Louws F J, Fulbright D W, Stephens C T, de Bruijn F J
Department of Botany and Plant Pathology, Michigan State University, East Lansing 48824.
Appl Environ Microbiol. 1994 Jul;60(7):2286-95. doi: 10.1128/aem.60.7.2286-2295.1994.
DNA primers corresponding to conserved motifs in bacterial repetitive (REP, ERIC, and BOX) elements and PCR were used to show that REP-, ERIC-, and BOX-like DNA sequences are widely distributed in phytopathogenic Xanthomonas and Pseudomonas strains. REP-, ERIC, and BOX-PCR (collectively known as rep-PCR) were used to generate genomic fingerprints of a variety of Xanthomonas and Pseudomonas isolates and to identify pathovars and strains that were previously not distinguishable by other classification methods. Analogous rep-PCR-derived genomic fingerprints were generated from purified genomic DNA, colonies on agar plates, liquid cultures, and directly from lesions on infected plants. REP, ERIC, and BOX-PCR-generated fingerprints of specific Xanthomonas and Pseudomonas strains were found to yield similar conclusions wtih regard to the identity of and relationship between these strains. This suggests that the distribution of REP-, ERIC, and BOX-like sequences in these strains is a reflection of their genomic structure. Thus, the rep-PCR technique appears to be a rapid, simple, and reproducible method to identify and classify Xanthomonas and Pseudomonas strains, and it may be a useful diagnostic tool for these important plant pathogens.
与细菌重复(REP、ERIC和BOX)元件中的保守基序相对应的DNA引物及聚合酶链式反应(PCR)被用于证明,类REP、类ERIC和类BOX的DNA序列广泛分布于植物病原黄单胞菌属和假单胞菌属菌株中。REP-PCR、ERIC-PCR和BOX-PCR(统称为rep-PCR)被用于生成多种黄单胞菌属和假单胞菌属分离株的基因组指纹图谱,并鉴定先前无法通过其他分类方法区分的致病型和菌株。类似的源自rep-PCR的基因组指纹图谱可从纯化的基因组DNA、琼脂平板上的菌落、液体培养物中直接生成,也可直接从受感染植物的病斑中生成。结果发现,特定黄单胞菌属和假单胞菌属菌株的REP-PCR、ERIC-PCR和BOX-PCR生成的指纹图谱,在这些菌株的身份及它们之间的关系方面得出了相似的结论。这表明这些菌株中类REP、类ERIC和类BOX序列的分布反映了它们的基因组结构。因此,rep-PCR技术似乎是一种快速、简单且可重复的方法,用于鉴定和分类黄单胞菌属和假单胞菌属菌株,并且它可能是这些重要植物病原体的一种有用的诊断工具。