文山石漠化地区豆科植物根瘤内生细菌多样性分析

Diversity analysis of endophytic bacterial separated from the leguminous plants nodules of Wenshan rocky desertification areas

【目的】开展石漠化地区豆科植物根瘤内生细菌多样性研究,为豆科植物在石漠化地区生态恢复与重建中的应用提供理论依据。【方法】采用YMA培养基、NA培养基对采集自文山石漠化地区6种豆科植物根瘤的内生细菌进行分离纯化,再对分离获得的内生细菌进行酶切去除冗余、ARDRA分析、16S r DNA序列同源性分析及生物学表型特征分析。【结果】从6种豆科植物根瘤中共分离获得135株根瘤内生细菌,分别隶属于7个属24个种,其中49株为假单胞菌属(6个种)、17株泛菌属(3个种)、25株肠杆菌属(6个种)、17株芽孢杆菌属(5个种)、14株类芽孢杆菌属(1个种)、10株根瘤菌属(2个种)、3株农杆菌属(1个种)。生物学表型特征分析显示,SWFU-W032菌株能在盐浓度为5%的培养基中生长;13株内生细可以在p H 12.0的培养基上生长;SWFU-W024、SWFU-W009菌株可以水解淀粉;SWFU-W011、SWFU-W015和SWFU-W020菌株具有抗生素合成能力;除SWFU-W060外的供试菌株均具有产IAA能力;SWFU-W045、SWFU-W046和SWFU-W070菌株具有产蛋白酶能力。【结论】文山石漠化地区豆科植物根瘤内生细菌具有丰富的多样性,其中存在一些具有较高抗性、较强耐受性和多种可促进植物生长的特异菌株,这些菌株可作为石漠化地区土壤改良和生态恢复等的资源应用。

[Objective]Diversity of endophytic bacterial separated from the leguminous plants nodules of Wenshan rocky areas was analyzed to provide theoretical references for the legume used in ecological restoration and reconstruction of rocky desertification area. [MethodlUsing YMA and NA medium, the endophytic bacteria of six legume nodules which were collected from Wenshan esertification region, were isolated and purified. On this basis, restriction enzyme digestion （removing redundant）, ARDRA, 16S rDNA sequence homology and biological phenotypic characteristics were analyzed. [ Result ]The results showed that 135 endophytic bacteria were identified as twenty four species, belonging to seven genera. Among them, 49 strains belonged to Pseudomonas（6 kinds）, 17 strains belonged to Pantoea （3 kinds）, 25 strains belonged to Enterobacter（6 kinds）, 17 strains belonged to Bacillus（5 kinds）, 14 strains belonged to Paenibacillus （1 kind）, 10 strains belonged to Rhizobium （2 kinds） and 3 strains belonged to Agrobacterium（lkind）. Phenotypic analysis showed that SWFU-W032 strain could grow in the 5% salt medium; 13 endophytic bacteria strains could grow in the pH 12.0 medium; SWFU-W024 and SWFU-WO09 strains hydrolyzed starch; SWFU-W011, SWFU-W015 and SWFU-W020 strains synthesized antibiotic; except SWFU-W60, all strains had the ability to produce IAA as well as SWFU-W045, SWFU- W046 and SWFU-W070 strains had the ability to produce protease. [Conclusion ]The endophytic bacteria of legume in Wenshan rocky desertification region are quite diversified. Some strains with higher resistance and strong tolerance can promote plant growth and these strains can be used for rocky desertification soil improvement and ecological restoration in rocky desertification areas.