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Wheat mature embryo tissue culture regeneration system optimized for high regeneration rate of transgenic genotype screening and preliminary study

Author: ZhangDongWu
Tutor: ZhaoHuiXian
School: Northwest University of Science and Technology
Course: Biochemistry and Molecular Biology
Keywords: Wheat Mature embryos Tissue culture Regeneration System Gene gun Genetic transformation
CLC: S512.1
Type: Master's thesis
Year: 2011
Downloads: 129
Quote: 1
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Abstract


Wheat is the most important food crops in China and one of the world on the use of transgenic technology for targeted improvement of wheat has attracted people's attention, it will gradually become a high-quality, high-yielding new varieties of wheat cultivation effective auxiliary ways. With rice, corn and other major food crops compared to wheat tissue culture regeneration difficult, and genetic transformation rate, which greatly retarded the transgenic technology in the cultivation of new varieties of wheat in the application pace. Therefore, improving the rate of tissue cultured wheat and wheat genetic transformation rate is an important foundation for transgenic research work. In this study area in recent years, the Huang-Huai Mai 19 fine wheat varieties and a spring wheat varieties as test materials, the previous studies based on the optimization and establishing a variety of mature embryos of wheat genotypes efficient regeneration system, including improving wheat more wound tissue quality, increase the frequency of dedifferentiation of mature embryos and regeneration, improving Rooting regeneration, etc.; On this basis, to be screened from the tested varieties with high regeneration rate of excellent genotypes; Finally, the high regeneration rate Wheat gene gun-mediated genetic transformation, trying to obtain transgenic plants. This study aimed to Huang-Huai wheat region transgenic research work has laid a good foundation. Made the following key findings: 1. Wheat mature embryo tissue culture regeneration system optimization: a longitudinal half-cut whole embryo callus induction, its frequency is greater than the longitudinal dedifferentiation full cut half-embryos, transverse semi-embryo, and scraping pieces of embryos and callus of good quality; wheat callus from mature embryos in the following three kinds of hormones, namely: B. IAA 1.0mg / L, ZT 2mg / L; E. 2,4 - D 0.1 mg / L, ZT 10mg / L; F. 6-BA 1mg / L, NAA 0.3mg / L, KT 4mg / L of medium differentiation and then generate a higher survival rate; regenerated seedlings without auxin 1/2MS better rooting medium. 2 genotypes with high regeneration rate: In the tested 20 wheat varieties, there are 12 varieties of wheat seedlings regenerated rate of 20% or more, which has three varieties including Xinong 928, 16 and West Ningchun 889 farmers re-generation seedling rate were as high as 77.6%, 52.7% and 36.9%; addition, B genotype differentiation medium for most callus differentiation culture has good versatility. In this study, the Huang-Huai wheat production area of ??wheat varieties were screened for high regeneration rate can be used as the Huang-Huai wheat varieties genetically modified wheat in the area of ??the receptor material. 3 particle bombardment-mediated transgenic wheat mature embryos preliminary study: the use of gene gun-mediated transformation method 928 for Xinong callus from mature embryos for genetic transformation of callus bombarded 1200, and ultimately get 28 regenerated plants ; wherein the gene has four Zm-RS of PCR positive plants are 14 Bar gene PCR positive plants; target gene conversion was 0.33%, Bar gene conversion was 1.2%. Bombarding the target distance 6cm suitable Xinong 928 mature embryos of transgenic research. This study shows that mature embryos Wesfarmers 928 is a very good conversion acceptor material that can be used for wheat genetic engineering.

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CLC: > Agricultural Sciences > Crop > Cereal crops > Wheat > Wheat
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