Large-scale analysis of soybean embryonic axis in five time points during germination
Ontology highlight
ABSTRACT: Purpose: This RNA-Seq study aims on elucidate the major trends in the transcriptional profile of soybean embryonic axes during germination. Methods: Soybean seeds were germinated in soaked cotton at 28ºC. In addition to dry seeds, seeds were harvested at 3, 6, 12, 24 hours after imbibition. Then the embryonic axes were separated from the cotyledons for RNA extraction. For each biological sample, 20 seeds were used. Results: Identification of genes and pathways involved in metabolism, hormone signaling and transcriptional regulation.
Project description:Purpose: A time-course transcriptome study to identify probable GA-responsive genes in soybean embryonic axes during seed germination. Methods: Seeds were germinated in the presence or absence of 200 µM PBZ. Seeds were germinated in 28°C temperature and 12/12h photoperiod (dark/light) and harvested at 12, 24 and 36 hours after imbibition (HAI). Three biological replicates were performed. Results: Identification of GA-responsive genes during germination in Glycine max.
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:To understand the molecular events underlying seed desiccation and germination, we performed transcriptome analysis of soybean (Glycine max) embryos at three seed developmental stages (early, mid and late desiccation), mature dry seeds, and three germination stages (3, 16 and 27 hour-after-imbibition).
Project description:What methylation changes are occurring during seed development largely remains unknown. To uncover the possible role of DNA methylation throughout all of seed development - from fertilization through dormancy and post-germination in soybean, we characterized the methylome of whole seeds representing the differentiation (GLOB and COT stages), maturation (early- [EM], mid- [B1] and late- [AA1] maturation stages), dormancy (DRY stage), and post-germination (seedling) phases of soybean seed development using Illumina sequencing. In addition, we characterized the methylome of the cotyledons of germinated seedling to examine methylation differences before and after germination.
Project description:To establish the basis for understanding molecular mechanism of seed germination response to temperature, we analyzed transcriptomes in freshly harvested dormant and dry stored after-ripened seeds. The after-ripened seeds started to show visible germination from 36h after the start of imbibition, and almost all the seeds germinated after 3 days. The freshly harvested seeds stayed dormant by imbibition at 26°C, and germination of the after-ripened seeds was almost completely inhibited at 34°C. Total RNA was prepared from 0 (dry), 6 and 24h imbibed seeds to find regulatory genes of seed dormancy and germination.
Project description:Background: To understand gene expression networks leading to functional properties of the soybean seed, we have undertaken a detailed examination of soybean seed development during the stages of major accumulation of oils, proteins, and starches, as well as the desiccating and mature stages, using microarrays consisting of up to 27,000 soybean cDNAs. Results: It was discovered that genes related to cell growth and maintenance processes, as well as energy processes like photosynthesis, decreased in expression levels as the cotyledons approached the mature, dry stage. Genes involved with some storage proteins had their highest expression levels at the stage of largest fresh weight. However, genes encoding many transcription factors and DNA binding proteins showed higher expression levels in the desiccating and dry seeds than in most of the green stages. Conclusions: Data on 27,000 cDNAs have been obtained over five stages of soybean development, including the stages of major accumulation of agronomically-important products, using microarrays. Of particular interest are the genes found to peak in expression at the desiccating and dry seed stages, such as those annotated as transcription factors, which may indicate the preparation of pathways that will be needed later in the early stages of imbibition and germination. Five stages of development of soybean cotyledons were studied. The first three, with fresh weights of 25-50mg, 75-100mg, and 400-500mg, were green in color. The last two, yellow-colored cotyledons with a fresh weight of 200-300mg and dry whole seeds approximately 100-200mg in weight, were desiccating and yellow/brown in color. The green stage of 100-200mg fresh weight cotyledons was used as a reference for all other stages. Four replicates, including two dye swaps, were made of each of the first three (green) stages. Two replicates, including one dye swap, were made of each of the last two (yellow/dry) stages. The results from three different platforms (GPL229, GPL1012, and GPL1013) were combined.
Project description:For production of soy-foods or supplements, imbibition of soybean seeds in the water is required step for generation of tofu, soy-milk, and other soy-products. With an aim to get new insight into effects of different imbibition temperature (4, 25, and 55℃), this study conducted integrated proteomics and metabolomics analysis of soybean seeds. For total proteome analysis, we applied TMT labeling based quantitative proteomics combined to FASP (Filter-Aided Sample Preparation) with high-throughput LC-MS/MS. A total of 2,616 proteins were identified out of which 801 proteins showed significantly difference of protein abundance (≥1.5 fold change, Benjamini-Hochberg FDR <0.05) among 4, 25, and 55℃ imbibition seeds. Functional analysis of identified proteins showed an increased abundance of proteins functioning as glycosyl hydrolase enzymes such as beta-glucosidase, alpha and beta-galactosidase, and alpha-mannosidase, or protease, and PTMs related enzymes as well. UPLC TOF-MS analysis showed around 20 fold increase in isoflavone aglycones (daidzein and genistein) while isoflavone glycosides (daidzin and genistin) were decreased in 55℃ imbibition seed, in agreement with proteomics results which we assume positively related to increase abundance of glycosyl hydrolase. A metabolomics analysis revealed 64 metabolites were significantly altered, for example, various free amino acids showed accumulation patterns by increased abundance of various protease enzymes and further confirmed the accumulation of isoflavone aglycones and degradation of raffinose and stachyose in 55℃ imbibition seeds. Based on these results, we recommend the use of 55℃ for soybean seed imbibition to increase the quality of soy-food products.
Project description:This series analyses germinating Lepidium sativum seeds with both temporal and spatial detail. This is a cross species microarray normalisation on Arabidopsis thaliana chips. Performed as part of the vSEED project Lepidium seeds were dissected into four compartments at seven time points during seed germination. The compartments were the micropylar endosperm (CAP), the non-micropylar endosperm (NME), the radicle and lower hypocotyl (RAD) and the cotyledons (COT). At testa and endosperm rupture the seeds were split into pre- and post- ruptured populations. Dry seeds were also sampled: as a whole seed (DRY), the radicle and endosperm part (DRYRC) and the dry seed part containing the non-micropylar endosperm and part of the cotyledons (DRYNMC).
Project description:au14-10_wd40 - effet of light on translatome of arabidopsis seeds during germination - Does light regulates germination via polysome association ? - At harvest seeds are dormant.They stay dormant if stored at -20°C.A.Th dormant seeds dont germinate at 25°C in darkness.Total RNA and polysomal RNA (polysomal fractions purified on sucrose gradients)were extracted from imbibed seeds for 20h at 22°C in darkness and light(3 biological replicates). Transcriptome and translatome are compared for light vs dark for 20h of imbibition. In silico comparison will allow to compare transcriptome and translatome for each type of sample.
Project description:Transcriptomic study of the impact of osmopriming on rape seeds (Brassica napus L.; cv 'Libomir') during priming process and after germination. The assays were replicated twice on two independent priming and germination experiments. Seeds were osmoprimed in PEG solution (-1.2 MPa osmotic potential) during 7 days, dried to initial moisture content and then germinated for 7 hours on water. The analysis during different phases of priming procedure (soaking and drying), after whole osmopriming process and germination were done.