photosystem I





chain XI precursor


755
drought-induced protein like
2106
photosystem II polypeptide,





putative


756
blue copper-binding protein, 15K
2107
photosystem II reaction



(lamin)

center 6.1 KD protein


757
calmodulin-like protein
2108
33 kDa polypeptide of





oxygen-evolving complex





(OEC) in photosystem II





(emb|CAA75629.1)


758
putative protein
2109
60S ribosomal protein





L11B


759
No function assigned by TIGR
2110
extA (emb|CAA47807.1)


760
alpha-mannosidase, putative
2111
zinc finger protein OBP4-





like


761
uncoupling protein (ucp/PUMP)
2112
sterol delta7 reductase


762
homeodomain-like protein
2113
putative RAS-related





protein, RAB11C


763
ribosomal protein S18,
2114
glucosyltransferase like



putative

protein


764
similar to SOR1 from the
2115
zinc finger protein (PMZ),



fungus Cercospora

putative



nicotianae


765
60S ribosomal protein L13,
2116
6,7-dimethyl-8-



BBC1 protein

ribityllumazine synthase





precursor


766
50S ribosomal protein L24,
2117
putative protein



chloroplast precursor


767
putative ribosomal protein
2118
osmotin precursor


768
unknown protein
2119
No function assigned by





TIGR


769
aspartate aminotransferase
2120
ferredoxin precusor isolog



(AAT1)


770
potassium channel protein
2121
GH3 like protein



AtKC


771
unknown protein
2122
non-specific lipid transfer





protein


772
peroxisomal targeting
2123
homeodomain transcription



signal type 2 receptor

factor (HAT9)


773
putative protein
2124
putative cytochrome P450





monooxygenase


774
Ras-related GTP-binding
2125
putative protein kinase



protein (ARA-4)


775
S-receptor kinase homolog
2126
putative protein



2 precursor


776
pathogenesis-related group
2127
glyceraldehyde-3-



5 protein, putative

phosphate dehydrogenase


777
Nitrilase 4 (sp|P46011)
2128
putative protein disulfide-





isomerase


778
biotin carboxyl carrier
2129
unknown protein



protein of acetyl-CoA



carboxylase precursor



(BCCP) (sp|Q42533)


779
photosystem I reaction
2130
beta-1,3-glucanase class I



centre subunit psaN

precursor



precursor (PSI-N)



(sp|P49107)


780
3(2),5-bisphosphate
2131
homeobox-leucine zipper protein



nucleotidase

HAT5 (HD-ZIP protein 5) (HD-





ZIP protein ATHB-1)


781
high affinity Ca2+
2132
putative cyclic nucleotide-



antiporter

regulated ion channel protein


782
putative cytoskeletal
2133
P II nitrogen sensing protein GLB I



protein


783
putative peroxidase
2134
H-protein promoter binding factor-





1 (gb|AAC24592.1)


784
respiratory burst oxidase
2135
GAST1-like protein



protein


785
beta-glucosidase
2136
cytochrome P450 GA3


786
calcium-dependent protein kinase
2137
putative protein



(pir||S71196)


787
phosphoinositide specific
2138
Myb-related transcription factor-



phospholipase C

like protein


788
similarity to S-domain receptor-
2139
putative phloem-specific lectin



like protein kinase, Zea mays


789
mitosis-specific cyclin 1b
2140
protein kinase-like protein


790
4-coumarate:CoA ligase 3
2141
unknown protein


791
transcription factor IIB (TFIIB)
2142
SCARECROW transcriptional





regulator-like


792
unknown protein
2143
unknown protein


793
hypothetical protein
2144
unknown protein


794
hypothetical protein
2145
putative protein


795
sugar transporter like protein
2146
calnexin homolog


796
putative trypsin inhibitor
2147
PP1/PP2A phosphatases





pleiotropic regulator PRL2


797
unknown protein
2148
xyloglucan endotransglycosylase,





putative


798
putative multispanning membrane
2149
putative calmodulin



protein


799
receptor-like kinase, putative
2150
spermine synthase (ACL5)


800
putative inosine-5-monophosphate
2151
snoRNA



dehydrogenase


801
inosine-5′-monophosphate
2152
photosystem I subunit V precursor,



dehydrogenase, putative

putative


802
amino acid permease 6
2153
putative potassium transporter



(emb|CAA65051.1)


803
NADPH-ferrihemoprotein
2154
Homeodomain-like protein



reductase (ATR2)


804
putative WRKY-type DNA binding
2155
putative protein



protein


805
putative ankyrin
2156
unknown protein


806
putative hexose transporter
2157
CALMODULIN-RELATED





PROTEIN 2, TOUCH-INDUCED





(TCH2)


807
aquaporin/MIP-like protein
2158
putative protein phosphatase 2C


808
Ser/Thr protein kinase isolog
2159
monosaccharide transport





protein, STP4


809
pectate lyase like protein
2160
hypothetical protein


810
putative 60S ribosomal protein L17
2161
unknown protein


811
putative protein
2162
hypothetical protein


812
unknown protein
2163
putative protein kinase


813
phenylalanine ammonia-lyase
2164
putative serine/threonine





protein kinase


814
putative cytochrome P450
2165
jasmonate inducible



monooxygenase

protein, putative


815
ARR1 protein, putative
2166
similar to several small





proteins (~100 aa) that are





induced by heat, auxin,





ethylene and wounding





such as Phaseolus aureus





indole-3-acetic acid





induced protein ARG





(SW: 32292)


816
putative bHLH transcription factor
2167
unknown protein


817
aminomethyltransferase-like
2168
MYB-like protein



precursor protein


818
purple acid phosphatase precursor
2169
putative protein kinase


819
AP2 domain containing
2170
unknown protein



protein, putative


820
ubiquitin-conjugating
2171
CLC-d chloride channel



enzyme E2-21 kD 1

protein



(ubiquitin-protein ligase 4)



(ubiquitin carrier protein 4)



(sp|P42748)


821
translation initiation factor
2172
cytochrome P450-like





protein


822
putative VAMP-associated
2173
putative glutathione S-



protein

transferase


823
spermidine synthase,
2174
putative mandelonitrile



putative

lyase


824
putative protein
2175
hypothetical protein


825
unknown protein
2176
putative trypsin inhibitor


826
AtKAP alpha
2177
male sterility 2-like protein





(emb|CAA68191.1)


827
glyceraldehyde-3-
2178
unknown protein



phosphate dehydrogenase,



putative


828
putative poly(A) binding
2179
unknown protein



protein


829
alpha-tubulin, putative
2180
putative protein


830
serine/threonine-specific
2181
putative peroxidase



protein kinase ATPK64



(pir||S20918)


831
putative aspartate-tRNA
2182
putative thromboxane-A



ligase

synthase


832
ras-related small GTP-
2183
putative cytochrome P450



binding protein RAB1c


833
cycloartenol synthase
2184
peroxidase ATP21a


834
No function assigned by
2185
unknown protein



TIGR


835
cytochrome P450
2186
putative glutathione S-





transferase


836
GTPase AtRAB8
2187
defender against cell death protein


837
3-phosphoserine
2188
AP2 domain containing protein,



phosphatase

putative


838
transcription factor CRC
2189
actin depolymerizing factor-like





protein


839
nuclear cap-binding
2190
putative calcium-dependent protein



protein; CBP20

kinase (U90439)



(gb|AAD29697.1)


840
chloroplast membrane
2191
phosphoribosylanthranilate



protein (ALBINO3)

transferase, putative


841
biotin holocarboxylase
2192
oligopeptide transporter, putative



synthetase


842
expansin AtEx6
2193
calmodulin-like protein


843
unknown protein
2194
putative protease inhibitor


844
mercaptopyruvate
2195
MAP kinase



sulfurtransferase, putative


845
putative thiosulfate
2196
DNA binding protein MybSt1,



sulfurtransferase

putative


846
dihydrolipoamide S-
2197
putative protein



acetyltransferase


847
auxin transport protein REH1,
2198
putative protein



putative


848
putative auxin transport protein
2199
unknown protein


849
apyrase (Atapy1)
2200
unknown protein


850
root cap 1 (RCP1)
2201
unknown protein


851
hypothetical protein
2202
putative protein


852
putative protein
2203
unknown protein


853
predicted protein of unknown
2204
unknown protein



function


854
hypothetical protein
2205
hypothetical protein


855
hypothetical protein
2206
uncharacterized protein


856
hypothetical protein
2207
putative protein


857
putative aldehyde dehydrogenase
2208
hypothetical protein


858
putative peroxidase
2209
peroxidase (emb|CAA66967.1)


859
UDP-glucose 4-epimerase-like
2210
putative flavonol 3-O-



protein

glucosyltransferase


860
indole-3-acetate beta-
2211
putative flavonol 3-O-



glucosyltransferase like protein

glucosyltransferase


861
putative beta-1,3-glucanase
2212
putative protein


862
disease resistance protein-like
2213
glycerol-3-phosphate





acyltransferase


863
putative respiratory burst oxidase
2214
putative beta-1,3-glucanase



protein B


864
ubiquitin-conjugating enzyme
2215
putative ethylene response element



UBC3

binding protein (EREBP)


865
cytoplasmic aconitate hydratase
2216
putative CONSTANS-like B-box





zinc finger protein


866
NADPH oxidoreductase, putative
2217
putative protein


867
PROTEIN TRANSPORT
2218
unknown protein



PROTEIN SEC61 GAMMA



SUBUNIT-like


868
putative protein
2219
putative trehalose-6-phosphate





phosphatase (AtTPPA)


869
unknown protein
2220
putative protein


870
60S acidic ribosomal protein P2
2221
putative protein


871
No function assigned by TIGR
2222
unknown protein


872
1,4-alpha-glucan branching
2223
unknown prptein



enzyme protein soform SBE2.2



precursor


873
calcium binding protein (CaBP-22)
2224
unknown protein


874
putative phosphoglucomutase
2225
hypothetical protein


875
shaggy-like protein kinase
2226
putative metal-binding



etha (EC 2.7.1.-)

protein


876
pyruvate decarboxylase
2227
putative



(gb|AAB16855.1)

phosphoribosylglycinamide





synthetase


877
hypothetical protein
2228
unknown protein


878
putative protein kinase
2229
putative protein


879
putative protein kinase
2230
unknown protein


880
putative leucine
2231
unknown protein



aminopeptidase


881
probable cytochrome P450
2232
putative beta-galactosidase


882
protein kinase 6-like protein
2233
putative protein kinase


883
arginine methyltransferase
2234
putative protein



(pam1)


884
MYB96 transcription
2235
putative protein



factor-like protein

phosphatase 2C


885
putative protein
2236
putative growth regulator





protein


886
metal ion transporter
2237
putative ABC transporter


887
No function assigned by
2238
chloride channel



TIGR

(emb|CAA70310.1)


888
flax rust resistance protein,
2239
adrenodoxin-like protein



putative


889
fructose-2,6-
2240
NAM (no apical meristem)-



bisphosphatase, putative

like protein


890
exonuclease RRP41
2241
putative transcription factor





MYB41


891
squamosa promoter binding
2242
Myb DNA binding protein-



protein-like 2

like



(emb|CAB56576.1)


892
putative squamosa-
2243
AtMYB84



promoter binding protein


893
O-acetylserine (thiol) lyase,
2244
photosystem II type I



putative

chlorophyll a/b binding





protein


894
snoRNA
2245
putative aspartic proteinase


895
snoRNA
2246
jasmonate inducible





protein, putative


896
ferredoxin-NADP+
2247
putative protein



reductase


897
H+-transporting ATP
2248
No function assigned by



synthase chain 9-like

TIGR



protein


898
photosystem I subunit III
2249
putative phosphatidylserine



precursor, putative

synthase


899
photosystem I subunit VI
2250
putative nicotianamine



precursor

synthase


900
auxin-binding protein 1
2251
lysine and histidine specific



precursor

transporter, putative


901
putative RAS superfamily GTP-
2252
putative protein



binding protein


902
disease resistance protein-like
2253
putative protein


903
protein kinase like protein
2254
putative sugar transporter protein


904
glucuronosyl transferase-like
2255
12S cruciferin seed storage protein



protein


905
putative homeodomain
2256
putative auxin-induced protein,



transcription factor

IAA17/AXR3-1


906
putative flavonol reductase
2257
putative cyclin D


907
putative protein
2258
farnesyl diphosphate synthase





precursor (gb|AAB49290.1)


908
salt-tolerance protein
2259
putative potassium transport





protein (TRH1)


909
40S ribosomal protein S30
2260
putative NPK1-related MAP kinase


910
putative bZIP transcription factor
2261
putative protein


911
putative protein
2262
putative ABC transporter


912
putative cinnamoyl CoA reductase
2263
putative DNA-directed RNA





polymerase subunit


913
unknown protein
2264
putative small nuclear





ribonucleoprotein E


914
putative RNA-binding protein
2265
unknown protein


915
phosphatidylinositol synthase
2266
reticuline oxidase-like protein



(PIS1)


916
unknown protein
2267
putative 1-aminocyclopropane-1-





carboxylate oxidase


917
hydroxyproline-rich glycoprotein
2268
similar to Mlo proteins from H.



homolog

vulgare


918
50S ribosomal protein L15,
2269
long-chain-fatty-acid—CoA ligase-



chloroplast precursor

like protein


919
unknown protein
2270
putative protein


920
putative YME1 ATP-dependant
2271
chromatin remodelling complex



protease

ATPase chain ISWI-like protein


921
unknown protein
2272
hypothetical protein


922
putative ribosomal protein L28
2273
latex-abundant protein, putative


923
unknown protein
2274
N-acetylornithine deacetylase-like





protein, fragment


924
putative protein
2275
putative DNA-binding protein


925
protein ch-42 precursor,
2276
putative anthranilate N-



chloroplast

hydroxycinnamoyl/benzoyltransfer





ase


926
protein serine/threonine kinase,
2277
putative DNA binding protein



putative


927
beta-VPE
2278
cytochrome P450-like protein


928
putative vacuolar sorting receptor
2279
putative DNA-binding protein


929
putative translation initiation factor
2280
putative peptide transporter



IF-2


930
predicted protein of unknown
2281
putative reticuline oxidase-like



function

protein


931
putative protein
2282
thioredoxin, putative


932
hypothetical protein
2283
nodulin-like protein


933
hypothetical protein
2284
UDP-galactose transporter-





like protein


934
phosphate transporter, putative
2285
putative fibrillin


935
No function assigned by
2286
unknown protein



TIGR


936
beta subunit of protein
2287
unknown protein



farnesyl transferase ERA1


937
putative glutamate
2288
unknown protein



decarboxylase


938
putative indole-3-acetate
2289
hypothetical protein



beta-glucosyltransferase


939
putative receptor-like
2290
glyceraldehyde 3-phosphate



protein kinase

dehydrogenase A subunit





(GapA)


940
UDP-galactose 4-
2291
predicted protein of



epimerase-like protein

unknown function


941
putative proliferating cell
2292
putative protein



nuclear antigen, PCNA


942
ubiquitin conjugating
2293
putative protein



enzyme E2 (UBC13)


943
cyclophilin (CYP2)
2294
myb-like protein


944
cystatin
2295
hypothetical protein



(emb|CAA03929.1)


945
putative alcohol
2296
putative U5 small nuclear



dehydrogenase

ribonucleoprotein, an RNA





helicase


946
acidic ribosomal protein p1
2297
unknown protein


947
glutathione transferase
2298
cinnamyl alcohol



AtGST 10

dehydrogenase-like



(emb|CAA10457.1)

protein


948
putative tropinone
2299
hypothetical protein similar



reductase

to extensin-like protein


949
ZIP4, a putative zinc
2300
unknown protein



transporter


950
unknown protein
2301
putative chlorophyll a/b





binding protein


951
putative protein
2302
probable plasma membrane





intrinsic protein 1c


952
putative protein
2303
hexokinase (ATHXK2)


953
putative C2H2-type zinc
2304
calcium-dependent protein



finger protein

kinase


954
putative RING zinc finger
2305
5′-adenylylphosphosulfate



protein

reductase, putative


955
putative microtubule-
2306
Erd1 protein precursor



associated protein

(sp|P42762)


956
unknown protein
2307
putative protein


957
putative protein
2308
putative protein


958
putative protein
2309
unknown protein



phosphatase-2c


959
V-ATPase subunit G (vag2
2310
BCS1 protein-like protein



gene)


960
hypothetical protein
2311
putative protein


961
unknown protein
2312
putative protein


962
unknown protein
2313
putative protein kinase


963
unknown protein
2314
indoleacetic acid (IAA)-inducible





gene (IAA7)


964
myrosinase-associated protein,
2315
ATP-dependent Clp protease



putative

regulatory subunit CLPX


965
hypothetical protein
2316
DNA-binding protein RAV1


966
hypothetical protein
2317
putative protein


967
No function assigned by TIGR
2318
hypothetical protein


968
unknown protein
2319
unknown protein


969
hypothetical protein
2320
unknown protein


970
LAX1/AUX1-like permease
2321
putative protein


971
putative UDP-N-
2322
putative thioredoxin reductase



acetylglucosamine--dolichyl-



phosphate N-



acetylglucosaminephosphotransfer



ase


972
chorismate mutase CM2
2323
unknown protein


973
inner mitochondrial membrane
2324
putative lectin



protein


974
DEF (CLA1) protein
2325
No function assigned by TIGR


975
decoy
2326
beta-fructosidase


976
citrate synthase
2327
chlorophyll a/b-binding protein





CP29


977
myosin
2328
photosystem I subunit PSI-E-like





protein


978
40S ribosomal protein S19
2329
peroxidase ATP8a


979
ripening-related protein-like
2330
putative fructose bisphosphate





aldolase


980
putative signal peptidase I
2331
zinc finger protein ATZF1,





putative


981
methionyl-tRNA synthetase
2332
DegP protease precursor



(AtcpMetRS)


982
ribosomal protein precursor-like
2333
transcription factor-like protein


983
50S ribosomal protein L21
2334
calcium-dependent protein kinase



chloroplast precursor (CL21)


984
putative MYB family transcription
2335
hypothetical protein



factor


985
cyclophilin-like protein
2336
putative protein


986
hypothetical protein
2337
glucose-1-phosphate





adenylyltransferase (APL3)


987
naringenin 3-dioxygenase like
2338
No function assigned by TIGR



protein


988
WD-repeat protein-like protein
2339
putative Eukaryotic initiation factor





4A


989
putative serine carboxypeptidase II
2340
No function assigned by TIGR


990
prenyltransferase, putative
2341
unknown protein


991
putative ligand-gated ion channel
2342
beta tubulin 1, putative



protein


992
clathrin adaptor medium chain
2343
one helix protein (OHP)



protein MU1B, putative


993
No function assigned by TIGR
2344
No function assigned by TIGR


994
putative Tal 1-like non-
2345
zinc finger protein 5, ZFP5



LTR retroelement protein


995
putative 3-isopropylmalate
2346
putative MYB family transcription



dehydrogenase

factor


996
3-isopropylmalate
2347
putative amino acid transporter



dehydratase, small subunit

protein


997
unknown protein
2348
putative potassium





transporter


998
unknown protein
2349
protein kinase (AFC2)


999
unknown protein
2350
putative protein


1000
hypothetical protein
2351
No function assigned by





TIGR


1001
putative protein
2352
putative ubiquitin-





conjugating enzyme E2


1002
No function assigned by
2353
unknown protein



TIGR


1003
putative beta-glucosidase
2354
cytochrome P450





monooxygenase (CYP71B3)


1004
putative pectate lyase A11
2355
putative myrosinase-





binding protein


1005
putative beta-glucosidase
2356
putative vacuolar sorting





receptor


1006
HD-Zip protein
2357
uridine diphosphate glucose





epimerase


1007
putative ubiquitin
2358
shaggy related protein



conjugating enzyme

kinase, ASK-GAMMA


1008
homeobox-leucine zipper
2359
ankyrin repeat protein



protein-like

EMB506


1009
cytochrome P450 like
2360
putative beta-alanine-



protein

pyruvate aminotransferase


1010
putative cysteine proteinase
2361
putative alcohol



inhibitor B (cystatin B)

dehydrogenase


1011
ethylene response sensor
2362
putative receptor-like



(ERS)

protein kinase


1012
putative SWH1 protein
2363
unknown protein


1013
putative glutathione S-
2364
putative methylmalonate



transferase

semi-aldehyde





dehydrogenase


1014
putative protein
2365
hypothetical protein


1015
unknown protein
2366
unknown protein


1016
putative protein
2367
peroxidase ATP13a



phosphatase 2C


1017
dnaJ protein homolog atj3
2368
putative glutathione





peroxidase


1018
ferredoxin
2369
squamosa promoter binding





protein-like 7


1019
hypothetical protein
2370
photosystem II core





complex protein, putative


1020
putative sugar transport
2371
snoRNA



protein, ERD6


1021
putative DnaJ protein
2372
photosystem I subunit X





precursor


1022
putative AP2 domain
2373
MYB transcription factor



transcription factor

(Atmyb2)


1023
putative protein
2374
putative PHD-type zinc finger





protein


1024
putative cyclin-dependent
2375
nuclear RNA binding protein A-



kinase regulatory subunit

like protein


1025
putative tropinone reductase
2376
unknown protein


1026
signal response protein (GAI)
2377
unknown protein


1027
putative steroid sulfotransferase
2378
unknown protein


1028
hypothetical protein
2379
putative amino-cyclopropane-





carboxylic acid oxidase (ACC





oxidase)


1029
nucleic acid binding protein-like
2380
hypothetical protein


1030
putative protein
2381
indole-3-acetate beta-





glucosyltransferase like protein


1031
blue copper binding protein
2382
predicted protein


1032
farnesylated protein (ATFP6)
2383
unknown protein


1033
unknown protein
2384
No function assigned by TIGR


1034
putative PCF2-like DNA binding
2385
putative photosystem I reaction



protein

center subunit IV


1035
teosinte branched1-like protein
2386
putative homeodomain





transcription factor


1036
putative protein
2387
putative purple acid phosphatase





precursor


1037
unknown protein
2388
No function assigned by TIGR


1038
unknown protein
2389
nitrate reductase 1 (NR1)


1039
2-oxoglutarate dehydrogenase, E1
2390
putative casein kinase II beta



component

subunit


1040
unknown protein
2391
pEARLI 1-like protein


1041
unknown protein
2392
putative protein


1042
CCAAT-binding transcription
2393
No function assigned by TIGR



factor subunit A (CBF-A)


1043
hypothetical protein
2394
unknown protein


1044
putative growth regulator protein
2395
putative cell wall-plasma





membrane disconnecting CLCT





protein (AIR1A)


1045
putative presenilin
2396
unknown protein


1046
putative expansin
2397
scarecrow-like 11-like


1047
ribosomal-like protein
2398
putative anthocyanidin synthase


1048
unknown protein
2399
putative AP2 domain transcription





factor


1049
unknown protein
2400
caffeoyl-CoA O-methyltransferase-





like protein


1050
putative protein
2401
unknown protein


1051
putative protein
2402
putative protein kinase


1052
unknown protein
2403
cytochrome P450-like protein


1053
unknown protein
2404
putative MADS-box protein ANR1


1054
unknown protein
2405
putative glutathione S-transferase


1055
unknown protein
2406
hypothetical protein


1056
unknown protein
2407
similar to gibberellin-





regulated proteins


1057
putative protein
2408
unknown protein


1058
putative protein
2409
putative sensory





transduction histidine





kinase


1059
argininosuccinate lyase (AtArgH)
2410
similar to late





embryogenesis abundant





proteins


1060
disease resistance protein homolog
2411
unknown protein


1061
aldehyde dehydrogenase like
2412
putative protein



protein


1062
GBF2, G-box binding factor
2413
putative ATP-dependent





RNA helicase


1063
CDPK-related kinase
2414
putative protein


1064
endo-1,4-beta-glucanase
2415
putative sucrose synthetase


1065
putative serine protease
2416
beta-fructofuranosidase 1


1066
serine/threonine-specific
2417
putative indole-3-acetate



kinase lecRK1 precursor, lectin

beta-glucosyltransferase



receptor-like


1067
putative MAP kinase
2418
hypothetical protein


1068
RNase L inhibitor-like
2419
DNA-directed RNA



protein

polymerase II, third largest subunit


1069
No function assigned by
2420
putative transcription factor



TIGR


1070
AP2 domain transcription
2421
homeobox-leucine zipper



factor

protein ATHB-5 (HD-zip protein





ATHB-5) (sp|P46667)


1071
polygalacturonase
2422
putative ftsH chloroplast



isoenzyme 1 beta subunit,

protease



putative


1072
putative lipid transfer
2423
replication protein A1-like



protein


1073
putative protein kinase
2424
hypothetical protein


1074
putative protein
2425
unknown protein


1075
ATP-dependent RNA
2426
unknown protein



helicase like protein


1076
putative cyclic nucleotide-
2427
putative methionine



regulated ion channel

aminopeptidase



protein


1077
COP1 like protein
2428
unknown protein


1078
putative peroxidase
2429
fatty acid elongase-like





protein (cer2-like)


1079
putative NAK-like ser/thr
2430
unknown protein



protein kinase


1080
putative cytochrome C
2431
putative disease resistance





response protein


1081
cytochrome c
2432
putative protein


1082
putative serine
2433
unknown protein



carboxypeptidase II


1083
acyl-(acyl carrier protein)
2434
putative protein



thioesterase


1084
DNA-binding factor,
2435
putative protein



putative


1085
MAP3K delta-1 protein
2436
unknown protein



kinase


1086
AtMlo-h1-like protein
2437
putative protein


1087
No function assigned by
2438
unknown protein



TIGR


1088
putative expansin
2439
unknown protein


1089
defender against cell death
2440
putative protein



protein, putative


1090
glycolate oxidase-like
2441
No function assigned by TIGR



protein


1091
putative ATP-dependent RNA
2442
MADS-box protein AGL14



helicase


1092
putative protein
2443
No function assigned by TIGR


1093
putative HMG protein
2444
peptidylprolyl isomerase


1094
squalene monooxygenase 2
2445
putative s-adenosylmethionine



(squalene epoxidase 2) (SE 2)

synthetase



(sp|O65403)


1095
eukaryotic peptide chain release
2446
peroxidase



factor subunit 1, putative


1096
auxin-induced protein-like
2447
ferrochelatase-I


1097
putative lipoamide dehydrogenase
2448
putative eukaryotic initiation factor





4, eIF4


1098
putative protein
2449
drought-inducible cysteine





proteinase RD21A precursor-like





protein


1099
unknown protein
2450
unknown protein


1100
putative oligopeptide transporter
2451
unknown protein


1101
putative translation elongation
2452
No function assigned by TIGR



factor ts


1102
putative CCAAT-binding
2453
No function assigned by TIGR



transcription factor subunit


1103
putative ABC transporter
2454
salt-inducible like protein


1104
putative superoxide-generating
2455
glucose-6-phosphate 1-



NADPH oxidase flavocytochrome

dehydrogenase


1105
aspartate kinase-homoserine
2456
3-hydroxy-3-methylglutaryl CoA



dehydrogenase-like protein

reductase (AA 1-592)


1106
putative bHLH transcription factor
2457
hypothetical protein


1107
putative geranylgeranyl transferase
2458
putative protein



type I beta subunit


1108
putative ARP2/3 protein complex
2459
putative putative 60S ribosomal



subunit p41

protein L17


1109
sulphite reductase
2460
putative inorganic pyrophosphatase


1110
putative auxin-regulated protein
2461
putative gamma-





glutamyltransferase


1111
transcription factor scarecrow-like
2462
heat shock transcription factor-



14, putative

like protein


1112
unknown protein
2463
mitochondrial chaperonin hsp60


1113
monooxygenase 2 (MO2)
2464
unknown protein


1114
putative amine oxidase
2465
putative zinc finger protein





identical to T10M13.22


1115
zinc finger protein, putative
2466
putative uridylyl transferase


1116
DNA-binding protein, putative
2467
nodulin-like protein


1117
putative protein
2468
putative B-box zinc finger protein


1118
putative protein
2469
No function assigned by TIGR


1119
Avr9 elicitor response like protein
2470
putative metalloproteinase


1120
putative protein
2471
putative cellular apoptosis





susceptibility protein


1121
hypothetical protein
2472
hypothetical protein


1122
putative nucleotide-sugar
2473
hypothetical protein



dehydratase


1123
UFD1 like protein
2474
scarecrow-like 13 (SCL13)


1124
putative trans-
2475
putative nucleoside



prenyltransferase

triphosphatase


1125
outward rectifying
2476
unknown protein



potassium channel KCO


1126
unknown protein
2477
No function assigned by





TIGR


1127
putative
2478
hypothetical protein



pectinacetylesterase


1128
putative protein
2479
putative phospholipase


1129
No function assigned by
2480
putative snRNP protein



TIGR


1130
unknown protein
2481
putative protein


1131
unknown protein
2482
putative lipase


1132
unknown protein
2483
putative nonsense-mediated





mRNA decay protein


1133
protein phosphatase
2484
No function assigned by



homolog (PPH1)

TIGR


1134
unknown protein
2485
protochlorophyllide





reductase precursor


1135
No function assigned by
2486
No function assigned by



TIGR

TIGR


1136
unknown protein
2487
trehalose-6-phosphate





synthase, putative


1137
unknown protein
2488
unknown protein


1138
unknown protein
2489
germin-like protein


1139
putative protein
2490
plastid protein


1140
unknown protein
2491
putative protein


1141
putative ubiquinol--
2492
hypothetical protein



cytochrome-c reductase


1142
unknown protein
2493
unknown protein


1143
contains similarity to high-
2494
unknown protein



glucose-regulated protein 8



GB:AAF08813 GI:6449083



from [Homo sapiens]


1144
unknown protein
2495
histone deacetylase-like





protein


1145
putative cis-Golgi SNARE
2496
unknown protein



protein


1146
unknown protein
2497
unknown protein


1147
glutamate-1-semialdehyde
2498
putative protein



aminotransferase


1148
No function assigned by
2499
putative protein



TIGR


1149
hypothetical protein
2500
No function assigned by





TIGR


1150
unknown protein
2501
putative zinc transporter





ZIP2-like


1151
unknown protein
2502
unknown protein


1152
unknown protein
2503
putative ribosomal-protein





S6 kinase (ATPK19)


1153
scarecrow-like 3
2504
unknown protein


1154
putative proline-rich protein
2505
unknown protein


1155
cytochrome c oxidoreductase like
2506
60S ribosomal protein L10A



protein


1156
putative
2507
putative protein



carboxymethylenebutenolidase


1157
unknown protein
2508
receptor protein kinase (IRK1),





putative


1158
unknown protein
2509
putative nematode-resistance





protein


1159
unknown protein
2510
tubulin alpha-5 chain-like protein


1160
unknown protein
2511
putative DNA-binding protein


1161
unknown protein
2512
unknown protein


1162
unknown protein
2513
putative RGA1, giberellin repsonse





modulation protein


1163
auxin-induced protein (IAA20)
2514
non phototropic hypocotyl 1-like


1164
50S ribosomal protein L4
2515
RING-H2 finger protein RHA1b


1165
putative DNA topoisomerase III
2516
putative myb-protein



beta


1166
No function assigned by TIGR
2517
hydroperoxide lyase (HPOL) like





protein


1167
isp4 like protein
2518
serine/threonine-protein kinase,





PK7


1168
putative protein kinase
2519
putative vacuolar proton-ATPase





subunit


1169
hypothetical protein
2520
putative polygalacturonase


1170
putative pyrophosphate--fructose-
2521
putative ribosomal protein L8



6-phosphate 1-phosphotransferase


1171
putative protein
2522
putative adenylate kinase


1172
putative protein
2523
germin-like protein (GLP10)


1173
putative protein
2524
putative chlorophyll a/b binding





protein


1174
unknown protein
2525
chloroplast single subunit DNA-





dependent RNA polymerase


1175
unknown protein
2526
putative protein


1176
putative protein
2527
hypothetical protein


1177
putative protein
2528
hypothetical protein


1178
unknown protein
2529
b-keto acyl reductase, putative


1179
unknown protein
2530
cellulose synthase catalytic subunit


1180
putative protein
2531
putative 1-aminocyclopropane-1-





carboxylate oxidase


1181
brassinosteroid insensitive 1 gene
2532
S-linalool synthase, putative



(BRI1)


1182
putative receptor protein kinase
2533
phosphoribosyl-ATP





pyrophosphohydrolase (At-IE)


1183
vacuolar-type H+-translocating
2534
disease resistance RPP5 like



inorganic pyrophosphatase

protein (fragment)


1184
protein kinase-like protein
2535
putative protein


1185
glycyl tRNA synthetase, putative
2536
beta-galactosidase like protein


1186
subtilisin proteinase-like
2537
putative translation





initiation factor eIF-2,





gamma subunit


1187
hypothetical protein
2538
ankyrin like protein


1188
cytochrome P450-like protein
2539
histone H2A-like protein


1189
cytochrome p450 like protein
2540
putative protein


1190
putative protein kinase
2541
salt-tolerance zinc finger





protein


1191
pectinesterase-like protein
2542
unknown protein


1192
putative receptor-like protein
2543
putative protein



kinase


1193
peroxidase ATP17a-like
2544
fructose-bisphosphate



protein

aldolase


1194
No function assigned by
2545
peroxidase



TIGR

(emb|CAA66964.1)


1195
cellulose synthase catalytic
2546
patatin-like protein



subunit-like protein


1196
RAS-related protein, RAB7
2547
salt-inducible protein





homolog


1197
putative aspartate
2548
hypothetical protein



aminotransferase


1198
cyclophilin
2549
xyloglucan endo-





transglycosylase-like





protein


1199
putative SF2/ASF splicing
2550
trihelix DNA-binding



modulator, Srp30

protein (GT2)


1200
putative cytochrome b5
2551
ubiquitin-conjugating





enzyme 16, putative


1201
glutamyl-tRNA reductase,
2552
homeobox protein



putative


1202
putative MADS-box protein
2553
envelope Ca2+-ATPase


1203
ammonium transport
2554
snap25a



protein (AMT1)


1204
No function assigned by
2555
putative annexin



TIGR


1205
putative beta-ketoacyl-CoA
2556
putative protein



synthase


1206
thaumatin-like protein
2557
homeodomain transcription





factor (ATHB-14)


1207
putative methionine
2558
heat shock protein, putative



aminopeptidase


1208
putative protein
2559
peroxidase ATP23a



phosphatase 2C


1209
kinase-like protein
2560
p68 RNA helicase, putative


1210
receptor-associated kinase
2561
potassium transporter,



isolog

putative


1211
mitochondrial ribosomal
2562
putative eukaryotic



protein S14

translation initiation factor 2 alpha





subunit, eIF2


1212
oleosin, 18.5K
2563
hypothetical protein


1213
chalcone isomerase
2564
carnitine racemase like





protein


1214
putative cyclin-dependent
2565
No function assigned by



kinase regulatory subunit

TIGR


1215
putative thaumatin-like
2566
unknown protein



protein


1216
putative two-component
2567
unknown protein



response regulator protein


1217
TATA binding protein-
2568
unknown protein



associated factor, putative


1218
predicted protein of
2569
serine/threonine kinase-like



unknown function

protein


1219
putative AP2 domain transcription
2570
peroxidase (emb|CAA66960.1)



factor


1220
brassinosteroid receptor kinase,
2571
putative protein



putative


1221
TINY-like protein
2572
hypothetical protein


1222
glucose-6-phosphate isomerase
2573
glycine-rich protein 2 (GRP2)


1223
putative protein
2574
unknown protein


1224
putative NAM (no apical
2575
berberine bridge enzyme-like



meristem)-like protein

protein


1225
unknown protein
2576
unknown protein


1226
putative nucleotide-binding protein
2577
putative WD-repeat protein


1227
bZIP transcription factor (POSF21)
2578
serine/threonine kinase-like





protein


1228
ubiquitin activating enzyme-like
2579
serine/threonine kinase-like



protein

protein


1229
telomere repeat-binding protein
2580
Cu2+-transporting ATPase-like





protein


1230
unknown protein
2581
translation initiation factor eIF4E


1231
mevalonate kinase
2582
O-methyltransferase-like protein


1232
putative protein
2583
translation initiation factor eIF3-





like protein


1233
hypothetical protein
2584
No function assigned by TIGR


1234
disease resistance RPP5 like
2585
unknown protein



protein


1235
putative protein
2586
hypothetical protein


1236
putative pectinesterase
2587
unknown protein


1237
Ttg1 protein (emb|CAB45372.1)
2588
unknown protein


1238
FUSCA PROTEIN FUS6
2589
glycine-rich protein like


1239
NHE1 Na+/H+ exchanger
2590
putative disease resistance protein


1240
No function assigned by TIGR
2591
putative Na+/Ca2+ antiporter


1241
Phospholipase like protein
2592
putative hydroxymethylglutaryl-





CoA lyase


1242
unknown protein
2593
putative





phosphoribosylaminoimidazole





carboxylase


1243
unknown protein
2594
SAR DNA-binding protein-like


1244
unknown protein
2595
response regulator, putative


1245
AUX1-like amino acid permease
2596
fibrillin precursor-like protein


1246
unknown protein
2597
beta-ketoacyl-CoA synthase





(FIDDLEHEAD)


1247
putative C2H2-type zinc finger
2598
lectin like protein



protein


1248
putative protein
2599
No function assigned by TIGR


1249
putative protein
2600
acidic endochitinase





(dbj|BAA21861.1)


1250
putative glucosyltransferase
2601
unknown protein


1251
putative lipase
2602
hypothetical protein


1252
putative protein
2603
predicted OR23 protein of





unknown function


1253
putative thioredoxin
2604
putative protein


1254
AIG2-like protein
2605
hypothetical protein


1255
short-chain alcohol dehydrogenase
2606
glycerol-3-phosphate



like protein

dehydrogenase


1256
hypothetical protein
2607
hypothetical protein


1257
putative protein
2608
tat-binding protein, putative


1258
putative protein
2609
putative protein


1259
glutathione peroxidase-
2610
putative trehalose-6-



like protein

phosphate phosphatase


1260
putative protein
2611
hypothetical protein


1261
putative disease resistance
2612
putative flavonol 3-O-



response protein

glucosyltransferase


1262
putative protein
2613
60S ribosomal protein L30


1263
senescence-associated
2614
putative auxin-induced



protein (SAG29)

protein


1264
glycolate oxidase, putative
2615
putative nonspecific lipid-





transfer protein precursor


1265
extensin-like protein
2616
AtRer1A


1266
putative protein
2617
putative aquaporin





(tonoplast intrinsic protein





gamma)


1267
unknown protein
2618
hypothetical protein


1268
putative disease resistance
2619
putative alanine acetyl



protein

transferase


1269
putative receptor-like
2620
putative NADP-dependent



protein kinase

glyceraldehyde-3-





phosphate dehydrogenase


1270
putative receptor-like
2621
putative DNA binding



protein kinase

protein


1271
basic chitinase
2622
putative cystathionine





gamma-synthase


1272
putative pectin
2623
unknown protein



methylesterase


1273
peroxidase ATP N
2624
malate oxidoreductase





(malic enzyme)


1274
class 2 non-symbiotic
2625
unknown protein



hemoglobin


1275
nitrate transporter
2626
cyclic nucleotide-gated





cation channel


1276
Ca2+/H+-exchanging
2627
glyoxalase II, putative



protein-like


1277
putative protein
2628
putative trypsin inhibitor


1278
hydroxynitrile lyase like
2629
unknown protein



protein


1279
putative AP2 domain
2630
unknown protein



transcription factor


1280
pectin methylesterase,
2631
unknown protein



putative


1281
putative protein
2632
nucleosome assembly protein I-like





protein


1282
beta-glucosidase-like
2633
membrane channel like protein



protein


1283
CCAAT box binding factor/
2634
anthocyanin2, putative



transcription factor Hap2a


1284
putative fibrillin
2635
TWIN SISTER OF FT (TSF)


1285
xyloglucan endo-
2636
putative myb-related transcription



transglycosylase

factor


1286
putative 10 kd chaperonin
2637
hypothetical protein


1287
No function assigned by TIGR
2638
putative RING zinc finger protein


1288
serine/threonine protein kinase
2639
amino acid transport protein AAT1



ATPK10


1289
putative lipase
2640
putative protein


1290
choline kinase GmCK2p-like
2641
putative protein



protein


1291
putative sugar transport protein,
2642
xanthine dehydrogenase



ERD6


1292
MYB27 protein-like
2643
xanthine dehydrogenase-like





protein


1293
DNA-binding protein, putative
2644
receptor protein kinase (IRK1),





putative


1294
similar to cold acclimation protein
2645
dehydrin-like protein



WCOR413 [Triticum aestivum]


1295
unknown protein
2646
unknown protein


1296
aquaporin (plasma membrane
2647
aldehyde dehydrogenase homolog,



intrinsic protein 2B)

putative


1297
No function assigned by TIGR
2648
Ran binding protein (AtRanBP1b)


1298
P-Protein-like protein
2649
putative squamosa-promoter





binding protein


1299
No function assigned by TIGR
2650
putative protein


1300
putative cytochrome P450
2651
kinesin like protein



monooxygenase


1301
putative cytochrome P450
2652
putative cellulose synthase



monooxygenase


1302
putative thioredoxin
2653
calmodulin (cam2)


1303
stromal ascorbate peroxidase
2654
fibrillarin-like protein


1304
ethylene responsive element
2655
putative transmembrane protein



binding factor-like protein

G5p



(AtERF6)


1305
auxin transport protein EIR1
2656
putative peroxidase



(gb|AAC39513.1)


1306
putative CONSTANS-like B-box
2657
putative SNF1-related protein



zinc finger protein

kinase


1307
putative protein kinase
2658
glutathione S-transferase, putative


1308
mitochondrial Lon protease
2659
unknown protein



homolog 1 precursor (sp|O64948)


1309
putative protein
2660
hypothetical protein


1310
heme activated protein, putative
2661
putative protein


1311
putative cytochrome P450
2662
phosphatidylinositol-4-phosphate





5-kinase isolog


1312
No function assigned by TIGR
2663
putative tyrosine decarboxylase


1313
putative lipase
2664
unknown protein


1314
putative protein
2665
SGP1 monomeric G-protein





(emb|CAB54517.1)


1315
putative sugar transporter protein
2666
putative serine





carboxypeptidase II


1316
putative sucrose transport protein,
2667
putative L5 ribosomal



SUC2

proteiii


1317
putative protein
2668
putative glucosyltransferase


1318
putative protein
2669
flavonoid 3,5-hydroxylase





like protein


1319
putative endochitinase
2670
putative protein


1320
putative acetone-
2671
putative protein



cyanohydrin lyase


1321
putative protein
2672
putative Fe (II)/ascorbate





oxidase


1322
calmodulin-like protein
2673
putative anthocyanin 5-





aromatic acyltransferase


1323
hypothetical protein
2674
casein kinase I


1324
cysteine proteinase like
2675
putative 2,3-



protein

bisphosphoglycerate-





independent





phosphoglycerate mutase


1325
heat shock protein 17.6-II
2676
putative glutathione S-





transferase TSI-1


1326
heat shock protein 18
2677
ATP-dependent RNA





helicase


1327
Arabidopsis mitochondrion-
2678
putative cytochrome P450



localized small heat shock



protein (AtHSP23.6-mito)


1328
unknown protein
2679
putative WD-40 repeat





protein


1329
putative WRKY-type DNA
2680
No function assigned by



binding protein

TIGR


1330
No function assigned by
2681
No function assigned by



TIGR

TIGR


1331
hypothetical protein
2682
putative protein


1332
putative integral membrane
2683
putative extensin



protein nodulin


1333
putative protein
2684
nodulin-26-like protein


1334
unknown protein
2685
RNA helicase





(emb|CAA09212.1)


1335
3-isopropylmalate
2686
predicted protein of



dehydratase, small subunit

unknown function


1336
unknown protein
2687
putative berberine bridge





enzyme


1337
putative homeodomain
2688
thioredoxin, putative



transcription factor


1338
unknown protein
2689
putative serine





carboxypeptidase I


1339
putative protein
2690
cytochrome P450-like





protein


1340
peroxidase ATP19a
2691
putative pyrophosphate-dependent





phosphofructokinase alpha subunit


1341
putative Na+/H+-
2692
putative flavonol



exchanging protein

glucosyltransferase


1342
putative auxin-regulated
2693
peroxidase ATP20a



protein

(emb|CAA67338.1)


1343
unknown protein
2694
TOPP8 serine/threonine protein





phosphatase type one


1344
unknown protein
2695
auxin regulated protein IAA18,





putative


1345
putative trehalose-6-
2696
putative WRKY-type DNA binding



phosphate synthase

protein


1346
putative lectin
2697
putative glucan synthase


1347
Mlo protein-like
2698
squalene monooxygenase


1348
unknown protein
2699
putative proline-rich protein


1349
ethylene response factor,
2700
G2484-1 protein



putative


1350
unknown protein
2701
heat shock protein 70 like protein


1351
unknown protein
2702
unknown protein




2703
unknown protein
















TABLE 2







ABIOTIC STRESS RESPONSIVE


GENE REGULATORY SEQUENCES












SEQ
REGULATORY
SEQ
REGULATORY



ID NO
REGION
ID NO
REGION
















1
2704
1353
4046



2
2705
1354
4047



3
2706
1355
4048



4
2707
1356
4049



5
2708
1357
4050



6
2709
1358
4051



7
2710
1359
4052



8
2711
1360
4053



9
2712
1361
4054



10
2713
1362
4055



11
2714
1363
4056



12
2715
1364
4057



13
2716
1365
4058



14
2717
1366
4059



15
2718
1367
4060



16
2719
1368
4061



17
2720
1369
4062



18
2721
1370
4063



19
2722
1371
4064



20
2723
1372
4065



21
2724
1373
4066



22
2725
1374
4067



23
2726
1375
4068



24
2727
1376
4069



25
2728
1377
4070



26
2729
1378
4071



27
2730
1379
4072



28
2731
1380
4073



29
2732
1381
4074



30
2733
1382
4075



31
2734
1383
4076



32
2735
1384
4077



33
2736
1385
4078



34
2737
1386
4079



35
2738
1387
4080



36
2739
1388
4081



37
2740
1389
4082



38
2741
1390
4083



39
2742
1391
4084



40
2743
1392
4085



41
2744
1393
4086



42
2745
1394
4087



43
NONE
1395
4088



44
2746
1396
4089



45
2747
1397
4090



46
2748
1398
4091



47
2749
1399
4092



48
2750
1400
4093



49
2751
1401
4094



50
2752
1402
4095



51
2753
1403
4096



52
2754
1404
4097



53
2755
1405
4098



54
2756
1406
4099



55
2757
1407
4100



56
2758
1408
4101



57
2759
1409
4102



58
2760
1410
4103



59
2761
1411
4104



60
2762
1412
4105



61
2763
1413
4106



62
2764
1414
4107



63
2765
1415
4108



64
2766
1416
4109



65
2767
1417
4110



66
2768
1418
4111



67
2769
1419
4112



68
2770
1420
4113



69
NONE
1421
4114



70
2771
1422
4115



71
2772
1423
4116



72
2773
1424
4117



73
2774
1425
4118



74
2775
1426
4119



75
2776
1427
4120



76
2777
1428
4121



77
2778
1429
4122



78
2779
1430
4123



79
2780
1431
4124



80
2781
1432
NONE



81
2782
1433
4125



82
2783
1434
4126



83
2784
1435
4127



84
2785
1436
4128



85
2786
1437
4129



86
2787
1438
4130



87
2788
1439
4131



88
2789
1440
4132



89
2790
1441
4133



90
2791
1442
4134



91
2792
1443
4135



92
2793
1444
4136



93
2794
1445
4137



94
2795
1446
4138



95
2796
1447
4139



96
2797
1448
4140



97
2798
1449
4141



98
2799
1450
4142



99
2800
1451
4143



100
2801
1452
4144



101
2802
1453
4145



102
2803
1454
4146



103
2804
1455
4147



104
2805
1456
4148



105
2806
1457
4149



106
2807
1458
4150



107
2808
1459
4151



108
2809
1460
4152



109
2810
1461
4153



110
2811
1462
4154



111
2812
1463
4155



112
2813
1464
4156



113
2814
1465
4157



114
2815
1466
4158



115
2816
1467
4159



116
2817
1468
4160



117
2818
1469
4161



118
2819
1470
4162



119
2820
1471
4163



120
2821
1472
4164



121
2822
1473
4165



122
2823
1474
4166



123
2824
1475
4167



124
2825
1476
4168



125
2826
1477
4169



126
2827
1478
4170



127
2828
1479
4171



128
2829
1480
4172



129
2830
1481
4173



130
2831
1482
4174



131
2832
1483
4175



132
2833
1484
4176



133
2834
1485
4177



134
2835
1486
4178



135
2836
1487
4179



136
2837
1488
4180



137
2838
1489
4181



138
2839
1490
4182



139
2840
1491
4183



140
2841
1492
4184



141
2842
1493
4185



142
2843
1494
4186



143
2844
1495
4187



144
NONE
1496
4188



145
2845
1497
4189



146
2846
1498
4190



147
2847
1499
4191



148
2848
1500
4192



149
2849
1501
4193



150
2850
1502
4194



151
2851
1503
4195



152
2852
1504
4196



153
2853
1505
4197



154
2854
1506
4198



155
2855
1507
4199



156
2856
1508
4200



157
2857
1509
4201



158
2858
1510
4202



159
2859
1511
4203



160
2860
1512
4204



161
2861
1513
4205



162
2862
1514
4206



163
2863
1515
4207



164
2864
1516
4208



165
2865
1517
4209



166
2866
1518
4210



167
2867
1519
4211



168
2868
1520
4212



169
2869
1521
4213



170
2870
1522
4214



171
2871
1523
4215



172
2872
1524
4216



173
2873
1525
4217



174
2874
1526
4218



175
2875
1527
4219



176
2876
1528
4220



177
2877
1529
4221



178
2878
1530
4222



179
2879
1531
4223



180
2880
1532
4224



181
2881
1533
4225



182
2882
1534
4226



183
2883
1535
4227



184
2884
1536
4228



185
2885
1537
4229



186
2886
1538
4230



187
2887
1539
4231



188
2888
1540
4232



189
2889
1541
4233



190
2890
1542
4234



191
2891
1543
4235



192
2892
1544
4236



193
2893
1545
4237



194
2894
1546
4238



195
2895
1547
4239



196
2896
1548
4240



197
2897
1549
4241



198
2898
1550
4242



199
2899
1551
4243



200
2900
1552
4244



201
2901
1553
4245



202
2902
1554
4246



203
2903
1555
4247



204
2904
1556
4248



205
2905
1557
4249



206
2906
1558
NONE



207
2907
1559
4250



208
2908
1560
4251



209
2909
1561
4252



210
2910
1562
4253



211
2911
1563
4254



212
2912
1564
4255



213
2913
1565
4256



214
2914
1566
4257



215
2915
1567
4258



216
2916
1568
4259



217
2917
1569
4260



218
2918
1570
4261



219
2919
1571
4262



220
2920
1572
4263



221
2921
1573
4264



222
2922
1574
4265



223
2923
1575
4266



224
2924
1576
4267



225
2925
1577
4268



226
2926
1578
4269



227
2927
1579
4270



228
2928
1580
4271



229
2929
1581
4272



230
2930
1582
4273



231
2931
1583
4274



232
2932
1584
4275



233
2933
1585
4276



234
2934
1586
4277



235
2935
1587
4278



236
2936
1588
4279



237
2937
1589
4280



238
2938
1590
4281



239
2939
1591
4282



240
2940
1592
4283



241
2941
1593
4284



242
2942
1594
4285



243
2943
1595
4286



244
2944
1596
4287



245
2945
1597
4288



246
2946
1598
4289



247
2947
1599
4290



248
2948
1600
4291



249
2949
1601
4292



250
2950
1602
4293



251
2951
1603
4294



252
2952
1604
4295



253
2953
1605
4296



254
2954
1606
4297



255
2955
1607
4298



256
2956
1608
4299



257
2957
1609
4300



258
2958
1610
4301



259
2959
1611
4302



260
2960
1612
4303



261
2961
1613
4304



262
2962
1614
4305



263
2963
1615
4306



264
2964
1616
4307



265
2965
1617
4308



266
2966
1618
4309



267
2967
1619
4310



268
2968
1620
4311



269
2969
1621
4312



270
2970
1622
4313



271
2971
1623
4314



272
2972
1624
4315



273
2973
1625
4316



274
2974
1626
4317



275
2975
1627
4318



276
2976
1628
4319



277
2977
1629
4320



278
2978
1630
4321



279
2979
1631
4322



280
2980
1632
4323



281
2981
1633
4324



282
2982
1634
4325



283
2983
1635
4326



284
2984
1636
4327



285
2985
1637
4328



286
2986
1638
4329



287
2987
1639
4330



288
2988
1640
4331



289
2989
1641
4332



290
2990
1642
4333



291
2991
1643
4334



292
2992
1644
4335



293
2993
1645
4336



294
2994
1646
4337



295
2995
1647
4338



296
2996
1648
4339



297
2997
1649
4340



298
2998
1650
4341



299
2999
1651
4342



300
3000
1652
4343



301
3001
1653
4344



302
3002
1654
4345



303
3003
1655
4346



304
NONE
1656
4347



305
3004
1657
4348



306
3005
1658
4349



307
3006
1659
4350



308
3007
1660
4351



309
3008
1661
4352



310
3009
1662
4353



311
3010
1663
NONE



312
3011
1664
4354



313
3012
1665
4355



314
3013
1666
4356



315
3014
1667
4357



316
3015
1668
4358



317
3016
1669
4359



318
3017
1670
4360



319
3018
1671
4361



320
3019
1672
4362



321
3020
1673
4363



322
3021
1674
4364



323
3022
1675
4365



324
3023
1676
4366



325
3024
1677
4367



326
3025
1678
4368



327
3026
1679
4369



328
3027
1680
4370



329
3028
1681
4371



330
3029
1682
4372



331
3030
1683
4373



332
3031
1684
4374



333
3032
1685
4375



334
3033
1686
4376



335
3034
1687
4377



336
3035
1688
4378



337
3036
1689
4379



338
3037
1690
4380



339
3038
1691
4381



340
3039
1692
4382



341
3040
1693
4383



342
3041
1694
4384



343
3042
1695
4385



344
3043
1696
4386



345
3044
1697
4387



346
3045
1698
4388



347
3046
1699
4389



348
3047
1700
4390



349
3048
1701
4391



350
3049
1702
4392



351
3050
1703
4393



352
3051
1704
4394



353
3052
1705
4395



354
3053
1706
4396



355
3054
1707
4397



356
3055
1708
4398



357
3056
1709
4399



358
3057
1710
4400



359
3058
1711
4401



360
3059
1712
NONE



361
3060
1713
4402



362
3061
1714
4403



363
3062
1715
4404



364
3063
1716
4405



365
3064
1717
4406



366
3065
1718
4407



367
3066
1719
4408



368
3067
1720
4409



369
3068
1721
4410



370
3069
1722
4411



371
3070
1723
4412



372
3071
1724
4413



373
3072
1725
4414



374
3073
1726
4415



375
3074
1727
4416



376
3075
1728
4417



377
3076
1729
4418



378
3077
1730
4419



379
3078
1731
4420



380
3079
1732
4421



381
3080
1733
4422



382
3081
1734
4423



383
3082
1735
4424



384
3083
1736
4425



385
3084
1737
4426



386
3085
1738
4427



387
3086
1739
4428



388
3087
1740
4429



389
3088
1741
4430



390
3089
1742
4431



391
3090
1743
4432



392
3091
1744
4433



393
3092
1745
4434



394
3093
1746
4435



395
3094
1747
4436



396
3095
1748
4437



397
3096
1749
4438



398
3097
1750
4439



399
3098
1751
4440



400
3099
1752
4441



401
3100
1753
4442



402
3101
1754
4443



403
3102
1755
4444



404
3103
1756
4445



405
3104
1757
4446



406
3105
1758
4447



407
3106
1759
4448



408
3107
1760
4449



409
3108
1761
4450



410
3109
1762
4451



411
3110
1763
4452



412
3111
1764
4453



413
3112
1765
4454



414
3113
1766
4455



415
3114
1767
4456



416
3115
1768
4457



417
3116
1769
4458



418
3117
1770
4459



419
3118
1771
4460



420
3119
1772
4461



421
3120
1773
4462



422
3121
1774
4463



423
3122
1775
4464



424
3123
1776
4465



425
3124
1777
4466



426
3125
1778
4467



427
3126
1779
4468



428
3127
1780
4469



429
3128
1781
4470



430
3129
1782
4471



431
3130
1783
4472



432
3131
1784
NONE



433
3132
1785
4473



434
3133
1786
4474



435
3134
1787
4475



436
3135
1788
4476



437
3136
1789
4477



438
3137
1790
4478



439
3138
1791
4479



440
3139
1792
4480



441
3140
1793
4481



442
3141
1794
4482



443
3142
1795
4483



444
3143
1796
4484



445
3144
1797
4485



446
3145
1798
4486



447
3146
1799
4487



448
3147
1800
4488



449
3148
1801
4489



450
3149
1802
4490



451
3150
1803
NONE



452
3151
1804
4491



453
3152
1805
4492



454
3153
1806
4493



455
3154
1807
4494



456
3155
1808
4495



457
3156
1809
4496



458
3157
1810
4497



459
3158
1811
4498



460
3159
1812
4499



461
3160
1813
4500



462
3161
1814
4501



463
3162
1815
4502



464
3163
1816
4503



465
3164
1817
4504



466
3165
1818
4505



467
3166
1819
4506



468
3167
1820
4507



469
3168
1821
4508



470
3169
1822
4509



471
3170
1823
4510



472
3171
1824
4511



473
3172
1825
4512



474
3173
1826
4513



475
3174
1827
4514



476
3175
1828
4515



477
3176
1829
4516



478
3177
1830
4517



479
3178
1831
4518



480
3179
1832
4519



481
3180
1833
4520



482
3181
1834
4521



483
3182
1835
4522



484
3183
1836
4523



485
3184
1837
4524



486
3185
1838
4525



487
3186
1839
4526



488
3187
1840
4527



489
3188
1841
4528



490
3189
1842
4529



491
3190
1843
4530



492
3191
1844
4531



493
3192
1845
4532



494
3193
1846
4533



495
3194
1847
4534



496
3195
1848
4535



497
3196
1849
4536



498
3197
1850
4537



499
3198
1851
4538



500
3199
1852
4539



501
3200
1853
4540



502
3201
1854
4541



503
3202
1855
4542



504
3203
1856
4543



505
3204
1857
4544



506
3205
1858
4545



507
3206
1859
4546



508
3207
1860
4547



509
3208
1861
4548



510
3209
1862
4549



511
3210
1863
4550



512
3211
1864
4551



513
3212
1865
4552



514
3213
1866
4553



515
3214
1867
4554



516
3215
1868
4555



517
3216
1869
4556



518
3217
1870
4557



519
3218
1871
4558



520
3219
1872
4559



521
3220
1873
4560



522
3221
1874
4561



523
3222
1875
4562



524
3223
1876
4563



525
3224
1877
4564



526
3225
1878
4565



527
3226
1879
4566



528
3227
1880
4567



529
3228
1881
4568



530
3229
1882
4569



531
3230
1883
4570



532
3231
1884
4571



533
3232
1885
4572



534
3233
1886
4573



535
3234
1887
4574



536
3235
1888
4575



537
3236
1889
4576



538
3237
1890
4577



539
3238
1891
4578



540
3239
1892
4579



541
3240
1893
4580



542
3241
1894
4581



543
3242
1895
4582



544
3243
1896
4583



545
3244
1897
NONE



546
3245
1898
4584



547
3246
1899
4585



548
3247
1900
4586



549
3248
1901
4587



550
3249
1902
4588



551
3250
1903
4589



552
3251
1904
4590



553
3252
1905
4591



554
3253
1906
4592



555
3254
1907
NONE



556
3255
1908
4593



557
3256
1909
4594



558
3257
1910
4595



559
3258
1911
4596



560
3259
1912
4597



561
3260
1913
4598



562
3261
1914
4599



563
3262
1915
4600



564
3263
1916
4601



565
3264
1917
4602



566
3265
1918
4603



567
3266
1919
4604



568
3267
1920
4605



569
3268
1921
4606



570
3269
1922
4607



571
3270
1923
4608



572
3271
1924
4609



573
3272
1925
4610



574
3273
1926
4611



575
3274
1927
4612



576
3275
1928
4613



577
3276
1929
4614



578
3277
1930
4615



579
3278
1931
4616



580
3279
1932
4617



581
3280
1933
4618



582
3281
1934
4619



583
3282
1935
4620



584
3283
1936
4621



585
3284
1937
4622



586
3285
1938
4623



587
3286
1939
4624



588
3287
1940
4625



589
3288
1941
4626



590
3289
1942
4627



591
3290
1943
4628



592
3291
1944
4629



593
3292
1945
4630



594
3293
1946
4631



595
3294
1947
4632



596
3295
1948
4633



597
3296
1949
4634



598
3297
1950
4635



599
3298
1951
4636



600
3299
1952
4637



601
3300
1953
4638



602
3301
1954
4639



603
3302
1955
4640



604
3303
1956
4641



605
3304
1957
4642



606
3305
1958
4643



607
3306
1959
4644



608
3307
1960
4645



609
3308
1961
4646



610
3309
1962
4647



611
3310
1963
4648



612
3311
1964
4649



613
3312
1965
4650



614
3313
1966
4651



615
3314
1967
4652



616
3315
1968
4653



617
3316
1969
4654



618
3317
1970
4655



619
3318
1971
4656



620
3319
1972
4657



621
3320
1973
4658



622
3321
1974
4659



623
3322
1975
4660



624
3323
1976
4661



625
3324
1977
4662



626
3325
1978
4663



627
3326
1979
4664



628
3327
1980
4665



629
3328
1981
4666



630
3329
1982
4667



631
3330
1983
4668



632
3331
1984
4669



633
3332
1985
4670



634
3333
1986
4671



635
3334
1987
4672



636
3335
1988
4673



637
3336
1989
4674



638
3337
1990
4675



639
3338
1991
4676



640
3339
1992
4677



641
3340
1993
4678



642
3341
1994
4679



643
3342
1995
4680



644
3343
1996
4681



645
3344
1997
4682



646
3345
1998
4683



647
3346
1999
4684



648
3347
2000
4685



649
3348
2001
4686



650
3349
2002
4687



651
3350
2003
4688



652
3351
2004
4689



653
3352
2005
4690



654
3353
2006
4691



655
3354
2007
4692



656
3355
2008
4693



657
3356
2009
4694



658
3357
2010
4695



659
3358
2011
4696



660
3359
2012
4697



661
3360
2013
4698



662
3361
2014
4699



663
3362
2015
4700



664
3363
2016
4701



665
3364
2017
4702



666
3365
2018
4703



667
3366
2019
4704



668
3367
2020
4705



669
3368
2021
4706



670
3369
2022
4707



671
3370
2023
4708



672
3371
2024
4709



673
3372
2025
4710



674
3373
2026
4711



675
3374
2027
4712



676
3375
2028
4713



677
3376
2029
4714



678
3377
2030
NONE



679
3378
2031
4715



680
3379
2032
4716



681
3380
2033
4717



682
3381
2034
4718



683
3382
2035
4719



684
3383
2036
4720



685
3384
2037
4721



686
3385
2038
4722



687
3386
2039
4723



688
3387
2040
4724



689
3388
2041
4725



690
3389
2042
4726



691
3390
2043
4727



692
3391
2044
4728



693
3392
2045
4729



694
3393
2046
4730



695
3394
2047
4731



696
3395
2048
4732



697
3396
2049
4733



698
3397
2050
4734



699
3398
2051
4735



700
3399
2052
4736



701
3400
2053
4737



702
3401
2054
4738



703
3402
2055
4739



704
3403
2056
4740



705
3404
2057
4741



706
3405
2058
4742



707
3406
2059
4743



708
3407
2060
4744



709
3408
2061
4745



710
3409
2062
4746



711
3410
2063
4747



712
3411
2064
4748



713
3412
2065
4749



714
3413
2066
4750



715
3414
2067
4751



716
3415
2068
4752



717
3416
2069
4753



718
3417
2070
4754



719
3418
2071
4755



720
3419
2072
4756



721
3420
2073
4757



722
3421
2074
4758



723
3422
2075
4759



724
3423
2076
4760



725
3424
2077
4761



726
3425
2078
4762



727
3426
2079
4763



728
3427
2080
4764



729
3428
2081
4765



730
3429
2082
4766



731
3430
2083
4767



732
3431
2084
4768



733
3432
2085
4769



734
3433
2086
4770



735
3434
2087
4771



736
3435
2088
4772



737
3436
2089
4773



738
3437
2090
4774



739
3438
2091
4775



740
3439
2092
4776



741
3440
2093
4777



742
3441
2094
4778



743
3442
2095
4779



744
3443
2096
4780



745
3444
2097
4781



746
3445
2098
4782



747
3446
2099
4783



748
3447
2100
4784



749
3448
2101
4785



750
3449
2102
4786



751
3450
2103
4787



752
3451
2104
4788



753
3452
2105
4789



754
3453
2106
4790



755
3454
2107
4791



756
3455
2108
4792



757
3456
2109
4793



758
3457
2110
4794



759
3458
2111
4795



760
3459
2112
4796



761
3460
2113
4797



762
3461
2114
4798



763
3462
2115
4799



764
3463
2116
4800



765
3464
2117
4801



766
3465
2118
4802



767
3466
2119
4803



768
3467
2120
4804



769
3468
2121
4805



770
3469
2122
4806



771
3470
2123
4807



772
3471
2124
4808



773
3472
2125
4809



774
3473
2126
4810



775
3474
2127
4811



776
3475
2128
4812



777
3476
2129
4813



778
3477
2130
4814



779
3478
2131
4815



780
3479
2132
4816



781
3480
2133
4817



782
3481
2134
4818



783
3482
2135
4819



784
3483
2136
4820



785
3484
2137
4821



786
3485
2138
4822



787
3486
2139
4823



788
3487
2140
4824



789
3488
2141
4825



790
3489
2142
4826



791
3490
2143
4827



792
3491
2144
4828



793
3492
2145
4829



794
3493
2146
4830



795
3494
2147
4831



796
3495
2148
4832



797
3496
2149
4833



798
3497
2150
4834



799
3498
2151
NONE



800
3499
2152
4835



801
3500
2153
4836



802
3501
2154
4837



803
3502
2155
4838



804
3503
2156
4839



805
3504
2157
4840



806
3505
2158
4841



807
3506
2159
4842



808
3507
2160
4843



809
3508
2161
4844



810
3509
2162
4845



811
3510
2163
4846



812
3511
2164
4847



813
3512
2165
4848



814
3513
2166
4849



815
3514
2167
4850



816
3515
2168
4851



817
3516
2169
4852



818
3517
2170
4853



819
3518
2171
4854



820
3519
2172
4855



821
3520
2173
4856



822
3521
2174
4857



823
3522
2175
4858



824
3523
2176
4859



825
3524
2177
4860



826
3525
2178
4861



827
3526
2179
4862



828
3527
2180
4863



829
3528
2181
4864



830
3529
2182
4865



831
3530
2183
4866



832
3531
2184
4867



833
3532
2185
4868



834
3533
2186
4869



835
3534
2187
4870



836
3535
2188
4871



837
3536
2189
4872



838
3537
2190
4873



839
3538
2191
4874



840
3539
2192
4875



841
3540
2193
4876



842
3541
2194
4877



843
3542
2195
4878



844
3543
2196
4879



845
3544
2197
4880



846
3545
2198
4881



847
3546
2199
4882



848
3547
2200
4883



849
3548
2201
4884



850
3549
2202
4885



851
3550
2203
4886



852
3551
2204
4887



853
3552
2205
4888



854
3553
2206
4889



855
3554
2207
4890



856
3555
2208
4891



857
3556
2209
4892



858
3557
2210
4893



859
3558
2211
4894



860
3559
2212
4895



861
3560
2213
4896



862
3561
2214
4897



863
3562
2215
4898



864
3563
2216
4899



865
3564
2217
4900



866
3565
2218
4901



867
3566
2219
4902



868
3567
2220
4903



869
3568
2221
4904



870
3569
2222
4905



871
3570
2223
4906



872
3571
2224
4907



873
3572
2225
4908



874
3573
2226
4909



875
3574
2227
4910



876
3575
2228
4911



877
3576
2229
4912



878
3577
2230
4913



879
3578
2231
4914



880
3579
2232
4915



881
3580
2233
4916



882
3581
2234
4917



883
3582
2235
4918



884
3583
2236
4919



885
3584
2237
4920



886
3585
2238
4921



887
NONE
2239
4922



888
3586
2240
4923



889
3587
2241
4924



890
3588
2242
4925



891
3589
2243
4926



892
3590
2244
4927



893
3591
2245
4928



894
NONE
2246
4929



895
NONE
2247
4930



896
3592
2248
NONE



897
3593
2249
4931



898
3594
2250
4932



899
3595
2251
4933



900
3596
2252
4934



901
3597
2253
4935



902
3598
2254
4936



903
3599
2255
4937



904
3600
2256
4938



905
3601
2257
4939



906
3602
2258
4940



907
3603
2259
4941



908
3604
2260
4942



909
3605
2261
4943



910
3606
2262
4944



911
3607
2263
4945



912
3608
2264
4946



913
3609
2265
4947



914
3610
2266
4948



915
3611
2267
4949



916
3612
2268
4950



917
3613
2269
4951



918
3614
2270
4952



919
3615
2271
4953



920
3616
2272
4954



921
3617
2273
4955



922
3618
2274
4956



923
3619
2275
4957



924
3620
2276
4958



925
3621
2277
4959



926
3622
2278
4960



927
3623
2279
4961



928
3624
2280
4962



929
3625
2281
4963



930
3626
2282
4964



931
3627
2283
4965



932
3628
2284
4966



933
3629
2285
4967



934
3630
2286
4968



935
NONE
2287
4969



936
3631
2288
4970



937
3632
2289
4971



938
3633
2290
4972



939
3634
2291
4973



940
3635
2292
4974



941
3636
2293
4975



942
3637
2294
4976



943
3638
2295
4977



944
3639
2296
4978



945
3640
2297
4979



946
3641
2298
4980



947
3642
2299
4981



948
3643
2300
4982



949
3644
2301
4983



950
3645
2302
4984



951
3646
2303
4985



952
3647
2304
4986



953
3648
2305
4987



954
3649
2306
4988



955
3650
2307
4989



956
3651
2308
4990



957
3652
2309
4991



958
3653
2310
4992



959
3654
2311
4993



960
3655
2312
4994



961
3656
2313
4995



962
3657
2314
4996



963
3658
2315
4997



964
3659
2316
4998



965
3660
2317
4999



966
3661
2318
5000



967
3662
2319
5001



968
3663
2320
5002



969
3664
2321
5003



970
3665
2322
5004



971
3666
2323
5005



972
3667
2324
5006



973
3668
2325
5007



974
3669
2326
5008



975
3670
2327
5009



976
3671
2328
5010



977
3672
2329
5011



978
3673
2330
5012



979
3674
2331
5013



980
3675
2332
5014



981
3676
2333
5015



982
3677
2334
5016



983
3678
2335
5017



984
3679
2336
5018



985
3680
2337
5019



986
3681
2338
5020



987
3682
2339
5021



988
3683
2340
NONE



989
3684
2341
5022



990
3685
2342
5023



991
3686
2343
5024



992
3687
2344
5025



993
3688
2345
5026



994
3689
2346
5027



995
3690
2347
5028



996
3691
2348
5029



997
3692
2349
5030



998
3693
2350
5031



999
3694
2351
5032



1000
3695
2352
5033



1001
3696
2353
5034



1002
3697
2354
5035



1003
3698
2355
5036



1004
3699
2356
5037



1005
3700
2357
5038



1006
3701
2358
5039



1007
3702
2359
5040



1008
3703
2360
5041



1009
3704
2361
5042



1010
3705
2362
5043



1011
3706
2363
5044



1012
3707
2364
5045



1013
3708
2365
5046



1014
3709
2366
5047



1015
3710
2367
5048



1016
3711
2368
5049



1017
3712
2369
5050



1018
3713
2370
5051



1019
3714
2371
NONE



1020
3715
2372
5052



1021
3716
2373
5053



1022
3717
2374
5054



1023
3718
2375
5055



1024
3719
2376
5056



1025
3720
2377
5057



1026
3721
2378
5058



1027
3722
2379
5059



1028
3723
2380
5060



1029
3724
2381
5061



1030
3725
2382
5062



1031
3726
2383
5063



1032
3727
2384
5064



1033
3728
2385
5065



1034
3729
2386
5066



1035
3730
2387
5067



1036
3731
2388
5068



1037
3732
2389
5069



1038
3733
2390
5070



1039
3734
2391
5071



1040
3735
2392
5072



1041
3736
2393
5073



1042
3737
2394
5074



1043
3738
2395
5075



1044
3739
2396
5076



1045
3740
2397
5077



1046
3741
2398
5078



1047
3742
2399
5079



1048
3743
2400
5080



1049
3744
2401
5081



1050
3745
2402
5082



1051
3746
2403
5083



1052
3747
2404
5084



1053
3748
2405
5085



1054
3749
2406
5086



1055
3750
2407
5087



1056
3751
2408
5088



1057
3752
2409
5089



1058
3753
2410
5090



1059
3754
2411
5091



1060
3755
2412
5092



1061
3756
2413
5093



1062
3757
2414
5094



1063
3758
2415
5095



1064
3759
2416
5096



1065
3760
2417
5097



1066
3761
2418
5098



1067
3762
2419
5099



1068
3763
2420
5100



1069
3764
2421
5101



1070
3765
2422
5102



1071
3766
2423
5103



1072
3767
2424
5104



1073
3768
2425
5105



1074
3769
2426
5106



1075
3770
2427
5107



1076
3771
2428
5108



1077
3772
2429
5109



1078
3773
2430
5110



1079
3774
2431
5111



1080
3775
2432
5112



1081
3776
2433
5113



1082
3777
2434
5114



1083
3778
2435
5115



1084
3779
2436
5116



1085
3780
2437
5117



1086
3781
2438
5118



1087
NONE
2439
5119



1088
3782
2440
5120



1089
3783
2441
5121



1090
3784
2442
5122



1091
3785
2443
NONE



1092
3786
2444
5123



1093
3787
2445
5124



1094
3788
2446
5125



1095
3789
2447
5126



1096
3790
2448
5127



1097
3791
2449
5128



1098
3792
2450
5129



1099
3793
2451
5130



1100
3794
2452
5131



1101
3795
2453
5132



1102
3796
2454
5133



1103
3797
2455
5134



1104
3798
2456
5135



1105
3799
2457
5136



1106
3800
2458
5137



1107
3801
2459
5138



1108
3802
2460
5139



1109
3803
2461
5140



1110
3804
2462
5141



1111
3805
2463
5142



1112
3806
2464
5143



1113
3807
2465
5144



1114
3808
2466
5145



1115
3809
2467
5146



1116
3810
2468
5147



1117
3811
2469
NONE



1118
3812
2470
5148



1119
3813
2471
5149



1120
3814
2472
5150



1121
3815
2473
5151



1122
3816
2474
5152



1123
3817
2475
5153



1124
3818
2476
5154



1125
3819
2477
5155



1126
3820
2478
5156



1127
3821
2479
5157



1128
3822
2480
5158



1129
3823
2481
5159



1130
3824
2482
5160



1131
3825
2483
5161



1132
3826
2484
5162



1133
3827
2485
5163



1134
3828
2486
5164



1135
3829
2487
5165



1136
3830
2488
5166



1137
3831
2489
5167



1138
3832
2490
5168



1139
3833
2491
5169



1140
3834
2492
5170



1141
3835
2493
5171



1142
3836
2494
5172



1143
3837
2495
5173



1144
3838
2496
5174



1145
3839
2497
5175



1146
3840
2498
5176



1147
3841
2499
5177



1148
3842
2500
5178



1149
3843
2501
5179



1150
3844
2502
5180



1151
3845
2503
5181



1152
3846
2504
5182



1153
3847
2505
5183



1154
3848
2506
5184



1155
3849
2507
5185



1156
3850
2508
5186



1157
3851
2509
5187



1158
3852
2510
5188



1159
3853
2511
5189



1160
3854
2512
5190



1161
3855
2513
5191



1162
3856
2514
5192



1163
3857
2515
5193



1164
3858
2516
5194



1165
3859
2517
5195



1166
3860
2518
5196



1167
3861
2519
5197



1168
3862
2520
5198



1169
3863
2521
5199



1170
3864
2522
5200



1171
3865
2523
5201



1172
3866
2524
5202



1173
3867
2525
5203



1174
3868
2526
5204



1175
3869
2527
5205



1176
3870
2528
5206



1177
3871
2529
5207



1178
3872
2530
5208



1179
3873
2531
5209



1180
3874
2532
5210



1181
3875
2533
5211



1182
3876
2534
5212



1183
3877
2535
5213



1184
3878
2536
5214



1185
3879
2537
5215



1186
3880
2538
5216



1187
3881
2539
5217



1188
3882
2540
5218



1189
3883
2541
5219



1190
3884
2542
5220



1191
3885
2543
5221



1192
3886
2544
5222



1193
3887
2545
5223



1194
3888
2546
5224



1195
3889
2547
5225



1196
3890
2548
5226



1197
3891
2549
5227



1198
3892
2550
5228



1199
3893
2551
5229



1200
3894
2552
5230



1201
3895
2553
5231



1202
3896
2554
5232



1203
3897
2555
5233



1204
3898
2556
5234



1205
3899
2557
5235



1206
3900
2558
5236



1207
3901
2559
5237



1208
3902
2560
5238



1209
3903
2561
5239



1210
3904
2562
5240



1211
3905
2563
5241



1212
3906
2564
5242



1213
3907
2565
5243



1214
3908
2566
5244



1215
3909
2567
5245



1216
3910
2568
5246



1217
3911
2569
5247



1218
3912
2570
5248



1219
3913
2571
5249



1220
3914
2572
5250



1221
3915
2573
5251



1222
3916
2574
5252



1223
3917
2575
5253



1224
3918
2576
5254



1225
3919
2577
5255



1226
3920
2578
5256



1227
3921
2579
5257



1228
3922
2580
5258



1229
3923
2581
5259



1230
3924
2582
5260



1231
3925
2583
5261



1232
3926
2584
5262



1233
3927
2585
5263



1234
3928
2586
5264



1235
3929
2587
5265



1236
3930
2588
5266



1237
3931
2589
5267



1238
3932
2590
5268



1239
3933
2591
5269



1240
3934
2592
5270



1241
3935
2593
5271



1242
3936
2594
5272



1243
3937
2595
5273



1244
3938
2596
5274



1245
3939
2597
5275



1246
3940
2598
5276



1247
3941
2599
NONE



1248
3942
2600
5277



1249
3943
2601
5278



1250
3944
2602
5279



1251
3945
2603
5280



1252
3946
2604
5281



1253
3947
2605
5282



1254
3948
2606
5283



1255
3949
2607
5284



1256
3950
2608
5285



1257
3951
2609
5286



1258
3952
2610
5287



1259
3953
2611
5288



1260
3954
2612
5289



1261
3955
2613
5290



1262
3956
2614
5291



1263
3957
2615
5292



1264
3958
2616
5293



1265
3959
2617
5294



1266
3960
2618
5295



1267
3961
2619
5296



1268
3962
2620
5297



1269
3963
2621
5298



1270
3964
2622
5299



1271
3965
2623
5300



1272
3966
2624
5301



1273
3967
2625
5302



1274
3968
2626
5303



1275
3969
2627
5304



1276
3970
2628
5305



1277
3971
2629
5306



1278
3972
2630
5307



1279
3973
2631
5308



1280
3974
2632
5309



1281
3975
2633
5310



1282
3976
2634
5311



1283
3977
2635
5312



1284
3978
2636
5313



1285
3979
2637
5314



1286
3980
2638
5315



1287
3981
2639
5316



1288
3982
2640
5317



1289
3983
2641
5318



1290
3984
2642
5319



1291
3985
2643
5320



1292
3986
2644
5321



1293
3987
2645
5322



1294
3988
2646
5323



1295
3989
2647
5324



1296
3990
2648
5325



1297
3991
2649
5326



1298
3992
2650
5327



1299
3993
2651
5328



1300
3994
2652
5329



1301
3995
2653
5330



1302
3996
2654
5331



1303
3997
2655
5332



1304
3998
2656
5333



1305
3999
2657
5334



1306
4000
2658
5335



1307
4001
2659
5336



1308
4002
2660
5337



1309
4003
2661
5338



1310
4004
2662
5339



1311
4005
2663
5340



1312
4006
2664
5341



1313
4007
2665
5342



1314
4008
2666
5343



1315
4009
2667
5344



1316
4010
2668
5345



1317
4011
2669
5346



1318
4012
2670
5347



1319
4013
2671
5348



1320
4014
2672
5349



1321
4015
2673
5350



1322
4016
2674
5351



1323
4017
2675
5352



1324
4018
2676
5353



1325
4019
2677
5354



1326
4020
2678
5355



1327
4021
2679
5356



1328
4022
2680
5357



1329
4023
2681
NONE



1330
NONE
2682
5358



1331
4024
2683
5359



1332
4025
2684
5360



1333
4026
2685
5361



1334
4027
2686
5362



1335
4028
2687
5363



1336
4029
2688
5364



1337
4030
2689
5365



1338
4031
2690
5366



1339
4032
2691
5367



1340
4033
2692
5368



1341
4034
2693
5369



1342
4035
2694
5370



1343
4036
2695
5371



1344
4037
2696
5372



1345
4038
2697
5373



1346
4039
2698
5374



1347
4040
2699
5375



1348
4041
2700
5376



1349
4042
2701
5377



1350
4043
2702
5378



1351
4044
2703
5379



1352
4045

















TABLE 3







COLD RESPONSIVE SEQUENCES








SEQ
AFFYMETRIX


ID NO:
ID NO:











1
11991_G_AT


2
11992_AT


3
11997_AT


4
11998_AT


5
12001_AT


6
12006_S_AT


7
12007_AT


8
12009_AT


9
12018_AT


10
12022_AT


11
12026_AT


12
12031_AT


13
12047_AT


14
12051_AT


15
12052_AT


16
12053_AT


17
12060_AT


18
12072_AT


19
12074_AT


20
12102_AT


21
12112_AT


22
12117_AT


23
12125_AT


24
12130_AT


25
12143_AT


26
12145_S_AT


27
12149_AT


28
12156_AT


29
12163_AT


30
12166_I_AT


31
12167_AT


32
12169_I_AT


33
12175_AT


34
12176_AT


35
12179_AT


36
12187_AT



15920_I_AT


37
12195_AT


38
12196_AT


39
12198_AT


40
12200_AT


41
12202_AT


42
12214_G_AT


43
12219_AT


44
12224_AT


45
12226_AT


46
12233_AT


47
12240_AT


48
12253_G_AT


49
12256_AT


50
12269_S_AT


51
12270_AT


52
12284_AT


53
12287_S_AT



17570_G_AT


54
12293_AT


55
12294_S_AT


56
12300_AT


57
12307_AT


58
12312_AT


59
12315_AT


60
12324_I_AT


61
12331_S_AT


62
12336_AT


63
12344_AT


64
12348_AT


65
12353_AT


66
12359_S_AT


67
12372_AT


68
12374_I_AT



12726_F_AT


69
12390_AT


70
12395_S_AT


71
12405_AT


72
12408_AT


73
12410_G_AT


74
12419_AT


75
12427_AT


76
12431_AT


77
12436_AT


78
12438_AT


79
12443_S_AT


80
12447_AT


81
12450_S_AT


82
12452_AT


83
12474_AT


84
12477_AT


85
12491_AT


86
12497_AT


87
12500_S_AT


88
12503_AT


89
12515_AT


90
12516_S_AT


91
12523_AT


92
12526_AT


93
12527_AT


94
12532_AT


95
12534_G_AT


96
12544_AT


97
12549_S_AT


98
12550_S_AT



17103_S_AT


99
12552_AT


100
12555_S_AT


101
12576_S_AT


102
12581_S_AT



16645_S_AT


103
12587_AT


104
12597_AT


105
12602_AT


106
12610_AT


107
12631_AT


108
12646_AT


109
12649_AT


110
12650_AT


111
12653_AT


112
12661_AT


113
12666_AT


114
12674_AT


115
12675_S_AT


116
12678_I_AT


117
12681_S_AT


118
12688_AT


119
12702_AT


120
12705_F_AT


121
12736_F_AT


122
12737_F_AT


123
12758_AT


124
12760_G_AT


125
12762_R_AT


126
12764_F_AT


127
12766_AT



15115_F_AT


128
12767_AT


129
12768_AT


130
12772_AT


131
12773_AT


132
12776_AT


133
12788_AT


134
12793_AT


135
12794_AT


136
12802_AT


137
12809_G_AT


138
12812_AT


139
12815_AT


140
12816_AT


141
12818_AT


142
12824_S_AT


143
12828_S_AT


144
12842_S_AT


145
12846_S_AT


146
12858_AT


147
12860_S_AT


148
12861_S_AT


149
12881_S_AT



17600_S_AT


150
12889_S_AT


151
12901_S_AT


152
12902_AT


153
12904_S_AT


154
12905_S_AT


155
12908_S_AT


156
12910_S_AT



16385_S_AT


157
12914_S_AT



15783_S_AT



17645_S_AT


158
12916_S_AT


159
12923_S_AT


160
12926_S_AT


161
12927_S_AT


162
12931_S_AT


163
12937_R_AT


164
12941_G_AT


165
12942_AT


166
12947_AT


167
12949_AT


168
12953_AT


169
12956_I_AT


170
12959_AT


171
12966_S_AT


172
12975_AT


173
12983_AT


174
12984_AT


175
12987_S_AT


176
12994_S_AT


177
13002_AT


178
13009_I_AT


179
13011_AT


180
13018_AT


181
13023_AT


182
13024_AT


183
13034_S_AT


184
13046_G_AT


185
13048_S_AT



13495_S_AT


186
13054_AT


187
13067_S_AT


188
13068_AT


189
13073_S_AT


190
13078_S_AT


191
13079_AT


192
13081_S_AT


193
13083_AT


194
13086_R_AT


195
13087_AT


196
13090_AT


197
13092_S_AT



16950_S_AT


198
13098_AT


199
13100_AT


200
13103_AT


201
13105_AT


202
13107_S_AT


203
13108_AT


204
13109_AT


205
13114_AT


206
13118_F_AT


207
13119_AT


208
13120_AT


209
13123_AT


210
13128_AT


211
13133_S_AT



17430_S_AT


212
13135_S_AT


213
13139_AT


214
13140_AT


215
13143_AT


216
13151_G_AT


217
13160_AT


218
13161_AT


219
13162_AT


220
13165_AT


221
13166_AT


222
13167_AT


223
13179_AT


224
13181_AT


225
13185_AT


226
13193_S_AT


227
13213_S_AT



16004_S_AT


228
13219_S_AT



20288_G_AT


229
13220_S_AT



13221_AT



18929_S_AT


230
13233_AT



14301_S_AT


231
13243_R_AT


232
13254_S_AT


233
13260_S_AT



15660_S_AT


234
13273_S_AT



16105_S_AT


235
13274_S_AT



17077_S_AT


236
13276_S_AT


237
13278_F_AT


238
13285_S_AT


239
13288_S_AT



17043_S_AT


240
13292_S_AT


241
13296_S_AT


242
13297_S_AT


243
13299_S_AT



15166_S_AT


244
13332_AT


245
13347_AT


246
13351_AT


247
13352_AT


248
13355_AT


249
13404_AT


250
13422_AT


251
13459_AT


252
13460_AT


253
13461_S_AT


254
13467_AT


255
13488_AT


256
13523_S_AT


257
13529_AT


258
13539_I_AT



14631_S_AT


259
13541_AT


260
13542_AT


261
13545_S_AT


262
13552_AT


263
13556_I_AT


264
13561_AT


265
13563_S_AT


266
13567_AT


267
13568_AT


268
13571_AT


269
13575_AT


270
13576_AT


271
13583_AT


272
13598_AT


273
13601_AT


274
13604_AT


275
13613_AT


276
13616_S_AT



16544_S_AT


277
13617_AT


278
13618_S_AT


279
13619_AT


280
13621_G_AT


281
13623_R_AT


282
13629_S_AT


283
13631_AT


284
13635_AT


285
13646_AT


286
13650_AT


287
13653_AT


288
13655_AT


289
13656_AT


290
13657_AT


291
13666_S_AT



17083_S_AT


292
13667_S_AT


293
13669_S_AT



17074_S_AT


294
13670_S_AT



15206_S_AT


295
13671_S_AT



16805_S_AT


296
13678_S_AT


297
13688_S_AT


298
13690_S_AT



16065_S_AT


299
13691_S_AT



16117_S_AT


300
13692_S_AT



16118_S_AT


301
13700_AT


302
13704_S_AT


303
13714_AT


304
13715_AT


305
13724_AT


306
13748_AT


307
13759_AT


308
13767_AT


309
13785_AT


310
13803_AT


311
13850_I_AT


312
13876_AT


313
13880_S_AT


314
13883_AT


315
13887_S_AT


316
13895_AT


317
13904_S_AT



18722_S_AT


318
13906_S_AT


319
13908_S_AT



18597_AT


320
13923_AT


321
13927_AT


322
13932_AT


323
13935_AT


324
13940_AT


325
13949_S_AT


326
13954_G_AT


327
13971_S_AT


328
13973_AT


329
13983_AT


330
13985_S_AT


331
13987_S_AT



18738_F_AT


332
13989_AT



20674_S_AT


333
14010_AT


334
14013_AT


335
14014_AT


336
14019_AT


337
14021_R_AT


338
14025_S_AT



18909_S_AT


339
14027_AT


340
14030_AT


341
14044_AT


342
14048_AT


343
14056_AT


344
14057_AT


345
14058_AT


346
14059_AT


347
14061_AT


348
14068_S_AT


349
14072_AT


350
14073_AT


351
14074_AT


352
14084_AT


353
14095_S_AT


354
14100_AT


355
14101_AT


356
14103_AT


357
14105_AT


358
14106_AT


359
14121_AT


360
14129_S_AT


361
14133_S_AT


362
14143_AT


363
14145_AT


364
14148_AT


365
14186_AT


366
14194_AT


367
14196_AT


368
14223_AT


369
14234_AT


370
14236_AT


371
14251_F_AT


372
14252_F_AT


373
14270_AT


374
14298_G_AT



17581_G_AT


375
14303_S_AT


376
14312_AT


377
14316_AT


378
14339_AT


379
14366_AT


380
14369_AT


381
14388_AT


382
14392_G_AT


383
14393_AT


384
14421_AT


385
14436_AT


386
14448_AT


387
14450_AT


388
14454_AT


389
14459_AT


390
14478_AT


391
14482_AT


392
14485_AT


393
14492_S_AT


394
14505_AT


395
14510_AT


396
14511_AT


397
14517_AT


398
14519_AT


399
14525_S_AT


400
14527_AT


401
14534_S_AT


402
14538_R_AT


403
14554_AT


404
14558_AT


405
14559_S_AT


406
14566_AT


407
14572_AT


408
14579_AT


409
14587_AT


410
14591_AT


411
14595_AT


412
14602_AT


413
14603_AT


414
14605_AT


415
14620_S_AT


416
14626_S_AT


417
14630_S_AT



16559_S_AT


418
14637_S_AT



17122_S_AT


419
14642_F_AT


420
14650_S_AT



15150_S_AT


421
14654_S_AT


422
14667_S_AT



18299_S_AT


423
14669_S_AT



16136_S_AT


424
14672_S_AT


425
14679_S_AT


426
14682_I_AT


427
14689_AT


428
14697_G_AT



16902_AT


429
14701_S_AT



14734_S_AT


430
14703_AT


431
14711_S_AT


432
14712_S_AT



20530_S_AT


433
14713_S_AT


434
14715_S_AT


435
14728_S_AT


436
14731_S_AT


437
14781_AT


438
14797_S_AT


439
14800_AT


440
14809_AT


441
14843_AT


442
14847_AT


443
14872_AT


444
14886_AT


445
14896_AT


446
14900_AT


447
14908_AT


448
14912_AT


449
14914_AT


450
14942_AT


451
14945_AT


452
14955_AT


453
14957_S_AT


454
14958_AT


455
14965_AT


456
14974_AT


457
14980_AT


458
14981_AT


459
14984_S_AT


460
14995_AT


461
15004_AT


462
15009_AT


463
15010_AT


464
15024_AT


465
15026_AT


466
15036_R_AT


467
15054_AT


468
15056_AT


469
15057_AT


470
15066_AT


471
15073_AT


472
15081_AT


473
15083_AT


474
15091_AT


475
15097_S_AT


476
15101_S_AT


477
15102_S_AT


478
15107_S_AT


479
15112_S_AT


480
15116_F_AT


481
15118_S_AT


482
15122_S_AT


483
15130_S_AT


484
15131_S_AT


485
15132_S_AT



17585_S_AT


486
15139_S_AT


487
15143_S_AT


488
15146_S_AT


489
15159_S_AT



15160_S_AT


490
15162_S_AT


491
15167_S_AT


492
15171_S_AT


493
15174_F_AT


494
15178_S_AT


495
15185_S_AT



18023_S_AT


496
15188_S_AT


497
15193_S_AT


498
15196_S_AT


499
15197_S_AT


500
15201_F_AT


501
15213_S_AT


502
15243_AT


503
15256_AT


504
15270_AT


505
15319_AT


506
15325_AT


507
15337_AT


508
15341_AT


509
15343_AT


510
15348_AT


511
15350_AT


512
15355_S_AT


513
15367_AT


514
15372_AT


515
15379_AT


516
15381_AT


517
15383_AT


518
15384_AT


519
15385_AT


520
15387_AT


521
15410_AT


522
15417_S_AT


523
15422_AT


524
15423_AT


525
15431_AT


526
15433_AT


527
15452_AT


528
15464_AT


529
15468_AT


530
15471_AT


531
15472_AT


532
15475_S_AT


533
15485_AT


534
15489_AT


535
15490_AT


536
15503_AT


537
15505_AT


538
15510_R_AT


539
15512_AT


540
15514_AT


541
15515_R_AT


542
15517_S_AT


543
15518_AT


544
15529_AT


545
15534_F_AT


546
15538_AT


547
15541_AT


548
15543_AT


549
15544_AT


550
15551_AT


551
15574_S_AT


552
15576_S_AT


553
15577_S_AT


554
15578_S_AT


555
15583_S_AT


556
15588_S_AT


557
15595_S_AT


558
15600_S_AT


559
15602_F_AT


560
15608_S_AT


561
15613_S_AT


562
15616_S_AT


563
15618_S_AT


564
15620_S_AT


565
15627_S_AT


566
15634_S_AT



16125_S_AT



18046_S_AT


567
15637_S_AT


568
15639_S_AT


569
15642_S_AT


570
15643_S_AT


571
15651_F_AT


572
15652_S_AT


573
15665_S_AT


574
15667_S_AT



18610_S_AT


575
15668_S_AT


576
15671_S_AT


577
15675_S_AT


578
15679_S_AT


579
15685_S_AT


580
15687_F_AT


581
15688_S_AT


582
15689_S_AT


583
15692_S_AT


584
15694_S_AT


585
15712_S_AT


586
15808_AT


587
15845_AT


588
15848_AT


589
15850_AT



20406_G_AT


590
15858_AT


591
15862_AT


592
15868_AT


593
15878_AT


594
15894_AT


595
15900_AT


596
15901_AT


597
15902_AT


598
15912_AT


599
15913_AT


600
15928_AT


601
15940_AT


602
15941_AT


603
15945_AT


604
15948_S_AT


605
15956_AT


606
15960_AT



16466_S_AT


607
15976_AT


608
15978_AT


609
15986_S_AT


610
15990_AT


611
16009_S_AT


612
16015_AT


613
16019_AT


614
16024_AT


615
16034_AT


616
16036_I_AT



18729_AT


617
16039_S_AT


618
16040_AT


619
16042_S_AT


620
16047_AT


621
16049_S_AT


622
16051_S_AT


623
16055_S_AT


624
16059_S_AT


625
16062_S_AT


626
16066_S_AT


627
16069_S_AT


628
16074_S_AT


629
16076_S_AT


630
16077_S_AT



17579_S_AT


631
16079_S_AT


632
16084_S_AT



17998_S_AT


633
16087_S_AT


634
16089_S_AT


635
16090_S_AT


636
16102_S_AT


637
16103_S_AT


638
16108_S_AT


639
16112_S_AT


640
16134_S_AT


641
16137_S_AT


642
16138_S_AT


643
16140_S_AT


644
16143_S_AT


645
16145_S_AT


646
16148_S_AT


647
16151_S_AT


648
16155_S_AT


649
16158_F_AT


650
16160_F_AT


651
16162_S_AT


652
16168_S_AT


653
16169_S_AT


654
16171_S_AT


655
16172_S_AT


656
16184_AT


657
16192_AT


658
16222_AT


659
16242_AT


660
16244_AT


661
16250_AT


662
16286_AT


663
16288_AT


664
16294_S_AT


665
16296_AT


666
16297_AT


667
16325_AT


668
16346_S_AT


669
16357_AT


670
16380_AT


671
16382_AT


672
16393_S_AT


673
16402_S_AT


674
16411_S_AT


675
16442_S_AT


676
16446_AT


677
16448_G_AT


678
16453_S_AT


679
16457_S_AT


680
16465_AT



16916_S_AT


681
16470_S_AT



18735_S_AT


682
16481_S_AT


683
16486_AT


684
16487_AT


685
16488_AT


686
16496_S_AT


687
16499_AT


688
16510_AT


689
16511_AT


690
16512_S_AT



18085_R_AT


691
16514_AT


692
16516_AT


693
16517_AT


694
16526_AT


695
16528_AT


696
16531_S_AT


697
16535_S_AT


698
16537_S_AT


699
16538_S_AT


700
16543_S_AT


701
16550_S_AT


702
16554_S_AT


703
16567_S_AT


704
16571_S_AT


705
16576_F_AT


706
16577_S_AT


707
16579_S_AT


708
16580_S_AT


709
16583_S_AT


710
16584_S_AT



18706_S_AT


711
16593_S_AT


712
16595_S_AT


713
16598_S_AT


714
16604_S_AT


715
16605_S_AT


716
16610_S_AT


717
16611_S_AT


718
16614_S_AT


719
16617_S_AT


720
16618_S_AT


721
16620_S_AT


722
16621_S_AT


723
16631_S_AT


724
16634_S_AT


725
16635_S_AT


726
16636_S_AT


727
16639_S_AT


728
16640_S_AT


729
16650_S_AT


730
16652_S_AT


731
16654_AT


732
16672_AT


733
16673_AT


734
16687_S_AT


735
16747_AT


736
16753_AT


737
16768_AT


738
16777_AT


739
16784_AT


740
16807_AT


741
16811_AT


742
16845_AT


743
16894_AT


744
16899_AT


745
16911_AT


746
16920_AT


747
16921_AT


748
16924_S_AT


749
16926_S_AT


750
16931_S_AT


751
16934_S_AT


752
16937_AT


753
16938_AT


754
16942_AT


755
16943_S_AT



18231_AT


756
16949_S_AT


757
16952_S_AT


758
16956_AT


759
16962_S_AT


760
16965_S_AT


761
16970_S_AT



18010_S_AT


762
16977_AT


763
16984_AT


764
16996_S_AT


765
16997_AT


766
17000_AT


767
17005_AT


768
17010_S_AT


769
17017_S_AT


770
17031_S_AT


771
17033_S_AT


772
17053_S_AT


773
17055_S_AT


774
17063_S_AT


775
17068_S_AT


776
17070_S_AT


777
17075_S_AT


778
17084_S_AT


779
17087_S_AT


780
17092_S_AT


781
17095_S_AT


782
17096_S_AT


783
17102_S_AT


784
17105_S_AT


785
17109_S_AT


786
17110_S_AT


787
17113_S_AT


788
17115_S_AT


789
17116_S_AT


790
17123_S_AT


791
17129_S_AT


792
17132_AT


793
17166_AT


794
17206_AT


795
17207_AT


796
17215_AT


797
17237_AT


798
17247_AT


799
17254_AT


800
17286_AT


801
17288_S_AT


802
17292_AT


803
17300_AT


804
17303_S_AT


805
17318_AT


806
17319_AT


807
17322_AT


808
17323_AT


809
17332_S_AT


810
17374_AT


811
17381_AT


812
17388_AT


813
17392_S_AT


814
17405_AT


815
17415_AT


816
17418_S_AT


817
17420_AT


818
17423_S_AT


819
17426_AT


820
17427_AT


821
17429_S_AT


822
17431_AT


823
17439_G_AT


824
17457_AT


825
17458_AT


826
17462_S_AT


827
17463_AT


828
17465_AT


829
17466_S_AT


830
17475_AT


831
17479_AT


832
17482_S_AT


833
17495_S_AT


834
17508_S_AT


835
17522_S_AT


836
17523_S_AT


837
17537_S_AT


838
17538_S_AT


839
17539_S_AT


840
17546_S_AT



18694_S_AT


841
17557_S_AT


842
17560_S_AT


843
17562_AT


844
17564_S_AT



19361_S_AT


845
17565_S_AT


846
17568_AT


847
17573_AT


848
17577_G_AT


849
17578_AT


850
17596_AT


851
17627_AT


852
17631_AT


853
17632_AT


854
17672_AT


855
17675_AT


856
17677_AT


857
17732_AT


858
17743_AT


859
17748_AT


860
17782_AT


861
17823_S_AT


862
17841_AT


863
17849_S_AT


864
17852_G_AT


865
17857_AT


866
17865_AT


867
17882_AT


868
17885_AT


869
17900_S_AT


870
17910_AT


871
17911_AT


872
17916_AT


873
17917_S_AT


874
17918_AT


875
17921_S_AT


876
17922_AT


877
17926_S_AT


878
17933_AT


879
17935_AT


880
17956_I_AT


881
17966_AT


882
17967_AT


883
17970_I_AT


884
17978_S_AT



20635_S_AT


885
17986_S_AT


886
17993_AT


887
18001_AT


888
18003_AT


889
18004_AT


890
18005_AT


891
18029_G_AT



18030_I_AT


892
18040_S_AT


893
18045_AT


894
18064_R_AT


895
18065_R_AT


896
18074_AT


897
18076_S_AT


898
18077_AT


899
18081_AT


900
18154_S_AT



18365_S_AT


901
18165_AT


902
18174_AT


903
18176_AT


904
18194_I_AT


905
18197_AT


906
18198_AT


907
18213_AT


908
18219_AT


909
18221_AT


910
18222_AT


911
18226_S_AT


912
18232_AT


913
18237_AT


914
18241_AT


915
18257_AT


916
18258_S_AT


917
18269_S_AT


918
18274_S_AT


919
18275_AT


920
18278_AT


921
18282_AT


922
18283_AT


923
18290_AT


924
18291_AT


925
18306_AT


926
18316_AT


927
18317_AT


928
18327_S_AT


929
18337_S_AT


930
18339_AT


931
18347_S_AT


932
18383_AT


933
18390_AT


934
18439_S_AT


935
18465_S_AT


936
18487_AT


937
18508_S_AT


938
18512_AT


939
18543_AT


940
18544_AT


941
18552_AT


942
18555_AT


943
18556_AT


944
18561_AT


945
18567_AT


946
18573_AT


947
18580_AT


948
18581_AT


949
18584_AT


950
18587_S_AT


951
18588_AT


952
18591_AT


953
18592_S_AT


954
18600_AT


955
18601_S_AT


956
18607_S_AT


957
18611_AT


958
18616_AT


959
18622_G_AT


960
18623_AT


961
18628_AT


962
18631_AT


963
18635_AT


964
18636_AT


965
18638_AT


966
18652_AT


967
18657_AT


968
18659_AT


969
18660_S_AT


970
18667_AT


971
18675_AT


972
18684_AT


973
18686_S_AT


974
18688_S_AT


975
18693_S_AT


976
18698_S_AT


977
18705_AT


978
18707_AT


979
18708_AT


980
18726_S_AT


981
18727_AT


982
18732_I_AT


983
18736_AT


984
18750_F_AT


985
18754_AT


986
18778_AT


987
18806_S_AT


988
18823_S_AT


989
18829_AT


990
18835_AT


991
18844_AT


992
18859_AT


993
18864_AT


994
18866_AT


995
18880_AT


996
18883_G_AT


997
18885_AT


998
18886_AT


999
18887_AT


1000
18888_AT


1001
18889_AT


1002
18892_S_AT


1003
18901_AT


1004
18911_AT


1005
18917_I_AT


1006
18939_AT


1007
18947_I_AT


1008
18950_AT


1009
18951_S_AT


1010
18954_AT


1011
18956_AT


1012
18959_AT


1013
18966_AT


1014
18974_AT


1015
18976_AT


1016
18980_AT


1017
18989_S_AT


1018
18994_AT


1019
19030_AT


1020
19039_AT


1021
19049_AT


1022
19083_AT


1023
19115_AT


1024
19117_S_AT


1025
19122_AT


1026
19125_S_AT


1027
19127_AT


1028
19130_AT


1029
19144_AT


1030
19157_S_AT


1031
19178_AT


1032
19190_G_AT


1033
19198_AT


1034
19202_AT


1035
19209_S_AT


1036
19211_AT


1037
19218_AT


1038
19222_AT


1039
19226_G_AT


1040
19229_AT


1041
19230_AT


1042
19232_S_AT


1043
19285_AT


1044
19326_AT


1045
19332_AT


1046
19346_AT


1047
19347_AT


1048
19362_AT


1049
19363_AT


1050
19364_AT


1051
19367_AT


1052
19373_AT


1053
19381_AT


1054
19382_AT


1055
19384_AT


1056
19401_AT


1057
19406_AT


1058
19413_AT


1059
19416_AT


1060
19426_S_AT


1061
19439_AT


1062
19441_S_AT


1063
19442_AT


1064
19448_S_AT


1065
19454_AT


1066
19462_S_AT


1067
19464_AT


1068
19470_AT


1069
19483_AT


1070
19489_S_AT


1071
19513_AT


1072
19548_AT


1073
19562_AT


1074
19563_S_AT


1075
19567_AT


1076
19581_AT


1077
19589_S_AT


1078
19595_S_AT


1079
19606_AT


1080
19623_AT


1081
19624_AT


1082
19627_S_AT


1083
19636_AT


1084
19652_AT


1085
19655_AT


1086
19657_S_AT


1087
19658_AT


1088
19660_AT


1089
19665_S_AT


1090
19667_AT


1091
19671_AT


1092
19677_AT


1093
19686_AT


1094
19689_AT


1095
19690_S_AT


1096
19695_AT


1097
19698_AT


1098
19700_S_AT


1099
19708_AT


1100
19717_AT


1101
19726_S_AT


1102
19744_AT


1103
19752_S_AT


1104
19759_AT


1105
19782_AT


1106
19803_S_AT


1107
19828_AT


1108
19831_I_AT


1109
19833_S_AT


1110
19834_AT


1111
19836_AT


1112
19841_AT


1113
19845_G_AT


1114
19854_AT


1115
19855_AT


1116
19866_AT


1117
19867_AT


1118
19870_S_AT


1119
19871_AT


1120
19872_AT


1121
19875_S_AT


1122
19876_AT


1123
19879_S_AT


1124
19881_AT


1125
19897_S_AT


1126
19903_AT


1127
19905_AT


1128
19906_AT


1129
19907_AT


1130
19910_AT


1131
19913_AT


1132
19920_S_AT


1133
19932_AT


1134
19939_AT


1135
19945_AT


1136
19947_AT


1137
19951_AT


1138
19956_AT


1139
19962_AT


1140
19963_AT


1141
19969_AT


1142
19970_S_AT


1143
19971_AT


1144
19972_AT


1145
19981_AT


1146
19990_AT


1147
19996_AT


1148
20003_S_AT


1149
20009_S_AT


1150
20013_AT


1151
20018_AT


1152
20024_S_AT


1153
20027_AT


1154
20045_AT


1155
20047_AT


1156
20048_AT


1157
20050_AT


1158
20051_AT


1159
20058_AT


1160
20067_AT


1161
20068_AT


1162
20069_AT


1163
20093_I_AT


1164
20099_AT


1165
20100_AT


1166
20113_S_AT


1167
20117_AT


1168
20123_AT


1169
20127_S_AT


1170
20129_AT


1171
20150_AT


1172
20154_AT


1173
20156_AT


1174
20165_AT


1175
20173_AT


1176
20178_S_AT


1177
20183_AT


1178
20188_AT


1179
20189_AT


1180
20197_AT


1181
20210_G_AT


1182
20213_AT


1183
20229_AT


1184
20232_S_AT


1185
20255_AT


1186
20257_AT


1187
20262_AT


1188
20275_AT


1189
20278_S_AT


1190
20282_S_AT


1191
20284_AT


1192
20293_AT


1193
20294_AT


1194
20312_S_AT


1195
20315_I_AT


1196
20330_S_AT


1197
20331_AT


1198
20350_S_AT


1199
20354_S_AT


1200
20355_AT


1201
20360_AT


1202
20363_AT


1203
20369_S_AT


1204
20378_G_AT


1205
20383_AT


1206
20384_AT


1207
20387_AT


1208
20393_AT


1209
20396_AT


1210
20399_AT


1211
20409_G_AT


1212
20412_S_AT


1213
20413_AT


1214
20439_AT


1215
20440_AT


1216
20444_AT


1217
20445_AT


1218
20449_AT


1219
20456_AT


1220
20462_AT


1221
20471_AT


1222
20474_AT


1223
20495_S_AT


1224
20499_AT


1225
20501_AT


1226
20511_AT


1227
20515_S_AT


1228
20516_AT


1229
20517_AT


1230
20518_AT


1231
20520_S_AT


1232
20536_S_AT


1233
20538_S_AT


1234
20539_S_AT


1235
20558_AT


1236
20561_AT


1237
20567_AT


1238
20571_AT


1239
20582_S_AT


1240
20586_I_AT


1241
20590_AT


1242
20592_AT


1243
20594_AT


1244
20608_S_AT


1245
20612_S_AT


1246
20616_AT


1247
20620_G_AT


1248
20637_AT


1249
20643_AT


1250
20649_AT


1251
20651_AT


1252
20654_S_AT


1253
20670_AT


1254
20684_AT


1255
20685_AT


1256
20693_AT


1257
20701_S_AT


1258
20704_AT


1259
20705_AT


1260
20715_AT


1261
20719_AT
















TABLE 4





2X UP IN COLD, ONLY




















11997_at
12688_at
13274_s_at
14145_at
15083_at
15639_s_at


11998_at
12701_i_at
13278_f_at
14170_at
15084_at
15641_s_at


12018_at
12702_at
13279_s_at
14186_at
15096_at
15660_s_at


12031_at
12719_f_at
13285_s_at
14196_at
15101_s_at
15665_s_at


12047_at
12726_f_at
13288_s_at
14227_at
15105_s_at
15687_f_at


12051_at
12736_f_at
13292_s_at
14234_at
15112_s_at
15694_s_at


12053_at
12754_g_at
13297_s_at
14250_r_at
15115_f_at
15712_s_at


12060_at
12762_r_at
13299_s_at
14270_at
15116_f_at
15783_s_at


12072_at
12766_at
13332_at
14298_g_at
15122_s_at
15808_at


12074_at
12767_at
13351_at
14303_s_at
15126_s_at
15837_at


12102_at
12768_at
13352_at
14312_at
15131_s_at
15850_at


12112_at
12773_at
13422_at
14339_at
15132_s_at
15862_at


12117_at
12788_at
13435_at
14388_at
15137_s_at
15868_at


12130_at
12802_at
13461_s_at
14393_at
15144_s_at
15878_at


12145_s_at
12860_s_at
13467_at
14511_at
15148_s_at
15901_at


12151_at
12861_s_at
13488_at
14525_s_at
15153_s_at
15912_at


12163_at
12879_s_at
13495_s_at
14527_at
15159_s_at
15920_i_at


12175_at
12891_at
13539_i_at
14534_s_at
15160_s_at
15941_at


12187_at
12914_s_at
13542_at
14554_at
15166_s_at
15945_at


12195_at
12927_s_at
13575_at
14566_at
15174_f_at
15960_at


12219_at
12947_at
13577_s_at
14579_at
15197_s_at
15990_at


12256_at
12956_i_at
13617_at
14591_at
15270_at
16001_at


12269_s_at
12966_s_at
13634_s_at
14595_at
15319_at
16009_s_at


12307_at
12974_at
13656_at
14600_at
15325_at
16010_s_at


12315_at
12987_s_at
13671_s_at
14631_s_at
15337_at
16034_at


12336_at
12994_s_at
13691_s_at
14635_s_at
15341_at
16036_i_at


12349_s_at
12998_at
13700_at
14679_s_at
15343_at
16039_s_at


12353_at
13002_at
13704_s_at
14691_at
15355_s_at
16040_at


12359_s_at
13018_at
13709_s_at
14697_g_at
15367_at
16042_s_at


12390_at
13023_at
13715_at
14709_at
15379_at
16047_at


12395_s_at
13046_g_at
13785_at
14711_s_at
15381_at
16049_s_at


12431_at
13054_at
13803_at
14728_s_at
15410_at
16051_s_at


12436_at
13086_r_at
13812_s_at
14731_s_at
15417_s_at
16062_s_at


12443_s_at
13087_at
13825_s_at
14797_s_at
15422_at
16079_s_at


12447_at
13100_at
13850_i_at
14809_at
15433_at
16087_s_at


12452_at
13109_at
13904_s_at
14843_at
15451_at
16090_s_at


12477_at
13119_at
13908_s_at
14847_at
15452_at
16117_s_at


12503_at
13120_at
13927_at
14872_at
15453_s_at
16118_s_at


12516_s_at
13128_at
13971_s_at
14886_at
15472_at
16137_s_at


12532_at
13134_s_at
13985_s_at
14896_at
15489_at
16155_s_at


12544_at
13140_at
14013_at
14897_at
15490_at
16162_s_at


12561_at
13143_at
14019_at
14900_at
15503_at
16184_at


12602_at
13167_at
14021_r_at
14956_s_at
15510_r_at
16192_at


12610_at
13172_s_at
14028_at
14958_at
15517_s_at
16222_at


12631_at
13178_at
14048_at
14965_at
15518_at
16244_at


12647_s_at
13179_at
14058_at
14984_s_at
15544_at
16250_at


12650_at
13181_at
14059_at
15004_at
15588_s_at
16260_at


12656_at
13187_i_at
14064_at
15010_at
15600_s_at
16286_at


12674_at
13209_s_at
14073_at
15036_r_at
15605_s_at
16296_at


12675_s_at
13219_s_at
14105_at
15040_g_at
15613_s_at
16297_at


12676_s_at
13221_at
14106_at
15046_s_at
15614_s_at
16342_at


12681_s_at
13243_r_at
14126_s_at
15057_at
15616_s_at
16367_i_at


12686_s_at
13260_s_at
14140_at
15073_at
15633_s_at
16411_s_at


16442_s_at
17077_s_at
17978_s_at
18885_at
19689_at
20412_s_at


16465_at
17102_s_at
17999_at
18887_at
19698_at
20413_at


16466_s_at
17109_s_at
18001_at
18888_at
19700_s_at
20432_at


16468_at
17113_s_at
18004_at
18889_at
19707_s_at
20433_at


16486_at
17123_s_at
18012_s_at
18901_at
19708_at
20456_at


16487_at
17128_s_at
18040_s_at
18907_s_at
19713_at
20462_at


16488_at
17129_s_at
18176_at
18917_i_at
19718_at
20471_at


16489_at
17132_at
18194_i_at
18939_at
19744_at
20511_at


16496_s_at
17166_at
18197_at
18947_i_at
19836_at
20515_s_at


16499_at
17206_at
18198_at
18949_at
19839_at
20517_at


16511_at
17237_at
18213_at
18954_at
19840_s_at
20518_at


16517_at
17300_at
18219_at
18959_at
19845_g_at
20529_at


16538_s_at
17319_at
18222_at
18974_at
19854_at
20536_s_at


16554_s_at
17322_at
18231_at
18976_at
19855_at
20538_s_at


16571_s_at
17332_s_at
18232_at
18980_at
19860_at
20539_s_at


16576_f_at
17381_at
18241_at
18989_s_at
19866_at
20576_at


16595_s_at
17388_at
18269_s_at
19019_i_at
19871_at
20582_s_at


16605_s_at
17392_s_at
18272_at
19049_at
19875_s_at
20586_i_at


16610_s_at
17408_at
18282_at
19083_at
19879_s_at
20608_s_at


16620_s_at
17424_at
18298_at
19130_at
19881_at
20649_at


16621_s_at
17429_s_at
18316_at
19156_s_at
19913_at
20651_at


16635_s_at
17457_at
18317_at
19178_at
19939_at
20684_at


16636_s_at
17458_at
18331_s_at
19190_g_at
19945_at
20685_at


16638_s_at
17466_s_at
18347_s_at
19199_at
19947_at
20699_at


16650_s_at
17477_s_at
18383_at
19202_at
19951_at
20705_at


16672_at
17482_s_at
18390_at
19209_s_at
19956_at
20715_at


16673_at
17538_s_at
18455_at
19211_at
19971_at


16687_s_at
17546_s_at
18465_s_at
19218_at
19976_at


16747_at
17562_at
18544_at
19229_at
19998_at


16753_at
17581_g_at
18555_at
19322_at
20003_s_at


16768_at
17627_at
18556_at
19326_at
20015_at


16805_s_at
17631_at
18560_at
19359_s_at
20027_at


16807_at
17632_at
18561_at
19367_at
20051_at


16845_at
17645_s_at
18571_at
19384_at
20068_at


16847_at
17672_at
18588_at
19389_at
20093_i_at


16896_s_at
17675_at
18597_at
19397_at
20117_at


16899_at
17677_at
18601_s_at
19406_at
20150_at


16902_at
17693_at
18611_at
19426_s_at
20156_at


16911_at
17732_at
18623_at
19441_s_at
20165_at


16914_s_at
17743_at
18635_at
19442_at
20257_at


16943_s_at
17748_at
18659_at
19470_at
20262_at


16956_at
17775_at
18660_s_at
19489_s_at
20275_at


16996_s_at
17782_at
18673_at
19562_at
20282_s_at


17010_s_at
17841_at
18694_s_at
19577_at
20288_g_at


17016_s_at
17852_g_at
18705_at
19589_s_at
20293_at


17032_s_at
17900_s_at
18708_at
19597_s_at
20315_i_at


17033_s_at
17901_at
18738_f_at
19611_s_at
20330_s_at


17043_s_at
17911_at
18750_f_at
19624_at
20360_at


17050_s_at
17921_s_at
18778_at
19657_s_at
20363_at


17055_s_at
17922_at
18829_at
19667_at
20369_s_at


17068_s_at
17933_at
18835_at
19671_at
20384_at


17071_s_at
17967_at
18866_at
19677_at
20393_at


17075_s_at
17970_i_at
18875_s_at
19686_at
20396_at
















TABLE 5





2X UP COLD 3 HR, ONLY

















12117_at



12145_s_at



12151_at



12163_at



12187_at



12256_at



12315_at



12349_s_at



12353_at



12359_s_at



12544_at



12602_at



12610_at



12676_s_at



12686_s_at



12701_i_at



12702_at



12719_f_at



12736_f_at



12754_g_at



12766_at



12767_at



12768_at



12773_at



12788_at



12879_s_at



12891_at



12947_at



12966_s_at



12974_at



12994_s_at



13002_at



13100_at



13140_at



13167_at



13172_s_at



13179_at



13187_i_at



13219_s_at



13260_s_at



13278_f_at



13279_s_at



13285_s_at



13288_s_at



13292_s_at



13297_s_at



13351_at



13352_at



13435_at



13467_at



13488_at



13495_s_at



13656_at



13671_s_at



13691_s_at



13785_at



13803_at



13825_s_at



13904_s_at



14013_at



14021_r_at



14028_at



14064_at



14126_s_at



14145_at



14170_at



14196_at



14250_r_at



14298_g_at



14303_s_at



14339_at



14527_at



14534_s_at



14554_at



14595_at



14635_s_at



14679_s_at



14691_at



14697_g_at



14709_at



14728_s_at



14809_at



14896_at



14965_at



14984_s_at



15046_s_at



15083_at



15096_at



15105_s_at



15115_f_at



15116_f_at



15122_s_at



15126_s_at



15131_s_at



15132_s_at



15137_s_at



15153_s_at



15159_s_at



15160_s_at



15197_s_at



15355_s_at



15379_at



15417_s_at



15422_at



15451_at



15452_at



15453_s_at



15489_at



15518_at



15588_s_at



15613_s_at



15614_s_at



15616_s_at



15639_s_at



15641_s_at



15660_s_at



15687_f_at



15694_s_at



15862_at



15868_at



15878_at



15901_at



16034_at



16039_s_at



16040_at



16042_s_at



16047_at



16062_s_at



16087_s_at



16117_s_at



16118_s_at



16162_s_at



16184_at



16222_at



16250_at



16411_s_at



16442_s_at



16465_at



16486_at



16488_at



16489_at



16517_at



16571_s_at



16605_s_at



16610_s_at



16620_s_at



16636_s_at



16650_s_at



16805_s_at



16845_at



16899_at



16914_s_at



16943_s_at



16996_s_at



17010_s_at



17043_s_at



17068_s_at



17109_s_at



17128_s_at



17237_at



17319_at



17392_s_at



17429_s_at



17477_s_at



17538_s_at



17581_g_at



17627_at



17672_at



17693_at



17782_at



17841_at



17900_s_at



17933_at



17978_s_at



18001_at



18012_s_at



18198_at



18219_at



18241_at



18269_s_at



18272_at



18282_at



18298_at



18383_at



18556_at



18588_at



18601_s_at



18611_at



18694_s_at



18708_at



18738_f_at



18778_at



18829_at



18835_at



18866_at



18875_s_at



18888_at



18907_s_at



18917_i_at



18939_at



18974_at



19190_g_at



19199_at



19202_at



19211_at



19384_at



19406_at



19426_s_at



19442_at



19470_at



19577_at



19597_s_at



19624_at



19657_s_at



19667_at



19845_g_at



19855_at



19866_at



19945_at



19951_at



19998_at



20003_s_at



20015_at



20051_at



20093_i_at



20117_at



20288_g_at



20360_at



20369_s_at



20384_at



20462_at



20471_at



20515_s_at



20538_s_at



20576_at



20608_s_at



20651_at



20685_at



20705_at

















TABLE 6





2X DOWN COLD, ONLY




















11991_g_at
12450_s_at
12881_s_at
13151_g_at
13621_g_at
14056_at


11992_at
12474_at
12889_s_at
13160_at
13623_r_at
14057_at


12001_at
12491_at
12901_s_at
13161_at
13629_s_at
14061_at


12006_s_at
12497_at
12902_at
13162_at
13631_at
14067_at


12007_at
12500_s_at
12904_s_at
13165_at
13635_at
14068_s_at


12009_at
12515_at
12905_s_at
13166_at
13646_at
14072_at


12022_at
12521_at
12908_s_at
13185_at
13650_at
14074_at


12023_s_at
12523_at
12910_s_at
13193_s_at
13652_at
14075_at


12026_at
12526_at
12916_s_at
13211_s_at
13653_at
14083_at


12037_at
12527_at
12923_s_at
13213_s_at
13655_at
14084_at


12052_at
12534_g_at
12926_s_at
13219_s_at
13657_at
14089_at


12125_at
12549_s_at
12931_s_at
13233_at
13666_s_at
14095_s_at


12143_at
12550_s_at
12937_r_at
13236_s_at
13667_s_at
14096_at


12149_at
12552_at
12941_g_at
13239_s_at
13669_s_at
14100_at


12156_at
12555_s_at
12942_at
13241_s_at
13670_s_at
14101_at


12166_i_at
12556_at
12949_at
13254_s_at
13672_s_at
14103_at


12167_at
12575_s_at
12953_at
13266_s_at
13678_s_at
14121_at


12169_i_at
12576_s_at
12958_at
13273_s_at
13679_s_at
14129_s_at


12176_at
12581_s_at
12959_at
13275_f_at
13688_s_at
14133_s_at


12179_at
12587_at
12966_s_at
13276_s_at
13690_s_at
14143_at


12196_at
12597_at
12975_at
13278_f_at
13691_s_at
14148_at


12198_at
12606_at
12983_at
13280_s_at
13692_s_at
14162_at


12200_at
12609_at
12984_at
13285_s_at
13714_at
14194_at


12202_at
12646_at
13002_at
13296_s_at
13724_at
14208_at


12212_at
12649_at
13009_i_at
13347_at
13748_at
14217_at


12214_g_at
12653_at
13011_at
13355_at
13751_at
14223_at


12224_at
12661_at
13014_at
13361_at
13759_at
14235_at


12226_at
12666_at
13024_at
13404_at
13767_at
14236_at


12233_at
12678_i_at
13034_s_at
13406_at
13789_at
14251_f_at


12240_at
12705_f_at
13041_s_at
13459_at
13876_at
14252_f_at


12253_g_at
12736_f_at
13048_s_at
13460_at
13880_s_at
14285_at


12270_at
12737_f_at
13067_s_at
13464_at
13883_at
14301_s_at


12278_at
12758_at
13068_at
13523_s_at
13887_s_at
14316_at


12284_at
12760_g_at
13073_s_at
13529_at
13895_at
14366_at


12287_s_at
12764_f_at
13078_s_at
13541_at
13906_s_at
14369_at


12293_at
12765_at
13079_at
13545_s_at
13919_at
14392_g_at


12294_s_at
12772_at
13081_s_at
13550_at
13923_at
14421_at


12300_at
12776_at
13083_at
13552_at
13932_at
14431_at


12312_at
12784_at
13090_at
13556_i_at
13935_at
14436_at


12315_at
12793_at
13092_s_at
13561_at
13940_at
14448_at


12324_i_at
12794_at
13098_at
13563_s_at
13949_s_at
14450_at


12331_s_at
12795_at
13103_at
13567_at
13954_g_at
14454_at


12344_at
12809_g_at
13105_at
13568_at
13973_at
14459_at


12348_at
12812_at
13107_s_at
13571_at
13983_at
14478_at


12353_at
12815_at
13108_at
13576_at
13989_at
14482_at


12372_at
12816_at
13114_at
13583_at
14010_at
14485_at


12374_i_at
12818_at
13118_f_at
13598_at
14014_at
14492_s_at


12405_at
12824_s_at
13123_at
13601_at
14015_s_at
14505_at


12408_at
12828_s_at
13124_at
13604_at
14016_s_at
14510_at


12410_g_at
12842_s_at
13133_s_at
13613_at
14025_s_at
14517_at


12419_at
12846_s_at
13135_s_at
13616_s_at
14027_at
14519_at


12427_at
12858_at
13139_at
13618_s_at
14030_at
14534_s_at


19438_at
12869_s_at
13146_s_at
13619_at
14044_at
14538_r_at


14558_at
15047_at
15512_at
15940_at
16357_at
16894_at


14559_s_at
15054_at
15514_at
15948_s_at
16380_at
16899_at


14572_at
15056_at
15515_r_at
15956_at
16382_at
16920_at


14584_at
15058_s_at
15529_at
15976_at
16385_s_at
16921_at


14587_at
15063_at
15534_f_at
15978_at
16393_s_at
16924_s_at


14595_at
15066_at
15538_at
15986_s_at
16402_s_at
16926_s_at


14602_at
15081_at
15541_at
16004_s_at
16417_s_at
16931_s_at


14603_at
15091_at
15543_at
16015_at
16442_s_at
16934_s_at


14605_at
15097_s_at
15551_at
16017_at
16446_at
16937_at


14620_s_at
15102_s_at
15574_s_at
16019_at
16448_g_at
16938_at


14626_s_at
15107_s_at
15576_s_at
16024_at
16453_s_at
16942_at


14630_s_at
15118_s_at
15577_s_at
16031_at
16457_s_at
16949_s_at


14637_s_at
15127_s_at
15578_s_at
16055_s_at
16470_s_at
16950_s_at


14640_s_at
15130_s_at
15581_s_at
16059_s_at
16481_s_at
16952_s_at


14642_f_at
15132_s_at
15583_s_at
16065_s_at
16510_at
16962_s_at


14650_s_at
15133_s_at
15591_s_at
16066_s_at
16512_s_at
16965_s_at


14654_s_at
15139_s_at
15595_s_at
16069_s_at
16514_at
16970_s_at


14667_s_at
15143_s_at
15602_f_at
16074_s_at
16516_at
16977_at


14668_s_at
15146_s_at
15606_s_at
16076_s_at
16523_s_at
16984_at


14669_s_at
15150_s_at
15608_s_at
16077_s_at
16526_at
16989_at


14672_s_at
15161_s_at
15616_s_at
16084_s_at
16528_at
16993_at


14673_s_at
15162_s_at
15618_s_at
16089_s_at
16531_s_at
16997_at


14675_s_at
15167_s_at
15620_s_at
16102_s_at
16535_s_at
17000_at


14679_s_at
15170_s_at
15627_s_at
16103_s_at
16537_s_at
17005_at


14681_g_at
15171_s_at
15634_s_at
16105_s_at
16543_s_at
17010_s_at


14682_i_at
15178_s_at
15637_s_at
16108_s_at
16544_s_at
17017_s_at


14689_at
15182_s_at
15642_s_at
16112_s_at
16550_s_at
17031_s_at


14701_s_at
15185_s_at
15643_s_at
16117_s_at
16559_s_at
17040_s_at


14703_at
15188_s_at
15646_s_at
16118_s_at
16567_s_at
17053_s_at


14712_s_at
15193_s_at
15651_f_at
16125_s_at
16577_s_at
17056_s_at


14713_s_at
15196_s_at
15652_s_at
16127_s_at
16579_s_at
17063_s_at


14715_s_at
15201_f_at
15667_s_at
16134_s_at
16580_s_at
17070_s_at


14734_s_at
15206_s_at
15668_s_at
16136_s_at
16583_s_at
17074_s_at


14781_at
15207_s_at
15670_s_at
16138_s_at
16584_s_at
17084_s_at


14800_at
15213_s_at
15671_s_at
16140_s_at
16593_s_at
17085_s_at


14856_s_at
15243_at
15675_s_at
16143_s_at
16598_s_at
17087_s_at


14882_at
15256_at
15679_s_at
16144_s_at
16603_s_at
17092_s_at


14908_at
15348_at
15685_s_at
16145_s_at
16604_s_at
17095_s_at


14912_at
15350_at
15688_s_at
16148_s_at
16611_s_at
17096_s_at


14914_at
15372_at
15689_s_at
16151_s_at
16614_s_at
17097_s_at


14924_at
15383_at
15692_s_at
16158_f_at
16617_s_at
17103_s_at


14942_at
15384_at
15775_at
16160_f_at
16618_s_at
17105_s_at


14945_at
15385_at
15776_at
16168_s_at
16620_s_at
17110_s_at


14955_at
15387_at
15845_at
16169_s_at
16631_s_at
17115_s_at


14957_s_at
15406_at
15848_at
16171_s_at
16634_s_at
17116_s_at


14974_at
15423_at
15858_at
16172_s_at
16639_s_at
17119_s_at


14980_at
15431_at
15866_s_at
16222_at
16640_s_at
17122_s_at


14981_at
15464_at
15894_at
16232_s_at
16652_s_at
17207_at


14995_at
15468_at
15900_at
16242_at
16654_at
17215_at


15009_at
15471_at
15901_at
16288_at
16777_at
17247_at


15018_at
15475_s_at
15902_at
16294_s_at
16784_at
17254_at


15024_at
15485_at
15913_at
16325_at
16811_at
17286_at


15026_at
15505_at
15928_at
16346_s_at
16893_at
17288_s_at


17292_at
17910_at
18337_s_at
18823_s_at
19382_at
19897_s_at


17303_s_at
17916_at
18339_at
18844_at
19401_at
19903_at


17305_at
17917_s_at
18365_s_at
18859_at
19402_at
19905_at


17318_at
17918_at
18402_at
18864_at
19406_at
19906_at


17323_at
17926_s_at
18439_s_at
18880_at
19413_at
19907_at


17374_at
17935_at
18487_at
18883_g_at
19416_at
19910_at


17405_at
17956_i_at
18508_s_at
18886_at
19429_at
19920_s_at


17415_at
17961_at
18512_at
18892_s_at
19432_s_at
19932_at


17418_s_at
17966_at
18543_at
18909_s_at
19439_at
19951_at


17420_at
17978_s_at
18552_at
18911_at
19448_s_at
19962_at


17423_s_at
17986_s_at
18567_at
18913_s_at
19454_at
19963_at


17426_at
17993_at
18573_at
18916_s_at
19462_s_at
19969_at


17427_at
17998_s_at
18580_at
18921_g_at
19464_at
19970_s_at


17430_s_at
18003_at
18581_at
18950_at
19469_at
19972_at


17431_at
18005_at
18584_at
18951_s_at
19483_at
19981_at


17439_g_at
18010_s_at
18587_s_at
18956_at
19484_s_at
19990_at


17442_i_at
18013_r_at
18590_at
18966_at
19513_at
19996_at


17449_s_at
18023_s_at
18591_at
18972_at
19548_at
19999_s_at


17462_s_at
18029_g_at
18592_s_at
18994_at
19563_s_at
20009_s_at


17463_at
18030_i_at
18600_at
19030_at
19567_at
20013_at


17465_at
18045_at
18601_s_at
19039_at
19581_at
20017_at


17475_at
18046_s_at
18607_s_at
19068_i_at
19595_s_at
20018_at


17479_at
18059_i_at
18610_s_at
19108_at
19606_at
20024_s_at


17495_s_at
18064_r_at
18611_at
19115_at
19623_at
20045_at


17508_s_at
18065_r_at
18616_at
19117_s_at
19627_s_at
20047_at


17522_s_at
18074_at
18622_g_at
19122_at
19636_at
20048_at


17523_s_at
18076_s_at
18628_at
19125_s_at
19641_at
20050_at


17529_s_at
18077_at
18631_at
19127_at
19652_at
20051_at


17537_s_at
18078_at
18636_at
19135_at
19655_at
20058_at


17539_s_at
18081_at
18638_at
19144_at
19658_at
20067_at


17543_s_at
18083_r_at
18652_at
19157_s_at
19660_at
20069_at


17555_s_at
18085_r_at
18657_at
19158_at
19665_s_at
20099_at


17557_s_at
18091_at
18667_at
19177_at
19667_at
20100_at


17560_s_at
18154_s_at
18675_at
19192_at
19690_s_at
20113_s_at


17564_s_at
18165_at
18684_at
19198_at
19695_at
20123_at


17565_s_at
18174_at
18686_s_at
19222_at
19717_at
20127_s_at


17568_at
18221_at
18688_s_at
19226_g_at
19726_s_at
20129_at


17570_g_at
18226_s_at
18693_s_at
19227_at
19752_s_at
20133_i_at


17573_at
18230_at
18698_s_at
19230_at
19759_at
20152_at


17577_g_at
18237_at
18706_s_at
19232_s_at
19782_at
20154_at


17578_at
18255_at
18707_at
19263_at
19789_s_at
20173_at


17579_s_at
18257_at
18726_s_at
19285_at
19803_s_at
20178_s_at


17585_s_at
18258_s_at
18727_at
19332_at
19828_at
20183_at


17596_at
18274_s_at
18732_i_at
19346_at
19831_i_at
20188_at


17600_s_at
18275_at
18735_s_at
19347_at
19833_s_at
20189_at


17823_s_at
18278_at
18736_at
19361_s_at
19834_at
20197_at


17840_s_at
18283_at
18738_f_at
19362_at
19835_at
20200_at


17849_s_at
18290_at
18747_f_at
19363_at
19841_at
20210_g_at


17857_at
18291_at
18754_at
19364_at
19867_at
20213_at


17865_at
18299_s_at
18782_at
19365_s_at
19870_s_at
20229_at


17882_at
18300_at
18789_at
19373_at
19871_at
20232_s_at


17885_at
18306_at
18806_s_at
19379_at
19872_at
20255_at


17902_s_at
18327_s_at
18814_at
19381_at
19876_at
20278_s_at


20284_at
20693_at


20288_g_at
20701_s_at


20294_at
20704_at


20312_s_at
20707_s_at


20331_at
20719_at


20335_s_at


20350_s_at


20354_s_at


20355_at


20369_s_at


20378_g_at


20383_at


20385_s_at


20387_at


20399_at


20409_g_at


20420_at


20429_s_at


20439_at


20440_at


20444_at


20445_at


20449_at


20474_at


20480_s_at


20495_s_at


20499_at


20501_at


20516_at


20520_s_at


20530_s_at


20538_s_at


20547_at


20558_at


20561_at


20567_at


20571_at


20590_at


20592_at


20594_at


20608_s_at


20612_s_at


20616_at


20620_g_at


20635_s_at


20637_at


20643_at


20654_s_at


20670_at


20674_s_at


20684_at


20685_at


20689_s_at
















TABLE 7







SALINE STRESS RESPONSIVE SEQUENCES








SEQ
AFFYMETRIX


ID NO:
ID NO:












2227
12011_S_AT



2228
12153_AT


2229
12180_AT


2230
12186_AT


2231
12216_AT


2232
12265_AT


2233
12335_AT


2234
12449_S_AT


2235
12470_AT


2236
12479_AT


2237
12487_AT


2238
12493_G_AT


2239
12562_AT


2240
12685_AT


2241
12704_F_AT


2242
12709_F_AT


2243
12734_F_AT


2244
12739_S_AT


2245
12750_S_AT


2246
12761_S_AT


2247
12813_AT


2248
12845_S_AT


2249
12946_AT


2250
13003_S_AT


2251
13052_S_AT


2252
13094_AT


2253
13142_AT


2254
13172_S_AT



17880_S_AT


2255
13198_I_AT


2256
13209_S_AT



16165_S_AT


2257
13229_S_AT


2258
13253_F_AT


2259
13344_S_AT


2260
13370_AT


2261
13387_AT


2262
13408_S_AT


2263
13429_AT


2264
13472_AT


2265
13526_AT


2266
13569_AT


2267
13614_AT


2268
13686_S_AT


2269
13718_AT


2270
13719_AT


2271
13902_AT


2272
13918_AT


2273
13944_AT


2274
13964_AT


2275
13993_S_AT


2276
14000_AT


2277
14003_AT


2278
14032_AT


2279
14043_AT


2280
14070_AT


2281
14267_AT


2282
14269_AT


2283
14418_AT


2284
14427_AT


2285
14501_AT


2286
14544_AT


2287
14546_S_AT


2288
14570_AT


2289
14596_AT


2290
14729_S_AT


2291
14874_AT


2292
14888_AT


2293
14951_AT


2294
14952_AT


2295
14959_AT


2296
14979_AT


2297
15006_AT


2298
15042_AT


2299
15049_AT


2300
15062_AT


2301
15108_S_AT


2302
15147_S_AT


2303
15175_S_AT


2304
15176_S_AT


2305
15186_S_AT



18696_S_AT


2306
15192_S_AT


2307
15208_S_AT


2308
15324_AT


2309
15371_AT


2310
15424_AT


2311
15463_AT


2312
15465_AT


2313
15497_S_AT


2314
15589_S_AT


2315
15636_S_AT


2316
15663_S_AT


2317
15770_AT


2318
15792_AT


2319
15855_AT


2320
15860_AT


2321
15891_AT


2322
15898_AT


2323
15909_AT


2324
15965_AT


2325
15969_S_AT








2326
15975_S_AT









2327
15995_S_AT



2328
15998_S_AT



18090_S_AT


2329
16028_AT


2330
16050_AT


2331
16060_S_AT


2332
16067_S_AT


2333
16072_S_AT


2334
16088_F_AT


2335
16273_AT


2336
16314_AT


2337
16413_S_AT


2338
16414_AT


2339
16426_AT


2340
16436_AT


2341
16455_AT


2342
16502_AT


2343
16548_S_AT


2344
16568_S_AT


2345
16582_S_AT


2346
16589_S_AT


2347
16594_S_AT


2348
16613_S_AT


2349
16651_S_AT


2350
16668_AT


2351
16820_AT


2352
16987_S_AT


2353
16995_AT


2354
17039_S_AT


2355
17273_AT


2356
17278_AT


2357
17433_AT


2358
17467_AT


2359
17566_AT


2360
17595_S_AT


2361
17744_S_AT


2362
17758_AT


2363
17864_AT


2364
17868_AT


2365
17876_AT


2366
17894_AT


2367
17942_S_AT


2368
18008_R_AT


2369
18027_AT


2370
18053_S_AT


2371
18062_AT


2372
18082_AT


2373
18121_S_AT


2374
18240_S_AT


2375
18248_S_AT


2376
18264_AT


2377
18276_AT


2378
18287_AT


2379
18310_AT


2380
18367_S_AT


2381
18506_AT


2382
18605_S_AT


2383
18618_S_AT


2384
18626_AT


2385
18666_S_AT


2386
18834_AT


2387
18847_AT


2388
18896_AT


2389
18899_S_AT


2390
18973_AT


2391
18983_S_AT


2392
18988_AT


2393
18998_S_AT


2394
19065_AT


2395
19119_I_AT



19121_AT


2396
19207_AT


2397
19220_AT


2398
19284_AT


2399
19315_AT


2400
19348_AT


2401
19403_S_AT


2402
19437_S_AT


2403
19502_AT


2404
19609_AT


2405
19645_AT


2406
19742_AT


2407
19863_AT


2408
19873_AT


2409
19891_AT


2410
20004_S_AT


2411
20053_AT


2412
20138_AT


2413
20193_AT


2414
20199_AT


2415
20220_AT


2416
20239_G_AT


2417
20297_AT


2418
20324_S_AT


2419
20353_AT


2420
20362_AT


2421
20389_AT


2422
20546_AT


2423
20600_AT


2424
20623_AT


2425
20629_AT


2426
20648_S_AT


2427
20668_AT
















TABLE 8





2X UP IN SALT, ONLY



















12037_at
14570_at
16190_at
18506_at
20648_s_at


12137_at
14578_s_at
16196_at
18605_s_at
20678_at


12153_at
14596_at
16273_at
18626_at
20686_at


12186_at
14646_s_at
16314_at
18666_s_at
20707_s_at


12216_at
14662_f_at
16413_s_at
18747_f_at


12268_at
14668_s_at
16414_at
18782_at


12449_s_at
14729_s_at
16417_s_at
18834_at


12470_at
14874_at
16455_at
18847_at


12476_at
14888_at
16548_s_at
18913_s_at


12487_at
14918_at
16582_s_at
18973_at


12493_g_at
14952_at
16589_s_at
18988_at


12609_at
14959_at
16594_s_at
18998_s_at


12685_at
14986_at
16613_s_at
19065_at


12704_f_at
15006_at
16651_s_at
19068_i_at


12709_f_at
15042_at
16668_at
19123_at


12734_f_at
15047_at
16690_g_at
19177_at


12739_s_at
15062_at
16762_at
19220_at


12750_s_at
15063_at
16820_at
19284_at


12761_s_at
15108_s_at
16873_i_at
19288_at


12819_at
15133_s_at
16987_s_at
19315_at


12845_s_at
15147_s_at
16989_at
19437_s_at


12946_at
15170_s_at
16995_at
19484_s_at


13142_at
15175_s_at
17039_s_at
19502_at


13198_i_at
15182_s_at
17040_s_at
19503_at


13229_s_at
15190_s_at
17400_s_at
19592_at


13275_f_at
15192_s_at
17425_s_at
19645_at


13344_s_at
15324_at
17433_at
19742_at


13370_at
15392_at
17467_at
19835_at


13408_s_at
15424_at
17490_s_at
19873_at


13464_at
15467_at
17529_s_at
19891_at


13472_at
15497_s_at
17543_s_at
19992_at


13526_at
15581_s_at
17566_at
20004_s_at


13614_at
15623_f_at
17595_s_at
20053_at


13652_at
15636_s_at
17744_s_at
20133_i_at


13679_s_at
15646_s_at
17758_at
20138_at


13751_at
15670_s_at
17855_at
20190_at


13918_at
15770_at
17864_at
20199_at


13919_at
15775_at
17876_at
20200_at


13944_at
15778_at
18008_r_at
20297_at


13964_at
15792_at
18013_r_at
20324_s_at


13987_s_at
15855_at
18024_s_at
20335_s_at


13993_s_at
15891_at
18027_at
20353_at


14000_at
15909_at
18053_s_at
20362_at


14032_at
15923_at
18078_at
20385_s_at


14043_at
15969_s_at
18082_at
20389_at


14052_at
15975_s_at
18090_s_at
20402_s_at


14067_at
15995_s_at
18091_at
20450_at


14070_at
15998_s_at
18121_s_at
20468_at


14269_at
16017_at
18264_at
20489_at


14285_at
16050_at
18276_at
20546_at


14427_at
16067_s_at
18300_at
20569_s_at


14501_at
16072_s_at
18367_s_at
20600_at


14540_at
16165_s_at
18471_at
20623_at
















TABLE 9





2X UP SALT, 3 HR ONLY




















12037_at
15042_at
16987_s_at
20004_s_at



12137_at
15047_at
16989_at
20053_at



12153_at
15062_at
17039_s_at
20133_i_at



12186_at
15063_at
17040_s_at
20138_at



12216_at
15108_s_at
17425_s_at
20190_at



12268_at
15133_s_at
17433_at
20199_at



12470_at
15147_s_at
17490_s_at
20200_at



12476_at
15170_s_at
17543_s_at
20220_at



12487_at
15175_s_at
17744_s_at
20362_at



12493_g_at
15182_s_at
17864_at
20385_s_at



12609_at
15190_s_at
17876_at
20389_at



12685_at
15192_s_at
18008_r_at
20489_at



12704_f_at
15324_at
18013_r_at
20546_at



12709_f_at
15424_at
18024_s_at
20623_at



12734_f_at
15467_at
18027_at
20648_s_at



12739_s_at
15497_s_at
18053_s_at
20678_at



12750_s_at
15623_f_at
18078_at
20707_s_at



12819_at
15636_s_at
18082_at



12946_at
15646_s_at
18090_s_at



13142_at
15670_s_at
18091_at



13229_s_at
15770_at
18121_s_at



13275_f_at
15775_at
18264_at



13370_at
15778_at
18276_at



13408_s_at
15792_at
18367_s_at



13464_at
15855_at
18471_at



13472_at
15891_at
18506_at



13614_at
15909_at
18605_s_at



13652_at
15923_at
18626_at



13679_s_at
15969_s_at
18666_s_at



13918_at
15975_s_at
18747_f_at



13919_at
15995_s_at
18782_at



13944_at
15998_s_at
18834_at



13987_s_at
16017_at
18847_at



13993_s_at
16050_at
18913_s_at



14000_at
16067_s_at
18973_at



14032_at
16072_s_at
18988_at



14043_at
16165_s_at
19065_at



14052_at
16196_at
19068_i_at



14067_at
16273_at
19123_at



14269_at
16314_at
19177_at



14285_at
16414_at
19220_at



14501_at
16417_s_at
19288_at



14540_at
16455_at
19315_at



14570_at
16548_s_at
19437_s_at



14596_at
16582_s_at
19484_s_at



14668_s_at
16589_s_at
19502_at



14729_s_at
16594_s_at
19503_at



14888_at
16613_s_at
19592_at



14918_at
16651_s_at
19645_at



14952_at
16668_at
19742_at



14959_at
16762_at
19835_at



14986_at
16820_at
19873_at



15006_at
16873_i_at
19891_at

















TABLE 10





2X DOWN SALT, ONLY



















12011_s_at
16046_s_at
20239_g_at



12180_at
16060_s_at
20433_at



12265_at
16088_f_at
20629_at



12335_at
16150_s_at
20668_at



12479_at
16166_s_at



12562_at
16316_at



12656_at
16340_at



12813_at
16367_i_at



13003_s_at
16426_at



13052_s_at
16427_at



13094_at
16436_at



13178_at
16489_at



13253_f_at
16502_at



13387_at
16568_s_at



13429_at
16638_s_at



13472_at
16646_s_at



13569_at
17273_at



13686_s_at
17278_at



13718_at
17567_at



13719_at
17868_at



13902_at
17880_s_at



14003_at
17894_at



14144_at
17901_at



14267_at
17942_s_at



14418_at
17960_at



14544_at
17999_at



14546_s_at
18062_at



14636_s_at
18240_s_at



14951_at
18248_s_at



14956_s_at
18267_at



14979_at
18279_s_at



14990_at
18287_at



15040_g_at
18310_at



15049_at
18351_s_at



15115_f_at
18455_at



15137_s_at
18560_at



15148_s_at
18571_at



15176_s_at
18618_s_at



15208_s_at
18896_at



15371_at
18899_s_at



15453_s_at
18967_s_at



15463_at
18983_s_at



15465_at
19119_i_at



15589_s_at
19121_at



15663_s_at
19207_at



15860_at
19348_at



15898_at
19403_s_at



15931_at
19609_at



15965_at
19742_at



15970_s_at
19826_at



15972_s_at
19863_at



16005_s_at
19883_at



16028_at
20193_at

















TABLE 11







OSMOTIC STRESS RESPONSIVE SEQUENCES








SEQ
AFFYMETRIX


ID NO:
ID NO:





2428
11994_AT


2429
12028_AT


2430
12033_AT


2431
12039_AT


2432
12068_AT


2433
12096_AT


2434
12110_AT


2435
12114_AT


2436
12135_AT


2437
12139_AT


2438
12189_AT


2439
12191_AT


2440
12211_AT


2441
12223_S_AT


2442
12366_S_AT



12869_S_AT


2443
12381_AT


2444
12406_S_AT


2445
12412_AT


2446
12453_AT


2447
12571_S_AT


2448
12662_AT


2449
12746_I_AT


2450
12774_AT


2451
12787_AT


2452
12847_AT


2453
12848_AT


2454
12895_AT


2455
12911_S_AT


2456
12920_AT



12921_S_AT


2457
13027_AT


2458
13059_AT


2459
13075_I_AT


2460
13180_S_AT


2461
13255_I_AT


2462
13270_AT



18167_S_AT


2463
13283_S_AT


2464
13382_AT


2465
13386_S_AT


2466
13433_AT


2467
13482_AT


2468
13732_AT


2469
13733_I_AT


2470
13842_AT


2471
13860_S_AT


2472
13868_AT


2473
13901_AT


2474
13933_AT


2475
13995_AT


2476
14062_AT


2477
14118_I_AT


2478
14141_AT


2479
14310_AT


2480
14354_AT


2481
14476_AT


2482
14513_S_AT


2483
14568_S_AT


2484
14604_AT


2485
14634_S_AT


2486
14660_S_AT


2487
14666_S_AT


2488
14686_S_AT



17464_AT


2489
14726_S_AT


2490
14848_S_AT


2491
14873_AT


2492
14883_AT


2493
15082_AT


2494
15121_S_AT



16014_S_AT


2495
15168_S_AT


2496
15271_AT


2497
15338_AT


2498
15418_AT


2499
15429_AT


2500
15548_AT


2501
15666_S_AT


2502
15672_S_AT


2503
15680_S_AT


2504
15867_AT


2505
15918_AT


2506
15999_S_AT


2507
16303_AT


2508
16363_AT


2509
16440_S_AT


2510
16458_S_AT


2511
16475_AT


2512
16513_S_AT


2513
16529_AT


2514
16547_S_AT


2515
16553_F_AT


2516
16563_S_AT


2517
16629_S_AT


2518
16797_AT


2519
16814_AT


2520
16832_AT


2521
16976_S_AT


2522
17007_AT


2523
17037_S_AT


2524
17054_S_AT


2525
17257_S_AT



18725_S_AT


2526
17270_AT


2527
17275_I_AT


2528
17376_AT


2529
17378_AT


2530
17468_AT


2531
17481_AT


2532
17511_S_AT


2533
17519_S_AT


2534
17815_S_AT


2535
17897_AT


2536
17923_S_AT


2537
17934_AT


2538
17937_S_AT


2539
17944_AT


2540
17958_AT


2541
18216_AT


2542
18227_AT


2543
18284_AT


2544
18301_S_AT


2545
18312_S_AT


2546
18326_S_AT


2547
18369_AT


2548
18411_AT


2549
18533_AT


2550
18576_S_AT


2551
18599_AT


2552
18640_AT


2553
18672_S_AT


2554
18720_S_AT


2555
18768_AT


2556
18877_AT


2557
18942_AT


2558
18945_AT


2559
18960_AT


2560
18965_AT


2561
19060_AT


2562
19164_G_AT


2563
19266_AT


2564
19366_S_AT


2565
19369_AT


2566
19371_AT


2567
19386_AT


2568
19412_AT


2569
19427_S_AT


2570
19622_G_AT


2571
19681_AT


2572
19819_S_AT


2573
19961_S_AT


2574
20002_AT


2575
20034_I_AT


2576
20062_AT


2577
20136_AT


2578
20223_AT


2579
20235_I_AT


2580
20401_AT


2581
20407_AT


2582
20470_AT


2583
20626_AT


2584
20631_S_AT


2585
20647_AT
















TABLE 12





2X UP IN MANNITOL, ONLY


















12039_at
16832_at



12068_at
16993_at



12139_at
17037_s_at



12212_at
17054_s_at



12278_at
17083_s_at



12366_s_at
17097_s_at



12453_at
17119_s_at



12556_at
17270_at



12575_s_at
17305_at



12746_i_at
17376_at



12848_at
17378_at



12869_s_at
17449_s_at



12920_at
17481_at



12921_s_at
17533_s_at



13041_s_at
17832_s_at



13059_at
17923_s_at



13241_s_at
17944_at



13255_i_at
18059_f_at



13270_at
18216_at



13382_at
18230_at



13406_at
18255_at



13433_at
18284_at



13550_at
18301_s_at



13672_s_at
18312_s_at



13716_at
18326_s_at



13842_at
18599_at



13933_at
18672_s_at



13995_at
18720_s_at



14062_at
18768_at



14075_at
18814_at



14162_at
18877_at



14208_at
18921_g_at



14217_at
18960_at



14235_at
19060_at



14310_at
19182_at



14431_at
19192_at



14513_s_at
19266_at



14584_at
19369_at



14604_at
19386_at



14673_s_at
19402_at



14856_s_at
19412_at



15207_s_at
19432_s_at



15338_at
19469_at



15406_at
19622_g_at



15418_at
19819_s_at



15591_s_at
19826_at



15666_s_at
20152_at



15680_s_at
20223_at



15866_s_at
20235_i_at



15918_at
20365_s_at



16340_at
20470_at



16553_f_at
20537_at



16797_at
20547_at

















TABLE 13





2X UP IN MANNITOL, 3 HR ONLY


















12039_at
17449_s_at



12068_at
17481_at



12139_at
17533_s_at



12212_at
17923_s_at



12278_at
17944_at



12366_s_at
18059_i_at



12453_at
18216_at



12556_at
18230_at



12575_s_at
18255_at



12746_i_at
18301_s_at



12848_at
18312_s_at



12869_s_at
18326_s_at



12920_at
18599_at



12921_s_at
18720_s_at



13041_s_at
18768_at



13059_at
18814_at



13241_s_at
18877_at



13382_at
18921_g_at



13406_at
18960_at



13433_at
19060_at



13550_at
19192_at



13672_s_at
19266_at



13933_at
19369_at



13995_at
19386_at



14062_at
19402_at



14075_at
19412_at



14162_at
19432_s_at



14217_at
19469_at



14310_at
19622_g_at



14431_at
19819_s_at



14513_s_at
20152_at



14584_at
20223_at



14604_at
20235_i_at



14673_s_at
20365_s_at



14856_s_at
20470_at



15207_s_at
20537_at



15338_at



15418_at



15591_s_at



15866_s_at



15918_at



16340_at



16553_f_at



16797_at



16832_at



17037_s_at



17054_s_at



17083_s_at



17097_s_at



17270_at



17305_at



17376_at



17378_at

















TABLE 14





2X DOWN IN MANNITOL, ONLY



















12028_at
14897_at
17958_at



12033_at
14918_at
18012_s_at



12110_at
15082_at
18227_at



12114_at
15084_at
18272_at



12189_at
15098_s_at
18331_s_at



12191_at
15105_s_at
18369_at



12211_at
15121_s_at
18411_at



12223_s_at
15126_s_at
18533_at



12268_at
15168_s_at
18576_s_at



12345_at
15271_at
18640_at



12381_at
15429_at
18696_s_at



12406_s_at
15548_at
18945_at



12412_at
15672_s_at
18949_at



12522_at
15753_at
18953_at



12571_s_at
15867_at
18965_at



12662_at
15999_s_at
19164_g_at



12787_at
16001_at
19322_at



12847_at
16021_s_at
19366_s_at



12895_at
16190_at
19371_at



12911_s_at
16260_at
19397_at



13027_at
16303_at
19427_s_at



13075_i_at
16363_at
19681_at



13221_at
16458_s_at
19707_s_at



13262_s_at
16468_at
19839_at



13283_s_at
16475_at
19961_s_at



13386_s_at
16513_s_at
19976_at



13447_s_at
16529_at
19998_at



13482_at
16563_s_at
20002_at



13634_s_at
16690_g_at
20034_i_at



13709_s_at
16814_at
20136_at



13732_at
16847_at
20382_s_at



13733_i_at
16927_s_at
20407_at



13812_s_at
16976_s_at
20529_at



13825_s_at
17007_at
20626_at



13860_s_at
17014_s_at
20631_s_at



13868_at
17016_s_at
20647_at



13901_at
17071_s_at
20699_at



14052_at
17090_s_at



14224_at
17257_s_at



14244_s_at
17275_i_at



14254_s_at
17424_at



14256_f_at
17464_at



14354_at
17468_at



14476_at
17511_s_at



14568_s_at
17519_s_at



14634_s_at
17525_s_at



14646_s_at
17645_s_at



14660_s_at
17741_at



14686_s_at
17815_s_at



14726_s_at
17897_at



14848_s_at
17899_at



14873_at
17934_at



14883_at
17937_s_at

















TABLE 15







COLD & OSOMOTIC STRESS RESPONSIVE SEQUENCES








SEQ
AFFYMETRIX


ID NO:
ID NO:





1699
12040_AT


1700
12048_AT


1701
12054_S_AT


1702
12077_AT


1703
12107_I_AT


1704
12113_AT


1705
12154_AT


1706
12171_AT


1707
12212_AT


1708
12278_AT


1709
12317_AT


1710
12325_AT


1711
12333_AT


1712
12345_AT


1713
12349_S_AT



14254_S_AT



14256_F_AT


1714
12356_AT


1715
12380_AT


1716
12392_AT


1717
12460_S_AT


1718
12556_AT


1719
12575_S_AT


1720
12686_S_AT


1721
12701_I_AT


1722
12754_G_AT


1723
12782_R_AT


1724
12784_AT


1725
12879_S_AT


1726
12891_AT



16817_S_AT


1727
12898_G_AT


1728
12974_AT


1729
12998_AT


1730
13041_S_AT


1731
13124_AT


1732
13134_S_AT


1733
13144_AT


1734
13147_AT


1735
13152_S_AT


1736
13187_I_AT



16981_S_AT


1737
13192_S_AT



17525_S_AT


1738
13212_S_AT


1739
13215_S_AT



16649_S_AT


1740
13241_S_AT


1741
13246_AT


1742
13262_S_AT


1743
13286_S_AT


1744
13324_AT


1745
13340_S_AT


1746
13361_AT


1747
13406_AT


1748
13441_S_AT


1749
13513_AT


1750
13550_AT


1751
13573_AT


1752
13577_S_AT


1753
13606_AT


1754
13609_AT


1755
13625_S_AT


1756
13626_AT


1757
13634_S_AT


1758
13672_S_AT



18916_S_AT


1759
13709_S_AT


1760
13736_AT


1761
13775_AT


1762
13810_AT


1763
13812_S_AT


1764
13825_S_AT


1765
14015_S_AT



14016_S_AT


1766
14029_AT


1767
14036_AT


1768
14051_AT


1769
14060_AT


1770
14064_AT


1771
14066_AT


1772
14075_AT


1773
14094_S_AT



19999_S_AT


1774
14096_AT


1775
14104_AT


1776
14123_S_AT


1777
14126_S_AT


1778
14131_AT


1779
14136_AT


1780
14139_AT



14140_AT


1781
14162_AT



14217_AT


1782
14178_AT


1783
14201_AT


1784
14208_AT


1785
14235_AT


1786
14242_S_AT


1787
14431_AT


1788
14480_AT


1789
14497_AT


1790
14553_AT


1791
14584_AT


1792
14600_AT


1793
14673_S_AT



19432_S_AT


1794
14681_G_AT


1795
14699_AT


1796
14751_AT


1797
14762_AT


1798
14828_S_AT


1799
14856_S_AT


1800
14882_AT


1801
14897_AT


1802
14978_AT


1803
14985_S_AT


1804
15031_AT


1805
15084_AT


1806
15096_AT


1807
15105_S_AT


1808
15110_S_AT


1809
15111_S_AT


1810
15120_S_AT


1811
15126_S_AT


1812
15142_S_AT


1813
15144_S_AT


1814
15184_S_AT


1815
15198_S_AT


1816
15203_S_AT


1817
15207_S_AT


1818
15240_AT


1819
15366_AT


1820
15398_AT


1821
15406_AT


1822
15448_AT


1823
15466_AT


1824
15481_AT


1825
15484_AT


1826
15549_AT


1827
15591_S_AT


1828
15606_S_AT


1829
15614_S_AT



16927_S_AT


1830
15629_S_AT


1831
15633_S_AT


1832
15641_S_AT



18012_S_AT


1833
15720_AT


1834
15815_S_AT


1835
15817_AT


1836
15837_AT


1837
15841_AT


1838
15866_S_AT



18255_AT


1839
15872_AT



18331_S_AT


1840
15892_AT


1841
15933_AT


1842
15947_AT


1843
15959_S_AT


1844
16001_AT


1845
16052_AT


1846
16161_S_AT


1847
16204_AT


1848
16232_S_AT


1849
16252_AT


1850
16260_AT


1851
16266_AT


1852
16299_AT


1853
16365_AT


1854
16468_AT


1855
16477_AT


1856
16491_AT


1857
16523_S_AT


1858
16566_S_AT


1859
16570_S_AT


1860
16688_AT


1861
16840_AT


1862
16847_AT


1863
16893_AT


1864
16896_S_AT


1865
16898_S_AT


1866
16912_S_AT


1867
16980_AT


1868
16993_AT


1869
17008_AT


1870
17012_S_AT


1871
17014_S_AT


1872
17016_S_AT


1873
17032_S_AT


1874
17050_S_AT



17051_S_AT


1875
17071_S_AT


1876
17090_S_AT



18690_S_AT


1877
17097_S_AT


1878
17104_S_AT


1879
17119_S_AT


1880
17160_AT


1881
17305_AT


1882
17424_AT


1883
17449_S_AT


1884
17452_G_AT


1885
17540_S_AT


1886
17552_S_AT


1887
17571_AT


1888
17589_AT


1889
17641_G_AT


1890
17741_AT



18098_AT


1891
17766_AT


1892
17873_S_AT


1893
17904_AT


1894
17920_S_AT


1895
17925_AT


1896
17943_AT


1897
18059_I_AT


1898
18230_AT


1899
18263_AT


1900
18272_AT


1901
18540_AT


1902
18608_AT


1903
18647_AT


1904
18662_S_AT


1905
18664_AT


1906
18695_S_AT


1907
18704_AT


1908
18814_AT


1909
18907_S_AT


1910
18921_G_AT


1911
18924_AT


1912
18949_AT



19707_S_AT


1913
18995_AT


1914
19017_AT


1915
19034_AT


1916
19063_AT


1917
19142_AT


1918
19158_AT


1919
19180_AT


1920
19187_AT


1921
19192_AT


1922
19195_AT


1923
19199_AT


1924
19231_AT


1925
19263_AT


1926
19308_AT


1927
19322_AT


1928
19365_S_AT


1929
19372_AT


1930
19389_AT


1931
19392_AT


1932
19397_AT


1933
19400_AT


1934
19402_AT


1935
19458_AT








1936
19469_AT


1937
19473_AT


1938
19597_S_AT


1939
19710_S_AT


1940
19830_AT


1941
19839_AT


1942
19840_S_AT


1943
19853_AT


1944
19860_AT


1945
19880_AT


1946
19889_AT


1947
19898_AT


1948
19914_AT


1949
19924_AT


1950
19949_AT


1951
19976_AT


1952
19998_AT


1953
20030_AT


1954
20151_AT


1955
20152_AT


1956
20187_AT


1957
20214_I_AT


1958
20269_AT


1959
20271_AT


1960
20273_AT


1961
20299_AT


1962
20323_AT


1963
20429_S_AT


1964
20457_AT


1965
20480_S_AT


1966
20529_AT


1967
20547_AT


1968
20555_S_AT


1969
20699_AT
















TABLE 16





2X UP IN MANNITOL & COLD, ONLY


















12345_at
17066_s_at



12784_at
17540_s_at



13153_r_at
17567_at



13212_s_at
17766_at



13215_s_at
17904_at



13246_at
17920_s_at



13262_s_at
17943_at



13361_at
18263_at



13625_s_at
18351_s_at



13764_at
18662_s_at



13810_at
18670_g_at



14015_s_at
18695_s_at



14016_s_at
18704_at



14060_at
18729_at



14096_at
18995_at



14123_s_at
19158_at



14139_at
19473_at



14219_at
19710_s_at



14248_at
19883_at



14254_s_at
19889_at



14256_f_at
20030_at



14609_at
20269_at



14636_s_at
20271_at



14681_g_at
20299_at



14699_at
20429_s_at



14704_s_at
20438_at



14828_s_at
20480_s_at



14882_at



15110_s_at



15184_s_at



15448_at



15629_s_at



15720_at



15846_at



15947_at



16161_s_at



16365_at



16427_at



16566_s_at



16570_s_at



16649_s_at



16688_at



16712_at



16817_s_at



16840_at



16893_at



16912_s_at



16916_s_at



16927_s_at



16981_s_at



17012_s_at



17014_s_at



17051_s_at

















TABLE 17





2X DOWN COLD & MANNITOL, ONLY



















12040_at
14553_at
17873_s_at



12048_at
14612_at
17925_at



12054_s_at
14751_at
18098_at



12077_at
14762_at
18540_at



12107_i_at
14978_at
18608_at



12113_at
14985_s_at
18647_at



12154_at
15031_at
18664_at



12171_at
15096_at
18690_s_at



12317_at
15111_s_at
18725_s_at



12325_at
15120_s_at
18924_at



12333_at
15142_s_at
19017_at



12356_at
15198_s_at
19034_at



12380_at
15203_s_at
19063_at



12392_at
15240_at
19141_at



12460_s_at
15366_at
19142_at



12686_s_at
15392_at
19180_at



12701_i_at
15398_at
19187_at



12782_r_at
15466_at
19195_at



12879_s_at
15481_at
19199_at



12898_g_at
15484_at
19231_at



12974_at
15549_at
19308_at



12998_at
15623_f_at
19372_at



13144_at
15815_s_at
19392_at



13147_at
15817_at
19400_at



13152_s_at
15841_at
19458_at



13192_s_at
15892_at
19597_s_at



13286_s_at
15933_at
19762_at



13324_at
15959_s_at
19830_at



13340_s_at
16052_at
19853_at



13441_s_at
16204_at
19869_at



13513_at
16252_at
19880_at



13573_at
16266_at
19898_at



13606_at
16299_at
19914_at



13609_at
16477_at
19924_at



13626_at
16491_at
19949_at



13736_at
16561_s_at
20151_at



13775_at
16645_s_at
20187_at



14029_at
16898_s_at
20214_i_at



14036_at
16980_at
20273_at



14051_at
17008_at
20323_at



14064_at
17104_s_at
20457_at



14066_at
17160_at
20555_s_at



14094_s_at
17317_at



14104_at
17400_s_at



14126_s_at
17452_g_at



14131_at
17477_s_at



14136_at
17500_s_at



14178_at
17552_s_at



14192_at
17571_at



14201_at
17572_s_at



14242_s_at
17589_at



14480_at
17641_g_at



14497_at
17855_at

















TABLE 18







COLD & SALINE STRESS RESPONSIVE SEQUENCES








SEQ
AFFYMETRIX


ID NO:
ID NO:





1970
12021_AT


1971
12037_AT


1972
12094_AT


1973
12098_AT


1974
12128_AT


1975
12148_AT


1976
12151_AT


1977
12357_S_AT


1978
12394_AT


1979
12472_S_AT


1980
12475_AT


1981
12482_S_AT


1982
12490_AT


1983
12505_S_AT


1984
12531_AT


1985
12540_S_AT


1986
12541_AT


1987
12577_AT


1988
12594_AT


1989
12629_AT


1990
12642_AT


1991
12656_AT


1992
12660_AT


1993
12712_F_AT


1994
12725_R_AT


1995
12745_AT


1996
12777_I_AT


1997
12790_S_AT


1998
12798_AT


1999
12801_AT


2000
12855_F_AT


2001
12887_S_AT


2002
12933_R_AT


2003
12951_AT


2004
13005_AT


2005
13015_S_AT


2006
13115_AT


2007
13178_AT


2008
13228_AT


2009
13236_S_AT



16646_S_AT


2010
13266_S_AT



15211_S_AT


2011
13275_F_AT


2012
13335_AT


2013
13362_S_AT


2014
13428_AT


2015
13464_AT


2016
13480_AT


2017
13538_AT


2018
13544_AT


2019
13549_AT


2020
13565_AT


2021
13580_AT


2022
13588_AT


2023
13649_AT


2024
13652_AT


2025
13679_S_AT


2026
13696_AT


2027
13702_S_AT


2028
13751_AT


2029
13919_AT


2030
13943_AT


2031
13950_S_AT


2032
14050_AT


2033
14055_S_AT



16166_S_AT


2034
14067_AT


2035
14078_AT


2036
14110_I_AT


2037
14144_AT


2038
14232_AT


2039
14285_AT


2040
14346_AT


2041
14432_AT


2042
14468_AT


2043
14479_AT


2044
14524_S_AT


2045
14608_AT


2046
14621_AT


2047
14635_S_AT



17128_S_AT


2048
14640_S_AT


2049
14643_S_AT


2050
14663_S_AT


2051
14668_S_AT


2052
14688_S_AT



18279_S_AT


2053
14737_S_AT


2054
14768_AT


2055
14875_AT


2056
14911_S_AT



17056_S_AT


2057
14924_AT


2058
14956_S_AT



15148_S_AT



18673_AT


2059
14964_AT


2060
15022_AT


2061
15040_G_AT


2062
15047_AT


2063
15063_AT


2064
15085_S_AT


2065
15123_S_AT


2066
15133_S_AT


2067
15137_S_AT


2068
15153_S_AT


2069
15170_S_AT


2070
15172_S_AT


2071
15182_S_AT


2072
15190_S_AT


2073
15241_S_AT


2074
15389_AT


2075
15453_S_AT


2076
15495_AT


2077
15496_AT


2078
15519_S_AT


2079
15562_AT


2080
15580_S_AT


2081
15582_S_AT


2082
15638_S_AT



18751_F_AT


2083
15646_S_AT


2084
15647_S_AT


2085
15654_S_AT


2086
15655_S_AT


2087
15658_S_AT


2088
15670_S_AT


2089
15775_AT


2090
15798_AT


2091
15930_AT


2092
15931_AT


2093
15949_S_AT


2094
16017_AT


2095
16053_I_AT


2096
16078_S_AT


2097
16086_S_AT


2098
16120_S_AT


2099
16126_S_AT


2100
16150_S_AT


2101
16159_S_AT


2102
16230_AT


2103
16306_AT


2104
16367_I_AT


2105
16417_S_AT



18083_R_AT


2106
16418_S_AT


2107
16423_AT


2108
16449_S_AT


2109
16484_S_AT


2110
16489_AT


2111
16565_S_AT


2112
16596_S_AT


2113
16600_S_AT


2114
16603_S_AT


2115
16638_S_AT


2116
16642_S_AT


2117
16763_AT


2118
16914_S_AT


2119
16968_AT


2120
16983_AT


2121
16989_AT


2122
17002_AT


2123
17015_S_AT


2124
17040_S_AT



18913_S_AT


2125
17232_AT


2126
17380_AT


2127
17394_S_AT



20640_S_AT


2128
17398_AT


2129
17448_AT


2130
17485_S_AT


2131
17490_S_AT


2132
17499_S_AT


2133
17505_S_AT


2134
17516_S_AT


2135
17529_S_AT


2136
17543_S_AT


2137
17593_R_AT



19858_S_AT


2138
17609_AT


2139
17698_AT


2140
17836_AT


2141
17886_AT


2142
17896_AT


2143
17901_AT


2144
17902_S_AT


2145
17913_S_AT


2146
17924_AT


2147
17954_S_AT


2148
17960_AT


2149
17991_G_AT



18967_S_AT


2150
17999_AT


2151
18057_I_AT


2152
18078_AT


2153
18091_AT


2154
18168_S_AT


2155
18252_AT


2156
18267_AT


2157
18300_AT


2158
18308_I_AT


2159
18328_AT


2160
18354_AT


2161
18402_AT


2162
18416_AT


2163
18455_AT


2164
18459_AT


2165
18571_AT


2166
18604_AT



19181_S_AT


2167
18644_AT


2168
18745_F_AT



19611_S_AT


2169
18782_AT


2170
18881_AT


2171
18904_S_AT


2172
18914_S_AT


2173
18963_AT


2174
19068_I_AT


2175
19078_AT


2176
19171_AT


2177
19177_AT


2178
19394_AT


2179
19411_AT


2180
19415_AT


2181
19466_S_AT


2182
19484_S_AT


2183
19549_S_AT


2184
19592_AT


2185
19633_AT


2186
19641_AT


2187
19669_AT


2188
19672_AT


2189
19684_AT


2190
19692_AT


2191
19746_AT


2192
19835_AT


2193
19848_S_AT


2194
19892_AT


2195
19904_AT


2196
19936_AT


2197
19974_S_AT


2198
19994_AT


2199
20005_S_AT


2200
20022_AT


2201
20032_AT


2202
20044_AT


2203
20049_AT


2204
20081_AT


2205
20133_I_AT


2206
20155_S_AT


2207
20163_S_AT


2208
20200_AT


2209
20296_S_AT


2210
20336_AT


2211
20341_AT


2212
20372_AT


2213
20385_S_AT


2214
20433_AT


2215
20489_AT


2216
20525_AT


2217
20543_AT


2218
20565_AT


2219
20570_AT


2220
20576_AT


2221
20577_AT


2222
20609_AT


2223
20646_AT


2224
20672_AT


2225
20707_S_AT


2226
20720_AT
















TABLE 19





2X UP IN SALT & COLD, ONLY

















12004_at



12098_at



12148_at



12251_at



12357_s_at



12394_at



12457_at



12505_s_at



12522_at



12541_at



12594_at



12606_at



12697_at



12745_at



12781_at



12798_at



12855_f_at



12945_at



12951_at



13005_at



13015_s_at



13115_at



13146_s_at



13335_at



13447_s_at



13480_at



13544_at



13549_at



13580_at



13649_at



13943_at



13950_s_at



14110_i_at



14144_at



14224_at



14432_at



14468_at



14479_at



14524_s_at



14640_s_at



14643_s_at



14735_s_at



14737_s_at



14768_at



14784_at



14924_at



15064_at



15127_s_at



15186_s_at



15189_s_at



15255_at



15389_at



15482_at



15495_at



15496_at



15519_s_at



15580_s_at



15582_s_at



15776_at



15798_at



15910_at



15931_at



15937_at



15949_s_at



15972_s_at



16048_at



16086_s_at



16120_s_at



16126_s_at



16150_s_at



16159_s_at



16230_at



16306_at



16418_s_at



16423_at



16449_s_at



16565_s_at



16603_s_at



16763_at



16968_at



16983_at



17002_at



17015_s_at



17019_s_at



17078_s_at



17232_at



17317_at



17394_s_at



17516_s_at



17585_s_at



17609_at



17698_at



17836_at



17896_at



17899_at



17902_s_at



17960_at



17963_at



18168_s_at



18252_at



18267_at



18308_i_at



18354_at



18402_at



18459_at



18484_at



18745_f_at



18904_s_at



18914_s_at



18929_s_at



18946_at



18963_at



19078_at



19137_at



19141_at



19411_at



19641_at



19672_at



19684_at



19692_at



19746_at



19762_at



19869_at



19894_at



19904_at



19936_at



19994_at



20005_s_at



20031_at



20044_at



20382_s_at



20406_g_at



20421_at



20525_at



20543_at



20565_at



20570_at



20640_s_at



20646_at



20720_at

















TABLE 20





2X DOWN IN COLD & SALT, ONLY



















12021_at
15123_s_at
19394_at



12094_at
15153_s_at
19415_at



12128_at
15172_s_at
19466_s_at



12151_at
15190_s_at
19549_s_at



12332_s_at
15211_s_at
19592_at



12472_s_at
15241_s_at
19633_at



12475_at
15437_at
19669_at



12482_s_at
15562_at
19848_s_at



12490_at
15638_s_at
19858_s_at



12531_at
15647_s_at
19878_at



12540_s_at
15654_s_at
19892_at



12577_at
15655_s_at
19974_s_at



12629_at
15658_s_at
20022_at



12642_at
15695_s_at
20032_at



12660_at
15846_at
20049_at



12676_s_at
15930_at
20081_at



12712_f_at
16053_i_at
20155_s_at



12725_r_at
16078_s_at
20163_s_at



12777_i_at
16229_at
20296_s_at



12790_s_at
16465_at
20336_at



12801_at
16484_s_at
20341_at



12887_s_at
16596_s_at
20365_s_at



12933_r_at
16600_s_at
20372_at



13153_r_at
16642_s_at
20489_at



13228_at
16914_s_at
20491_at



13362_s_at
17027_s_at
20576_at



13428_at
17066_s_at
20577_at



13538_at
17083_s_at
20609_at



13565_at
17128_s_at
20672_at



13588_at
17380_at



13696_at
17398_at



13702_s_at
17448_at



13716_at
17485_s_at



13764_at
17490_s_at



14050_at
17499_s_at



14055_s_at
17505_s_at



14069_at
17514_s_at



14078_at
17593_r_at



14232_at
17886_at



14346_at
17913_s_at



14608_at
17924_at



14609_at
17954_s_at



14621_at
17991_g_at



14635_s_at
18057_i_at



14663_s_at
18069_at



14688_s_at
18328_at



14691_at
18416_at



14704_s_at
18604_at



14875_at
18644_at



14911_s_at
18881_at



14964_at
19171_at



15022_at
19181_s_at



15085_s_at
19182_at

















TABLE 21







OSMOTIC & SALINE STRESS RESPONSIVE SEQUENCES








SEQ
AFFYMETRIX


ID NO:
ID NO:





2586
12126_S_AT


2587
12137_AT


2588
12227_AT


2589
12239_AT


2590
12268_AT


2591
12369_AT


2592
12476_AT


2593
12484_G_AT


2594
12494_AT


2595
12644_AT


2596
12645_AT


2597
12796_S_AT


2598
12819_AT


2599
12841_AT


2600
12852_S_AT



19455_S_AT


2601
13084_AT


2602
13171_AT


2603
13174_R_AT


2604
13596_AT


2605
13807_AT


2606
13977_AT


2607
13999_AT


2608
14052_AT


2609
14293_AT


2610
14335_AT


2611
14486_AT


2612
14506_AT


2613
14518_AT


2614
14540_AT


2615
14578_S_AT


2616
14646_S_AT


2617
14662_F_AT



15962_S_AT


2618
14901_AT


2619
14918_AT


2620
14986_AT


2621
15053_S_AT


2622
15179_S_AT


2623
15252_G_AT


2624
15280_AT


2625
15467_AT


2626
15607_S_AT


2627
15625_S_AT


2628
15703_I_AT


2629
15827_AT


2630
15863_AT


2631
15923_AT


2632
15946_S_AT


2633
16005_S_AT


2634
16073_F_AT


2635
16114_S_AT


2636
16127_S_AT



18744_F_AT


2637
16190_AT


2638
16196_AT


2639
16236_G_AT



19531_AT


2640
16310_AT


2641
16316_AT


2642
16334_S_AT


2643
16335_AT


2644
16340_AT


2645
16450_S_AT


2646
16500_AT


2647
16524_AT


2648
16533_AT


2649
16690_G_AT


2650
16762_AT


2651
16819_AT


2652
16873_I_AT


2653
16972_AT


2654
16991_AT


2655
17099_S_AT


2656
17339_AT


2657
17397_S_AT


2658
17419_AT


2659
17460_AT


2660
17554_S_AT


2661
17939_AT


2662
18013_R_AT



18178_S_AT


2663
18024_S_AT


2664
18032_I_AT


2665
18054_AT


2666
18151_AT


2667
18281_AT


2668
18445_AT


2669
18520_AT


2670
18583_AT


2671
18663_S_AT


2672
18753_S_AT


2673
18876_AT


2674
18938_G_AT


2675
18971_AT


2676
18977_AT


2677
18981_AT


2678
19099_AT


2679
19196_AT


2680
19376_AT


2681
19409_AT


2682
19503_AT


2683
19826_AT


2684
19847_S_AT


2685
19930_AT


2686
19992_AT


2687
20096_AT


2688
20108_AT


2689
20256_S_AT


2690
20290_S_AT


2691
20298_AT


2692
20305_AT


2693
20322_AT


2694
20333_AT


2695
20402_S_AT


2696
20424_AT


2697
20446_S_AT


2698
20450_AT


2699
20468_AT


2700
20569_S_AT


2701
20639_AT


2702
20678_AT


2703
20686_AT
















TABLE 22





2X UP IN SALT & MANNITOL, ONLY


















12126_s_at
17548_s_at



12227_at
17554_s_at



12369_at
17961_at



12521_at
18032_i_at



12644_at
18054_at



12645_at
18151_at



12724_f_at
18167_s_at



12795_at
18281_at



12796_s_at
18520_at



12841_at
18663_s_at



12852_s_at
18744_f_at



12958_at
18753_s_at



13014_at
18789_at



13174_r_at
18876_at



13211_s_at
18909_s_at



13596_at
18938_g_at



13640_at
18977_at



13789_at
19099_at



13977_at
19108_at



13999_at
19135_at



14069_at
19227_at



14083_at
19376_at



14089_at
19429_at



14293_at
19455_s_at



14675_s_at
19531_at



15053_s_at
19789_s_at



15058_s_at
19878_at



15252_g_at
20017_at



15280_at
20096_at



15437_at
20256_s_at



15607_s_at
20290_s_at



15625_s_at
20305_at



15827_at
20322_at



15863_at
20333_at



15880_at
20420_at



16005_s_at
20424_at



16031_at
20689_s_at



16073_f_at



16316_at



16334_s_at



16335_at



16450_s_at



16500_at



16524_at



16533_at



16597_s_at



16819_at



17085_s_at



17099_s_at



17339_at



17419_at



17442_i_at



17514_s_at

















TABLE 23





2X DOWN IN MANNITOL & SALT, ONLY


















12239_at
20108_at



12251_at
20298_at



12476_at
20421_at



12484_g_at
20432_at



12494_at
20446_s_at



12561_at
20639_at



12647_s_at



12719_f_at



12819_at



12841_at



13084_at



13171_at



13172_s_at



13435_at



13807_at



14250_r_at



14335_at



14486_at



14506_at



14518_at



14901_at



15046_s_at



15179_s_at



15451_at



15703_i_at



15946_s_at



16014_s_at



16114_s_at



16310_at



16342_at



16712_at



16762_at



16972_at



16991_at



17397_s_at



17408_at



17460_at



17775_at



17939_at



18445_at



18583_at



18751_f_at



18971_at



18981_at



19156_s_at



19196_at



19359_s_at



19409_at



19503_at



19713_at



19718_at



19847_s_at



19930_at

















TABLE 24







COLD, OSMOTIC & SALINE RESPONSIVE SEQUENCES








SEQ
AFFYMETRIX


ID NO:
ID NO:





1262
12004_AT


1263
12023_S_AT


1264
12078_AT


1265
12115_AT


1266
12118_AT


1267
12150_AT


1268
12251_AT


1269
12271_S_AT


1270
12276_AT


1271
12332_S_AT



13211_S_AT


1272
12338_AT


1273
12400_AT


1274
12430_AT


1275
12457_AT


1276
12521_AT


1277
12522_AT


1278
12530_AT


1279
12536_S_AT


1280
12538_AT


1281
12561_AT


1282
12574_AT



19019_I_AT


1283
12595_AT


1284
12606_AT


1285
12609_AT


1286
12622_AT


1287
12630_AT


1288
12647_S_AT


1289
12676_S_AT


1290
12697_AT


1291
12698_AT


1292
12719_F_AT


1293
12724_F_AT



15871_S_AT



16597_S_AT


1294
12749_AT


1295
12765_AT


1296
12769_AT


1297
12781_AT


1298
12785_AT


1299
12792_S_AT


1300
12795_AT


1301
12805_S_AT


1302
12857_AT


1303
12883_S_AT


1304
12909_S_AT



16539_S_AT


1305
12932_S_AT



15605_S_AT


1306
12945_AT


1307
12958_AT


1308
12964_AT


1309
12968_AT


1310
12972_AT


1311
12989_S_AT


1312
13004_AT


1313
13014_AT


1314
13025_AT


1315
13036_AT


1316
13099_S_AT


1317
13136_AT


1318
13146_S_AT



13239_S_AT


1319
13153_R_AT


1320
13159_AT


1321
13176_AT


1322
13217_S_AT



17500_S_AT


1323
13225_S_AT



15997_S_AT


1324
13230_S_AT



15972_S_AT


1325
13279_S_AT



17477_S_AT


1326
13280_S_AT



20301_S_AT


1327
13282_S_AT



17027_S_AT


1328
13426_AT


1329
13432_AT


1330
13435_AT


1331
13447_S_AT


1332
13474_AT


1333
13511_AT


1334
13546_AT


1335
13547_S_AT


1336
13548_AT


1337
13555_AT


1338
13587_AT


1339
13595_AT


1340
13610_S_AT


1341
13627_AT


1342
13640_AT


1343
13645_AT


1344
13647_AT


1345
13706_S_AT



19701_S_AT


1346
13716_AT



18228_AT


1347
13725_AT


1348
13764_AT


1349
13771_AT


1350
13789_AT


1351
13916_AT


1352
13965_S_AT


1353
13967_AT


1354
14028_AT


1355
14039_AT


1356
14046_AT


1357
14049_AT


1358
14069_AT


1359
14077_AT


1360
14080_AT


1361
14083_AT


1362
14089_AT


1363
14090_I_AT


1364
14097_AT


1365
14116_AT


1366
14151_AT



14219_AT


1367
14170_AT


1368
14172_AT


1369
14192_AT


1370
14224_AT


1371
14227_AT


1372
14244_S_AT



14245_AT



14645_S_AT



15974_G_AT


1373
14248_AT


1374
14250_R_AT


1375
14367_AT


1376
14381_AT


1377
14384_AT


1378
14398_S_AT


1379
14487_AT


1380
14582_AT


1381
14597_AT


1382
14609_AT


1383
14612_AT



19267_S_AT


1384
14614_AT


1385
14636_S_AT


1386
14644_S_AT



14658_S_AT



14659_S_AT



15964_S_AT


1387
14675_S_AT


1388
14691_AT



14709_AT


1389
14704_S_AT



15846_AT


1390
14705_I_AT


1391
14733_S_AT


1392
14735_S_AT


1393
14779_AT


1394
14784_AT


1395
14923_AT


1396
14947_AT


1397
14950_AT


1398
14990_AT


1399
14998_AT


1400
15005_S_AT


1401
15018_AT


1402
15045_AT


1403
15046_S_AT


1404
15052_AT


1405
15058_S_AT


1406
15064_AT


1407
15088_S_AT


1408
15098_S_AT


1409
15103_S_AT


1410
15109_S_AT


1411
15124_S_AT


1412
15127_S_AT


1413
15145_S_AT


1414
15154_S_AT


1415
15161_S_AT


1416
15189_S_AT


1417
15214_S_AT


1418
15255_AT


1419
15356_AT


1420
15357_AT


1421
15364_AT


1422
15392_AT


1423
15403_S_AT


1424
15437_AT


1425
15451_AT


1426
15476_AT


1427
15482_AT


1428
15483_S_AT


1429
15521_S_AT


1430
15522_I_AT


1431
15531_I_AT


1432
15573_AT


1433
15581_S_AT


1434
15586_S_AT


1435
15594_S_AT


1436
15609_S_AT


1437
15611_S_AT


1438
15621_F_AT


1439
15623_F_AT


1440
15669_S_AT


1441
15695_S_AT


1442
15702_S_AT


1443
15753_AT


1444
15761_AT


1445
15776_AT


1446
15778_AT


1447
15839_AT


1448
15842_AT


1449
15857_S_AT


1450
15859_AT


1451
15880_AT


1452
15886_AT


1453
15906_S_AT


1454
15910_AT


1455
15937_AT


1456
15957_AT


1457
15970_S_AT


1458
15985_AT


1459
16010_S_AT



16011_S_AT



17078_S_AT


1460
16021_S_AT


1461
16031_AT


1462
16038_S_AT


1463
16045_S_AT


1464
16046_S_AT


1465
16048_AT


1466
16061_S_AT


1467
16082_S_AT


1468
16111_F_AT


1469
16115_S_AT


1470
16141_S_AT


1471
16144_S_AT


1472
16163_S_AT


1473
16173_S_AT


1474
16229_AT


1475
16298_AT


1476
16301_S_AT


1477
16322_AT


1478
16342_AT


1479
16351_AT


1480
16412_S_AT


1481
16422_AT


1482
16427_AT


1483
16438_AT


1484
16474_S_AT


1485
16482_S_AT


1486
16485_S_AT



18052_S_AT


1487
16493_AT


1488
16534_S_AT


1489
16555_S_AT


1490
16561_S_AT



17572_S_AT


1491
16592_S_AT


1492
16615_S_AT


1493
16637_S_AT


1494
16692_AT


1495
16712_AT


1496
16789_AT


1497
16818_S_AT


1498
16971_S_AT


1499
17018_S_AT


1500
17019_S_AT


1501
17029_S_AT


1502
17041_S_AT


1503
17047_S_AT


1504
17066_S_AT


1505
17085_S_AT


1506
17089_S_AT


1507
17179_AT


1508
17180_AT


1509
17228_AT


1510
17252_AT


1511
17317_AT


1512
17338_AT


1513
17384_AT


1514
17387_S_AT


1515
17400_S_AT


1516
17407_S_AT


1517
17408_AT


1518
17413_S_AT


1519
17416_AT


1520
17425_S_AT


1521
17440_I_AT


1522
17442_I_AT


1523
17473_AT


1524
17484_AT


1525
17514_S_AT


1526
17520_S_AT


1527
17533_S_AT


1528
17548_S_AT



19614_AT


1529
17549_S_AT


1530
17555_S_AT


1531
17567_AT


1532
17654_AT


1533
17693_AT


1534
17697_AT


1535
17722_AT


1536
17752_AT


1537
17755_AT


1538
17775_AT


1539
17832_S_AT


1540
17840_S_AT


1541
17843_S_AT


1542
17855_AT


1543
17860_AT


1544
17869_AT


1545
17888_AT


1546
17899_AT


1547
17929_S_AT


1548
17930_S_AT


1549
17932_S_AT


1550
17936_S_AT



18670_G_AT


1551
17957_AT


1552
17961_AT


1553
17962_AT


1554
17963_AT


1555
17971_S_AT


1556
17975_AT



18742_F_AT


1557
18016_R_AT


1558
18069_AT


1559
18122_AT


1560
18140_AT


1561
18199_AT


1562
18224_S_AT


1563
18225_AT


1564
18235_AT


1565
18259_S_AT


1566
18265_AT


1567
18270_AT1568



18280_AT


1569
18289_AT


1570
18296_AT


1571
18298_AT


1572
18314_I_AT


1573
18318_AT


1574
18325_AT


1575
18351_S_AT


1576
18471_AT


1577
18482_S_AT


1578
18484_AT


1579
18560_AT


1580
18564_AT


1581
18590_AT


1582
18594_AT


1583
18595_AT


1584
18596_AT


1585
18629_S_AT


1586
18637_AT


1587
18661_AT


1588
18668_AT


1589
18699_I_AT


1590
18747_F_AT



18789_AT


1591
18761_AT


1592
18833_AT


1593
18875_S_AT


1594
18894_AT


1595
18936_AT


1596
18946_AT


1597
18953_AT


1598
18955_AT


1599
18972_AT


1600
19008_S_AT


1601
19108_AT


1602
19123_AT


1603
19135_AT


1604
19137_AT


1605
19141_AT


1606
19152_AT


1607
19156_S_AT


1608
19182_AT


1609
19186_S_AT


1610
19214_AT


1611
19216_AT


1612
19227_AT


1613
19243_AT


1614
19288_AT


1615
19359_S_AT


1616
19368_AT


1617
19379_AT


1618
19380_S_AT


1619
19398_AT


1620
19421_AT


1621
19424_AT


1622
19429_AT


1623
19430_AT


1624
19450_AT


1625
19457_AT


1626
19467_AT


1627
19516_AT


1628
19545_AT


1629
19564_AT


1630
19577_AT


1631
19593_AT


1632
19602_AT


1633
19618_AT


1634
19638_AT


1635
19640_AT


1636
19646_S_AT


1637
19656_S_AT


1638
19670_AT


1639
19696_AT


1640
19713_AT


1641
19718_AT


1642
19722_S_AT


1643
19749_AT


1644
19755_AT


1645
19762_AT


1646
19789_S_AT


1647
19815_AT


1648
19843_AT


1649
19869_AT


1650
19878_AT


1651
19883_AT


1652
19894_AT


1653
19926_AT


1654
19944_AT


1655
19968_AT


1656
19977_AT


1657
19982_AT


1658
19987_AT


1659
19991_AT


1660
20015_AT


1661
20017_AT


1662
20031_AT


1663
20040_AT


1664
20042_S_AT


1665
20060_AT



20438_AT


1666
20089_AT


1667
20118_AT


1668
20144_AT


1669
20149_AT


1670
20179_AT


1671
20190_AT


1672
20194_AT


1673
20219_AT


1674
20245_S_AT


1675
20263_AT


1676
20308_S_AT


1677
20335_S_AT


1678
20338_AT


1679
20345_AT


1680
20365_S_AT


1681
20382_S_AT


1682
20390_S_AT


1683
20395_AT


1684
20420_AT


1685
20421_AT


1686
20432_AT


1687
20437_AT


1688
20442_I_AT


1689
20463_S_AT


1690
20491_AT


1691
20537_AT


1692
20573_AT


1693
20636_AT


1694
20638_AT


1695
20641_AT


1696
20658_S_AT


1697
20689_S_AT


1698
20698_S_AT
















TABLE 25





2X UP IN COLD, SALT & MANNITOL




















12023_s_at
14733_s_at
17047_s_at
19640_at



12332_s_at
14923_at
17179_at
19646_s_at



12530_at
14990_at
17180_at
19656_s_at



12536_s_at
15005_s_at
17252_at
19701_s_at



12574_at
15018_at
17384_at
19843_at



12595_at
15052_at
17407_s_at
19944_at



12698_at
15088_s_at
17484_at
19982_at



12749_at
15098_s_at
17520_s_at
19987_at



12765_at
15103_s_at
17555_s_at
19991_at



12769_at
15145_s_at
17572_s_at
20042_s_at



12785_at
15154_s_at
17722_at
20060_at



12857_at
15161_s_at
17752_at
20118_at



12964_at
15214_s_at
17840_s_at
20144_at



12972_at
15356_at
17843_s_at
20149_at



12989_s_at
15521_s_at
17860_at
20179_at



13004_at
15573_at
17929_s_at
20194_at



13025_at
15586_s_at
17936_s_at
20245_s_at



13036_at
15609_s_at
17962_at
20390_s_at



13099_s_at
15611_s_at
18052_s_at
20437_at



13136_at
15621_f_at
18069_at
20463_s_at



13176_at
15669_s_at
18122_at
20491_at



13220_s_at
15695_s_at
18199_at
20641_at



13225_s_at
15753_at
18259_s_at
20658_s_at



13230_s_at
15761_at
18280_at



13239_s_at
15857_s_at
18289_at



13426_at
15871_s_at
18314_i_at



13474_at
15964_s_at
18318_at



13548_at
15970_s_at
18325_at



13555_at
15974_g_at
18482_s_at



13595_at
15997_s_at
18590_at



13627_at
16011_s_at
18594_at



13645_at
16021_s_at
18595_at



13647_at
16038_s_at
18596_at



13706_s_at
16046_s_at
18629_s_at



13965_s_at
16082_s_at
18661_at



13967_at
16111_f_at
18668_at



14080_at
16115_s_at
18699_i_at



14090_i_at
16127_s_at
18722_s_at



14097_at
16141_s_at
18936_at



14116_at
16144_s_at
18953_at



14151_at
16163_s_at
18955_at



14172_at
16236_g_at
18972_at



14192_at
16301_s_at
19008_s_at



14244_s_at
16322_at
19152_at



14245_at
16422_at
19186_s_at



14367_at
16474_s_at
19214_at



14398_s_at
16482_s_at
19368_at



14582_at
16485_s_at
19379_at



14614_at
16555_s_at
19380_s_at



14644_s_at
16561_s_at
19421_at



14645_s_at
16592_s_at
19545_at



14658_s_at
16637_s_at
19614_at



14659_s_at
17041_s_at
19638_at

















TABLE 26





2X DOWN IN COLD, MANNITOL & SALT, ONLY




















12078_at
15189_s_at
17869_at
20015_at



12115_at
15357_at
17888_at
20040_at



12118_at
15364_at
17930_s_at
20089_at



12150_at
15403_s_at
17932_s_at
20190_at



12271_s_at
15476_at
17957_at
20219_at



12276_at
15483_s_at
17963_at
20263_at



12338_at
15522_i_at
17971_s_at
20301_s_at



12400_at
15531_i_at
17975_at
20308_s_at



12430_at
15594_s_at
18016_r_at
20338_at



12538_at
15702_s_at
18140_at
20345_at



12622_at
15778_at
18224_s_at
20395_at



12630_at
15839_at
18225_at
20442_i_at



12792_s_at
15842_at
18228_at
20537_at



12805_s_at
15859_at
18235_at
20573_at



12883_s_at
15872_at
18265_at
20636_at



12909_s_at
15880_at
18270_at
20638_at



12932_s_at
15886_at
18296_at
20698_s_at



12968_at
15906_s_at
18298_at



13159_at
15957_at
18471_at



13217_s_at
15985_at
18564_at



13279_s_at
16045_s_at
18637_at



13282_s_at
16061_s_at
18742_f_at



13432_at
16173_s_at
18761_at



13511_at
16298_at
18833_at



13546_at
16351_at
18875_s_at



13547_s_at
16412_s_at
18894_at



13587_at
16438_at
18946_at



13610_s_at
16493_at
19123_at



13640_at
16534_s_at
19216_at



13725_at
16539_s_at
19243_at



13771_at
16615_s_at
19267_s_at



13916_at
16692_at
19288_at



14028_at
16789_at
19398_at



14039_at
16818_s_at
19424_at



14046_at
16971_s_at
19430_at



14049_at
17018_s_at
19450_at



14077_at
17029_s_at
19457_at



14170_at
17089_s_at
19467_at



14227_at
17228_at
19516_at



14248_at
17338_at
19564_at



14381_at
17387_s_at
19577_at



14384_at
17413_s_at
19593_at



14487_at
17416_at
19602_at



14597_at
17425_s_at
19618_at



14705_i_at
17440_i_at
19670_at



14709_at
17473_at
19696_at



14779_at
17533_s_at
19722_s_at



14947_at
17549_s_at
19749_at



14950_at
17654_at
19755_at



14998_at
17693_at
19815_at



15045_at
17697_at
19926_at



15109_s_at
17755_at
19968_at



15124_s_at
17832_s_at
19977_at

















TABLE 27





2X ROOT SPECIFIC (COLD, SALINE


& OSMOTIC STRESSES)




















11997_at
14069_at
16052_at
18327_s_at



12004_at
14072_at
16053_i_at
18597_at



12051_at
14073_at
16105_s_at
18607_s_at



12072_at
14097_at
16161_s_at
18636_at



12150_at
14139_at
16165_s_at
18663_s_at



12151_at
14235_at
16298_at
18782_at



12166_i_at
14250_r_at
16334_s_at
18885_at



12219_at
14578_s_at
16422_at
18888_at



12315_at
14582_at
16427_at
18942_at



12332_s_at
14640_s_at
16440_s_at
18955_at



12374_i_at
14643_s_at
16442_s_at
19060_at



12482_s_at
14644_s_at
16468_at
19108_at



12515_at
14658_s_at
16488_at
19135_at



12522_at
14659_s_at
16511_at
19137_at



12538_at
14711_s_at
16529_at
19195_at



12571_s_at
14900_at
16553_f_at
19263_at



12574_at
14924_at
16568_s_at
19376_at



12609_at
14990_at
16914_s_at
19406_at



12678_i_at
15018_at
16965_s_at
19432_s_at



12698_at
15022_at
16981_s_at
19835_at



12749_at
15107_s_at
16989_at
19836_at



12760_g_at
15116_f_at
17033_s_at
19840_s_at



12765_at
15120_s_at
17066_s_at
19841_at



12768_at
15124_s_at
17085_s_at
19843_at



12769_at
15131_s_at
17252_at
19926_at



12772_at
15132_s_at
17376_at
19972_at



12777_i_at
15137_s_at
17378_at
19977_at



12958_at
15184_s_at
17388_at
19991_at



12989_s_at
15188_s_at
17415_at
20034_i_at



13015_s_at
15208_s_at
17429_s_at
20042_s_at



13134_s_at
15252_g_at
17463_at
20189_at



13146_s_at
15343_at
17485_s_at
20194_at



13172_s_at
15389_at
17490_s_at
20200_at



13178_at
15392_at
17567_at
20214_i_at



13179_at
15448_at
17585_s_at
20239_g_at



13187_i_at
15503_at
17595_s_at
20262_at



13211_s_at
15531_i_at
17840_s_at
20269_at



13239_s_at
15594_s_at
17860_at
20294_at



13273_s_at
15609_s_at
17880_s_at
20312_s_at



13297_s_at
15623_f_at
17894_at
20382_s_at



13549_at
15639_s_at
17896_at
20396_at



13604_at
15670_s_at
17899_at
20432_at



13629_s_at
15680_s_at
17911_at
20444_at



13706_s_at
15859_at
17935_at
20446_s_at



13714_at
15900_at
17961_at
20480_s_at



13751_at
15923_at
18024_s_at
20586_i_at



13895_at
15962_s_at
18122_at
20612_s_at



13933_at
15964_s_at
18222_at
20672_at



13967_at
15965_at
18224_s_at
20686_at



13985_s_at
15975_s_at
18252_at
20689_s_at



14028_at
15985_at
18255_at



14030_at
16001_at
18269_s_at



14058_at
16048_at
18270_at

















TABLE 28





2X LEAF SPECIFIC (COLD, SALINE


& OSMOTIC STRESSES)


















12169_i_at
16136_s_at



12186_at
16172_s_at



12187_at
16316_at



12211_at
16385_s_at



12212_at
16455_at



12214_g_at
16485_s_at



12270_at
16512_s_at



12645_at
16547_s_at



12754_g_at
16548_s_at



12774_at
16629_s_at



12793_at
16673_at



12796_s_at
16899_at



12910_s_at
17010_s_at



12916_s_at
17018_s_at



12953_at
17054_s_at



13090_at
17095_s_at



13124_at
17097_s_at



13335_at
17273_at



13550_at
17394_s_at



13567_at
17420_at



13568_at
17449_s_at



13596_at
17600_s_at



13614_at
17843_s_at



13678_s_at
17913_s_at



13719_at
17966_at



14014_at
18003_at



14096_at
18081_at



14118_i_at
18560_at



14369_at
18588_at



14478_at
18626_at



14513_s_at
18644_at



14540_at
18666_s_at



14596_at
18742_f_at



14733_s_at
18977_at



14986_at
18994_at



15045_at
19227_at



15097_s_at
19373_at



15098_s_at
19834_at



15145_s_at
19867_at



15153_s_at
19998_at



15154_s_at
20062_at



15182_s_at
20199_at



15203_s_at
20256_s_at



15372_at
20284_at



15521_s_at
20437_at



15581_s_at
20442_i_at



15621_f_at
20450_at



15642_s_at
20468_at



15776_at
20547_at



15910_at
20635_s_at



16017_at



16046_s_at



16115_s_at

















TABLE 29





2X TRANSCRIPTION (COLD, SALINE


& OSMOTIC STRESSES)



















12068_at
15665_s_at
19836_at



12166_i_at
15679_s_at
19860_at



12374_i_at
15720_at
19866_at



12392_at
15871_s_at
19898_at



12431_at
16072_s_at
20262_at



12450_s_at
16073_f_at
20335_s_at



12503_at
16105_s_at
20362_at



12536_s_at
16111_f_at
20424_at



12540_s_at
16127_s_at
20437_at



12541_at
16534_s_at
20456_at



12587_at
16582_s_at
20515_s_at



12594_at
16589_s_at
20635_s_at



12595_at
16747_at



12704_f_at
17019_s_at



12705_f_at
17129_s_at



12709_f_at
17160_at



12712_f_at
17520_s_at



12719_f_at
17538_s_at



12724_f_at
17555_s_at



12725_r_at
17609_at



12726_f_at
17896_at



12734_f_at
17971_s_at



12736_f_at
17975_at



12737_f_at
17978_s_at



12812_at
18121_s_at



12949_at
18167_s_at



12951_at
18197_at



12966_s_at
18222_at



13023_at
18318_at



13034_s_at
18576_s_at



13087_at
18629_s_at



13270_at
18738_f_at



13273_s_at
18742_f_at



13432_at
18744_f_at



13555_at
18745_f_at



13688_s_at
18747_f_at



13714_at
18750_f_at



13965_s_at
18751_f_at



13987_s_at
18789_at



14003_at
18834_at



14144_at
18942_at



14178_at
19083_at



14223_at
19202_at



14235_at
19209_s_at



14303_s_at
19232_s_at



14393_at
19315_at



14553_at
19489_s_at



14781_at
19611_s_at



15046_s_at
19646_s_at



15053_s_at
19707_s_at



15214_s_at
19722_s_at



15510_r_at
19744_at



15638_s_at
19755_at

















TABLE 30





2X PHOSPHATES (COLD, SALINE & OSMOTIC STRESSES)

















12470_at



12556_at



13128_at



13135_s_at



13180_s_at



13192_s_at



13193_s_at



13587_at



13995_at



14335_at



15073_at



15171_s_at



15240_at



15586_s_at



15641_s_at



15651_f_at



15990_at



16232_s_at



16576_f_at



16753_at



17423_s_at



17525_s_at



17537_s_at



17929_s_at



17954_s_at



18012_s_at



18308_i_at



18616_at



18847_at



18936_at



18980_at



19243_at



19263_at



19638_at



19883_at



19932_at



20333_at



20393_at



20570_at

















TABLE 31





2X KINASES (COLD, SALINE & OSMOTIC STRESSES)



















12253_g_at
16059_s_at
20144_at



12270_at
16087_s_at
20219_at



12271_s_at
16088_f_at
20223_at



12276_at
16125_s_at
20232_s_at



12278_at
16137_s_at
20235_i_at



12284_at
16140_s_at
20282_s_at



12300_at
16143_s_at
20298_at



12307_at
16144_s_at
20396_at



12353_at
16160_f_at
20439_at



12357_s_at
16171_s_at
20462_at



12390_at
16357_at



12394_at
16412_s_at



12395_s_at
16568_s_at



12408_at
16570_s_at



12452_at
16571_s_at



12477_at
16584_s_at



12490_at
16651_s_at



12497_at
16652_s_at



12532_at
16672_at



12697_at
16818_s_at



12901_s_at
16840_at



12902_at
17068_s_at



12958_at
17122_s_at



12959_at
17252_at



13068_at
17323_at



13246_at
17475_at



13324_at
17752_at



13332_at
17921_s_at



13362_s_at
17933_at



13370_at
17935_at



13550_at
18013_r_at



14030_at
18046_s_at



14048_at
18122_at



14194_at
18176_at



14196_at
18316_at



14217_at
18455_at



14459_at
18459_at



14603_at
18482_s_at



14637_s_at
18543_at



14686_s_at
18706_s_at



15005_s_at
18782_at



15175_s_at
18924_at



15270_at
19117_s_at



15475_s_at
19437_s_at



15497_s_at
19442_at



15577_s_at
19458_at



15616_s_at
19464_at



15633_s_at
19469_at



15634_s_at
19562_at



15668_s_at
19655_at



15680_s_at
19749_at



15798_at
19854_at



16034_at
19904_at

## Claims

1. An isolated polynucleotide, comprising SEQ ID NO: 1034.

2. A recombinant nucleic acid molecule, comprising the isolated polynucleotide of claim 1 operatively linked to a heterologous nucleotide sequence.

3. A vector, comprising the polynucleotide of claim 1.

4. The vector of claim 3, which is an expression vector.

5. A cell containing the isolated polynucleotide of claim 1.

6. The cell of claim 5, which is a plant cell.