bluuebunny/crossref_metadata_2025_split huggingface dataset:
Developmental and Thermal Regulation of the Maize Heat Shock Protein, HSP101
Authors: Young Todd E. , Ling Jun , Geisler-Lee C. Jane , Tanguay Robert L. , Caldwell Christian , Gallie Daniel R. 
Published: November 2008

Abstract
The plant heat stress protein, Hsp101, and the yeast ortholog, Hsp104, are required to confer thermotolerance in plants and yeast (Saccharomyces cerevisiae), respectively. In addition to its function during stress, Hsp101 is developmentally regulated in plants although its function during development is not known. To determine how the expression of Hsp101 is regulated in cereals, we investigated the Hsp101 expression profile in developing maize (Zea mays). Hsp101 protein was most abundant in the developing tassel, ear, silks, endosperm, and embryo. It was less abundant in the vegetative and floral meristematic regions and was present at only a low level in the anthers and tassel at anthesis, mature pollen, roots, and leaves. As expected, heat treatment resulted in an increase in the level of Hsp101 protein in several organs. In expanding foliar leaves, husk leaves, the tassel at the premeiosis stage of development, or pre-anthesis anthers, however, the heat-mediated increase in protein was not accompanied by an equivalent increase in mRNA. In contrast, the level of Hsp101 transcript increased in the tassel at anthesis following a heat stress without an increase in Hsp101 protein. In other organs such as the vegetative and floral meristematic regions, fully expanded foliar leaves, the young ear, and roots, the heat-induced increase in Hsp101 protein was accompanied by a corresponding increase in Hsp101 transcript level. However, anthers at anthesis, mature pollen, developing endosperm, and embryos largely failed to mount a heat stress response at the level of Hsp101 protein or mRNA, indicating that Hsp101 expression is not heat inducible in these organs. In situ RNA localization analysis revealed that Hsp101 mRNA accumulated in the subaleurone and aleurone of developing kernels and was highest in the root cap meristem and quiescent center of heat-stressed roots. These data suggest an organ-specific control of Hsp101 expression during development and following a heat stress through mechanisms that may include posttranscriptional regulation.
Xylem Water Content and Wood Density in Spruce and Oak Trees Detected by High-Resolution Computed Tomography
Authors: Fromm Jörg H. , Sautter Irina , Matthies Dietmar , Kremer Johannes , Schumacher Peter , Ganter Carl 
Published: November 2008

Abstract
Elucidation of the mechanisms involved in long-distance water transport in trees requires knowledge of the water distribution within the sapwood and heartwood of the stem as well as of the earlywood and latewood of an annual ring. X-ray computed tomography is a powerful tool for measuring density distributions and water contents in the xylem with high spatial resolution. Ten- to 20-year-old spruce (Picea abies L. KARST.) and oak (Quercus robur) trees grown in the field were used throughout the experiments. Stem and branch discs were collected from different tree heights, immediately deep frozen, and used for the tomographic determinations of spatial water distributions. Results are presented for single-tree individuals, demonstrating heartwood and sapwood distribution throughout their entire length as well as the water relations in single annual rings of both types of wood. Tree rings of the sapwood show steep water gradients from latewood to earlywood, whereas those of the heartwood reflect water deficiency in both species. Although only the latest two annual rings of the ringporous species are generally assumed to transport water, we found similar amounts of water and no tyloses in all rings of the oak sapwood, which indicates that at least water storage is important in the whole sapwood.
Conditional Circadian Regulation of PHYTOCHROME AGene Expression
Authors: Hall Anthony , Kozma-Bognár László , Tóth Réka , Nagy Ferenc , Millar Andrew J. 
Published: November 2008

Abstract
The phytochrome photoreceptors and the circadian clock control many of the same developmental processes, in all organs and throughout the growth of Arabidopsis plants. Phytochrome A (phyA) provides light input signals to entrain the circadian clock. The clock is known to rhythmically regulate its light input pathway, so we tested rhythmic regulation of phyA, using transgenic plants carrying aPHYA promoter fusion to the luciferase reporter (PHYA:LUC). We provide the first images ofLUC activity with subcellular resolution in intact tissue. PHYA transcription and the accumulation of all three PHYA mRNAs were indeed clock controlled.PHYA is expressed throughout the seedling, so we tested whether circadian rhythms were observed in allPHYA-expressing organs and whether the rhythms were autonomously controlled by each organ. In contrast to our previous results using other clock controlled genes, the rhythmic pattern ofPHYA expression varied markedly among isolated organs and between isolated organs and intact plants. High-amplitude rhythms were maintained for many days in isolated leaves in darkness, whereas the leaves of intact plants rapidly lost rhythmicity. Wounding the leaves of intact plants had no effect. The rhythmic pattern ofPHYA expression is not organ autonomous but depends upon the physical continuity or isolation of the rhythmic tissues, consistent with the presence of a transmitted signal that controls the overt expression of circadian rhythms without necessarily affecting the underlying clock. A circadian system might be present in most, if not all, plant cells, but its effect on intracellular rhythms can be controlled by supracellular signaling.
The Maize CRINKLY4 Receptor Kinase Controls a Cell-Autonomous Differentiation Response
Authors: Becraft Philip W. , Kang Suk-Hoon , Suh Sang-Gon 
Published: November 2008

Abstract
The maize (Zea mays)CRINKLY4 (cr4) gene encodes a receptor-like kinase that controls a variety of cell differentiation responses, particularly in the leaf epidermis and in the aleurone of the endosperm. In situ hybridization indicated that the cr4 transcript is present throughout the shoot apical meristem and young leaf primordia. A genetic mosaic analysis was conducted to test whether CR4 signal transduction directly regulated the cellular processes associated with differentiation or whether differentiation was controlled through the production of a secondary signal. Genetic mosaics were created using γ-rays to induce chromosome breakage in a cr4/Cr4+ heterozygote. The mutant cr4 allele was marked with the albino mutation,Oy-700. Breakage and loss of the chromosome arm carrying the wild-type alleles created a sector of albino, cr4mutant tissue in an otherwise normal leaf. Analysis of such sectors indicated that cr4 functions cell autonomously to regulate cell morphogenesis, implying that CR4 signal transduction regulates cell differentiation through strictly intracellular functions and not the production of secondary intercellular signals. However, several sectors altered cell patterning in wild-type tissue adjacent to the sectors, suggesting that cr4 mutant cells are defective in the production of other lateral signals.
The ArabidopsisSomatic Embryogenesis Receptor Kinase 1Gene Is Expressed in Developing Ovules and Embryos and Enhances Embryogenic Competence in Culture
Authors: Hecht Valérie , Vielle-Calzada Jean-Philippe , Hartog Marijke V. , Schmidt Ed D.L. , Boutilier Kim , Grossniklaus Ueli , de Vries Sacco C. 
Published: November 2008

AbstractWe report here the isolation of the Arabidopsis SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE 1 (AtSERK1) gene and we demonstrate its role during establishment of somatic embryogenesis in culture. The AtSERK1 gene is highly expressed during embryogenic cell formation in culture and during early embryogenesis. The AtSERK1 gene is first expressed in planta during megasporogenesis in the nucleus of developing ovules, in the functional megaspore, and in all cells of the embryo sac up to fertilization. After fertilization, AtSERK1 expression is seen in all cells of the developing embryo until the heart stage. After this stage, AtSERK1 expression is no longer detectable in the embryo or in any part of the developing seed. Low expression is detected in adult vascular tissue. Ectopic expression of the full-length AtSERK1 cDNA under the control of the cauliflower mosaic virus 35S promoter did not result in any altered plant phenotype. However, seedlings that overexpressed theAtSERK1 mRNA exhibited a 3- to 4-fold increase in efficiency for initiation of somatic embryogenesis. Thus, an increased AtSERK1 level is sufficient to confer embryogenic competence in culture.
Biochemical Characterization of the Arabidopsis Biotin Synthase Reaction. The Importance of Mitochondria in Biotin Synthesis
Authors: Picciocchi Antoine , Douce Roland , Alban Claude 
Published: November 2008

Abstract
Biotin synthase, encoded by the bio2 gene in Arabidopsis, catalyzes the final step in the biotin biosynthetic pathway. The development of radiochemical and biological detection methods allowed the first detection and accurate quantification of a plant biotin synthase activity, using protein extracts from bacteria overexpressing the Arabidopsis Bio2 protein. Under optimized conditions, the turnover number of the reaction was &gt;2 h−1 with this in vitro system. Purified Bio2 protein was not efficient by itself in supporting biotin synthesis. However, heterologous interactions between the plant Bio2 protein and bacterial accessory proteins yielded a functional biotin synthase complex. Biotin synthase in this heterologous system obeyed Michaelis-Menten kinetics with respect to dethiobiotin (K m = 30 μm) and exhibited a kinetic cooperativity with respect toS-adenosyl-methionine (Hill coefficient = 1.9;K 0.5 = 39 μm), an obligatory cofactor of the reaction. In vitro inhibition of biotin synthase activity by acidomycin, a structural analog of biotin, showed that biotin synthase reaction was the specific target of this inhibitor of biotin synthesis. It is important that combination experiments using purified Bio2 protein and extracts from pea (Pisum sativum) leaf or potato (Solanum tuberosum) organelles showed that only mitochondrial fractions could elicit biotin formation in the plant-reconstituted system. Our data demonstrated that one or more unidentified factors from mitochondrial matrix (pea and potato) and from mitochondrial membranes (pea), in addition to the Bio2 protein, are obligatory for the conversion of dethiobiotin to biotin, highlighting the importance of mitochondria in plant biotin synthesis.
Effects of Natural Intensities of Visible and Ultraviolet Radiation on Epidermal Ultraviolet Screening and Photosynthesis in Grape Leaves
Authors: Kolb Christiane A. , Käser Martin A. , Kopecký Jiri , Zotz Gerhard , Riederer Markus , Pfündel Erhard E. 
Published: November 2008

AbstractGrape (Vitis vinifera cv Silvaner) vine plants were cultivated under shaded conditions in the absence of ultraviolet (UV) radiation in a greenhouse, and subsequently placed outdoors under three different light regimes for 7 d. Different light regimes were produced by filters transmitting natural radiation, or screening out the UV-B (280–315 nm), or screening out the UV-A (315–400 nm) and the UV-B spectral range. During exposure, synthesis of UV-screening phenolics in leaves was quantified using HPLC: All treatments increased concentrations of hydroxycinnamic acids but the rise was highest, reaching 230% of the initial value, when UV radiation was absent. In contrast, UV-B radiation specifically increased flavonoid concentrations resulting in more than a 10-fold increase. Transmittance in the UV of all extracted phenolics was lower than epidermal UV transmittance determined fluorimetrically, and the two parameters were curvilinearly related. It is suggested that curvilinearity results from different absorption properties of the homogeneously dissolved phenolics in extracts and of the non-homogeneous distribution of phenolics in the epidermis. UV-B-dependent inhibition of maximum photochemical yield of photosystem II (PSII), measured as variable fluorescence of dark-adapted leaves, recovered in parallel to the buildup of epidermal screening for UV-B radiation, suggesting that PSII is protected against UV-B damage by epidermal screening. However, UV-B inhibition of CO2assimilation rates was not diminished by efficient UV-B screening. We propose that protection of UV-B inactivation of PSII is observed because preceding damage is efficiently repaired while those factors determining UV-B inhibition of CO2 assimilation recover more slowly.
Isolation and Characterization of Mutants Defective in Seed Coat Mucilage Secretory Cell Development in Arabidopsis
Authors: Western Tamara L. , Burn Joanne , Tan Wei Ling , Skinner Debra J. , Martin-McCaffrey Luke , Moffatt Barbara A. , Haughn George W. 
Published: November 2008

Abstract
In Arabidopsis, fertilization induces the epidermal cells of the outer ovule integument to differentiate into a specialized seed coat cell type producing extracellular pectinaceous mucilage and a volcano-shaped secondary cell wall. Differentiation involves a regulated series of cytological events including growth, cytoplasmic rearrangement, mucilage synthesis, and secondary cell wall production. We have tested the potential of Arabidopsis seed coat epidermal cells as a model system for the genetic analysis of these processes. A screen for mutants defective in seed mucilage identified five novel genes (MUCILAGE-MODIFIED [MUM]1–5). The seed coat development of these mutants, and that of three previously identified ones (TRANSPARENT TESTA GLABRA1, GLABRA2, and APETALA2) were characterized. Our results show that the genes identified define several events in seed coat differentiation. Although APETALA2is needed for differentiation of both outer layers of the seed coat,TRANSPARENT TESTA GLABRA1, GLABRA2, andMUM4 are required for complete mucilage synthesis and cytoplasmic rearrangement. MUM3 and MUM5may be involved in the regulation of mucilage composition, whereasMUM1 and MUM2 appear to play novel roles in post-synthesis cell wall modifications necessary for mucilage extrusion.
Rapid Isolation of Monoclonal Antibodies. Monitoring Enzymes in the Phytochelatin Synthesis Pathway
Authors: Li Yujing , Kandasamy Muthugapatti K. , Meagher Richard B. 
Published: November 2008

Abstract
Genomics projects have identified thousands of interesting new genes whose protein products need to be examined at the tissue, subcellular, and molecular levels. Furthermore, modern metabolic engineering requires accurate control of expression levels of multiple enzymes in complex pathways. The lack of specific immune reagents for characterization and monitoring of these numerous proteins limits all proteomic and metabolic engineering projects. We describe a rapid method of isolating monoclonal antibodies that required only sequence information from GenBank. We show that large synthetic peptides were highly immunogenic in mice and crude protein extracts were effective sources of antigen, thus eliminating the time-consuming step of purifying the target proteins for antibody production. A case study was made of the three-enzyme pathway for the synthesis of phytochelatins. Enzyme-linked immunosorbent assays and western blots with the recombinant proteins in crude extracts demonstrated that the monoclonal antibodies produced to synthetic peptides were highly specific for the different target proteins, gamma-glutamyl cysteine synthetase, glutathione synthetase, and phytochelatin synthase. Moreover, immunofluorescence localization studies with antibacterial γ-glutamyl cysteine synthetase and antiglutathione synthetase antibodies demonstrated that these immune reagents reacted strongly with their respective target proteins in chemically fixed cells from transgenic plants. This approach enables research to progress rapidly from the genomic sequence of poorly characterized target genes, to protein-specific antibodies, to functional studies.
Effects of Phosphorylation on Phosphoenolpyruvate Carboxykinase from the C4 Plant Guinea Grass
Authors: Walker Robert P. , Chen Zhi-Hui , Acheson Richard M. , Leegood Richard C. 
Published: November 2008

Abstract
In the C4 plant Guinea grass (Panicum maximum), phosphoenolpyruvate carboxykinase (PEPCK) is phosphorylated in darkened leaves and dephosphorylated in illuminated leaves. To determine whether the properties of phosphorylated and non-phosphorylated PEPCK were different, PEPCK was purified to homogeneity from both illuminated and darkened leaves. The final step of the purification procedure, gel filtration chromatography, further separated phosphorylated and non-phosphorylated forms. In the presence of a high ratio of ATP to ADP, the non-phosphorylated enzyme had a higher affinity for its substrates, oxaloacetate and phosphoenolpyruvate. The activity of the non-phosphorylated form was up to 6-fold higher when measured at low substrate concentrations. Comparison of proteoloytically cleaved PEPCK from Guinea grass, which lacked its N-terminal extension, from yeast (Saccharomyces cerevisiae), which does not possess an N-terminal extension, and from the C4 plantUrochloa panicoides, which possesses an N-terminal extension but is not subject to phosphorylation, revealed similar properties to the non-phosphorylated full-length form from Guinea grass. Assay of PEPCK activity in crude extracts of Guinea grass leaves, showed a large difference between illuminated and darkened leaves when measured in a selective assay (a low concentration of phosphoenolpyruvate and a high ratio of ATP to ADP), but there was no difference under assay conditions used to estimate maximum activity. Immunoblots of sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels showed no difference in the abundance of PEPCK protein in illuminated and darkened leaves. There were no light/dark differences in activity detected in maize (Zea mays) leaves, in which PEPCK is not subject to phosphorylation.
Arabidopsis Dynamin-Like 2 That Binds Specifically to Phosphatidylinositol 4-Phosphate Assembles into a High-Molecular Weight Complex in Vivo and in Vitro
Authors: Kim Yong-Woo , Park Dae-Sup , Park Seung-Cheol , Kim Sung Hee , Cheong Gang-Won , Hwang Inhwan 
Published: November 2008

Abstract
Arabadopsis dynamin-like (ADL) 2, a member of the high-molecular weight (M r) dynamin family found in Arabidopsis, has been shown to be targeted to the plastid. In the chloroplast, most of the ADL2 was present in the fraction containing the envelope membranes when analyzed by suborganellar fractionation. Sucrose gradient and gel filtration experiments showed that when associated with membranes, ADL2 existed as a high-M r complex, whereas the soluble form existed as a monomer. The recombinant ADL2 expressed inEscherichia coli was present as a high-M r form and showed higher GTPase activity at a low NaCl concentration, whereas ADL2 existed as a low-M r form with a low level of GTPase activity at a high NaCl concentration. Electron microscopy studies revealed that the purified recombinant ADL2 formed spiral-coiled structures or rings. In the presence of guanosine-5′-O-(3-thio)triphosphate, these structures were transformed into a long rod structure. In contrast, in the presence of GDP, these structures disassembled into oligomers that were shown to be tetramer with 4-fold symmetry. Finally, a lipid-binding assay revealed that recombinant ADL2 purified fromE. coli bound specifically to phosphatidylinositol 4-phosphate. Together, these results demonstrated that the biochemical properties of ADL2 were very similar to those of dynamin and other related proteins. Based on this similarity, we propose that ADL2 may be involved in vesicle formation at the chloroplast envelope membrane.
Translocation and Utilization of Fungal Storage Lipid in the Arbuscular Mycorrhizal Symbiosis
Authors: Bago Berta , Zipfel Warren , Williams Rebecca M. , Jun Jeongwon , Arreola Raoul , Lammers Peter J. , Pfeffer Philip E. , Shachar-Hill Yair 
Published: November 2008

Abstract
The arbuscular mycorrhizal (AM) symbiosis is responsible for huge fluxes of photosynthetically fixed carbon from plants to the soil. Carbon is transferred from the plant to the fungus as hexose, but the main form of carbon stored by the mycobiont at all stages of its life cycle is triacylglycerol. Previous isotopic labeling experiments showed that the fungus exports this storage lipid from the intraradical mycelium (IRM) to the extraradical mycelium (ERM). Here, in vivo multiphoton microscopy was used to observe the movement of lipid bodies through the fungal colony and to determine their sizes, distribution, and velocities. The distribution of lipid bodies along fungal hyphae suggests that they are progressively consumed as they move toward growing tips. We report the isolation and measurements of expression of an AM fungal expressed sequence tag that encodes a putative acyl-coenzyme A dehydrogenase; its deduced amino acid sequence suggests that it may function in the anabolic flux of carbon from lipid to carbohydrate. Time-lapse image sequences show lipid bodies moving in both directions along hyphae and nuclear magnetic resonance analysis of labeling patterns after supplying 13C-labeled glycerol to either extraradical hyphae or colonized roots shows that there is indeed significant bidirectional translocation between IRM and ERM. We conclude that large amounts of lipid are translocated within the AM fungal colony and that, whereas net movement is from the IRM to the ERM, there is also substantial recirculation throughout the fungus.
Circadian Clock-Regulated Expression of Phytochrome and Cryptochrome Genes in Arabidopsis
Authors: Tóth Réka , Kevei Éva , Hall Anthony , Millar Andrew J. , Nagy Ferenc , Kozma-Bognár László 
Published: November 2008

AbstractMany physiological and biochemical processes in plants exhibit endogenous rhythms with a period of about 24 h. Endogenous oscillators called circadian clocks regulate these rhythms. The circadian clocks are synchronized to the periodic environmental changes (e.g. day/night cycles) by specific stimuli; among these, the most important is the light. Photoreceptors, phytochromes, and cryptochromes are involved in setting the clock by transducing the light signal to the central oscillator. In this work, we analyzed the spatial, temporal, and long-term light-regulated expression patterns of the Arabidopsis phytochrome (PHYA toPHYE) and cryptochrome (CRY1 andCRY2) promoters fused to the luciferase (LUC +) reporter gene. The results revealed new details of the tissue-specific expression and light regulation of the PHYC and CRY1 and2 promoters. More importantly, the data obtained demonstrate that the activities of the promoter::LUC + constructs, with the exception ofPHYC::LUC +, display circadian oscillations under constant conditions. In addition, it is shown by measuring the mRNA abundance of PHY andCRY genes under constant light conditions that the circadian control is also maintained at the level of mRNA accumulation. These observations indicate that the plant circadian clock controls the expression of these photoreceptors, revealing the formation of a new regulatory loop that could modulate gating and resetting of the circadian clock.