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Method for directly transforming cotton seed embryos by utilizing agrobacterium tumefaciens

A cotton seed, Agrobacterium tumefaciens technology, applied in the fields of botanical equipment and methods, applications, angiosperms/flowering plants, etc., can solve the problems such as the limitation of plant quality improvement by transgenic technology, the high incidence of deformed seedlings, and the complicated transformation methods. , to achieve the effect of low survival rate, high incidence rate and easy operation of direct transplanting

Inactive Publication Date: 2011-11-02
COTTON RES INST SHANXI ACAD OF AGRI SCI
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Problems solved by technology

However, due to the limitations of plant transgenic methods and receptor materials, the use of transgenic technology to improve plant quality has been limited, especially for some difficult-to-transform plant materials such as cotton.
At present, in the process of cultivating new cotton varieties using transgenic technology, there are problems such as cumbersome and inefficient transformation methods, high incidence of deformed seedlings, and low survival rate of transplanting.

Method used

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  • Method for directly transforming cotton seed embryos by utilizing agrobacterium tumefaciens
  • Method for directly transforming cotton seed embryos by utilizing agrobacterium tumefaciens
  • Method for directly transforming cotton seed embryos by utilizing agrobacterium tumefaciens

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Embodiment 1

[0027] Embodiment 1, the Agrobacterium genetic transformation of cotton

[0028] 1. Construction of recombinant Agrobacterium tumefaciens containing target DNA

[0029] The target gene TaNHX2 (Genbank No. AY040246, Sequence Listing Sequence 1) was recombined into the plant binary expression vector pBin438 (Li Taiyuan, Tian Yingchuan, Qin Xiaofeng, etc. Research on highly efficient insect-resistant transgenic tobacco [J]. Chinese Science (Series B), 1994, 24 (3): 276-282.) Between the Sal I and Kpn I restriction sites behind the CaMV 35S promoter, the recombinant vector pTaNHX2 ( figure 1 ). The selectable marker gene in pTaNHX2 is the kanamycin resistance gene NPT II.

[0030] pTaNHX2 was transformed into Agrobacterium tumefaciens GV3101 (Agrobacterium tumefaciens) to obtain recombinant Agrobacterium tumefaciens GV3101 / TaNHX2 containing TaNHX2.

[0031] 2. Preparation, inoculation and co-cultivation of explants

[0032] Take Jinmian No. 7 (Jinmian No. 7 was bred by the Lin...

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Abstract

The invention discloses a method for directly transforming cotton seed embryos by utilizing agrobacterium tumefaciens. The method provided by the invention is used for transforming target DNA (Deoxyribonucleic Acid) into a cotton cell through utilizing mediation of meloidogynosis agrobacterium tumefaciens and comprises the following steps of: (1) inoculating a recombinant meloidogynosis agrobacterium tumefaciens strain containing the target DNA into a damaged sections of the cotton seed embryos which do not have seed skin and are scratched; then carrying out co-culturing on the inoculated cotton seed embryos to integrate the T-DNA (Transfer-Deoxyribonucleic Acid) of the recombinant meloidogynosis agrobacterium tumefaciens with the chromosomes of the cotton; and (2) directly inoculating the co-cultured seed embryos obtained in the step (1) into a seedling growing culture medium to be cultured to obtain a seedling with roots, stems and cotyledons. The method provided by the invention solves the problem of the limitation of the explant type in the cotton genetic transformation, can be used for reducing the incidence rate of deformed seedlings and improving the transplanting survival rate and has the advantages of simpleness and convenience for operation, short transforming period, low cost, high transforming efficiency and higher application value.

Description

technical field [0001] The invention relates to a method for directly transforming cotton seed embryos by using Agrobacterium. Background technique [0002] Since the first transgenic plant came out in 1983, transgenic plant research has played a huge role in cultivating disease-resistant plants, insect-resistant plants, improving crop quality and yield, and improving crop stress resistance. However, due to the limitations of plant transgenic methods and receptor materials, the use of transgenic technology to improve plant quality has been limited, especially for some difficult-to-transform plant materials such as cotton. At present, in the process of cultivating new cotton varieties using transgenic technology, there are problems such as cumbersome transformation methods and low efficiency, high incidence of deformed seedlings, and low survival rate of transplanting. Contents of the invention [0003] The purpose of the present invention is to provide a method for cultiv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00
Inventor 张慧军武宗信石跃进韩满仓王丹
Owner COTTON RES INST SHANXI ACAD OF AGRI SCI
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