| 小麦苗枯病菌的ITS分析及PCR检测 |
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| 引用本文： | 葛芸英,郭坚华.小麦苗枯病菌的ITS分析及PCR检测[J].植物病理学报,2003,33(3):198-202. |
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| 作者姓名： | 葛芸英 郭坚华 |
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| 作者单位： | 南京农业大学植物保护学院植物病理系, 南京 210095 |
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| 基金项目： | 国家自然科学基金 (39970 4 81 )资助 |
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| 摘 要： | 小麦苗枯病菌(Clavibacter fangii,Cf)是引起小麦细菌性苗枯病的病原,本研究用16S~23S rDNA间的内源转录间隔区(internally transcribed spacer,ITS)序列通用引物L1(5'-AGTCGTAACAAGGTAGCCGT-3')和L2(5'-GTGCCAAGGCATCCACC-3')扩增Cf和其它相关细菌的基因组DNA;并对其PCR产物进行回收、克隆和测序,将所获序列和其它已报道的细菌ITS序列进行多重比较后设计出Cf的特异性引物I1(5'-TGCCAAGTCACACTGAGACGA-3')和I2(5'-CAATGATCTACCACCCTCCGA-3')。此引物可以从Cf中扩增出351bp的特异性片段,而其余参试的21个细菌PCR反应结果均为阴性。该方法可以应用于小麦苗枯病菌的快速、可靠检测。此外,本研究对多种植物病原棒形杆菌的ITS序列进行比较研究,发现其具有一定的分类意义。
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| 关 键 词： | 小麦苗枯病菌 ITS分析 分类 检测 |
| 文章编号： | 0412-0914(2003)03-0198-05 |
| 修稿时间： | 2002年7月18日 |
Analysis of ITS sequence and PCR protocol for the detection of Clavibacter fangii, the causal agent of wheat seedling wilting disease |
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| GE Yun ying,GUO Jian hua.Analysis of ITS sequence and PCR protocol for the detection of Clavibacter fangii, the causal agent of wheat seedling wilting disease[J].Acta Phytopathologica Sinica,2003,33(3):198-202. |
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| Authors: | GE Yun ying GUO Jian hua |
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| Institution: | Department of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China |
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| Abstract: | A polymerase chain reaction (PCR) protocol was developed to specifically detect Clavibacter fangii (Cf), the causal agent of wheat seedling wilting disease. Generic PCR products from the internally transcribed spacer (ITS) region of 16S-23S ribosomal DNA of Cf and other two related bacteria were cloned and sequenced. Based on a multiple sequences alignment among these obtained sequences, a pair of Cf specific PCR primers I1(5 ' TGCCAAGTCACACTGAGACGA 3 ')/I2(5 ' CAATGATCTACCACCCTCCGA 3 ') were designed. These two oligonucleotides allowed specific amplification of a 351 bp DNA product from genomic DNA samples of Cf strains,but not from other 21 bacterial isolates tested. The PCR protocol provides a rapid and reliable tool for routine detection and identification of Cf. In addition, the classification implication of ITS sequence homology was first found in bacteria by comparing sequences from Clavibacter spp. and Rathayibacter tritici . |
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| Keywords: | Clavibacter fangii ITS analysis classification detection |
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