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Method for obtaining transgene wheat by using gene gun, and special culture medium

A technology of transgenic wheat and basic medium, applied in horticultural methods, botanical equipment and methods, applications, etc., can solve the problems of somatic cell variation, long transformation cycle, low transformation efficiency, etc., and achieve broad application prospects, transformation cycle Short, transformation stable and reproducible effects

Active Publication Date: 2014-07-02
SUZHOU QI BIODESIGN BIOTECHNOLOGY CO LTD
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AI Technical Summary

Problems solved by technology

[0003] Although some foreign companies and laboratories have been able to use the gene gun method to obtain stable genetically modified wheat so far, they generally have low transformation efficiency, long transformation cycle, many somatic cell variations, and serious escape phenomena, which are only applicable to "Bobwhite" and "Fielder". Technical defects and deficiencies of foreign varieties

Method used

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  • Method for obtaining transgene wheat by using gene gun, and special culture medium
  • Method for obtaining transgene wheat by using gene gun, and special culture medium
  • Method for obtaining transgene wheat by using gene gun, and special culture medium

Examples

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Embodiment 1

[0050] Embodiment 1, utilize gene gun to obtain transgenic wheat

[0051] Using the young embryos of wheat varieties Shi 4185, Zhongmai 1, Jinmai 45, Luohan 7, Jishi 02-1, Yangmai 19, and Kenong 199 as explants, the bar gene The plasmid pAHC20 was transferred into wheat cells, and the transformation method and results were as follows:

[0052] 1. Preparation of culture medium

[0053] 1. Hypertonic medium: add 90g / L mannitol, 5.0mg / L 2,4-D and 30g / L sucrose to the MS basic medium (Table 1) to adjust the pH to 5.8. Add phytogel at a final concentration of 3 g / L, and autoclave at 121° C. for 20 minutes to make a solid medium.

[0054] 2. Callus induction medium: Add 2,4-D with a final concentration of 2.0mg / L, 0.5mg / L copper sulfate, and 0.5g / L hydrolyzed casein to MS basic medium (Table 1), 30g / L sucrose, adjust the pH to 5.8, add plant gel with a final concentration of 3g / L, and autoclave at 121°C for 20min to make a solid medium.

[0055] 3. Differentiation medium: Add ki...

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Abstract

The present invention discloses a method for obtaining transgene wheat by using a gene gun, and a special culture medium. According to the present invention, wheat immature embryo is adopted as explant, a hypertonic culture medium provided by the present invention is directly used to carry out a hypertonic treatment without pre-culture, gene gun bombardment is performed, and the special culture medium provided by the present invention is adopted to carry out a hypertonic treatment, callus induction, differentiation and rooting culture after bombardment, wherein a screening agent is used during the differentiation and rooting stages and is not used during the callus induction state so as to significantly improve the wheat gene transformation efficiency, wherein the wheat gene transformation efficiency can be up to 36%; the method has advantages of short transformation period (the time from immature embryo taking to obtaining of the regenerated plant having root, stem and leaf is only 8-10 weeks), high transformation stability and high repeatability; and the effective approach is provided for wheat genetic improvement industrialization, and the broad application prospects are provided.

Description

technical field [0001] The invention relates to a method for obtaining transgenic wheat by using a gene gun and a special culture medium thereof. Background technique [0002] Wheat is an important food crop widely grown all over the world, and it is related to the economic crisis and food security of the country. China is the world's largest wheat producer. In 2010, after corn and rice, the output of wheat in China reached 2.24 million tons. At present, efforts to improve wheat yield and quality through conventional breeding methods have been difficult, especially in terms of tapping the potential of yield per mu and ensuring the stability of annual yield. Since Vasil et al. introduced the gus / bar gene into wheat and obtained the first transgenic wheat in 1992, the research on genetic improvement and breeding in wheat has made great progress. In 2008, the state added the genetic improvement of wheat to major transgenic projects, and is actively and steadily promoting the...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N5/04A01H4/00A01H5/00
Inventor 高彩霞张康刘金星
Owner SUZHOU QI BIODESIGN BIOTECHNOLOGY CO LTD
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