Method for in-situ transgenosis of corn mature embryo

Abstract

The invention relates to a method for in-situ transgenosis of corn mature embryos. The method comprises the following steps: obtaining germs of mature embryos needed for in-situ transgenosis of the corn mature embryos; treating the germs to expose growing points and slightly damaging the growing points; carrying out in-situ transgenosis in the growing points of the corn by utilizing impregnating fluid; coculturing impregnated cells at the growth points and agrobactrium tumefaciens to enable Bar genes to be integrated into corn chromosomes by virtue of T-DNA (triplex-deoxyribonucleic acid) integration of the agrobactrium tumefaciens and rejuvenating the Bar genes; carrying out herbicide screening to initially obtain a herbicide resistant plant for in-situ transgenosis and transplanting the plant; obtaining genetically modified corn seeds by selfing pollination of a PCR (polymerase chain reaction) positive plant of the in-situ genetically modified corn. According to the method provided by the invention, transgenosis is carried out on the growth points of the corn mature embryos and the method of spraying or coating herbicide on corn lamina to screen transgenic plants and carrying out molecular detection to further verify the PCR positive plants is adopted. The method provided by the invention is simple and efficient and can secure a conversion rate of 4%, thus being of great significance for genetic engineering and inheritance breeding of corn.

Description

technical field [0001] The invention relates to a method for transgenicing plants in genetic engineering, in particular to a method for transgenicing mature corn embryos, in particular to a method for in situ transgenic corn mature embryos. Background technique [0002] Transferring exogenous genes or modified endogenous genes into plants to change a certain characteristic of plants is an important way to improve crop quality. Since the world's first transgenic plant, the transgenic tobacco, was first bred in 1983, scientists have tried different transgenic methods, introducing exogenous genes or endogenous genes modified in vitro into plants, improving plant traits, and explored A variety of plant transgenic methods. At present, there are two main types of gene transfer methods in maize: genetic transformation mediated by Agrobacterium tumefaciens and transformation methods of direct DNA introduction, the latter mainly including gene gun method, pollen tube passage method,...

AI Technical Summary

Problems solved by technology

However, using mature corn embryos as materials, that is, transgenic corn mature embryos, there are few reports of transgenic development by obtaining embryo tips, bud points, etc.

The former is mostly limited by time, and the operation is cumbersome. It needs to rely on the mature corn regeneration system, so the cost is high and the cycle is long.

Compared with the former, the latter method is simple to operate and is not greatly affected by the seasons. It can be carried out throughout the year and is a better method for transgenic corn. Low, many chimeras, and low seed setting rate

Images