A method for improving transgenic efficiency of cotton

Abstract

The invention provides a method for improving the transgenic efficiency of cotton. The method comprises the following steps: a. Infecting the shoot tip of the aseptic cotton seedling with the plant expression vector containing the target gene by using the Agrobacterium-mediated method, and the infected Cotton shoot tips are placed on the co-cultivation medium for co-cultivation; b. After co-cultivation, the cotton shoot tips are transferred to the screening medium for cultivation; both the co-cultivation medium and the screening medium contain a concentration of 0.3mg / L of KT and a concentration of 0.2mg / L of IAA. Utilize the method of the present invention to carry out aseptic seedling culture time 2-3 days, co-culture 2 days, screening culture 15-20 days, rooting culture 20 days, transformation cycle is no more than 2 months, and positive transformation rate can reach 8 after screening. %.

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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