Expression and function of HD2‐type histone deacetylases in Arabidopsis development
Changhe ZhouHélène LabbéSunandini SridhaLi WangLining TianMarysia Latoszek‐GreenZhen YangDaniel C. BrownBrian MikiKeqiang Wu
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Summary HD2 (histone deacetylase) proteins are plant‐specific histone deacetylases (HDACs). The Arabidopsis genome contains four HD2 genes, namely HD2A , HD2B , HD2C , and HD2D . We have previously demonstrated that HD2A, HD2B, and HD2C can repress transcription directly by targeting to promoters in planta . Here, we show that the N‐terminal conserved motif (EFWG) and histidine 25 (H25), a potential catalytic residue, were important for the gene repression activity of HD2A . In situ hybridization indicated that HD2A , HD2B , and HD2C were expressed in ovules, embryos, shoot apical meristems, and primary leaves. Furthermore, all three genes were strongly induced during the process of somatic embryogenesis. HD2D mRNA was only detected in the stems and flowers with young siliques and may have adopted different functions. Using green fluorescent protein (GFP) fusions, we demonstrated that HD2A, HD2B, and HD2C accumulated in the nuclei of Arabidopsis cells. Overexpression of 35S::GFP–HD2A in transgenic Arabidopsis plants generated pleiotropic developmental abnormalities, including abnormal leaves, delayed flowering, and aborted seed development. The data showed that normal pattern of HD2 expression was essential for normal plant development and that HD2A, HD2B, and HD2C may be needed for embryogenesis and embryo development. Reverse transcriptase (RT)‐PCR analysis revealed that a number of genes involved in seed development and maturation were repressed in the 35S::GFP–HD2A plants, supporting a role of HD2A in the regulation of gene expression during seed development.Keywords:
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豆荚粉碎是是细胞分化和分离的协调行动的结果的一个 agronomical 特点。在 Arabidopsis，豆荚粉碎被在 ALCATRAZ (ALC ) 的一个复杂基因网络控制，基本 helix-loop-helix 家庭的一个成员，在水果裂开期间为房间分离是批评的。此处，我们经由酵母报导交往 ALC PROTEIN1 (ACI1 ) 的鉴定二混血儿的屏幕。ACI1 与一个充满离氨酸的领域和 C 终端编码原子蛋白质充满丝氨酸的领域。ACI1 主要在 siliques 在整个阀门的植物和中果皮在维管系被表示。我们的数据证明 ACI1 在由双分子的荧光互补试金示威了的原子核在酵母房间并且在植物房间与 ALC 的 N 终端部分强烈交往。ACI1 和 ALC 分享一个重叠表示模式，建议他们多半在植物一起工作一。然而，没有可检测的显型被 RNA 干扰技术与减少的 ACI1 表示在植物发现，建议 ACI1 可能是冗余的。总起来说，这些数据显示 ALC 可以与 ACI1 和它的相当或相同的事物交往在 Arabidopsis 在水果裂开期间控制房间分离。
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This chapter contains sections titled: Introduction Morphology of the Arabidopsis silique Determining the boundary between valve and replum: valve margin genes The making of valves and replum requires repression of valve margin genes Suppressors of the rpl phenotype: setting up territories A model for patterning the mediolateral axis of the Arabidopsis silique Auxin: a signaling molecule for the mediolateral axis? A biotechnological view Acknowledgements References
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Abstract To understand the molecular mechanism regulating meristem development in the monocot rice (Oryza sativa), we describe here the isolation and characterization of three floral organ number4 (fon4) alleles and the cloning of the FON4 gene. The fon4 mutants showed abnormal enlargement of the embryonic and vegetative shoot apical meristems (SAMs) and the inflorescence and floral meristems. Likely due to enlarged SAMs, fon4 mutants produced thick culms (stems) and increased numbers of both primary rachis branches and floral organs. We identified FON4 using a map-based cloning approach and found it encodes a small putatively secreted protein, which is the putative ortholog of the Arabidopsis (Arabidopsis thaliana) CLAVATA3 (CLV3) gene. FON4 transcripts mainly accumulated in the small group of cells at the apex of the SAMs, whereas the rice ortholog of CLV1 (FON1) is expressed throughout the SAMs, suggesting that the putative FON4 ligand might be sequestered as a possible mechanism for rice meristem regulation. Exogenous application of the peptides FON4p and CLV3p corresponding to the CLV3/ESR-related (CLE) motifs of FON4 and CLV3, respectively, resulted in termination of SAMs in rice, and treatment with CLV3p caused consumption of both rice and Arabidopsis root meristems, suggesting that the CLV pathway in limiting meristem size is conserved in both rice and Arabidopsis. However, exogenous FON4p did not have an obvious effect on limiting both rice and Arabidopsis root meristems, suggesting that the CLE motifs of Arabidopsis CLV3 and FON4 are potentially functionally divergent.
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Arabidopsis thaliana was grown from seed to seed wholly in microgravity on the International Space Station. Arabidopsis plants were germinated, grown, and maintained inside a growth chamber prior to returning to Earth. Some of these seeds were used in a subsequent experiment to successfully produce a second (back-to-back) generation of microgravity-grown Arabidopsis. In general, plant growth and development in microgravity proceeded similarly to those of the ground controls, which were grown in an identical chamber. Morphologically, the most striking feature of space-grown Arabidopsis was that the secondary inflorescence branches and siliques formed nearly perpendicular angles to the inflorescence stems. The branches grew out perpendicularly to the main inflorescence stem, indicating that gravity was the key determinant of branch and silique angle and that light had either no role or a secondary role in Arabidopsis branch and silique orientation. Seed protein bodies were 55% smaller in space seed than in controls, but protein assays showed only a 9% reduction in seed protein content. Germination rates for space-produced seed were 92%, indicating that the seeds developed in microgravity were healthy and viable. Gravity is not necessary for seed-to-seed growth of plants, though it plays a direct role in plant form and may influence seed reserves.
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Arabidopsis Argonaute10 (AGO10) specifically sequesters miR165 and miR166 and antagonizes their activity, thus regulating shoot apical meristem (SAM) development. However, where and when this sequestration acts is currently unclear. We show here that AGO10 represses miR165/166 activity in the embryo proper during early embryogenesis, through the apical and central regions of mature embryos, and eventually in the entire adaxial domain and vasculature of the cotyledons and leaf primordia. These locations are essentially identical to regions expressing PHABULOSA and REVOLUTA, mRNA targets of miR165/166. The Arabidopsis genome contains nine MIR165/166 genes. Sequestration of miR165/166 by the MIR165b, MIR166a, MIR166b, and MIR166g promoters efficiently rescues the SAM defect in ago10 mutants. Comparison of the expression patterns of AGO10 and the four MIR165/166 members suggests that AGO10 quenches the non-cell-autonomous activity of any miR165/166 that moves into AGO10-expressing niches. Thus, this study provides insight into how the spatiotemporal regulation of AGO10-miR165/166 activity affects SAM development.
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Gibberellins (GAs) form a complex family of diterpenoid compounds, some of which are potent endogenous regulators of plant growth and development. Pumpkin seeds contain GA-oxidases with unique catalytic properties resulting in GAs of unknown function for plant development. In order to understand their potential to achieve changes in GA levels and their role for plant development, we produced transgenic Arabidopsis expressing sense or antisense copies of pumpkin genes encoding GA-oxidases; CmGA7ox, CmGA3ox1, and CmGA2ox1 as well as sense copies of CmGA20ox1. Arabidopsis plants were transformed with the pumpkin genes downstream of a strong constitutive promoter cassette (E12-35S-Omega). The expression levels of pumpkin GA-oxidases were determined by competitive RT-PCR. Over-expression of CmGA7ox in Arabidopsis resulted in seedlings with elongated roots, taller plants that flower earlier and developed more siliques in long day. Similarly, over-expression of CmGA3ox1 resulted in elongated phenotypes and an increase in bioactive GA4, indicating that both enzymes catalyse limiting steps in the GA biosynthesis of Arabidopsis. In contrast, over-expression of CmGA20ox1 in Arabidopsis resulted in shorter hypocotyls and internodes, less siliques and flowering was delayed with reduced GA4 levels and increased levels of the inactive GA17 and GA25. Severe dwarfed plants were obtained by over-expressing CmGA2ox1 in Arabidopsis and associated with a decrease in the levels of bioactive GA4 and an increase inactive product GA34 in comparison to control plants. The phenotype of plants expressing antisense copies did not change compared to the wild type plants. Modifying GA-biosynthesis using this approach may be useful for controlling plant development in agricultural and horticultural important species.
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With wild type and PAtJ70:GUS transgenic Arabidopsis,tissue-specific expression of Arabidopsis J-domain protein AtJ70 was investigated using GUS staining and real-time PCR.The results show that the AtJ70 gene expresses throughout roots,stems,leaves,flowers and fruits.The expression level of the AtJ70 gene is the highest in flowers and leaves,followed by siliques and roots.The AtJ70 gene has little expression in stems.In addition,with AtJ70-mGFP4 transgenic Arabidopsis,subcellular localization of AtJ70 protein was tested using Confocal.The result shows that AtJ70 mainly targets to the nucleus.
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In order to gain a better understanding of the function of SKIP16, a gene encoding the F-box protein of the SKP1-CULLIN-F-BOX PROTEIN (SCF) complex, on the reproductive development of Arabidopsis thaliana, we conducted an experiment with transgenic plants containing the gene ASK1:SKIP16. We did this to see the effect of overexpression of SKIP16 on the plants to further begin to understand the role of SKIP16 on Arabidopsis development since the ASK1 promoter ubiquitously drives high expression levels of SKIP16. While it is known that SCF complexes impact growth and development in plants, the function of SKIP16 is unknown. We observed that at the T1 generation, two of the ASK1:SKIP16 plants had only one terminal flower on each inflorescence stem while many other plants produced either infertile siliques or siliques with reduced fertility. These observations indicate that SKIP16 plays an important role in Arabidopsis reproductive development. Further research on the function of SKIP16 may yield crucial information about how SCF F-box protein regulates plant development in Arabidopsis and other plants.
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Glycerol-3-phosphate acyltransferases (GPATs) play an important role in glycerolipid biosynthesis, and are mainly involved in oil production, flower development, and stress response. However, their roles in regulating plant height remain unreported. Here, we report that Arabidopsis GPAT1 is involved in the regulation of plant height. GUS assay and qRT-PCR analysis in Arabidopsis showed that GPAT1 is highly expressed in flowers, siliques, and seeds. A loss of function mutation in GPAT1 was shown to decrease seed yield but increase plant height through enhanced cell length. Transcriptomic and qRT-PCR data revealed that the expression levels of genes related to gibberellin (GA) biosynthesis and signaling, as well as those of cell wall organization and biogenesis, were significantly upregulated. These led to cell length elongation, and thus, an increase in plant height. Together, our data suggest that knockout of GPAT1 impairs glycerolipid metabolism in Arabidopsis, leading to reduced seed yield, but promotes the biosynthesis of GA, which ultimately enhances plant height. This study provides new evidence on the interplay between lipid and hormone metabolism in the regulation of plant height.
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