生物类(生物科学等)专业毕业论文--RLK基因遗传转化植株的筛选与鉴定.docVIP

RLK基因遗传转化植株的筛选与鉴定 生物技术专业 指导教师 摘要：番茄青枯病是严重影响番茄生产的病害之一，有“植物中的癌症”之称。植物体中类受体蛋白激酶(Receptor Likekinase RLK)有PCR和RT-PCR技1代进行转基因后的分子鉴定。结果显示，三种转基因番茄经特异引物PCR后均能产生特异性条带，而且D-RLK-6号经RT-PCR后D-RLK-6号的CaRLK基因能够顺利表达。此研究将为番茄青枯病的防治提供重要的理论支撑。 关键词：番茄；CaRLK；分子鉴定；PCR ；RT-PCRThe Selection and Identification of CaRLK Transgenic Tomatoes Plants Abstract：Tomato bacterial wilt is a serious disease affecting tomato production and it is called the Cancer of Plant. The Receptor Likekinase (RLK) in plants can increase their resistance function. In this study, we used molecular cloning technology to transfer the CaRLK from pepper to tomato, hoping to obtain bacterial wilt-resistant tomato plants. In addition to using traditional methods, such as morphological comparison, we mainly used PCR and RT-PCR to identify the three varieties of transgenic tomato plants of T1 generation of D-RLK-2, D-RLK-6 and D-RLK-10. ResuLts showed that three transgenic plants after the process of PCR by specific primers were able to produce specific bands, and that the D-RLK-6 produced specific band after the process of RT-PCR by specific primers, suggesting that the CaRLK gene had been successfully transferred into transgenic tomato genome. This study will provide important theoretical support for the prevention and control of tomato bacterial wilt. Key words ： Lycopersicon escuLentumMill；CaRLK；Molecular identification；PCR；RT-PCR 番茄(Lycopersicon escuLentumMill)40多种[1]，其中青枯病是严重影响番茄产量和质量的病害之一。 植物青枯病菌具有广泛的寄主能够侵染54个科的450(Ralstonia solanacearun)引20%-30%，发病严重时，田块病死株率可达80%以1-6年20世纪80年Tsai等[6]、康耀卫等[7] 、罗宽等[8]及董春等[9]利用60Co辐射、紫外线诱变、转座子诱变及自发突变体等方法获得了无致病力青枯菌，并且取得了一定的抗性效果。随着分子标记技术的快速发展和番茄基因连锁图谱的日趋饱和，番茄的一些重要病害的抗病基因得到精确定位，这不但可以为建立完善的番茄抗病基因的辅助选择体系提供可能， 而且为分离和克隆抗病基因提供了可能。到目前为止，对两套材料发展的群体应用RFLP、AFLP、RAPD(Receptor Protein Kinase RPK)是动(Receptor-Likekinase RLK)。RLK在植现在已经从拟南芥、水稻和小麦等高等植物中克隆出了大量的RLKs基因，发现它们对植物生长发育过程均起着十分重要的调控。如拟南芥中抗细菌性叶斑病密切相关的FLS2基因；水稻中与抗白叶枯病Xa21基因；小麦中参与抗白粉病反应的TaLRK基因等[]。植物通过RLKs基因的可逆磷酸化参与抗病防御过程。一般都会经历3个过程首先，病原信号分子