# Patent

## Title
Inbred sweet corn line CRAUGSH2W-89

## Abstract

Broadly this invention provides inbred corn line ZS09247. The methods for producing a corn plant by crossing the inbred line ZS09247 are encompassed by the invention. Additionally, the invention relates to the various parts of inbred ZS09247 including culturable cells. This invention relates to hybrid corn seeds and plants produced by crossing the inbred line ZS09247 with at least one other corn line.

## Background
This invention is in the field of corn breeding, specifically relating to an inbred corn line designated ZS09247.

The original maize plant was indigenous to the Western Hemisphere. The plants were weedlike and only through the efforts of early breeders was a cultivated crop species developed. The physical traits of maize are such that self pollination or cross pollination can occur. Each plant has a separate male and female flower, the tassel and ear, respectively. Natural pollination occurs when wind transfers pollen from tassel to the silks on the corn ears. This type of pollination contributed to the wide variation of maize varieties present in the Western Hemisphere.

The development of a planned breeding program for maize only occurred in the last century. Originally, maize was an open pollinated variety having heterogeneous genotypes. The maize farmer selected uniform ears from the yield of these genotypes and reserved them for planting the next season. The result was a field of maize plants that were segregating for a variety of traits. This type of maize selection lead to at most incremental increases in seed yield.

Large increases in seed yield were the result of the development of hybrid corn varieties in planned breeding programs. Hybrids were developed by selecting corn lines and selfing these lines for several generations to develop homozygous pure inbred lines and crossing selected inbred lines with unrelated inbred lines to produce hybrid progeny (F1). Inbred lines can be difficult to produce since the inbreeding process in corn decreases the vigor. However, when two inbred lines are crossed, the hybrid plant evidences greatly increased vigor compared to open pollinated segregating maize plants. An important factor of the homozygosity and the homogeneity of the inbred lines is that the hybrid from any cross will always be the same, and can be reproduced.

The ultimate objective of the commercial maize seed companies is to produce high yielding, agronomically sound plants which perform well in certain regions or areas of the Corn Belt. To produce these types of hybrids, the companies must develop inbreds which carry needed traits into the hybrid combination. Hybrids are not uniformly adapted for the Corn Belt, but are specifically adapted for regions of the Corn Belt. Northern regions of the Corn Belt require shorter season hybrids than do southern regions of the Corn Belt. Hybrids that grow well in Colorado and Nebraska soils may not flourish in rich Illinois soil. Thus, a variety of major agronomic traits are important in hybrid combination for the various Corn Belt regions, and have an impact on hybrid performance.

Inbred line development and hybrid testing have been emphasized in the past half century in commercial maize production as a means to increase hybrid performance. Inbred development is usually done by pedigree selection. Pedigree selection can be selection in an F.sub.2 population produced from a planned cross of two genotypes (often elite inbred lines), or selection of progeny of synthetic varieties, open pollinated, composite, or backcross populations. This type of selection is effective for highly inheritable traits, but other traits, for example, yield requires replicated test crosses at a variety of stages for accurate selection.

Maize breeders select for a variety of traits in inbreds that impact hybrid performance along with selecting for acceptable parental traits. Such traits include yield potential in hybrid combination; dry down; maturity; grain moisture at harvest; greensnap; resistance to root lodging; resistance to stalk lodging; grain quality; disease and insect resistance; ear and plant height; performance in different soil types such as: low level of organic matter, clay, sand, black, high pH, low pH; performance in: wet environments, drought environments, and no tillage conditions. These traits appear to be governed by a complex genetic system that makes selection and breeding of an inbred line extremely difficult. Even if an inbred, in hybrid combination, has excellent yield (a desired characteristic), it may not be useful because it fails to have acceptable parental traits such as seed yield, seed size, pollen production, good silks, plant height, etc.

To illustrate the difficulty of breeding and developing inbred lines, the following example is given. Two inbreds compared for similarity of 29 traits differed significantly for 18 traits between the two lines. If 18 simply inherited single gene traits were polymorphic with gene frequencies of 0.5 in the parental lines, and assuming independent segregation (as would essentially be the case if each trait resided on a different chromosome arm), then the specific combination of these traits as embodied in an inbred would only be expected to become fixed at a rate of one in 262,144 possible homozygous genetic combinations. Selection of the specific inbred combination is also influenced by the specific selection environment on many of these 18 traits which makes the probability of obtaining this one inbred even more remote. Thus, the general procedure of producing a non segregating F.sub.1 generation and self pollinating to produce a F.sub.2 generation that segregates for traits does not easily lead to a useful inbred. Great care and breeder expertise must be used in selection of breeding material to continue to increase yield and agronomics of inbreds and resultant commercial hybrids.

The present invention relates to an inbred corn line ZS09247. Specifically, this invention relates to plants and seeds of this line. Additionally, this relates to a method of producing hybrid seed corn from this inbred. More particularly, this invention relates to the unique combination of traits in corn line ZS09247.

Generally then, the present invention includes an inbred corn seed designated ZS09247. This seed produces a corn plant.

The invention also includes the tissue culture of regenerable cells of ZS09247 wherein the tissue regenerates plants having the genotype of ZS09247. The tissue culture is selected from the group consisting of leaves, pollen, embryos, roots, root tips, anthers, silk, flowers, kernels, ears, cobs, husks and stalks, and cells and protoplasts thereof.

The invention extends to hybrid seed produced by planting, in pollinating proximity, seeds of corn inbred lines ZS09247 and another inbred line; cultivating corn plants resulting from said planting; preventing pollen production by the plants of one of the inbred lines; allowing natural cross pollinating to occur between said inbred lines; and harvesting seeds produced on plants of the inbred. The hybrid seed produced by hybrid combination of plants of inbred corn seed designated ZS09247 and plants of another inbred line. Hybrid plants grown from this hybrid seed.

The invention further includes a method of hybrid F1 production. A first generation (F1) hybrid corn plant produced by the process of planting, in pollinating proximity, seeds of corn inbred lines ZS09247 and another inbred line; cultivating corn plants resulting from said planting; preventing pollen production by the plants of one of the inbred lines; allowing natural cross pollinating to occur between said inbred lines; harvesting seeds produced on plants of the inbred; and growing a harvested seed.

A tissue culture of the regenerable cells of hybrid plants produced with use of ZS09247 genetic material. A tissue culture of the regenerable cells of the corn plant produced by the method described above.

In the description and examples which follow, a number of terms are used. In order to provide a clear and consistent understanding of the specifications and claims, including the scope to be given such terms, the following definitions are provided.

BL MOIST

The moisture percentage of the grain at black layer, i.e., when 50% of the plants per plot have reached physiological maturity.

COLD GERM

Cold Germ is a measurement of seed germination under cold soil conditions. Data is reported as percent of seed germinating.

ECB

European corn borer a maize eating insect. ECBI is the first brood generation of European corn borers. ECBII is the second generation of European corn borers.

EMERGE

The number of emerged plants per plot (planted at the same seedling rate) collected when plants have two fully developed leaves.

GI

This is a selection index which provides a single quantitative measure of the worth of a hybrid based on four traits. Yield is the primary trait contributing to index values. The GI value is calculated by combining stalk lodging, root lodging, yield and dropped ears according to the attached mathematical formula:

GLS

Gray Leaf Spot (Cercospora Zeae) disease rating. This is rated on a 1-9 scale with a "1" being very susceptible, and a "9" being very resistant.*

GW

Goss' Wilt (Corynebacterium nebraskense). This is rated on a 1-9 scale with a "1" being very susceptible, and a "9" being very resistant.*

HEATP10

The number of Growing Degree Units (GDU's) or heat units required for an inbred line or hybrid to have approximately 10% percent of the plants shedding pollen. This trait is measured from the time of planting. Growing Degree Units are calculated by the Barger Method where the GDU's for a 24 hour period are: ##EQU1## The highest maximum temperature used is 86.degree. F. and the lowest minimum temperature used is 50.degree. F. For each inbred or hybrid it takes a certain number of GDU's to reach various stages of plant development.

HEATBL

The number of GDU's after planting when approximately 50 percent of the inbred or hybrid plants in a plot have grain which has reached physiological maturity (black layer).

HEATPEEK

The number of GDU's after planting of an inbred when approximately 50 percent of the plants show visible tassel extension.

HEATP50 or HTP50

The number of GDU's required for an inbred or hybrid to have approximately 50 percent of the plants shedding pollen. Growing Degree Units are calculated by the Barger Method as shown in the HEATP10 definition.

HEATP90

The number of GDU's accumulated from planting when the last 100 percent of plants in an inbred or hybrid are still shedding enough viable pollen for pollination to occur. Growing Degree Units are calculated by the Barger Method as shown in the HEATP10 definition.

HEATS10

The number of GDU's required for an inbred or hybrid to have approximately 10% percent of the plants with silk emergence of at least 0.5 inches. Growing Degree Units are calculated by the Barger Method as shown in the HEATP10 definition.

HEATS50 or HTS50

The number of GDU's required for an inbred or hybrid to have approximately 50 percent of the plants with silk emergence of at least 0.5 inches. Growing Degree Units are calculated by the Barger Method as shown in the HEATP10 definition.

HEATS90

The number of GDU's required for an inbred or hybrid to have approximately 90 percent of the plants with silk emergence of at least 0.5 inches. Growing Degree Units are calculated by the Barger Method as shown in the HEATP10 definition.

MDMV.sub.A

Maize Dwarf Mosaic Virus strain A. The corn is rated on a 1-9 scale with a "1" being very susceptible, and a "9" being very resistant.*

MDMV.sub.B

Maize Dwarf Mosaic Virus strain B. This is rated on a 1-9 scale with a "1" being very susceptible and a "9" being very resistant.*

MOISTURE

The average percentage grain moisture of an inbred or hybrid at harvest time.

NLB

Northern Leaf Blight (Exserohilum turcicum) disease rating. This is rated on a 1-9 scale with a "1" being very susceptible, and a "9" being very resistant.*

PCT TILLER

The total number of tillers per plot divided by the total number of plants per plot.

PLANT

This term includes plant cells, plant protoplasts, plant cell tissue cultures from which corn plants can be regenerated, plant calli, plant clumps, and plant cells that are intact in plants or parts of plants, such as embryos, pollen, flowers, kernels, ears, cobs, leaves, husks, stalks, roots, root tips, anthers, silk and the like.

PLANT HEIGHT

The distance in centimeters from ground level to the base of the tassel peduncle.

RM

Predicted relative maturity based on the moisture percentage of the grain at harvest. This rating is based on known set of checks and utilizes standard linear regression analyses and is referred to as the Minnesota Relative Maturity Rating System.

SHED

The volume of pollen shed by the male flower rated on a 1-9 scale where a "1" is a very light pollen shedder, a "4.5" is a moderate shedder, and a "9" is a very heavy shedder.

SLB

Southern Leaf Blight (Bipolaris maydis) disease rating. This is rated on a 1-9 scale with a "1" being very susceptible, and a "9" being very resistant.*

TWT

The measure of the weight of grain in pounds for a one bushel volume adjusted for percent grain moisture.

VIGOR

Visual rating of 1 to 9 made 2-3 weeks post-emergence where a "1" indicates very poor early plant development, and a "9" indicates superior plant development.

WARM GERM

A measurement of seed germination under ideal (warm, moist) conditions. Data is reported as percent of seeds germinating.

YIELD (YLD)

Actual yield of grain at harvest adjusted to 15.5% moisture. Measurements are reported in bushels per acre.

% DROPPED EARS (DE)

The number of plants per plot which dropped their primary ear divided by the total number of plants per plot.

% LRG FLAT

Percentage by weight of shelled corn that passes through a 26/64 inch round screen and a 14/64 inch slot screen, but does not pass through a screen with 20.5/64 inch round openings.

% LRG ROUND

Percentage by weight of shelled corn that passes through a 26/64 inch round screen, but does not pass through a 14/64 inch slot screen or a screen with 20.5/64 inch round openings.

% MED FLAT

Percentage by weight of shelled corn that passes through a 20.5/64 inch round screen and a 13/64 inch slotted screen, but does not pass through a screen with 17/64 inch round openings.

% MED ROUND

Percentage by weight of shelled corn that passes through a 20.5/64 inch round screen, but does not pass through a 13/64 inch slot screen or a screen with 17/64 inch round openings.

% SML FLAT

Percentage by weight of shelled corn that passes through a 17/64 inch round screen and a 12/64 inch slotted screen, but does not pass through a screen with 15/64 inch round openings.

% SML ROUND

Percentage by weight of shelled corn that passes through a 17/64 inch round screen, but does not pass through a 12/64 inch slotted screen or a screen with 15/64 inch round openings.

% ROOT LODGE (RL)

Percentage of plants per plot leaning more that degrees from vertical divided by total plants per plot.

% STALK LODGE (SL)

Percentage of plants per plot with the stalk broken below the primary ear node divided by the total plants per plot.

*Resistant--on a scale of 1-9 with 9 evidencing the trait most strongly: 1-2.9 ratings are susceptible, 3-5.9 ratings are intermediate, and 6-9 ratings are resistant.

ZS09247 can be used as a male, having acceptable pollen shed characteristics. When placed in hybrid combination with females, this inbred forms a workhorse hybrid with steady yield potential in all environments. In hybrid combinations this inbred can be used as a female but has a tendency toward silk balling. Due to this inbred's exceptional male characteristics it is infrequently used as a female. This inbred in hybrid combination, is a robust hybrid exhibiting excellent plant intergrity even in years that causes other lines intergrity problems and emergence.

The inbred has shown uniformity and stability within the limits of environmental influence for all the traits as described in the Variety Description Information (Table 1) that follows. Most of the data in the Variety Description information was collected at Slater, Iowa or other Garst research stations.

The inbred has been self-pollinated for a sufficient number of generations to give inbred uniformity. During plant selection in each generation, the uniformity of plant type was selected to ensure homozygosity and phenotypic stability. The line has been increased in isolated farmland environments with data on uniformity and agronomic traits being observed to assure uniformity and stability. No variant traits have been observed or are expected in ZS09247.

The best method of producing the invention, ZS09247 which is substantially homozygous, is by planting the seed of ZS09247 and self-pollinating or sib pollinating the resultant plant in an isolated environment, and harvesting the resultant seed or the resultant pollen. The hybrid containing ZS09247 is best produced by planting the inbred ZS09247 and an appropriate crossing line in an isolated environment, detasseling one inbred and cross-pollinating with the pollen of the other inbred and harvesting the resultant seed or the resultant pollen. Alternatively, the pollen of ZS09247 can be stored and then applied to an inbred to form hybrid seed. This seed can be planted to form the hybrid plant which then forms grain that is segegrating. __________________________________________________________________________ ZS09247 VARIETY DESCRIPTION INFORMATION Type: Dent, well adapted for Central and Eastern regions of the cornbelt GRM: ZS09247 has a GRM 105-107. Maturity: Days       Heat Limits .about.71-82 1456-1517 From planting to 50% of plants in silk .about.69-79 1415-1424 From planting to 50% of plants in pollen .about.3-4       From 10% to 90% pollen shed DISEASE RESISTANCE Northern leaf blight = 7.0 Eyespot = 1.0 BW (Bacterial Wilt) = 4.0 An Inbred comparable to ZS09247 is ZS01202. __________________________________________________________________________ 1997 PVP TRAITS: PROJECT SN INBRED ZS09247 N MEAN STD. T-STAT PROB 95% CI __________________________________________________________________________ EAR HEIGHT(CM)        15 54.33 6.75 31.19 0.0000 (50.92, 57.75) LENGTH OF PRIMARY EAR LEAF(CM) 15 92.20 3.30 108.2 0.0000 (90.53, 93.87) WIDTH OF PRIMARY EAR LEAF(CM) 15 9.43 0.43 88.37 0.0000 (9.22, 9.63) TOP EAR INTERNODE(CM) 15 14.05 2.20 24.73 0.0000 (12.93, 15.16) DEGREE OF LEAF ANGLE  15 27.47 5.60 18.98 0.0000 (24.63, 30.30) # OF EARS PER PLANT   16 1.87 0.35 20.55 0.0000 (1.69, 2.04) # OF LEAVES ABOVE TOP EAR 15 6.07 0.26 91.00 0.0000 (5.94, 6.20) # OF PRIMARY LATERAL TASSEL BRANCHES 15 4.47 1.55 11.14 0.0000 (3.68, 5.25) TASSEL LENGTH(CM)     15 40.27 5.54 28.17 0.0000 (37.47, 43.07) TASSEL BRANCH ANGLE   15 47.87 6.24 29.69 0.0000 (44.71, 51.03) # OF TILLER PER PLANTS 15 0.00 0.00        (0.00, 0.00) __________________________________________________________________________ INBRED SN: ZS09247 __________________________________________________________________________ #3  MATURITY DAYS HEATUNITS 82  1517   FROM PLANTING TO 50% OF PLANTS IN SILK 79  1424   FROM PLANTING TO 50% OF PLANTS IN POLLEN 4          FROM 10% TO 90% POLLEN SHED #4  PLANT DATA 4          ANTHOCYANIN OF BRACE ROOTS: 1 = ABSENT 2 = FAINT 3 = MODERATE 4 = DARK #5  LEAF COLOR/DATA 3/DARK GREEN LEAF COLOR **MUNSELL CODE-5GY 4/4 6          LEAF SHEATH PUBESCENCE (1 = NONE TO 9 = PEACH FUZZ) 4          MARGINAL WAVES (1 = NONE TO 9 = MANY) 4          LONGITUDINAL CREASES (1 = NONE TO 9 = MANY) #6  TASSEL COLOR/DATA 5          POLLEN SHED (0 = STERILE TO 9 = HEAVY SHEDDER) 7/YELLOW   ANTHER COLOR **MUNSELL CODE-5Y 8/8 2&17/MGRN/PUR GLUME COLOR **MUNSELL CODE-5GY 6/6 W/S 5R 4/8 1          BAR GLUME: 1 = ABSENT 2 = PRESENT __________________________________________________________________________

The purity and homozygosity of inbred ZS09247 is constantly being tracked using isozyme genotypes as shown in Table 2.

Isozyme Genotypes for ZS09247

Isozyme data were generated for inbred corn line ZS09247 according to procedures known and published in the art. The data in Table 2 gives the electrophoresis data on ZS09247. __________________________________________________________________________ ELECTROPHORESIS RESULTS FOR ZS09247 INBRED ACP1 ACP4 ADH MDH1 MDH2 PGD1 PGD2 PH1 PGM IDH __________________________________________________________________________ ZS09247 11  00  22 22  22  11  11  11 22 11 __________________________________________________________________________

Inbred and Hybrid Performance of ZS09247

The traits and characteristics of inbred corn line ZS09247 are listed to compare with other inbreds and/or in hybrid combination ZS09247 data shows the characteristics and traits of importance, giving a snapshot of ZS09247.

Table 3A compares inbred ZS09247 with inbred ZS01202. ZS09247 has significantly lower ear height placement than ZS01202. ZS09247 evidences significantly better emergence than ZS01202. Inbred ZS09247 yields significantly less than ZS01202. __________________________________________________________________________ PAIRED INBRED COMPARISON DATA __________________________________________________________________________ PCT    PLANT EAR          EAR   PCT YEAR  INBRED VIGOR EMERGE TILLER HEIGHT HEIGHT SHED   QUALITY BARREN __________________________________________________________________________ OVERALL ZS09247 7.1   90.8         160.0 63.9  4.7 ZS1202 5.7   80.2         172.5 75.3  5.3 # EXPTS 7     7            7     7     3 DIFF   1.5   10.6         12.5  11.4  0.7 PROB   0.054* 0.003***     0.106 0.019** 0.184 __________________________________________________________________________ YEAR  INBRED HEATP10 HEATP50 HEATP90 HEATS10 HEATS50 HEATS90 __________________________________________________________________________ OVERALL ZS09247 1406  1448  1530   1467  1508  1551 ZS1202 1414  1461  1566   1459  1496  1543 # EXPTS 3     3     3      3     3     3 DIFF   8     13    36     8     12    8 PROB   0.631 0.493 0.300  0.668 0.529 0.808 __________________________________________________________________________ BL     % ROOT % STALK % DROPPED YEAR  INBRED HEATPEEK HEATBL MOIST  LODGE LODGE EARS   MOISTURE YIELD __________________________________________________________________________ OVERALL ZS09247 1352                                  11.3  60.1 ZS1202 1309                                  11.8  72.8 # EXPTS 3                                     7     7 DIFF   43                                    0.5   12.7 PROB   0.193                                 0.118 0.065* __________________________________________________________________________ WARM  COLD  % LRG  % LRG % MED % MED  % SML % SML YEAR  INBRED GERM  GERM  ROUND  FLAT  ROUND FLAT   ROUND FLAT __________________________________________________________________________ OVERALL ZS09247 91.9  74.5               56.2  12.1 ZS1202 93.1  76.2               35.5  22.4 # EXPTS 7     7                  7     7 DIFF   1.2   1.7                20.7  10.3 PROB   0.245 0.755              0.000*** 0.000*** __________________________________________________________________________ *.05 < PROB <= .10 **.01 < PROB <= .05 ***.00 < PROB <= .01

Table 4 shows the GCA (general combining ability) estimates of ZS09247 compared with the GCA estimates of other inbreds. The estimates show the general combining ability is weighted by the number of experiment/location combinations in which the specific hybrid combination occurs. The interpretation of the data for all traits is that a positive comparison is a practical advantage. A negative comparison is a practical disadvantage. The general combining ability of an inbred is clearly evidenced by the results of the general combining ability estimates. This data compares the inbred parent in a number of hybrid combinations to a group of "checks". The check data is from other companies' hybrids, particularly the leader in the industry and Garst's commercial products and pre-commercial hybrids which were grown in the same sets and locations.

Table 4 shows ZS09247 crossed in hybrid combinations. ZS09247 shows standability, root strength, test weight beyond the checks. ______________________________________ N      FI     YM   GI   I     YLD  MST ______________________________________ ZS09247 XR = 86     1.2    -0.2 1.2  -0.5  -2.0 0.0 ______________________________________ % SL    % RL   % DE  TWT   POP   RM ______________________________________ ZS09247 XR = 0.7     1.3    0.1   0.2   10    107 ______________________________________

Table 5 shows ZS09247, in a hybrid combination, in comparison with the plants in the environment around it at the same location. ZS09247 in hybrid combination yields well in low to medium yielding environments. In these yielding environments ZS09247, in hybrid combination, out yields ZS01202 as a hybrid. In high yielding environments, ZS09247 hybrid tends to show less aggressive yields compared to the environment and similar yields in hybrid combination to ZS01202/X. ______________________________________ YIELD RESPONSE ______________________________________ 1.  HYBRID     YIELD ______________________________________ Environment 75     100     125   150  175   200 ZS09247/X  95     116     137   158  179   201 ______________________________________ 2.  HYBRID     YIELD ______________________________________ Environment 75     100     125   150  175   200 ZS01202/X  77     102     127   152  176   201 ______________________________________

Table 6A shows the advantage ZS09247 hybrid has compared to one commercially available Garst hybrid. The ZS09247 hybrid is significantly better in yield, is better in yield/moisture advantage compared to hybrid 8692IT. These two hybrids carry a common inbred. PAIRED HYBRID COMPARISON DATA __________________________________________________________________________ % ROOT % STALK % DROPPED TEST YEAR  HYBRID LODGE LODGE EARS   WEIGHT MOISTURE YIELD GI __________________________________________________________________________ OVERALL ZS09247/X 0.1   3.0  0.0    52.9 21.9  168.2 181.2 8692IT 0.1   4.9  0.0    53.7 20.4  153.0 172.0 # EXPTS 28    28   28     26   28    28   28 DIFF  0.1   1.8  0.0    0.8  1.5   15.2 9.1 PROB  0.637 0.026** 0.292  0.006*** 0.000*** 0.000*** 0.000*** __________________________________________________________________________ YEAR  INBRED MATURITY Y M  FI __________________________________________________________________________ OVERALL ZS09247/X   7.8  131 8692IT      7.6  125 # EXPTS     28   28 DIFF        0.2  5.7 PROB        0.198 0.001*** __________________________________________________________________________ *.05 < PROB <= .10 **.01 < PROB <= .05 ***.00 < PROB <= .01

Table 6B shows the advantages and disadvantages generated by comparison of the agronomic data of the two hybrids. ZS09247/X brings vigor, plant integrity, and staygreen into the hybrid package. ______________________________________ ZS09247/x vs. 8692IT AGRONOMIC DATA Advantage of ZS09247/x over 86921T PLANT     STAY- HYBRID N      ESTAND   EMG  VIGOR INTEGRITY GREEN ______________________________________ 54     -2.5     0.0  1.0   1.0     1.7 ______________________________________

The inbred ZS09247 can be employed as a male in a hybrid production field. This inbred is a vigorous line. ZS09247, in hybrid combination, produces hybrids that have excellent yield potential across low environments. ZS09247 inbred has good moisture. This inbred, when in hybrid combination, shows yield even in stress conditions, and late season plant integrity.

The present invention can by mutagenisis be transformed into an isogenic line with the brown mid rib mutation, low phytic acid mutation, the imalzethaphryl (IT) mutation, and the various starch mutations such as ae, waxy, dull, and the like. The use of EMS as a mutating agent chemical permits single point mutations in corn has been known and used since at least the early seventies. Other mutating chemicals are also capable of causing these type of mutations and generating a line that is isogenic or nearly isogenic to the present invention.

The present invention can also be transformed to carry well known transgenic genes (as well as less well known genes) such as the bacillus thuringiensis genes that encode for an insect resistant crystalline protein for example Cry genes like Cry V, Cry lab, Cry lac, Cry 9c. The present invention can also be transformed to carry herbicide genes such as ESPS, bar, pat, and the like. The regeneration of transformed cells is now a precise science which generates plants that are characterized by the original plant characteristics with the added benefit of the transgenes trait. The use of pollen transformation as a transformation method eliminates the need for regeneration of the cells and appears to produce lines which are nearly isogenic to the present invention while additionally carrying the characteristic expressed by the added transgene.

The foregoing is set forth by way of example and is not intended to limit the scope of the invention.

This invention also is directed to methods for producing a corn plant by crossing a first parent corn plant with a second parent corn plant wherein the first or second parent corn plant is an inbred corn plant from the line ZS09247. Further, both first and second parent corn plants can come from the inbred corn line ZS09247. A variety of breeding methods can be selected depending on the mode of reproduction, the trait, the condition of the germplasm. Thus, any such methods using the inbred corn line ZS09247 are part of this invention: selfing, backcrosses, hybrid production, crosses to populations, haploid and anther culturing and the like.

Various culturing techniques known to those skilled in the art, such as haploid, transformation, and a host of other conventional and unconventional methods are within the scope of the invention. All plants and plant cells produced using inbred corn line ZS09247 are within the scope of this invention. The invention encompasses the inbred corn line used in crosses with other, different, corn inbreds to produce (F1) corn hybrid seeds and plants. This invention includes cells which upon growth and differentiation produce corn plants having the physiological and morphological characteristics of the inbred line ZS09247.

Duncan, from at least 1985-1988 produced literature on plant regeneration from callus. Both inbred and hybrid callus have resulted in regenerated plants at excellent efficiency rates. Somatic embryogenesis has been performed on various maize tissue such as glume which before the 1980's was considered unusable for this purpose. The prior art clearly teaches the regeneration of plants from various maize tissues.

European Patent Application, publication 160,390, describes tissue culture of corn which can be used by those skilled in the art. Corn tissue culture procedures are also described in the literature as early as 1982. Various culturing techniques known to those skilled in the art, such as haploid, (stock six is a method that has been in use for twenty years and is well known to those with skill in the art), transformation, and a host of other conventional and unconventional methods are within the scope of the invention. All plants and plant cells produced using the inbred corn line are within the scope of this invention. The term transgenic plant refers to plants having exogenous genetic sequences which are introduced into the genome of a plant by a transformation method and the progeny thereof.

Transformation Methods--are means for integrating new genetic coding sequences into the plant's genome by the incorporation of these sequences into a plant through man's assistance.

Though there are a large number of known methods to transform plants, certain types of plants are more amenable to transformation than are others. Tobacco is a readily transformable plant. The basic steps of transforming plants including monocots are known in the art. These steps are concisely outlined in U.S. Pat. No. 5,484,956 "Fertile Transgenic Zea mays Plants Comprising Heterologous DNA Encoding Bacillus Thuringiensis Endotoxin" issued Jan. 16, 1996 and U.S. Pat. No. 5,489,520 "Process of Producing Fertile Zea mays Plants and Progeny Comprising a Gene Encoding Phosphinothricin Acetyl Transferase" issued Feb. 6, 1996.

Plant cells such as maize can be transformed by a number of different techniques. Some of these techniques which have been reported on and are known in the art include maize pollen transformation (See University of Toledo 1993 U.S. Pat. No. 5,177,010); Biolistic gun technology (See U.S. Pat. No. 5,484,956); Whiskers technology (See U.S. Pat. Nos. 5,464,765 and 5,302,523); Electroporation; PEG on Maize; Agrobacterium (See 1996 article on transformation of maize cells in Nature Biotechnology, Volume 14, June 1996) along with numerous other methods which may have slightly lower efficiency rates then those listed. Some of these methods require specific types of cells and other methods can be practiced on any number of cell types.

The use of pollen, cotyledons, meristems and ovum as the target issue can eliminate the need for extensive tissue culture work. However, the present state of the technology does not provide very efficient use of this material.

Generally, cells derived from meristematic tissue are useful. Zygotic embryos can also be used. Additionally, the method of transformation of meristematic cells of cereal is also taught in the PCT application WO96/04392. Any of the various cell lines, tissues, plants and plant parts can and have been transformed by those having knowledge in the art. Methods of preparing callus from various plants are well known in the art and specific methods are detailed in patents and references used by those skilled in the art. Cultures can be initiated from most of the above identified tissue. The only true requirement of the transforming material is that it can form a transformed plant. The transgenic gene can come from various non-plant genes (such as; bacteria, yeast, animals, viruses) along with being from animal or plants.

The DNA used for transformation of these plants clearly may be circular, linear, double or single stranded. Usually, the DNA is in the form of a plasmid. The plasmid usually contains regulatory and/or targeting sequences which assists the expression of the gene in the plant. The methods of forming plasmids for transformation are known in the art. Plasmid components can include such items as: leader sequences, transit polypeptides, promoters, terminators, genes, introns, marker genes, etc. The structures of the gene orientations can be sense, antisense, partial antisense, or partial sense: multiple gene copies can be used.

The regulatory promoters employed can be constitutive such as CaMv35S (usually for dicots) and polyubiquitin for monocots or tissue specific promoters such as CAB promoters, etc. The prior art includes but is not limited to octopine synthase, nopaline synthase, CaMv19S, mannopine synthase promoters. These regulatory sequences can be combined with introns, terminators, enhancers, leader sequences and the like in the material used for transformation.

The isolated DNA is then transformed into the plant. Many dicots can easily be transformed with Agrobacterium. Some monocots are more difficult to transform. As previously noted, there are a number of useful transformation processes. The improvements in transformation technology are beginning to eliminate the need to regenerate plants from cells. Since 1986, the transformation of pollen has been published and recently the transformation of plant meristems have been published. The transformation of ovum, pollen, and seedlings meristem greatly reduce the difficulties associated with cell regeneration of different plants or genotypes within a plant can present. Duncan, from at least 1985-1988 produced literature on plant regeneration from callus. Both inbred and hybrid callus have resulted in regenerated plants. Somatic embryogenesis has been performed on various maize tissue which was considered unusable for this purpose. The prior art clearly teaches the regeneration of plants from various maize tissues.

The most common method of transformation is referred to as gunning or microprojectile bombardment. This biolistic process has small gold coated particles coated with DNA shot into the transformable material. Techniques for gunning DNA into cells, tissue, callus, embryos, and the like are well known in the prior art.

After the transformation of the plant material is complete, the next step is identifying the cells or material which has been transformed. In some cases, a screenable marker is employed such as the beta-glucuronidase gene of the uidA locus of E. coli. Then, the transformed cells expressing the colored protein are selected for either regeneration or further use. In many cases, the transformed material is identified by a selectable marker. The putatively transformed material is exposed to a toxic agent at varying concentrations. The cells which are not transformed with the selectable marker that provides resistance to this toxic agent die. Cells or tissues containing the resistant selectable marker generally proliferate. It has been noted that although selectable markers protect the cells from some of the toxic affects of the herbicide or antibiotic, the cells may still be slightly effected by the toxic agent by having slower growth rates. If the transformed material was cell lines then these lines are regenerated into plants. The cell's lines are treated to induce tissue differentiation. Methods of regeneration of cellular maize material are well known in the art since early 1982. European Patent Application, publication 160,390, describes tissue culture of corn which can be used by those skilled in the art. The plants from either the transformation process or the regeneration process or crossed to either such plants or a progeny of such plants are transgenic plants.

Various techniques known to those skilled in the art, such as haploid, transformation, and a host of other conventional and unconventional methods are within the scope of the invention. All plants and plant cells produced using inbred corn line ZS09247 are within the scope of this invention. The invention encompasses the inbred corn line used in crosses with other, different, corn inbreds to produce (F1) corn hybrid seeds and plants with the characteristics that make good hybrids. This invention includes cells which upon growth and differentiation produce corn plants having the physiological and morphological characteristics of the inbred line ZS09247.

A deposit of at least 2500 seeds of the inbred seed of this invention is maintained by Garst, 2369 330th Street, Slater, Iowa 50244. Access to this deposit will be available during the pendency of this application to the Commissioner of Patents and Trademarks and persons determined by the Commissioner to be entitled thereto upon request. All restrictions on availability to the public of such material will be removed upon issuance of any claims in this application by depositing at least 2500 seeds of this invention at the American Type Culture Collection, Rockville, Md. The deposit of at least 2500 seeds will be from inbred seed taken from the deposit maintained by Garst. The ATCC deposit will be maintained in that depository, which is a public depository, for a period of 30 years, or 5 years after the last request, or for the enforceable life of the patent, whichever is longer, and will be replaced if it becomes nonviable during that period.

Information on some ZS designations may be available from the PVP office.

Accordingly, the present invention has been described with some degree of particularity directed to the preferred embodiment of the present invention. It should be appreciated, though, that the present invention is defined by the following claims construed in light of the prior art so that modifications or changes may be made to the preferred embodiment of the present invention without departing from the inventive concepts contained herein.

## Claims

1. Inbred corn seed designated ZS09247, some of said seed deposited in the ATCC and designated accession number X.

2. A corn plant produced by the seed of claim 1.

3. A tissue culture of regenerable cells of ZS09247 of claim 2 wherein the tissue regenerates plants having the genotype of ZS09247.

4. A tissue culture according to claim 3, the tissue culture selected from the group consisting of leaves, pollen, embryos, roots, root tips, anthers, silk, flowers, kernels, ears, cobs, husks and stalks, and cells and protoplasts thereof.

5. A corn plant having all of the physiological and morphological characteristics of ZS09247 regenerated from the tissue culture of claim 3.

6. Hybrid seed produced by: corn inbred lines ZS09247 and another inbred line, one of said inbred lines not releasing pollen; and

7. Hybrid seed produced by hybrid combination of plants of inbred corn seed designated ZS09247 in claim 1 and plants of another inbred line.

8. Hybrid plants grown from seed of claim 7.

9. A first generation (F1) hybrid corn plant produced by using ZS09247 according to claim 1 the process of:

10. A tissue culture of the regenerable cells of the corn plant of claim 8.

11. A tissue culture of the regenerable cells of the corn plant of claim 9.

12. An inbred corn plant with all of the phenotypic, physiological and morphological characteristics of inbred corn line of claim 2.

13. A plant according to claim 2, including in the plant at least one transgenic gene, wherein said transgenic gene is selected from the group consisting of: Cry V, Cry lab, Cry lac, Cry 9c, ESPS, bar and pat.

14. A seed according to claim 1, including at least one transgenic gene, wherein said transgenic gene is selected from the group consisting of: Cry V, Cry lab, Cry lac, Cry 9c, ESPS, bar and pat.

15. Hybrid seed containing at least one transgenic gene said seed produced by hybrid combination of plants of inbred corn seed designated ZS09247 in claim 14 and plants of another inbred line.

16. A plant according to claim 2, including in the plant at least one mutant gene.

17. A seed according to claim 1, including at least one mutant gene.

18. Hybrid seed containing at least one mutant gene said seed produced by hybrid combination of plants of inbred corn seed designated ZS09247 in claim 17 and plants of another inbred line.

19. A corn plant according to claim 2 having all of the morphological and physiological traits of ZS09247.