Project description:3 samples, 2 reps each. comparison of wildtype cotyledon to RNAioleosin transgenic Using RNAi, the seed oil body protein 24-kDa oleosin has been suppressed in transgenic soybeans. The endoplasmic reticulum (ER) forms micro-oil bodies about 50 nm in diameter that coalesce with adjacent oil bodies forming a hierarchy of oil body sizes. The oil bodies in the oleosin knockdown form large oil body-ER complexes with the interior dominated by micro-oil bodies and intermediate-sized oil bodies, while the peripheral areas of the complex are dominated by large oil bodies. The complex merges to form giant oil bodies with onset of seed dormancy that disrupts cell structure. The transcriptome of the oleosin knockdown shows few changes compared to wild-type. Proteomic analysis of the isolated oil bodies of the 24-kDa oleosin knockdown shows the absence of the 24-kDa oleosin and the presence of abundant caleosin and lipoxygenase. The formation of the micro-oil bodies in the oleosin knockdown is interpreted to indicate a function of the oleosin as a surfactant.
Project description:Soybean is one of the most economically important crops in the world. The cotyledon is the nutrient storage area in seeds, and it is critical for seed quality and yield. Cotyledon mutants are important for the genetic dissection of embryo patterning and seed development.Here, we characterised a soybean curled-cotyledon (cco) mutant. Compared with wild-type (WT), the entire embryos of cco mutant resembled the “tail of swallow”. To explore the molecular mechanisms underlying soybean cotyledon development, we executed RNA-Seq using the Illumina HiSeq2000 system. Pods at 7DAF collected from WT and cco were analyzed by RNA-seq.
Project description:Seeds are comprised of three major 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 part undergoes distinct developmental programs during seed development. What methylation changes occur 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 cotyledon stage seeds, the seed coat, embryonic cotyledons, and embryonic axis, using Illumina sequencing. Illumina sequencing of bisulfite-converted genomic DNA from three parts of soybean cotyledon stage seeds: seed coat (COT-SC), embryonic cotyledons (COT-COT), and embryonic axis (COT-AX).
Project description:We microdissected each compartment from 8-micron paraffin sections using the Leica LMD6000 system to identify all genes active in different compartments of a soybean seed containing a cotyledon-stage embryo. Experiment Overall Design: cotyledon-stage seed compartments were isolated using the Leica LMD6000 system. Total RNA was amplified and hybridized with Affymetrix Soybean Genome Arrays.
Project description:Soybean is one of the most economically important crops in the world. The cotyledon is the nutrient storage area in seeds, and it is critical for seed quality and yield. Cotyledon mutants are important for the genetic dissection of embryo patterning and seed development.Here, we characterised a soybean curled-cotyledon (cco) mutant. Compared with wild-type (WT), the entire embryos of cco mutant resembled the “tail of swallow”. To explore the molecular mechanisms underlying soybean cotyledon development, we executed RNA-Seq using the Illumina HiSeq2000 system.
Project description:To understand the processes governing soybean seed development, we profiled the mRNA transcriptome and chromatin accessibility of cotyledon-stage seeds at the single-nucleus level.
Project description:We performed small-RNA profiling of soybean seed regions and sub-regions, to gain new insights into the regulatory processes required for seed differentiation. Eight subregions of the Cotyledon stage of the soybean seeds were analyzed. Two biological replicates were collected for each subregions.
Project description:We microdissected each compartment from 8-micron paraffin sections using the Leica LMD6000 system to identify all genes active in different compartments of a soybean seed containing a cotyledon-stage embryo. Keywords: cell type comparison