Method for cultivating starch-content-increased transgenic plant through multi-gene transformation

Abstract

The invention discloses a method for cultivating a starch-content-increased transgenic plant through multi-gene transformation. According to the transgenic plant cultivation method provided by the invention, a selection marker gene, an adenosine diphosphate glucose pyrophosphorylase small subunit gene, an adenosine diphosphate glucose pyrophosphorylase large subunit gene, a sucrose synthase gene, and a granule-bound starch synthase gene are introduced into a target plant, such that a transgenic plant is obtained. The total starch content of the transgenic plant is higher than that of the target plant. As a result of experiment of the invention, a plurality of genes can be simultaneously transferred in one-time through one time of gene gun bombardment, such that transgenic material cultivation period and work load can be greatly shortened. Specifically, the 4 genes related to starch metabolism are simultaneously transferred in, such that the total starch content of a transgenic regenerated plant is higher than that of wild-type plant.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for cultivating transgenic plants with increased starch content by using multigene transformation. Background technique [0002] In 1977, Ackermann et al first used Ri plasmid to transform tobacco cells to obtain regenerated plants, creating a history of plant transgenics. Since then, a variety of technologies have been invented and applied to plant transgenesis, mainly including Agrobacterium-mediated method, gene gun method, protoplast method and pollen tube passage method, as well as virus-mediated method, electric shock method, microinjection method, Sonophoresis, imbibition, pollen carrying method, liposome method, laser microbeam puncture method, ion beam method, etc., by applying these methods, a large number of plant species have been successfully transformed and transgenic plants have been produced. These transformation methods generally introduce a target gene int...

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Problems solved by technology

[0003] In order to improve the yield or other agronomic traits of crops, it is often necessary to introduce several genes into recipient plants. The common transgenic method can only adopt the strategy of transferring one gene each time in multiple times, and the transformation efficiency is low. Therefore, Some multi-gene transformation methods have been produced. One is to connect multiple gene expression cassettes in series to the T-DNA region when constructing the transformation vector. This technology is relatively cumbersome to construct vectors. Due to the limitation of the capacity of Ti plasmids, it is impossible to construct sequences containing more than 50Kb of inserts. The second is to construct different exogenous genes into the T-DNA regions of different expression vectors, and then transfer the expression vectors containing different genes into the same Agrobacterium cell for multi-gene transformation. The ability of Agrobacterium to accommodate multiple plasmids needs to be considered

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