Introduced The Cold-resistance Related Genes AtGoLS3 And CBF3 Into Stylossanthes Guianensis CV. And Arabidopsis Thaliana

Till now, Stylosanthes guianensis cv. is the best tropical pasture in the world said as "the star of the tropical pasture ". This pasture is suitable to grow in warm and moist tropical conditions, in the area of the south of latitude 23 degrees north and annual rainfalls more than 1000 millimeters. It's easily injured by low temperature. It grows well at 25~28℃. The output of seed is seriously affected when the temperature is lower than 19℃. In this research, we introduced the function and regulatory genes related to cold-resistance into Stylosanthes guianensis cv. by genetic engineering techniques. It will be benefit to improve the cold-tolerance and expand the growth scope of Stylosanthes guianensis cv.CBF3,AtGolS3, which relate to cold tolerance, were isolated from Arabidopsis thaliana by RT-PCR. The plant expression vectors of pVKH-35S-CBF3-pA,pVKH-35S-AtGolS3-pA were constructed by inserting CBF3,AtGolS3 into the vector pVKH respectively. Also through vector pRT101, AtGolS3 was inserted to pVKH-35S-CBF3-pA, and constructed the bi-expression vector pVKH-35S-CBF3-pA-35S-AtGolS3-pA, which the two genes have their respective promoter and pA. The recombinant plasmids are introduced into Stylosanthes guianensis cv. and Arabidopsis thaliana by using the Agrobacterium-mediated transformation method.In this paper, the Stylosanthes guianensis cv. Reyan No.5 was used as experiment material. After 4 days on the callus medium, the explants were infected with Agrobacterium tumefaciens and then put them on the callus medium in the dark. The co-cultivated explants were transferred into selective medium, which included carbenicillin 500mg/L cefazolin 200mg/L and hygromycin 40mg/L. The shoots were cut off and transferred into root inducible medium. After 20 days, the rooted plantlets were transferred into pots. Transgenic plants were obtained through Hygromycin resistance, we have...