Pigment Analysis Of The Seed Coat Color Transcription Factor Gene TT2 Genetic Transformation System Optimization In Brassic Juncea

Rape seed oil content is an important goal of breeding quality. In the same genetic background, the oil content of yellow-seeded generally higher than black seeds, brown seeds, and yellow-seeded with species of thin, less pigment, little impurity, cellulose content is low, the oil is clear and transparent, the advantages of higher protein content. This study is to optimize the efficiency of Brassica juncea genetic transformation system, by constructing the expression vector TT2 genetic transformation of Brassica juncea, identification of its function.In this study, Brassica juncea, Sichuan yellow seed sterile hypocotyls as explants to study the different stages of aseptic explants callus formation, hypocotyl upper, middle and lower segments of the callus, pre-culture medium and differentiation medium, hormone ratio on hypocotyl regeneration. The results showed that 3-5-day sterile formation of callus. Hypocotyl callus on the fastest section of the formation, differentiation rate, reaching 41.33 percent. Under the second, the slowest mid-callus formation, and it will turn brown. Hypocotyl in the MS+6-BA 1 mg/L+2,4-D 1.5 mg/ L pre-culture medium and then switching 2d MS+2,4-D 2 mg/L+6-BA 0.5 mg/L of callus induction medium to induce callus. Interference with the vector containing the TT2 gene Agrobacterium Brassica juncea hypocotyl, differentiation medium MS+6-BA 2mg/ L induced differentiation, transgenic plants with normal morphology, flowering, fruit, T0 plants for seed coat color is yellow. By floral-dip transformation method, harvest a large number of seeds, seed coat color is still yellow, the next step to be detected.Using butanol-hydrochloric acid method of seed coat proanthocyanidins content, Sichuan, seeded by the water bath before the solution was orange, orange remains after the water bath. NILs NILA and NILB and purple leaf mustard, orange solution before bath, water bath, the solution becomes purple. This shows that the purple leaf mustard and near-isogenic seed coat contains cyanidin (cyanidin), or geranium (pelargonidin) pigment derivatives, Sichuan yellow seeds do not contain these substances. Studies show that proanthocyanidins and is closely related to seed coat color.