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An Efficient Method for the Isolation of Highly Purified RNA from Seeds for Use in Quantitative Transcriptome Analysis
Published on: January 11, 2017
Gene expression at early stages of Brassica napus seed development as revealed by transcript profiling of
Jinzhuo Dong1, Wilf A Keller, Wei Yan
1Plant Biotechnology Institute, National Research Council Canada, 110 Gymnasium Place, S7N 7R9, Saskatoon, Saskatchewan, Canada.
Abstract:
Approximately 5000 plaques derived from a Brassica napus L. (canola) seed-cDNA library representing 15 days after pollination (DAP) were differentially screened for highly expressed genes at the early stages of seed development. Analysis of 104 differentially expressed sequence tags revealed 54 unique genes, of which 33 had putative homologues described in Arabidopsis thaliana (L.) Heynh. or B. napus. These encoded diverse proteins, ranging from proteins of unknown function to metabolic enzymes and proteins associated with cell structure and development. Twenty-five genes were only expressed in seeds, and 11 of these started to express as early as 5 or 10 DAP. The majority of the seed-specific genes that are expressed at early stages of seed development encoded proteins with high similarity to hypothetical Arabidopsis proteins. Tissue-specificity determined by Northern analysis revealed that four seed-specific genes were expressed only in seed coats and another five in both embryos and seed coats. Analysis of transcript profiles of seed-abundant as well as seed-specific genes, and their expression patterns, implies that the B. napus seed is undergoing an active cell proliferation during 10-20 DAP, while establishing metabolic networks for subsequent seed maturation.
