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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (3): 240-244.doi: 10.11924/j.issn.1000-6850.2011-3133

所属专题： 生物技术； 烟草种植与生产

• 生物技术科学 • 上一篇    下一篇

农杆菌介导脂滴包被蛋白基因8-3转化烟草的研究

牛伯庆 汪文静 宋洪英 谢响明   

  • 收稿日期:2011-10-27 修回日期:2011-12-12 出版日期:2012-01-25 发布日期:2012-01-25
  • 基金资助:

    国家转基因生物新品种培育科技重大专项

Study of Transformation of Tobacco with Perilipin Gene 8-3 Mediated by Agrobacterium tumefaciens

  • Received:2011-10-27 Revised:2011-12-12 Online:2012-01-25 Published:2012-01-25

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摘要：

为开发新的抗菌核病基因，针对抗病相关的脂滴包被蛋白基因8-3，设计了分别带有酶切位点Xba I和BamH I的一对引物，从克隆载体pMD19-T-8-3中扩增到基因8-3，双酶切8-3并连接到植物表达载体pBI 121上；将重组质粒导入根癌农杆菌(Agrobacterium tumefaciens) LBA4404中，通过其介导将8-3基因转化到烟草中；获得了35株抗性转化株，PCR及测序分析鉴定阳性植株为21株，RT-PCR检测到基因8-3在其中能够表达；通过接种核盘菌和草酸处理试验，表明8-3基因可使烟草提高了草酸耐受水平，从而对菌核病具备一定的抗性。本实验研究了潜在功能基因8-3的抗病性，为植物抗菌核病的研究奠定了基础。

关键词: 台地园, 台地园, 中国古典园林, 景观

Abstract:

To find new resistance genes and study perilipin gene 8-3 related to Sclerotinia sclerotiorum, a couple of primers containing restriction enzyme sites of Xba I and BamH I were designed to clone the 8-3 from cloning vector pMD19-T-8-3, and the product was digested and lignased with plant expression vector pBI121. The pBI121-8-3 was transformed into LBA4404 and the 8-3 gene was transformed into Nicotiana tabacum 89 mediated by Agrobacterium tumefaciens. 35 resistant lines were obtained, 21 of which were identified as transgenic plants by PCR and sequencing. The further identification of RT-PCR was confirmed that the 8-3 was expressed in the transgenic plants. The transgenic tobacco infected with Sclerotinia sclerotiorum and oxalic acid showed that 8-3 could improve the plants' tolerance level to oxalic acid and enhance the resistance to Sclerotinia sclerotiorum to a certain extent, which laid the foundation for the study of 8-3 gene and plant resistance to Sclerotinia sclerotiorum.