Non-embryo transgenic process for preparing transgenic cotton

Abstract

The present invention provides a kind of transgenic cotton preparing process. The process includes a bacterially culturing cotton seedling, adopting the hypocotyl piece of the seedling with the growing point retained as the explant, longitudinally cutting the growing point, infecting the explant with agrobacterium carrying exogenous gene, organ differentiating to form test tube plantlet, and grafting transplantation. The process features that the agrobacterium infected cell or tissue is cultured into seedling directly through organ differentiation without forming embryonic tissue. The present invention can shorten the exogenous gene transforming period of cotton obviously, and has no limitation from the genotype of acceptor material, high transforming efficiency, simple operation, low cost and few mutations except that caused by the exogenous gene.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a method for genetic transformation of cotton. Background technique [0002] Agrobacterium is a Gram-negative bacterium ubiquitous in soil, which can chemotactically infect the injured parts of most dicotyledonous plants under natural conditions and induce crown galls or hairy roots. There is a section of T-DNA in the cells of Agrobacterium tumefaciens and Agrobacterium rhizogenes. After Agrobacterium enters the cells by infecting plant wounds, it can insert T-DNA into plant genes. Therefore, Agrobacterium is a natural plant genetic transformation system. People insert the target gene into the modified T-DNA region, and realize the transfer and integration of the exogenous gene to the plant cell through the infection of Agrobacterium. Transgenic plants are then regenerated through cell and tissue culture techniques. Agrobacterium-mediated transformation...

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Problems solved by technology

[0003] However, the conventional Agrobacterium-mediated transformation method has its disadvantages, as it must go through dedifferentiation and redifferentiation of cells to form embryogenic tissues and embryogenic organs (embryogenic callus, embryoid bodies and somatic embryos). process, its result is: (1) cycle is long, conventional method from the Agrobacterium infection explant with exogenous gene to test-tube seedling transplanting and surviving generally needs 8~12 months, before two years switch to test-tube seedling grafting, other The cycle still needs 6 to 10 months; (2) The receptor material is limited, that is, there is a so-called "genotype" restriction problem; (3) The variation other than the foreign gene is large, that is, in addition to the expression of the foreign gene, some It is derived from the variation in the process of tissue culture (mainly callus formation, cell dedifferentiation, redifferentiation, etc.)

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