Transcription factor AtDOF4.2 regulates shoot branching and seed coat formation in Arabidopsis
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ABSTRACT: Plant-specific DOF-type transcription factors regulate various biological processes. Here, we characterized a silique-abundant gene AtDOF4.2 for its functions in Arabidopsis. AtDOF4.2 is localized in the nuclear region and has transcriptional activation activity in both yeast and plant protoplast assays. The Thr-Met-Asp motif in AtDOF4.2 is essential for its activation. AtDOF4.2-overexpressing plants exhibit an increased branching phenotype, and the mutation of Thr-Met-Asp motif in AtDOF4.2 significantly reduces the branching in transgenic plants. AtDOF4.2 may achieve this function through the upregulation of three branching-related genes, AtSTM, AtTFL1 and AtCYP83B1. The seeds of an AtDOF4.2-overexpressing plant show collapse-like morphology in epidermal cells of the seed coat. Mucilage contents and the concentration and composition of mucilage monosaccharides are significantly changed in the seed coat of transgenic plants. AtDOF4.2 may exert its effects on the seed epidermis through the direct binding and activation of the cell wall loosening-related gene AtEXPA9. The dof4.2 mutant did not exhibit changes in branching or its seed coat; however, the silique length and seed yield were increased. AtDOF4.4, which is a close homolog of AtDOF4.2, also promotes shoot branching and affects silique size and seed yield. Manipulation of these genes should have a practical use in the improvement of agronomic traits in important crops. Two-week-old seedlings (aerial part) of Col-0 and AtDOF4.4-overexpressing transgenic lines 4.4-1 and 4.4-5, grown on MS, were used for extraction of total RNA and subjected to chip analysis using Agilent Arabidopsis Oligo Microarray (4X44K) in SHANGHAIBIO CORPORATION (www.ebioservice.com; Shanghai, China).
Project description:Plant-specific DOF-type transcription factors regulate various biological processes. Here, we characterized a silique-abundant gene AtDOF4.2 for its functions in Arabidopsis. AtDOF4.2 is localized in the nuclear region and has transcriptional activation activity in both yeast and plant protoplast assays. The Thr-Met-Asp motif in AtDOF4.2 is essential for its activation. AtDOF4.2-overexpressing plants exhibit an increased branching phenotype, and the mutation of Thr-Met-Asp motif in AtDOF4.2 significantly reduces the branching in transgenic plants. AtDOF4.2 may achieve this function through the upregulation of three branching-related genes, AtSTM, AtTFL1 and AtCYP83B1. The seeds of an AtDOF4.2-overexpressing plant show collapse-like morphology in epidermal cells of the seed coat. Mucilage contents and the concentration and composition of mucilage monosaccharides are significantly changed in the seed coat of transgenic plants. AtDOF4.2 may exert its effects on the seed epidermis through the direct binding and activation of the cell wall loosening-related gene AtEXPA9. The dof4.2 mutant did not exhibit changes in branching or its seed coat; however, the silique length and seed yield were increased. AtDOF4.4, which is a close homolog of AtDOF4.2, also promotes shoot branching and affects silique size and seed yield. Manipulation of these genes should have a practical use in the improvement of agronomic traits in important crops.
Project description:Tissues were isolated from the globular seed using laser microdissection. At least two bioreplicates are included for each tissue. Tissues examined are : the embryo proper (EP), micropylar endosperm (MCE), peripheral endosperm (PEN), chalazal endosperm (CZE), chalazal proliferating tissue (CPT), chalazal seed coat (CZSC), inner seed coat (ISC), and outer seed coat (OSC).
Project description:Transcriptional profiling during Arabidopsis seed coat development at 3 key developmental timepoints by using 2 mutant lines and their wild types. The data provides a globe view of seed coat development in arabidopsis can be used for identification of new gene candidates for seed coat development.
Project description:In addition to the evolutionarily-conserved Ca2+ sensor, calmodulin (CaM), plants possess a large family of CaM-related proteins (CMLs). Using a cml39 loss-of-function mutant, we investigated the roles of CML39 in Arabidopsis and discovered a range of phenotypes across developmental stages and in different tissues. In mature plants, loss of CML39 results in shorter siliques, reduced seed number per silique, and reduced number of ovules per pistil. We also observed changes in seed development, germination, and seed coat properties in cml39 mutants in comparison to wild-type plants. Using radicle emergence as a measure of germination, cml39 mutants showed more rapid germination than wild-type plants. In marked contrast to wild-type seeds, the germination of developing, immature cml39 seeds was not sensitive to cold-stratification. In addition, germination of cml39 seeds was less sensitive than wild-type to inhibition by ABA or by treatments that impaired gibberellic acid biosynthesis. Tetrazolium red staining indicated that the seed-coat permeability of cml39 seeds is greater than that of wild-type seeds. RNA sequencing analysis of cml39 seedlings suggests that changes in chromatin modification may underlie some of the phenotypes associated with cml39 mutants, consistent with previous reports that orthologs of CML39 participate in gene silencing. Aberrant ectopic expression of transcripts for seed storage proteins in 7-day old cml39 seedlings was observed, suggesting mis-regulation of early developmental programs. Collectively, our data support a model where CML39 serves as an important Ca2+ sensor during ovule and seed development, as well as during germination and seedling establishment.
Project description:Identification of differentially expressed genes in seeds and silique walls at the seed-filling stage in Brassica napus through transcriptional profiling Two tissues, three biological replicates, one biological duplicate with two technical replicates
Project description:Transcriptional profiling during Arabidopsis seed coat development at 3 key developmental timepoints by using 2 mutant lines and their wild types. The data provides a globe view of seed coat development in arabidopsis can be used for identification of new gene candidates for seed coat development. 3 seed coat development stages, 4 lines (2 wild type + 2 mutants) of arabidopsis were sampled. 4 biological replicates.
Project description:The seed coat of black (iRT) soybean with the dominant R allele begins to accumulate cyanic pigments at the transition stage of seed development (300 – 400 mg fresh seed weight), whereas the brown (irT) nearly-isogenic seed coat with the recessive r allele lacks cyanic pigments at all stages of seed development. We used microarrays to determine global gene expression differences between black (iRT) and brown (irT) soybean seed coats at the transition stage of seed development (300 – 400 mg fresh seed weight).
Project description:Seeds are comprised of three majors parts of distinct parental origin: the seed coat, embryo, and endosperm. The maternally-derived seed coat is important for nurturing and protecting the seeds during development. By contrast, the embryo and the endosperm are derived from a double fertilization event, where one sperm fertilizes the egg to form the diploid zygote and the other sperm fertilizes the central cell to form the triploid endosperm. Each seed parts undergo distinct developmental programs during seed development. What methylation changes occurring in the different seed parts, if any, remains unknown. To uncover the possible role of DNA methylation in different parts of the seed, we characterized the methylome of three major parts of an early maturation stage seed: seed coat, embryonic cotyledons, and embryonic axis using Illumina sequencing.