利用农杆菌介导法将抗草甘膦基因转入玉米自交系H99的研究

doi:10.12190/j.issn.2096-1197.2020.02.04

Abstract

Abstract: 【Objective】Breeding herbicide-resistant maize cultivars by transgenic method to solve the problem of grass damage.【Methods】Genetic transformation was conducted on immature embryos of maize inbred line of H99 by agrobacterium-mediated method, which was carry out single factor test, the different size of immature embryo, concentration of agrobacterium and co-culture time were set respectively, which was compared and analyzed based on GUS transient expression rate in transgenic embryos.【Results】The results showed that the GUS transient expression rate was the highest when the size of the immature embryo was 1.5-2.0 mm, the concentration of agrobacterium OD₆₀₀ was 0.4, the co-culture time was 3 d and the infection time was 5 min. PCR detection proved that 7 positive transgenic seedlings were successfully obtained, and phenotypic identification also demonstrated that transgenic seedlings were capable of resistance to glufosinate.【Conclusion】This study provides a reference for the optimization of improving maize transgenic technology system and the breeding of new herbicide-resistant varieties.

Key words: Maize, Glyphosate resistance gene(EPSPS), Agrobacterium mediated method, Genetic transformation, GUS staining

CLC Number:

Q943.2

HAN Ping′an, WU Xinrong, SUN Ruifen, SHI Haibo, XUE Chunlei, CHANG Yue, TANG Kuangang, LIANG Yahui, REN Qinyong, LI Xiaodong. Genetic transformation system of maize inbred line H99 with agrobacterium-mediated glyphosate resistance gene[J].Journal of Northern Agriculture, 2020, 48(2): 21-26.

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Genetic transformation system of maize inbred line H99 with agrobacterium-mediated glyphosate resistance gene

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