Improved transforming method of wheat young ears mediated by agrobacterium tumefaciens and application thereof
An Agrobacterium-mediated, wheat-based technology, applied in the fields of agricultural biology and plant genetic engineering, to achieve the effect of making up for the cumbersome transformation operations
Inactive Publication Date: 2011-11-02
SHANDONG AGRICULTURAL UNIVERSITY
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[0013] The technical effects of the present invention can overcome the above-mentioned defects, and provide a simple and easy method for high-efficiency transgenic wheat, which makes up for the shortcomings of the current cumbersome transformation operations of wheat
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[0019] The present invention will be described in detail below in conjunction with specific embodiments.
[0020] Wheat Antisense TaCKX1 Gene Introduction
[0021] 1.1 Experimental materials and expression vectors
[0022] Materials: Wheat Jimai 20.
[0023] Vector: a plant expression vector with antisense TaCKX1 gene, and the screening marker gene is NptII gene.
[0024] 1.2 Test method
[0025] 1.2.1 Gene introduction
[0026] The wheat transgenic operation was carried out in the experimental area.
[0027] The transformation period is about 10-15 days before wheat heading, when the flag leaves of wheat are all pulled out, and the distance between leaf rings is about 2-5 cm. At this time wheat is in the meiosis of flower development. Young spikes enclosed in flag leaves. The whole ear injection process can be carried out in the field or in the greenhouse without operating under sterile conditions.
[0028] Activation and resuspension of Agrobacterium containing the t...
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The invention relates to an improved transforming method of wheat young ears mediated by agrobacterium tumefaciens, belonging to the field of agricultural biotechnology and plant gene engineering. The method provided by the invention mainly comprises the following steps of: after planting the wheat, selecting an appropriate transforming period, wherein the transforming period is selected to be about 10-15 days before the head sprouting period of the wheat when flag leaves of the wheat all spring up and the annular distance of the leaves is about 2-5 cm; activating and resuspending agrobacterium tumefaciens bacterial liquid containing the target genes and then injecting the liquid into young ears covered by the flag leaves, wherein the whole young ear injection process can be carried out in a land for growing field crops or a greenhouse and does not need to be operated in an aseptic condition. The method provided by the invention has low technical requirements on the operators, is easyto be mastered and can be used for simply, fast and efficiently obtaining a transgenic plant; the method provided by the invention has the advantages of excluding the influence of the somaclonal variation on the transgenic plant, overcoming the limitation of the wheat gene type on the transformation of the agrobacterium tumefaciens and having important effects and a wide application prospect on identification of the wheat gene function and genetic engineering breeding.
Description
technical field [0001] The invention relates to an improved Agrobacterium-mediated wheat ear transformation method and application thereof, belonging to the fields of agricultural biotechnology and plant genetic engineering. Background technique [0002] Since the 1990s, the application of transgenic technology in crop genetic improvement has become a focus of attention. Transgenic technology can break the boundaries of species and realize the transfer of genes between different species. [0003] At present, the commonly used wheat transgenic methods can be roughly divided into two categories: one is the transformation method that does not depend on tissue culture, such as pollen tube passage method, microinjection method, etc.; the other is the transformation method that relies on tissue culture, including gene gun method, Agrobacterium-mediated method, electroporation method, PEG method, etc. The most widely used methods are particle gun method and Agrobacterium-mediated...
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Inventor 封德顺王洪刚田纪春马信尹娜张盈
Owner SHANDONG AGRICULTURAL UNIVERSITY