作　　者： ; ; ; ; ; ;

机构地区： 石河子大学农学院

出　　处： 《新疆农业科学》 2011年第10期1773-1778,共6页

摘　　要： 【目的】初步获知棉花NBS-LRR类基因的生物学意义,构建干扰载体。【方法】以抗枯萎病棉花中棉12号为材料,用霍格兰营养液培养至三叶期,进行枯萎病菌接菌诱导处理,以不接菌为对照。利用cDNA-AFLP技术筛选差异基因片段,在GenBank中进行序列同源性比对,找到抗枯萎病基因片段,该基因片段的序列与GenBank中报道的序列同源性达到72%。将此基因片段以正向和反向插入到pPZP35S,经限制性酶切和质粒PCR鉴定。【结果】已成功构建了干扰载体。采用农杆菌侵染法将干扰载体转入烟草中。【结论】为研究棉花NBS-LRR类基因的功能打下基础,为通过转基因技术深入研究该基因在棉花抗病机理创造条件。 [ Objective] The purpose of this program was to get an initial knowledge of the biological significance of the gene and construct interfering carriers. [ Method] In this study, resistance to Fusarium wilt of cotton in 12 was taken as the experimental material which was cultured with Hoagland nutrient solution to the three - leaf stage, and then the induction of Fusarium inoculation treatment was done with non - inoculation as control, cDNA- AFLP was used to screen the differential gene fragments in order to have a sequence homology comparison in the GeneBank and find the gene that is resistant to Fusarium wilt. The sequence of the gene fragment that we found reached 72% homology with the one reported in the GeneBank. This found gene fragment was inserted into pPZP35S in forward and reverse direction and identified by restriction digestion and plasmid PCR. [ Result ] The interference carrier has been successfully built. Using Agrobacterium infection into tobacco law will interfere with the carrier, which laid the foundation for studying cotton NBS - LRR class gene. [ Conclusion ] This study created condition for the adoption of transgenic technology in - depth study of the mechanism of disease resistance genes in cotton.

关 键 词： 棉花 枯萎病 干扰 烟草转化

领　　域： [农业科学] [农业科学]