Jana T K, Singh N K, Koundal K R, Sharma T R
National Research Centre on Plant Biotechnology, Indian Agricultural Research Institute, Pusa, New Delhi, India.
Can J Microbiol. 2005 Feb;51(2):159-64. doi: 10.1139/w04-122.
Forty isolates of Macrophomina phaseolina, a pathogen causing charcoal dry root rot of soybean, cotton, and chickpea, were genetically characterized with universal rice primers (URP; primers derived from DNA repeat sequences in the rice genome) using polymerase chain reaction (URP-PCR). Out of 12 URPs used in this study, 5 primers were effective in producing polymorphic fingerprint patterns from the DNA of M. phaseolina isolates. Three primers (URP-2F, URP-6R, and URP-30F) were quite informative and produced high levels of polymorphism among the isolates of M. phaseolina. Analysis of the entire fingerprint profiles using unweighted pair-group method with arithmetic averages (UPGMA) clearly differentiated M. phaseolina isolates obtained from soybean, cotton, and chickpea hosts into specific groups. In this study, we found for the first time transferability and use of PCR primers derived from plant genomes to generate host-specific fingerprint profiles of M. phaseolina, a broad host range plant pathogenic fungus. These results demonstrate that URPs are sensitive and technically simple to use for assaying genetic variability in M. phaseolina populations.
引起大豆、棉花和鹰嘴豆炭腐根腐病的病原菌菜豆壳球孢(Macrophomina phaseolina)的40个分离株,使用通用水稻引物(URP;源自水稻基因组中DNA重复序列的引物)通过聚合酶链反应(URP-PCR)进行了遗传特征分析。在本研究使用的12个URP中,有5个引物能够有效地从菜豆壳球孢分离株的DNA中产生多态性指纹图谱。三个引物(URP-2F、URP-6R和URP-30F)信息量很大,在菜豆壳球孢分离株中产生了高水平的多态性。使用算术平均数的非加权配对组方法(UPGMA)对整个指纹图谱进行分析,清楚地将从大豆、棉花和鹰嘴豆寄主中获得的菜豆壳球孢分离株分为特定的组。在本研究中,我们首次发现源自植物基因组的PCR引物具有可转移性,并可用于生成寄主范围广泛的植物病原真菌菜豆壳球孢的寄主特异性指纹图谱。这些结果表明,URP对于分析菜豆壳球孢群体的遗传变异性而言灵敏且技术上易于使用。