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Method for Agrobacterium-mediated genetic transformation of sugarcane growth points

An Agrobacterium-mediated, genetic transformation technology, applied in the field of Agrobacterium-mediated genetic transformation of sugarcane growth points, can solve problems such as shortening and callus regeneration difficulties, and achieve the goals of shortening time, improving infectivity, and improving transformation efficiency Effect

Active Publication Date: 2020-11-06
GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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Problems solved by technology

[0003] The purpose of the present invention is to provide a method for the genetic transformation of sugarcane growth points mediated by Agrobacterium, solve the technical problem of difficult callus regeneration after Agrobacterium transformation in the existing sugarcane cultivation, shorten the time for obtaining transformed plants, and improve the transformation efficiency

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  • Method for Agrobacterium-mediated genetic transformation of sugarcane growth points

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[0038] In order to make the object, technical solution and advantages of the present invention clearer, preferred embodiments are given to further describe the present invention in detail. However, it should be noted that many of the details listed in the specification are only for readers to have a thorough understanding of one or more aspects of the present invention, and these aspects of the present invention can be implemented even without these specific details.

[0039] A method for Agrobacterium-mediated genetic transformation of sugarcane growing points, said method comprising the steps of:

[0040] 1.1 Plant materials, strains, plasmids

[0041] The material for genetic transformation is the self-cultivated sweet variety ‘Guitang 35 (GT35)’ of the Institute of Sugarcane, Academy of Agricultural Sciences, Guangxi Zhuang Autonomous Region. Escherichia coli strain JM109, Agrobacterium tumefaciens strain EHA105 and plasmid pCAMBIA3301, which are preserved in our laborato...

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Abstract

The invention discloses a method for Agrobacterium-mediated genetic transformation of sugarcane growth points, and belongs to the technical field of sugarcane cultivation. The method comprises the following steps: selecting a genetically transformed sugarcane material, a strain and plasmid, preparing a callus induction medium M1, a differentiation medium M2, a rooting medium M3, a transformation medium MR and a co-medium M4, inducing and differentiating sugarcane callus, preparing an Agrobacterium engineering bacterial solution, using an Agrobacterium-mediated method to perform genetic transformation on the sugarcane growth points, and testing transgenic plants. The method shortens the time for sugarcane to obtain transformed plants, and improves the transformation efficiency. Through theAgrobacterium-mediated transformation technology, a pCAMBIA3301 plant expression vector is introduced into GT35 sterile seedlings, a total of 353 resistant plants are obtained after screening by a screening medium, the resistant plants are tested by PCR, 333 plants are positive, a positive rate reaches to 94.33%, and thereby the result indicates that the foreign gene is successfully introduced into sugarcane.

Description

technical field [0001] The invention relates to the technical field of sugarcane cultivation, in particular to a method for genetic transformation of sugarcane growth points mediated by Agrobacterium. Background technique [0002] Since sugarcane is an aneuploid allopolyploid crop, the genetic background is complex, and the offspring of hybrids between varieties are widely separated. It is difficult to clarify its genetic law by conventional genetic methods, and it is difficult to combine various excellent targets by conventional cross-breeding methods. Characters are integrated, and molecular breeding is an important means to obtain superior varieties of sugarcane. At present, the main methods used in the genetic transformation of sugarcane are Agrobacterium-mediated method and particle gun bombardment method. Whether it is Agrobacterium-mediated method or gene gun bombardment method, the recipients of the transformation are embryogenic callus. These traditional methods al...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84A01H4/00A01H5/00A01H6/46
CPCA01H4/001A01H4/008C12N15/8205C12N15/8274
Inventor 秦翠鲜黄东亮陈忠良汪淼廖芬周丽李傲梅周建辉
Owner GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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