Method for establishing agrobacterium tumefacien mediated castor genetic transformation system
A genetic transformation system and Agrobacterium-mediated technology are applied in the field of establishing an Agrobacterium-mediated castor genetic transformation system, which can solve the problems of in-depth research on castor transgenic, useless finding, and failure to establish a genetic transformation system, and the cost is achieved. Low cost, easy operation, simple process effect
Inactive Publication Date: 2012-06-27
INNER MONGOLIA UNIV FOR THE NATITIES
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Problems solved by technology
This method is simple and easy, and avoids the cumbersome process of tissue culture, but its disadvantage is that most of the obtained transgenic plants are chimeras
It can be seen that the research on castor-bean transgenesis is still not in-depth, and an efficient and stable genetic transformation system has not been established, let alone a relatively mature method for establishing a castor-bean genetic transformation system.
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[0016] The general idea of the present invention is: aiming at the problems existing in the prior art, the castor cotyledon leaf nodes that have not been used in castor bean transgenic experiments are used as receptors instead, mediated by Agrobacterium, and kanamycin is used as a screening agent , using cephalosporin as a fungicide, through pre-cultivation, Agrobacterium infection, co-cultivation, fungus removal, screening and bud rooting, etc., to obtain castor plants with heritable target genes, especially castor plants that can be used for transgenic research material system.
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The invention relates to a plant genetic transformation method in the field of bioengineering, and relates to a method for establishing an agrobacterium tumefacien mediated castor genetic transformation system. The method is characterized in that castor cotyledonary nodes are adopted as acceptors; culturing, agrobacterium tumefacien infecting, co-culturing, bacteria separating, screening and seedling rooting are carried out, such that castor progenies with descendible target genes are obtained. The method provided by the invention is advantaged in simple procedure, optimized parameters, convenient operations, and low cost. With the method, a castor genetic transformation system can be established in a relatively short period of time. According to PCR detections, the effects of the obtained resistant seedlings are good. Therefore, a basis is established for simple and highly efficient castor transgene researches.
Description
technical field [0001] The invention relates to a plant gene transformation method in the field of bioengineering, that is, a method for establishing an Agrobacterium-mediated castor plant genetic transformation system. Background technique [0002] There are very few reports on castor transgenic research at home and abroad. Sujatha M et al. reported for the first time in 2005 that the Agrobacterium-mediated method was used to genetically transform castor hypocotyls, and the herbicide resistance gene bar was constructed on the expression vector. Using the herbicide Basta as a screening agent, transgenic castor plants were obtained. MalathiB et al. reported in 2006 that transgenic plants were obtained using the same method. The disadvantage of this technique is that it takes a long time. Mckeon T A etc. applied for a patent in 2003, and infected the whole plant, bud tip and hypocotyl, flower bud of castor with Agrobacterium. Wherein: the transformation of the whole plant i...
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Inventor 黄凤兰陈永胜孟凡娟李国瑞彭木陈丽丽
Owner INNER MONGOLIA UNIV FOR THE NATITIES