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中国农学通报 ›› 2010, Vol. 26 ›› Issue (6): 35-38.

所属专题: 生物技术 园艺

• 生物技术科学 • 上一篇    下一篇

Ri质粒介导双价抗真菌病基因转化番茄的初步研究

王金杰1,王志英1,徐香玲2,张海玲3   

  • 收稿日期:2009-10-29 修回日期:2009-11-19 出版日期:2010-03-20 发布日期:2010-03-20
  • 基金资助:

    从碱草中克隆抗逆基因脱宽番茄种质资源的研究

Study on Genetic Transformation of Tomato Expressing Bivalent Anti-fungal Disease Gene Mediated by Ri Plasmid

  • Received:2009-10-29 Revised:2009-11-19 Online:2010-03-20 Published:2010-03-20

摘要:

以番茄新品种T431、1911、0701为材料,以9天苗龄的叶片为受体,通过农杆菌Ri质粒介导,将几丁质酶基因和?-1,3葡聚糖酶基因转化番茄。共获得再生植株100株,经PCR技术检测,导入几丁质酶基因和?-1,3葡聚糖酶基因的植株分别为27株和13株,转化率为27% 和13%;两个目的基因同时导入的植株10株,转化率为10%。PCR-Southern检测呈阳性,表明外源基因已经整合到番茄基因组中。对9株双转化的植株进行RT-PCR检测,几丁质酶基因和?-1,3葡聚糖酶基因的阳性植株分别为5株和4株,证明转化的基因已表达。

关键词: 铜, 铜, 锰, 荷斯坦种公牛, 抗氧化指标

Abstract:

New tomato varioties T431, 1911 and 0701 were used as the materials, chitinase gene and β-1,3-glucanase gene were transformed into tomatoes by Agrobacterium-mediated transformation. A total of 100 regeneration plants were obtained. In these regenerated plants, 27 and 13 plants appeared respectively to be positive in PCR test with chitinase gene and β-1,3-glucanase gene primer. The transferring frequency of foreign gene was 27% and 13% separately. If the two genes were done simultaneously, 10 plants appeared to be positive in PCR test with a transferring frequency of 10%. PCR-Southern analysis of these plants showed that they produced positive hybridization ,indicating that the chitinase gene and β-1,3-glucanase gene have been conveyed and integrated into genome of these plants.9 plants chosen from double-gene transgenic plants were taken RT-PCR analysis,5 and 4 plants appeared respectively to be positive in 9 plants, which proved that the chitinase gene and β-1,3-glucanase gene acquired stable expression in transgenic plants.