Project description:High temperature stress results in yield loss and alterations to seed composition during seed filling in oilseed rape (Brassica napus). However, the mechanism underlying this heat response is poorly understood. In this study, we employed a microarray analysis with silique walls and seeds from the developing siliques (20 days after flowering) of Brassica napus that had undergone heat stress. Two-condition experiment, control vs heat stress, 2 time points
Project description:High temperature stress results in yield loss and alterations to seed composition during seed filling in oilseed rape (Brassica napus). However, the mechanism underlying this heat response is poorly understood. In this study, we employed a microarray analysis with silique walls and seeds from the developing siliques (20 days after flowering) of Brassica napus that had undergone heat stress. Overall design: Two-condition experiment, control vs heat stress, 2 time points
Project description:Time course of gene expression profiles during seed development and maturation in Brassica napus were studied using Combimatrix Brassica microarray. The time course expression of 90K Brassica napus EST contigs were measured at 8 developing seed stages of 10, 15, 20, 25, 30, 35, 40 and 45 DAF (days after flowering) using single color microarray
Project description:Compared to ordinary rapeseed, high-oleic acid rapeseed has higher levels of monounsaturated fatty acids and lower levels of saturated fatty acid and polyunsaturated fatty acids, and thus is of high nutritional and health value. In addition, high-oleic acid rapeseed oil imparts cardiovascular protective effects. Based on these properties, high-oleic acid oil crops have been extensively investigated and cultivated. In this study, we employed a microarray analysis with high oleic acid line and low oleic acid line from the developing seeds (27 days after flowering) of Brassica napus. Overall design: High oleic acid line vs. low oleic acid line, 2 replicates.
Project description:Understanding the regulation of lipid metabolism is vital for genetic engineering of Brassica napus (B. napus) to increase oil yield or modify oil composition. We report the application of Illumina Hiseq 2000 for transcriptome profiling of seeds of B. napus at different developmental stages, which may uncover the dynamic changes in lipid metabolism and reveal key genes involved in lipid biosynthesis and degradation. Total RNA from developing seeds at 2, 4, 6, and 8 weeks after pollination (WAP) were isolated and sequenced separately. The gene expression levels of all samples were quantified and normalized by the DESeq normalization. We found that the biosynthesis of fatty acids is a dominant cellular process from 2 to 6 WAP, while the degradation mainly happens after 6 WAP. Two genes, encoding for acetyl-CoA carboxylase and acyl-ACP desaturase, might be critical for fatty acid biosynthesis in oil rape seeds. This study provides insight into the mechanism underlying lipid metabolism and reveals candidate genes that are worthy of further investigation for their values in genetic engineering of B. napus. Whole Transcriptome profiling of developing Brassica napus seeds at 2, 4, 6, 8 WAP by RNA sequencing using Illumina HiSeq 2000.
Project description:Gene expression profiles during seed development and fatty acid (FA) metabolism, as well as the relevant regulation, of Brassica napus were studied through multiple high-throughput genomic approaches. Serial Analysis of Gene Expression (SAGE) using seed materials obtained a total of 68,718 tags, of which 23,897 were unique and 503 tags were functionally identified, and revealed the transcriptome of approximately 35,000 transcripts in B. napus developing seeds. Further, ~22,000 independent ESTs were obtained by large-scale sequencing using immature embryos at different stages, and 8343 uni-ESTs and 3355 full-length cDNAs were identified respectively, resulting in the systemic identification of B. napus FA biosynthesis-related genes. Gene expression profiles were further studied employing cDNA chip hybridization to reveal the global regulatory network of FA metabolism in developing seeds. Together with the analysis on FA amounts and composition, it was shown that 17-21 days after pollination (DAP) was a crucial stage for transition of seed to sink tissue. High expressions of FA biosynthesis-related genes and transition of FA components are mainly at stages 21 DAP or 21-25 DAP respectively. In addition, compared to Arabidopsis, more critical roles of starch metabolism are detected for B. napus seed FA metabolism and storage components accumulation. Crucial effects of starch metabolism, carbon flux, oxidative pentose phosphate pathway (OPPP), photosynthesis, and other regulators in FA metabolism were discussed. Keywords: Brassica napus, immature seed, SAGE, EST, cDNA microarray Overall design: Brassica napus immature seeds at 5 and 9 days after pollination (DAP) were collected and used to construct SAGE library. Rapeseed embryos at different developmental stages were observed and two cDNA libraries were constructed using materials at 3-9 day after pollination(DAP) or 11-19 DAP. Gene expression profiles during seed development were further detailed studied via cDNA microarray. cDNA inserts of ESTs were PCR amplified and used to generate the glass-based cDNA chips. Immature seeds at different developmental(7, 9, 12, 17, 19, 21, 25, 31DAP) stages were used for hybridization, and the seed of 3 day after pollination was used as control.
Project description:To broadly identify genes regulated by Transparent Testa16 in Brassica napus In order to broadly identify genes regulated by BnTT16s, microarray technology was employed to compare gene expression levels in developing seeds (2-DAP) of Bntt16 RNAi and wild-type plants.
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:Background: Auxin/Indoleacetic acid (Aux/IAA) genes participate in the auxin signaling pathway and play key roles in plant growth and development. Although the Aux/IAA gene family has been identified in many plants, within allotetraploid Brassica napus little is known. Results: In this study, a total of 119 Aux/IAA genes were found in the genome of B. napus. They were distributed non-randomly across all 19 chromosomes and other non-anchored random scaffolds, with a symmetric distribution in the A and C subgenomes. Evolutionary and comparative analysis revealed that 111 (94.1%) B. napus Aux/IAA genes were multiplied due to ancestral Brassica genome triplication and recent allotetraploidy from B. rapa and B. oleracea. Phylogenetic analysis indicated seven subgroups containing 29 orthologous gene sets and two Brassica-specific gene sets. Structures of genes and proteins varied across different genes but were conserved among homologous genes in B. napus. Furthermore, analysis of transcriptional profiles revealed that the expression patterns of Aux/IAA genes in B. napus were tissue dependent. Auxin-responsive elements tend to be distributed in the proximal region of promoters, and are significantly associated with early exogenous auxin up-regulation. Conclusions: The Aux/IAA gene family were identified and analyzed comprehensively in the allotetraploid B. napus genome. This analysis provides a deeper understanding of diversification of Aux/IAA gene family and will facilitate further dissection of Aux/IAA gene function in B. napus. Overall design: Examination of the auxin response of gene exoression in Brassica napus seedling