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[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 research aimed to study the ISSR marker technology of disease-resistance genes in Medicago sativa. [Method] ISSR molecular marker technique was used in combination with cluster analysis method to screen the molecular markers linked to the genes of anti-sclerotinia in 5 resistant plants and 7 susceptible plants. A total of 94 plants from F1 hybrid F1 were tested for resistance. [Result] Out of the 93 ISSR primers, 35 primers could produce clear and stable amplified bands. Among them, 6 primers could produce 9 specific bands between resistant and susceptible DNA pools. The results of resistance test showed that 825-1400, 831-1480, 850-1800, 858-1600, 869-1900 and 888-1400 could be used as ISSR markers for the resistance gene of S. sclerotiorum. [Conclusion] The results laid the foundation for the in-depth study of the localization, cloning and transgene of disease-resistance genes in Medicago sativa.