Genome-Wide Small RNA Profiling of a Soybean Seed Compartment
ABSTRACT: We report the genome-wide small RNA of soybean early maturation seed coat parenchyma compartment soybean early maturation seeds using Illumina high-throughput sequencing technology. Illumina sequencing of small RNA from early maturation seed coat parenchyma compartment and early-maturation stage whole seeds
Project description:We used laser capture microdissection (LCM) to capture soybean seed compartments and profiled the transcriptomes of several compartments using next-generation sequencing. We profiled the transcriptomes of the embryo-proper and the suspensor region from globular stage embryos and the seed coat parenchyma layer of early maturation stage seeds using the Illumina GAIIx system. Illumina sequencing of transcripts from laser-captured embryo proper and suspensor of globular stage embryo and seed coat parenchyma layer of the early maturation stage seed.
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. Seventeen subregions of the Early Maturation stage of the soybean seeds were analyzed. Two biological replicates were collected for each subregions.
Project description:How epigenetics is involved in the transition from seed maturation to seed germination largely remains elusive. To uncover the possible role of epigenetics in gene expression during the transition from seed maturation to seed germination in soybean, the transcriptome of cotyledons from four stages of soybean seed maturation and germination, including mid-late maturation, late maturation, seed dormancy and seed germination, were profiled by Illumina sequencing. For the genes that are quantitatively regulated at the four stages, two antagonistic epigenetic marks, H3K4me3 and H3K27me3, together with the binding of RNA polymerase II, were investigated at the four stages by chromatin immunoprecipitation (ChIP). For 10 out of 16 genes examined, the relative enrichment of histone modification marks (H3K4me3 and H3K27me3) and RNA polymerase II binding on their promoter regions correlates well with their relative expression levels at four stages, suggesting the involvement of epigenetics in transcriptional regulation. A striking finding is that seed germination-specific genes start to show open chromatin (H3K4me3) during late seed maturation although their transcripts do not accumulate, which is further supported by RNA polymerase II binding. Together, our results provide the first evidence that seed germination genes can be primed for transcription (open chromatin and RNA polymerase II binding) during seed maturation, highlighting that the transition from seed maturation to seed germination starts at late seed maturation stages at both the genetic and epigenetic levels. Illumina sequencing of transcripts from cotyledons of mid-maturation (B1 stage) and late maturation (AA1 stage) seeds, whole dry seeds, and cotyledons of seedlings six days after imbibition.
Project description:What methylation changes are occurring in different compartments of early maturation stage seed largely remains unknown. To uncover the possible role of DNA methylation in different compartments of early maturation stage seed, we characterized the methylome of two major compartments in embryonic cotyledon: cotyledon abaxial parenchyma (EM-COT-ABPY) and cotyledon adaxial parenchyma (EM-COT-ADPY) using Illumina sequencing. Illumina sequencing of bisulfite-converted genomic DNA from cotyledon abaxial parenchyma (EM-COT-ABPY) and cotyledon adaxial parenchyma (EM-COT-ADPY) compartments.
Project description:Five degradome libraries were constructed from three different seed developmental stages. Separate degradome libraries were constructed for seed coat and cotyledons to identify the tissue specific miRNAs and their potential targets. Sequencing and analysis of degradome libraries gives identification of 183 different targets for 80 known soybean miRNAs. We found 30 cotyledon specific, 18 seed coat specific and 32 miRNAs found in both tissues irrespective of the developmental stages. One interesting observation is that we found more miRNA targets in late seed developmental stages than earlier stages. Additionally, we have validated four different auxin response factor genes as targets for gma-miR160 via RNA ligase mediated 5′ rapid amplification of cDNA ends (RLM-5′RACE). GO analysis indicated the enrichment of miRNA target genes in seed development. Construction of degradome libraries using cotyledons and seed coats from 3 different developmental stages
Project description:We report the genome-wide transcriptome of soybean seeds across several stages of seed development and the entire life cycle using Illumina high-throughput sequencing technology. Specifically, we profiled whole seeds containing globular-stage, heart-stage, cotyledon-stage, and early maturation-stage embryos. We also profiled dry soybean seeds, and vegetative and reproductive tissues including leaves, roots, stems, seedlings, and floral buds. Illumina sequencing of transcripts from whole seeds at five stages of seed development (globular, heart, cotyledon, early-maturation, dry), and vegetative (leaves, roots, stems, seedlings) and reproductive (floral buds) tissues.
Project description:What methylation changes are occurring in different parts of early maturation stage seed largely remains unknown. To uncover the possible role of DNA methylation in different parts of early maturation stage seed, we characterized the methylome of seed coats,cotyledons, and the embryonic seed axis using Illumina sequencing. seed coats, cotyledon, and axis
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 early maturation-stage embryo. Experiment Overall Design: early maturation-stage seed compartments were isolated using the Leica LMD6000 system. Total RNA was amplified and hybridized with Affymetrix Soybean Genome Arrays.
Project description:What methylation changes are occurring during seed development largely remains unknown. To uncover the possible role of DNA methylation during the transition from seed differentiation to maturation and dormany in soybean, we characterized the methylome of whole seeds representing the differentiation (GLOB stage), maturation (early- (EM), mid- (B1) and late- (AA1) maturation stages), and dormancy (DRY stage) phases of soybean seed development using Illumina sequencing. In addition, we characterized the methylome of the mid-maturation stage embryonic axis (B1-AX) to examine methylation differences, if any, between an embryonic region compared to the whole seed. Illumina sequencing of bisulfite-converted genomic DNA from globular stage (GLOB), early-maturation stage (EM), mid-maturation stage (B1), and late-maturation stage (AA1) seeds, dormancy stage (DRY) and mid-maturation embryonic axis (B1-AX).
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. Illumina sequencing of bisulfite-converted genomic DNA from three parts of an mid-maturation (B1) stage seed: seed coat (B1-SC), embryonic cotyledons (B1-COT), and embryonic axis (B1-AX).