玫瑰根系AMF 18S rRNA基因克隆文库构建及分析

摘要

[Objective] The AMF diversity and composition structure of roses root were studied in this paper.[Method] Three'Rosa Carola'plant were randomly collected from germplasm resources gardon of Kunming college.The roots DNA were extracted using CTAB method and AMF partial 18S rRNA gene were amplified by Nested-PCR.The DNA mixture was used to construct AMF clone library.Finally,thirty positive clones were chosen for sequencing and analyzed.Meanwhile,clone library was evaluated and analyzed in this paper.[Result] Coverage C value of the library was 70.0 ％,but the rarefaction curve was not saturated,and the number of library clones should be increased.The Shannon index and Simpson index was 1.432 and 0.25,respectively,and the species richness was 4.0.Sequences with ＞98 ％ similarity were assigned to the same OTUs.Totally,30 sequences were divided into 15 OTUs and all of them belonged to Glomus.Among them,OTU1 was the main type of library,OTU2-OTU9 represented the common types,while OTU10-OTU15 were rare types.10 OTUs which clustered separately represented the Glomus novel types.[Conclusion] The AMF found in the roots of'Rosa Carola'were all Glomus species.Among them,there were 1 main type,8 common types and 6 rare types.The classification status of all clones was not clear.%[目的]本文研究了玫瑰根系的AMF多样性和组成结构.[方法]从昆明学院种质资源圃随机采集玫瑰‘卡罗拉’3株,采用CTAB法提取根系DNA,运用巢式PCR扩增AM真菌部分18S rRNA基因区域,扩增产物混合后构建克隆文库,最终确定30个阳性克隆子测序,并对文库进行评价和分析.[结果]文库Coverage C值为80％,但Rarefaction曲线不够饱和,应加大文库克隆子数目;文库多样性参数香农指数和辛普森指数分别为1.432和0.25,物种丰富度为4.0;以98％为界,30条序列被划分为15个OTU,均为Glomus的种类;其中OTU1是文库中的主要类型,OTU2～ OTU9代表了常见类型,而OTU10 ～OTU15为稀有类型;有10个OTU在系统进化树上单独聚类,代表着Glomus较为新颖的种类.[结论]在玫瑰根系发现的15个OTU全为Glomus的种类,其中主要类型1种,常见类型8种,稀有类型6种,具体分类地位均不清楚.