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Wheat mature embryo callus gene gun genetic transformation method

A genetic transformation method and callus technology, applied in the field of agricultural biology, can solve the problems of difficult induction, low transformation efficiency, poor regeneration characteristics, etc., and achieve the effect of improving the efficiency of genetic transformation and molecular breeding of wheat, and high genetic transformation efficiency

Inactive Publication Date: 2014-04-09
NORTHWEST A & F UNIV
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Problems solved by technology

[0003] In view of the defects or deficiencies in the above-mentioned prior art, the object of the present invention is to provide a method for efficiently inducing, regenerating and transforming wheat mature embryo callus. And the optimization of the process of cultivation and transformation technology has obtained technical effects similar to those of wheat immature embryo callus induction, regeneration and transformation efficiency, and overcomes the difficulty in induction of mature wheat embryo callus, poor regeneration characteristics, low transformation efficiency and the transformation of receptors. Defects that tissue sampling is affected by growing seasons and time periods

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  • Wheat mature embryo callus gene gun genetic transformation method
  • Wheat mature embryo callus gene gun genetic transformation method

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Embodiment Construction

[0042] This embodiment provides a method for gene gun genetic transformation of mature wheat embryo callus, comprising the following steps:

[0043] 1) Selection and pretreatment of mature embryos:

[0044] Select wheat seeds with full grains that were harvested or stored for about one year in the current year, washed twice with sterile water, disinfected with 70% alcohol for 5 minutes under aseptic conditions, and then soaked in 0.1% mercuric chloride solution. Soak the seeds in medium for 15 minutes, rinse with sterile water 4 times;

[0045] The sterilized wheat seeds were pretreated in 10mg / L thiadiazole urea (TDZ) solution at room temperature of 25°C for 15h;

[0046] 2) Inoculation and callus induction:

[0047] The pretreated wheat seeds were sterilized again with 0.1% mercuric chloride solution for 10 minutes, washed 4 times with sterile water, stripped mature embryos under aseptic conditions, and inoculated on mature embryo callus induction medium to induce callus ...

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Abstract

The invention discloses a wheat mature embryo callus gene gun genetic transformation method. By pretreatment of wheat mature embryo culture, addition and organic matters, and optimization of cultivation and transformation technology processes, technical effects which are close to wheat young embryo callus induction, regeneration and transformation efficiencies are obtained. Compared with reported mature embryo callus gene gun genetic transformation technologies, the method has higher transformation efficiency, the induction, differentiation and transformation capabilities of the wheat mature embryo callus are close to that of young embryo callus, and technical problems of low induction rate, regeneration frequency and transformation efficiency of callus in previous wheat mature embryo callus gene gun genetic transformation are solved. Wheat genetic transformation is free from seasons and periods of recipient material selection. High-efficiency transformation study on mature embryos of seeds harvested in the current year or stored for one year in any time is possible. Wheat genetic transformation and molecule breeding efficiencies are raised.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and in particular relates to a gene gun genetic transformation method of mature wheat embryo callus. Background technique [0002] Wheat is one of the important food crops. Since Vasil et al. introduced the GUS / Bar gene into the wheat immature embryo callus in 1992 and obtained the first transgenic wheat through the regeneration of transformed cells, the research on transgenic wheat has made great progress. Although wheat Many transgenic methods have been reported, but microprojectile bombardment is still the most effective technology for genetic transformation of wheat. At present, the main recipient of wheat biolistic genetic transformation is the callus induced by wheat immature embryos about 13 days old, but the source of this important explant is affected by the wheat growth season and time period, so it is very inconvenient to apply. Wheat mature embryos mainly come from harvested...

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Application Information

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IPC IPC(8): C12N15/82A01H4/00
Inventor 陈耀锋王丽李春莲
Owner NORTHWEST A & F UNIV
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