Studies On Transformation Of Ac/Ds Gene Of Chinese Cabbage By Agrobacterium Tumefaciens-Mediated Methods

The Chinese cabbage is a kind of popular vegetable. Although great progress has been achieved in Chinese cabbage breeding in the recent years, improving its quantity and quality will still be the main task with the increasing population. Searching for interesting genes for breeding by identification and appraisal of plant gene function is of great importance. Studying the mutants to find out the function of related genes is one of the most effective methods in exploring redients the genes. So it is urgent to study the function of gene of Chinese cabbage. The construction of Chinese cabbage mutant library can not only provide us the useful materials in studying the function of the gene, but also bring us Chinese cabbage with new phenotype , important materials and germplasm for genetic breeding. Research results are mainly as follows:1. A high effective regeneration system has been established.Chinese cabbage number 6 of four-days cotyledon, is placed on the polarization medium (MS+5mg/L6-BA+0.5mg/LNAA+2mg/AgNO3). The number of shoot differentiation is best of all.2. The activation tagging of expression vector is built promted by CaMV35S according to plasmid of pUbiTs and pBi121.3. The expression vector of p35STs and pDsBar1300H are transformed into Chinese cabbage number 6 with via Agrobacterium-mediated method, and the system is optimized.(1).The suitable antibiotics and their concentration are studied for the selection of transformants. The suitable concentration of the filter, Kanamicin is 10mg/L, Hygromycin is 5mg/L; the suitable concentration of Carbenicillin disodium salt is 500mg/L.(2). The experiment shows that: The cotyledon with 1~2mm of petiole regeneration frequency is very high; The co-cultivation time is 2 days; the pre-culturing time is 36h.The highest frequency is observed when explants are immersed in bacterium suspension with OD600 value about 0.3 for 5 minutes.4. Transgenic T0 progeny of 30 are checked by PCR and PCR-Southern methods.The results prove the existence of target genes in transformation plants of Chinese cabbage number 6 .