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Transformation of Brassica Juncea L. with RNAi Vector of ANS Gene and Construction of Vector for Expression of DFR Gene

Author: YangLiu
Tutor: LiuZhongSong
School: Hunan Agricultural University
Course: Crop Genetics and Breeding
Keywords: Brassica juncea Agrobacterium-mediated transformation ANS DFR Seed coat color
CLC: S565.4
Type: Master's thesis
Year: 2009
Downloads: 35
Quote: 1
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Abstract


Brassica juncea germplasm resources are very rich in China, more than half are the genetically stable yellow-seeded varieties. It is of great significance to make full use of the germplasm resources of Yellow-seeded Brassica juncea, and innovate germplasm through the useful genes transferring in rape breeding. A large number of genes were involved in seed coat color formation in Oilseed Rape. Three genes, DFR、ANS、BAN, were demonstrated to be the key genes in controlling seed coat color. The effects of factors such as infection time of Agrobacterium, hormone proportions in pre-culture medium and differentiation medium on regeneration of Cotyledon petiole explants of Sichuan Yellow, Purple-leaf Mustard and their near-isogenic lines aseptic seedling were studied. The highest transformation efficiency was at 10min of infection. Transformation efficiency declined, followed by browning phenomenon, when infection time prolonged. The highest differentiation rate, 78.9 percent, of cotyledon petiole in the 6-BA 2.0 mg / L differentiation medium was achieved after 2 days’ pre-culture in the MS + 6-BA 1 mg / L + 2 ,4-D 1.5 mg / L pre-culture medium. Brassica juncea transformation was mediated by Agrobacterium which contained ANS gene interference vector, 4 PPT-resistant regenerated plants were survived. The result by PCR showed that ANS gene had been integrated into the genome of transgenic plants. All the transformants were normal in morphology. The seed coat color of T0 generation was black. Expression vector of DFR was constructed and subsequently introduced into the Agrobacterium tumefaciens strain EHA105 with freeze-thaw method, which provided foundations for analysis of the function of the gene.

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CLC: > Agricultural Sciences > Crop > Economic crops > Oil crops > Rapeseed ( Brassica )
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