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草业学报 ›› 2009, Vol. 18 ›› Issue (6): 51-58.

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Chi基因的克隆及转基因马铃薯植株的获得

张茹1,2,李金花1,3,柴兆祥3,张俊莲1,2,张金文2, 王蒂1,2*   

  1. 1.甘肃省作物遗传与种质创新重点实验室,甘肃 兰州 730070;
    2.甘肃农业大学农学院,
    甘肃 兰州 730070;
    3.甘肃农业大学草业学院,甘肃 兰州 730070
  • 收稿日期:2009-05-04 出版日期:2009-12-20 发布日期:2009-12-20
  • 作者简介:张茹(1978-),女,甘肃兰州人,在读博士。E-mail: chouruer@163.com
  • 基金资助:
    转基因抗逆耐盐及抗病马铃薯新品系培育研究(GNSW-2006-01),马铃薯优质高效配套生产技术研究与示范(2006BAD21B053)和马铃薯镰刀菌干腐病原学及抗病种质资源筛选研究(GKLAU0603)项目资助

Cloning of a chitinase gene and the acquisition of transgenic potato plants

ZHANG Ru1,2, LI Jin-hua1,3, CHAI Zhao-xiang3, ZHANG Jun-lian1,2, ZHANG Jin-wen2 , WANG Di1,2   

  1. 1.Gansu Key Lab of Crop improvement and Germplasm Enhancement, Gansu Agricultural University,
    Lanzhou 730070, China;
    2.College of Agronomy, Gansu Agricultural University, Lanzhou 730070,
    China;
    3.College of Grassland Science, Gansu Agricultural University, Lanzhou 730070, China
  • Received:2009-05-04 Online:2009-12-20 Published:2009-12-20

摘要: 为了提高马铃薯对真菌病害的抗性,本试验进行了马铃薯转基因抗病育种的初步研究。通过PCR扩增,从生防木霉菌Trichoderma atroviride的cDNA中克隆到一个内切几丁质酶基因Chi,构建了植物表达载体p33ECR-Chi。通过农杆菌介导的转化方法将Chi基因转化到马铃薯栽培品种“费乌瑞它”和“夏波蒂”试管薯片中。经侵染和共培养后,用卡那霉素和羟苄青霉素筛选抗性芽,共获得18株植株,对所得转化植株进行PCR检测,结果12株为阳性,占67%。本试验为利用基因工程技术提高马铃薯的生防能力奠定了基础。

Abstract: The antagonistic ability of potato against fungal disease was enhanced by acquiring transgenic potato plants via gene engineering. A chitinase gene was cloned via PCR products from cDNA of Trichoderma atroviride. This gene was recombined into the plant expression vector p33ECR and transferred into micro tubers from flasks of two varieties (Favorita and Shepody) of potato via Agrobacterium tumefaciens-mediated transformation. After infection and co-culture, 18 transformed plants were acquired through Kanamycin and Carbenicillin screening tests. After PCR tests for transgenic plants, 67% of the plants (12 plantlets) were positive. This will give support for obtaining high biologically effective potato varieties in future research.

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