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甘蔗宿根矮化病菌PCR检测及目的片段核苷酸序列分析
作者姓名:沈万宽  周国辉  邓海华  周凌云
作者单位:广州甘蔗糖业研究所 广州510316(沈万宽,邓海华),华南农业大学植物病毒研究室 广州510642(周国辉,周凌云)
基金项目:广东省科技攻关计划;引进国际先进农业科技计划(948计划)
摘    要:根据甘蔗宿根矮化病(RatoonStuntingDisease,RSD)病原细菌(Leifsoniaxylisubsp.xyli,Lxx)16S-23SrDNA基因间隔区核苷酸序列设计的一对特异性引物,建立了甘蔗宿根矮化病PCR检测方法;同时对PCR扩增的目的片段核苷酸序列进行测定与分析。结果表明广东湛江蔗区RSD病菌16S-23SrDNA基因间隔区核苷酸序列与巴西、澳大利亚及美国路易斯安娜州RSD株系基因组相应区段核苷酸同一率接近100%,病菌高度的同源。而与近缘的木质部赖氏杆菌狗牙根变种(Leifsoniaxylisubsp.Cynodontis,Lxc)和形态上相近的马铃薯环腐病菌(Clavibactermichiganensissubsp.Michiganensis)基因组相应区段核苷酸同一率较低,存在明显的分化。

关 键 词:甘蔗  宿根矮化病  序列分析

Detection of Sugarcane Ratoon Stunting Disease Pathogen with Polymerase Chain Reaction (PCR) and Nucleotide Sequence Analysis
Authors:Shen Wankuan  Zhou Guohui  Deng Haihu  Zhou Linyun
Institution:1Guangzhou Sugarcane Industry Research Institute, Guangzhou 510316;2.Laboratory of Plant virology, South China Asricultural University, Guangzhou 510642
Abstract:PCR technique was established for detection of Leifsonia xyli subsp. xyli(Lxx.), the causal agent of sugarcane ratoon stunting disease(RSD), with specific primers based on the intergenic transcribed spacer(ITS) region of 16S-23S ribosomal DNA of Lxx.By sequence comparison of intergenic transcribed spacer(ITS) region of 16S-23S rDNA, it was revealed that RSD pathogen was highly conversat. Lxx guangdong isolate shares almost 100% nucleotide identity with Brazil,Australia and USA isolates. However,Lxx ITS sequences share only low identity with related close(Leifsonia xyli subsp. Cynodontis,Lxc) and morphologic similar bacteria(Clavibacter michiganensis subsp.michiganensis).
Keywords:PCR
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