应用克隆文库构建法研究番茄根系AMF多样性

从昆明学院种植基地随机采取番茄根系样品,CTAB法提取DNA,运用巢式PCR扩增目的片段,获得产物连接转化入大肠杆菌JM109构建AMF 18S rRNA部分基因克隆文库,随机挑选50个克隆子,从中排除6个假阳性,经ARDRA分析产生了13个差异图谱,测序分析最终确定11个AMF序列。结果表明：文库的Coverage C值高达95.2%,且Rarefaction曲线亦趋于饱和;11个序列与免培养的Glomus属克隆序列相似度较高,代表了本属的不同AMF种类;其中seq1和seq9所代表的地表球囊霉Glomus versiforme和摩西球囊霉Glomus mosseae是侵染番茄根系的优势AM...

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Bibliographic Details
Published in西南农业学报 Vol. 28; no. 1; pp. 323 - 328
Main Author 任禛 夏体渊 张永福 韩丽 陈丽娟 汪颖 尹敏
Format Journal Article
LanguageChinese
Published 昆明学院/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 2015
Subjects
18S
AMF
Nested-PCR
rDNA
系统发育分析
Phylogenetic analysis
18S rDNA
AMF
Nested-PCR
系统发育分析
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ISSN1001-4829
DOI10.16213/j.cnki.scjas.2015.01.060

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Summary:从昆明学院种植基地随机采取番茄根系样品,CTAB法提取DNA,运用巢式PCR扩增目的片段,获得产物连接转化入大肠杆菌JM109构建AMF 18S rRNA部分基因克隆文库,随机挑选50个克隆子,从中排除6个假阳性,经ARDRA分析产生了13个差异图谱,测序分析最终确定11个AMF序列。结果表明：文库的Coverage C值高达95.2%,且Rarefaction曲线亦趋于饱和;11个序列与免培养的Glomus属克隆序列相似度较高,代表了本属的不同AMF种类;其中seq1和seq9所代表的地表球囊霉Glomus versiforme和摩西球囊霉Glomus mosseae是侵染番茄根系的优势AMF类型;seq3、seq10和seq11均与Glomus属免培养克隆子聚集在一起;而seq2、seq4、seq5、seq6、seq7和seq8间亲缘关系较近,共同聚集在一个分支上。
Bibliography:51-1213/S
Tomato roots samples were collected randomly from Kunming College planting base. Roots DNA was isolated by CTAB method.Nested PCR was carried out for 18 S rRNA gene fragments. The PCR product was ligated with p MD18-T vector and transformed into E. coli JM109 to construct clone library. 50 clones were picked out randomly and 6 false-positive clones were excluded. 13 different OTUs were generated by ARDRA（ Amplified Ribosomal DNA Restriction Analysis） and 11 AMF sequences were obtained by DNA sequence. The Coverage C value of this library was 95. 2 % and the Rarefaction curve showed a tendency to saturation. 11 sequences showed high similarity with uncultured Glomus sequences which represented different AMF. Seq1 and seq9 which represented Glomus mosseae and Glomus versiforme respectively were the predominant AMF in tomato roots. Seq3,seq10 and seq11 got together with uncultured Glomus sequences. The relationship of Seq2,Seq4,Seq5,seq6,seq7 and seq8 were close and they got together in a branch.
AMF; 18
ISSN:1001-4829
DOI:10.16213/j.cnki.scjas.2015.01.060