Agrobacterium tumefaciens-mediated sugarcane callus efficient genetic transformation method

A genetic transformation method, Agrobacterium-mediated technology, applied in the field of high-efficiency genetic transformation of sugarcane callus mediated by Agrobacterium, can solve the problems of difficulty in improving the overall transformation efficiency, plant callus damage and death, increased workload, etc., to achieve improved The effects of survival rate and differentiation ability, improvement of bacterial transformation activity, and improvement of accuracy
CN114774464AActive Publication Date: 2022-07-22SUGARCANE RES INST OF YUNNAN ACADEMY OF AGRI SCI
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
SUGARCANE RES INST OF YUNNAN ACADEMY OF AGRI SCI
Publication Date
2022-07-22

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Abstract

The invention discloses an efficient genetic transformation method for agrobacterium-mediated sugarcane calluses, and belongs to the field of tissue culture. According to the invention, by enhancing the Agrobacterium tumefaciens Vir gene induction activation level and the bacterial liquid dip dyeing strength of sugarcane calluses, the occurrence frequency of transgenic events is greatly improved; an antioxidant is added into subsequent co-culture, screening and differential culture media to inhibit callus browning caused by agrobacterium-enhanced dip dyeing, so that the survival rate and the differentiation capacity of the dip-dyed calluses are obviously improved; a double-marker screening system with a green fluorescent protein marker EGFP gene and a herbicide glufosinate-ammonium resistance marker Bar gene at the same time is adopted, positive transgenic calluses are visually and nondestructively screened by using an EGFP marker, positive transgenic differentiated seedlings are screened by using Bar marker resistance, the accuracy of transgenic event screening is remarkably improved, and the screening efficiency is improved. And finally, the efficient genetic transformation of the agrobacterium tumefaciens mediated sugarcane callus is realized.
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Description

technical field

[0001] The invention relates to the field of tissue culture, in particular to a method for efficient genetic transformation of sugarcane callus mediated by Agrobacterium. Background technique

[0002] Sugarcane is the most important sugar crop and an important source of bioethanol in my country and the world. The continuous genetic improvement of sugarcane varieties is the foundation and key to the healthy development of the industry. However, sugarcane is a highly heterozygous allopolyploid and aneuploid vegetatively propagated crop with complex genetic background and huge genome, making conventional cross-breeding very difficult. At the same time, problems such as difficulty in inducing flowering of sugarcane parents and lack of flowering period further limit the efficiency of trait improvement in cross-breeding. In contrast, genetic engineering breeding can achieve targeted genetic improvement of plant traits without causing large-scale genetic recombina...

Claims

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