Study On Genetic Transformation Of BcDREB1 Gene In Chinese Cabbage

Chinese cabbage is one vegetables originated from our country, occuping an important position in annual supplies of vegetable. With the development of biotechnology, the improvement of Chinese cabbage varieties using genetic engineering technology has become one of interesting research topics currently. In this experiment the BcDREB1 gene cloned from Chinese cabbage was transferred into Chinese cabbage and Arabidopsis thaliana respectively by agrobacterium-mediated genetic transformation technology. And the transformation plants were obtained, which were verified the resistance function with the BcDREB1 gene of Chinese cabbage. This will lay a foundation for resistance molecular breeding in Chinese cabbage. The main research results are as follows:1 Through analyzing the effects of different genotypes, explant types, hormone combinations and pre-cultivation conditions on the shoot regeneration, the regeneration system with high frequency in Chinese cabbage was established, in which the optimal genotype was inbred line'85-1', the optimal explant was cotyledon with petioles, pre-cultivation condition was 0.5 mg/L 2,4-d 1d, the optimal differentiation medium was MS + 0.7 mg/LTDZ + 0.5 mg/LNAA + 5.0 mg/L AgNO3.2 The genetic transformation system of Chinese cabbage was established and optimized through screening the time with agrobacterium infection, the Acetosyringone concentrations and Kanamycin resistance. During the process of genetic transformation in Chinese cabbbage, we selected the optimal conditions of screening resistance buds with kanamycin concentration of 10mg/L, agrobacterium-mediated infection time of 5 min, co-cultivation of 2d. And we found that it is useful for transformation by adding Acetosyringone with 150 mg/L concentration during the process of co-cultivation.3 Fifteen transgenic plants were identified by GUS-histochemical staining, GUS-PCR detection of T0 transgenic plants obtained from the antibiotics screening. The resistance to salt of Chinese cabbage transgenic plants was significantly increased by testing cold resistance with -5℃and osmotic stress resistance with 250mmol/LNacl. 4 Using Floral-dip method, the gene BcDREB1 of the Chinese cabbage was transferred into Arabidopsis thaliana, Twenty five T0 generation plants and 10 T1 generation plants were obtained by screening the antibiotics, in which 8 transgenic plants were identified by GUS-histochemical staining and GUS-PCR detection and T2 generation seeds were harvested. The function in resistance to salt and cold of Chinese cabbage gene BcDREB1 was verified by testing resistance to the salt and cold in Arabidopsis thaliana transformation plants.