四川地区大豆根际土壤豆科植物根瘤菌捕获及多样.doc

四川地区结瘤大豆根际土壤中紫云英、苜蓿和三叶草根瘤菌的多样性分析* 李艳梅 钟宇舟 谭渊 何中山 徐雯芳 陈强 余秀梅** 四川农业大学资源环境学院微生物系 成都 610041 摘 要 [目的]分析四川地区结瘤大豆根际土壤是否存在与紫云英、苜蓿和三叶草共生结瘤的根瘤菌及其多样性, 从而为豆科植物的在四川地区种植与轮作提供依据。[方法]本研究从四川盆地采集了不同种植模式下结瘤大豆根际土壤样品31份；利用紫云英、苜蓿和三叶草进行盆栽捕获试验以获得共生根瘤，从根瘤中分离纯化出根瘤菌，对其做表型和分子鉴定；将根瘤菌回接到紫云英、苜蓿、三叶草和大豆中，以检测根瘤菌的寄主范围。[结果]苜蓿和三叶草分别在6个土壤样品中共生结瘤，而紫云英只在2个土样中共生结瘤，并从共生根瘤中分离出14株根瘤菌；16S基因序列的相似性分析表明这14株根瘤菌均属于根瘤菌属（Rhizobium），且16S基因的系统发育树分析揭示它们分属于根瘤菌属的不同种；回接实验结果表明这些根瘤菌都只能让捕获豆科植物结瘤，而不能让其他三种豆科植物结瘤。[结论]四川地区结瘤大豆根际土壤中能与紫云英 （Rhizobium astragali）、苜蓿 （Rhizobium meliloti）和三叶草 （Rhizobium leguminosarum var. trifolii）共生根瘤菌资源较少，且其与大豆根瘤菌（Bradyrhizobium japonicum）分属于不同的属；根瘤菌的回接实验进一步证明了根瘤菌的寄主专一性，且这些根瘤菌并不是大豆根瘤菌。 关键词 根瘤菌；大豆根际土壤；紫云英；苜蓿；三叶草 Analysis for the diversity of rhizobia nodulating Astragalus sinicus, Medicago sativa and Trifolium repens in nodulated soybean rhizosphere soil in Sichuan LI Yanmei, ZHONG Yuzhou, TAN Yuan, HE Zhongshan, XU Wenfang, CHEN Qiang YU Xiumei** Department of Microbiology, College of Resource and Environmental Sciences, Sichuan Agricultural University, Chengdu611130, China Abstract: Objective: The research aimed to analyze whether rhizobia nodulating with Astragalus sinicus, Medicago sativa and Trifolium repens appear around nodulated soybean rhizosphere in Sichuan and the diversity of the rhizobia. which should apply some useful basis information for planting leguminous plants in Sichuan. Methods: Thirty-one rhizosphere soil samples of nodulated soybean with different planting style were collected from Sichuan basin. Three leguminous plants of Astragalus sinicus, Medicago sativa and Trifolium repens were planted in the 31 soil samples to capture rhizobia. Rhizobia strains were isolated from different root nodules, and then phenotype and molecule identification were performed for them. To test nodulating host scope, rhizobia isolates was used to inoculate for Astragalus sinicus, Medicago sativa and Trifolium repens and Glycine max. Result: Medicago sativa and Trifolium repens were nod