Method for carrying out agrobacterium tumefaciens mediated banana genes by employing liquid co-culture system

Abstract

The invention discloses a method for converting the meloidogynosis agrobacterium lead banana gene with the fluid co culture system, which comprises the following steps: stewing the banana ECS in the meloidogynosis agrobacterium and the black condition for a spell of time; proceeding with the co culture in the banana ECS liquid culture medium; transferring the banana ECS to the idiosome directly in order to induce and grow; culturing on the selective culture medium; transferring the mature resistant idiosome to the blastema germination culture medium; culturing in the light-black alternative condition until the idiosome is germinated; getting sprout; transferring the sprout to the basis screen selective culture medium; acquiring the whole conversion plant by culturing in the light-black alternative condition. The invention changes the co culture condition, which solves the brown problem of banana ECS in the co culture process, and shortens the cycle of converting the plant.

Description

technical field [0001] The invention belongs to the technical field of banana genetic transformation, in particular to a method for utilizing a liquid co-cultivation system to mediate banana gene transformation by Agrobacterium tumefaciens. Background technique [0002] At present, pests and diseases are becoming more and more rampant, banana production is facing a severe test, and there is an urgent need to cultivate new varieties with good quality and disease resistance. Because cultivated bananas are highly sterile and have no seeds, it is difficult to obtain new varieties through traditional cross-breeding methods, and using genetic engineering technology to obtain disease-resistant transgenic bananas is one of the effective breeding methods. The establishment of a stable and efficient banana gene transformation system suitable for all genotypes is the premise of banana genetic engineering breeding. Current research shows that banana ECS is the most ideal transformation...

AI Technical Summary

Problems solved by technology

Current research shows that banana ECS is the most ideal transformation recipient, but some banana varieties cannot be genetically improved due to the deficiencies of existing technologies. Agrobacterium tumefaciens mediates exogenous gene transformation into banana embryogenic cell suspensions, The prior art of ECS) is: after the banana ECS is soaked in the Agrobacterium tumefaciens bacterium liquid, transfer to the semi-solid medium that has added acetosyringone and co-cultivate for 3 or 6 days, the Agrobacterium tumefaciens infection after Banana ECS is inoculated on the semi-solid proliferation medium supplemented with corresponding antibiotics for screening of transformed cells. After culturing for about 2 to 3 months, the screened resistant clones are transferred to somatic embryo induction and development culture supplemented with corresponding antibiotics. On the base, transfer the mature resistant somatic embryos to the somatic embryo germination medium, and finally transfer the germinated somatic embryos to the rooting medium supplemented with the corresponding antibiotics for rooting selection and culture to obtain complete transgenic plants

There are mainly two deficiencies: (1) ECS of some banana varieties and Agrobacterium tumefaciens are co-cultured on semi-solid medium, which is very easy to brown and cause transformation failure; (2) after co-cultivation, Agrobacterium tumefaciens Bacillus-infected banana ECS was first inoculated on semi-solid proliferation medium for screening of transformed cells, prolonging the cycle of obtaining transformed plants

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