Dissertation > Agricultural Sciences > Gardening > Vegetable gardening > Chinese cabbage

Transgenic Methods of Chinese Cabbage:Agrobacterium-mediated Transformation by In Vitro Tissue Culture and In Planta Infiltration

Author DaiDaPeng
Tutor CaoMingQing
School Capital Normal University
Course Biology, genetics
Keywords Heading Chinese cabbage Chinese cabbage Transgenic Agrobacterium tumefaciens Vitro transformation Penetrate transformed in situ
CLC S634.034
Type Master's thesis
Year 2001
Downloads 199
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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 .

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