Method for improving cotton transgenosis efficiency

Abstract

A provided method for improving cotton transgenosis efficiency comprises the following steps: a, employing an agrobacterium-mediated method to infect cotton aseptic-seedling shoot tips with a plant expression carrier containing a target gene, and placing the infected cotton shoot tips in a co-culture medium for culture; and b, after co-culture is performed, transferring the cotton shoot tips to a selective medium for culture. The co-culture medium and the selective medium both contain KT with a concentration of 0.3 mg / L and IAA with a concentration of 0.2 mg / L. By utilizing the method, the aseptic seedling culture time is 2-3 days, the co-culture time is 2 days, the selective culture time is 15-20 days, the rooting culture time is 20 days, the conversion period does not exceed 2 months, and the positive conversion rate reaches 8% after screening is performed.

Description

technical field [0001] The invention relates to the technical field of cotton transgenes, in particular to a method for improving the efficiency of cotton transgenes. Background technique [0002] Significant progress has been made in the genetic improvement of cotton using genetic engineering technology. In 1987, Umbeck et al first reported the transfer of kanamycin-resistant gene into cotton by Agrobacterium-mediated method (Umbeck P. Genetically transformed cotton (Gosspium hirsutum L.) Plants. Bio / Teehnology, 1987, 5: 263-266). The traditional genetic transformation method has a long transformation period, strong genotype dependence of plant regeneration, high deformity rate of regenerated seedlings, and low survival rate of transplanting, which has become a bottleneck restricting the research of cotton transgenic breeding. [0003] Cotton shoot tips have the characteristics of direct differentiation and regeneration, once transformed, the formation of leaves and roots ...

AI Technical Summary

Problems solved by technology

However, this technical solution is mainly an in vitro shoot tip transformation method, without the process of cotton cell and tissue culture, the probability of chimerism increases, and the inheritance cannot be stable, and the offspring do not conform to the Mendelian inheritance law

However, many cotton transgenic methods (such as by hypocotyl transformation) through cell and tissue culture, because the hypocotyl will be induced and differentiated through callus, the transformation cycle is longer, generally 6-9 months ( Liu Mingyue, Optimization of Agrobacterium-mediated Cotton Genetic Transformation System, 2011, page 9), and is limited by the genotype of the transformed recipient

There are also gene gun transformation methods in which the shoot tip is the receptor, but compared with the Agrobacterium transformation method, the cost is high, and transient expression and chimerism occur

There are other Agrobacterium-mediated shoot tip transformation methods that show that it takes 3-4 months from co-cultivation to obtain resistant strains, which is a long period (Xu Dawei et al., 2011, China Agricultural Science Bulletin)

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