野生大豆抗感大豆孢囊线虫材料内生细菌多样性分析  

Endophytic bacterial diversity of wild soybean(Glycine soja) varieties with different resistance to soybean cyst nematode(Heterodera glycines)

作　　者：武云鹏[1,2] 史凤玉[1] M.Imran Hamid 张瑞敬 朱英波[1] 

机构地区：[1]河北科技师范学院生命科技学院,河北秦皇岛066600 [2]中国科学院微生物研究所,真菌学国家重点实验室,北京100101 [3]张家口市植物保护植物检疫站,河北张家口075000

出　　处：《微生物学报》2014年第8期926-935,共10页Acta Microbiologica Sinica

基　　金：河北省自然科学基金项目(C2014407021);中国博士后科学基金(20100480028);河北科技师范学院博士启动基金(2007YB010)~~

摘　　要：【目的】对抗感野生大豆材料根内生细菌的多样性进行比较分析,为研究野生大豆内生细菌与大豆孢囊线虫之间的相互关系奠定基础。【方法】在野生大豆抗大豆孢囊线虫3号生理小种筛选基础上,利用扩增核糖体DNA限制性分析(Amplified ribosomal DNA restriction analysis,ARDRA)和16S rDNA克隆文库测序相结合的方法,对抗感野生大豆根系内生细菌多样性及群落结构进行分析。【结果】野生大豆根内生细菌分属于6大类群,其中变形菌门(Proteobacteria)和厚壁菌门(Firmicutes)为优势类群,相对丰度分别为46.8%和13.6%,另外有少量的放线菌门(Actinobacteria)、拟杆菌门(Bacteroidetes)、酸杆菌门(Acidobacteria)、异常球菌-栖热菌门(Deincoccus-Thermus)和古细菌(Archaea),18.8%克隆序列与环境中未培养细菌的16S rDNA序列有较高的相似性。野生大豆高抗材料内生细菌的多样性比高感材料更为丰富,且抗感材料内生细菌优势菌群存在明显差异,中慢生根瘤菌(Mesorhizobium tamadayense)、肠杆菌(Enterobacter ludwigii)和巨大芽孢杆菌(Bacillus megaterium)为野生大豆高抗材料特有的可操作分类单元(Operational Taxonomic Units,OTUs)中的优势种群。【结论】研究结果表明抗感野生大豆根内生细菌的优势种群存在明显差异,而内生细菌的优势种群与大豆孢囊线虫的相互关系正在深入分析研究。[ Objective] The aim of this study was to investigate endophytic bacterial diversity of wild soybean varieties with different resistance to soybean cyst nematode（Heterodera glycines） , for deciphering the interactions of soybean cyst nematode with endophytic bacteria. [ Methods] After screening wild soybean varieties against race 3 of H. glycines, we investigated endophytic bacterial diversity in root tissues of wild soybean varieties with different resistance to H. glycines using 16S rDNA cloning library and amplified ribosomal DNA restriction analysis. [ Results] Endophytic bacteria of wild soybean root belonged to 6 bacterial groups, the clones belonging to group Proteobacteria and Firmicutes were the endophyte dominants in wild soybean with 46. 8% and 13. 6% of total clones, respectively. Actinobacteria, Bacteroidetes, Acidobacteria, Deincoccus-Thermus and Archaea were less represented. 18.8% of clone sequences were similar to those of uncultured bacteria in the environment. The bacterial diversity was higher in H. glycines-Resistant than -Susceptible wild soybean varieties, and the dominant group was different between H. glycines-Resistant and -Susceptible wild soybean varieties. Mesorhizobium tamadayense, Enterobacter ludwigii and Bacillus megaterium were the main bacterial groups in special operational taxonomic units （OTUs）of H. glycines-Resistant wild soybean variety. [ Conclusions] By 16S rDNA cloning library and amplified ribosomal DNA restriction analysis, the diversity of dominant group of endophytic bacteria in root tissues has difference among H. glycines-Resistant and -Susceptible wild soybean varieties.

关 键 词：大豆孢囊线虫 野生大豆材料 内生细菌 扩增核糖体DNA限制性分析 分类操作单元 

分 类 号：S565.1[农业科学—作物学]