玫瑰根系AMF 18S rRNA基因克隆文库构建及分析

[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...

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Published in西南农业学报 Vol. 30; no. 9; pp. 2113 - 2118
Main Author 任禛 尹敏 夏体渊 陈泽斌 尹利方 韩丽 靳松 王定康
Format Journal Article
LanguageChinese
Published 昆明学院农学院/云南省高等学校都市型现代农业工程研究中心,云南昆明,650214%云南大学医学院,云南昆明,650091 2017
Subjects
丛枝菌根真菌
克隆文库
玫瑰‘卡罗拉’
系统发育树
评价和分析
丛枝菌根真菌
克隆文库
Rosa Carola
系统发育树
Phylogenetic tree
玫瑰‘卡罗拉’
评价和分析
AMF
Clone library
Evaluation and analysis
Online AccessGet full text
ISSN1001-4829
DOI10.16213/j.cnki.scjas.2017.9.032

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Summary:[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。
Bibliography:51-1213/S
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 andAMF partial 18S rRNA gene were amplified by Nested-PCR. The DNA mixture was used to construct AMF clone library. Finally, thirty posi-five clones were chosen for sequencing and analyzed. Meanwhile, e/one library was evaluated and analyzed in this paper. [ Result1 CoverageC value of the library was 70.0 %, but the rarefaction curve was not saturated, and the number of library clones should be increased. TheShannon index and Simpson index was 1. 432 and 0.25, respectively, and the species richness was 4. O. Sequences with 〉98 % similaritywere assigned to the same OTUs. Totally, 30 sequences were divided into 15 OTUs and all of them belonged to Glomus. Among them, OTU1was the main type of library, OTU2 - OTU9 represented the common types, while OTUI
ISSN:1001-4829
DOI:10.16213/j.cnki.scjas.2017.9.032