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许昌泡桐丛枝病植原体的分子鉴定
引用本文:李婷婷,史梦蝶,张宁,赵文军,孙现超.许昌泡桐丛枝病植原体的分子鉴定[J].河南农业科学,2013,42(2).
作者姓名:李婷婷  史梦蝶  张宁  赵文军  孙现超
作者单位:1. 西南大学植物保护学院,重庆,400716
2. 中国检验检疫科学研究院动植物检疫研究所,北京,100029
基金项目:质检公益性行业科研专项,重庆市自然科学基金项目,农业部公益性行业科研专项
摘    要:对采自河南许昌的表现丛枝症状的泡桐样品进行了病原鉴定,并通过序列同源性分析确定了其分类地位。采用16SrDNA基因的通用引物R16mF2/R16mR1、R16F2n/R16R2和延伸因子(EF-Tu)tuf基因的通用引物fTufu/rTufu,对样品总DNA进行PCR扩增,分别得到了约1.2kb和840bp的特异条带。经克隆测序,并在NCBI网站比对分析,确定所得序列分别为植原体特定的16SrDNA和tuf基因序列。将测得序列与已报道的翠菊黄化组植原体序列进行同源性比对,并构建系统进化树,显示河南许昌的泡桐丛枝病植原体属于16SrⅠ-D亚组。

关 键 词:泡桐丛枝病  植原体  16S  rDNA基因  延伸因子tuf基因  系统进化分析

Molecular Detection of Phytoplasma Associated with Paulownia Witches' Broom in Xuchang
LI Ting-ting , SHI Meng-die , ZHANG Ning , ZHAO Wen-jun , SUN Xian-chao.Molecular Detection of Phytoplasma Associated with Paulownia Witches' Broom in Xuchang[J].Journal of Henan Agricultural Sciences,2013,42(2).
Authors:LI Ting-ting  SHI Meng-die  ZHANG Ning  ZHAO Wen-jun  SUN Xian-chao
Abstract:In this study,the pathogenic agent in paulownia leaves showing witches' broom symptoms collected from Xuchang,Henan province was identified.Universal phytoplasma primer pairs,R16mF2/R16mR1,R16F2n/R16R2 and fTufu/rTufu were used to amplify 16S rDNA and elongation factor(EF-Tu) tuf gene by PCR,respectively.Specific fragments about 1.2 kb and 840 bp were yielded in the end.After cloning,sequencing and analysis,the obtained fragments were separately identified as the specific 16S rDNA and tuf fragments of phytoplasma.BIAST in NCBI and phylogenetic analysis showed that the phytoplasma causing paulownia witches' broom in Xuchang(PaWB-Xuchang) belonged to 16SrⅠ-D group.
Keywords:paulownia witches'broom  phytoplasma  16S rDNA gene  elongation factor tuf gene  phylogenetic analysis
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