基于18SrRNA序列的云南大叶千斤拔与细叶千斤拔根内丛枝菌根真菌鉴定

目的:调查云南省大叶千斤拔(Flemingia macrophylla)与细叶千斤拔(Flemingia lineata)根内丛枝菌根真菌(Arbuscular Mycorrhizal Fungi,AMF)群落结构多样性。方法:使用巢式-PCR、克隆、限制性片段长度多态性(Restriction Fragment Length Polymorphism,RFLP)分析及测序技术。结果:共获得558个含有丛枝菌根真菌18S rRNA片段的克隆子,经RFLP分析后得到83个RFLP类型,DNA序列分析可将其划分为23个可操作分类单元(Operational Taxonomic Units,OTUs),分属于5个科,Glomeraceae为优势类群。在MaarjAM数据库中进行比对后,23个OTUs可鉴定为18个虚拟分类分子种,分布于13种不同的生境。经统计分析大叶千斤拔与细叶千斤拔根内丛枝菌根真菌群落组成比较,差异有统计学意义(P<0.001)。结论:比较大叶千斤拔与细叶千斤拔根内丛枝菌根真菌群落差异,结合丛枝菌根真菌在生态系统中的分布特点,为种植千斤拔属植物的选址提供有力地环境指标数据,并为筛选促生菌株提供依据。

Identification of Arbuscular Mycorrhizal Fungi of Flemingia macrophylla and Flemingia lineata in Yunnan, Based on 18S rRNA Sequences

To investigate the community diversify of arbuscular mycorrhizal fungi(AMF) of Flemingia macrophylla and Flemingia lineata.Methods:The nested-PCR, cloning, RFLP analyzing and sequencing techniques were used.Results:A total of 558 clones containing 18S rRNA genes from AMF were obtained.After RFLP analysis, 83 different RFLP types were obtained, which can be divided into 23 Operational Taxonomic Units (OTUs) by DNA sequence analysis.They belonged to 5 subjects, and Glomeraceae was the dominant group.After they were compared in MaarjAM database, 23 OTUs could be identified as 18 molecular virtual taxon, scattered in 13 different habitats.By statistical analysis, AMF communities were significantly different between F.macrophylla and F.lineata roots, the difference was statistically significant (P<0.001).Conclusion:Compared with the difference of AMF community in F.macrophylla and F.lineata roots, combining with the distribution features of AMF species in ecosystem, valuable environmental indicator data to choose site for the Flemingia plantation is provided.