Transgenic Methods of Chinese Cabbage:Agrobacterium-mediated Transformation by In Vitro Tissue Culture and In Planta Infiltration
|School||Capital Normal University|
|Keywords||Heading Chinese cabbage Chinese cabbage Transgenic Agrobacterium tumefaciens Vitro transformation Penetrate transformed in situ|
The take five kinds of Chinese cabbage leaf explants , eight kinds of experiments used hormone proportion genotypes containing lmg / l of 6- BA , NAA lmg / l, AgNO 3 < / sub > 3mg / l medium can obtain a higher rate of regeneration but the genotypes regeneration ability . Made a preliminary study of a number of factors that may affect the regeneration , the results showed that genotype is the main factor affecting the ability to regenerate ; the AgNO 3 < / sub > added significantly improve the regeneration rate , but in the range of 0.5 ~ 9mg / l , the leaf regeneration rate did not change significantly . Drawn parts of the and drawn time of the renewable . Transformed material , the use of the highest regenerative force genotype \pre- cultured for 3 days, 2-3 days of culture , co-culture medium pH 5.2 optimum transformation conditions . Use good transformation system by Agrobacterium tumefaciens -mediated transformation, the TuMV-NIa and LMV-CP , two anti- virus genes into heading Chinese cabbage , PCR and PCR-Southern blot confirmed that the purpose of the gene has been integrated into the plant genome . Although GUS expression rates of up to about 60%, but the ultimate objective gene transformation frequency of only 0.67%. Discussed the huge difference between the two , and put forward recommendations to solve this problem . Infiltrated by in situ conversion of cabbage inflorescence incision and inflorescence and ultimately anti Basta plants from six batches of screening T 1 < / sub > generation seed . PCR and PCR-Southern blot confirmed that the purpose of the gene has been integrated into the cabbage genome . Pumping the removal of the vacuum process , so that the conversion operation tends to be more simplified, but further studies are needed to improve the conversion efficiency .