紫花苜蓿菌核病抗病基因ISSR标记初步研究

Primary study on ISSR Molecular Makers of resistant Gene against Sclerotinia trifoliorum in Medicago sativa L.

[目的]研究紫花苜蓿菌核病抗病基因的ISSR标记技术。[方法]运用ISSR分子标记技术,结合集群分离分析法对5株抗病植株和7株感病植株进行抗菌核病基因连锁的分子标记筛选;用叶片离体接种法对高抗83号×高感4号杂交F1代的94个植株进行抗性验证。[结果]在93个ISSR引物中,有35个引物能够产生清晰稳定的扩增条带,其中6个引物能在抗病、感病DNA池间产生9个特异性条带。抗性验证试验结果显示:825-1400、831-1480、850-1800、858-1600、866-1900、888-1400可以作为苜蓿菌核病抗性基因的ISSR分子标记。[结论]研究结果为紫花苜蓿菌核病抗病基因的定位、克隆、转基因等深入研究奠定了基础。

[Objective]The paper was to study ISSR molecular makers of resistant gene against Sclerotinia trifoliorum in Medicago sativa L.[Method]Using inter-simple sequence repeat（ISSR） molecular maker technology,combined with bulked-segregant analysis（BSA） method,the molecular makers for gene linkage with resistance against S.trifoliorum were screened from 5 resistant plants and 7 susceptible plants.Leaf inoculation method in vitro was used to carry out resistant verification on 94 hybrid plants in F1 generation of high resistant No.83×high susceptible No.4.[Result]Among 93 ISSR primers,35 primers could produce clear and stable amplification bands,and six of them could produce 9 specific bands between resistant and susceptible DNA pools.Resistance verification result showed that 825-1400,831-1480,850-1800,858-1600,866-1900,888-1400 could be used as ISSR molecular makers of the resistant gene against S.trifoliorum in M.sativa.[Conclusion]The results provided basis for the further research on mapping,cloning and genetically modified of resistant gene against S.trifoliorum in M.sativa.