New taxonomic markers for identification of Rhizobium leguminosarum and discrimination between closely related species
New taxonomic markers for identification of Rhizobium leguminosarum and discrimination between closely related species
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DOI：
10.1007/s00203-008-0447-6
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发表时间：
2009-03-01
影响因子：
2.8
通讯作者：
Skorupska, Anna Maria
中科院分区：
文献类型：
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作者：
Janczarek, Monika;Kalita, Michal;Skorupska, Anna Maria
根瘤菌，产生物种特异性外多糖（eps），包括一个非常多样化的土壤细菌群，能够与豆科植物建立固氮共生关系。基于豆科根瘤菌EPS合成基因序列，建立了一种灵敏、可靠的根瘤菌鉴别方法。为鉴定豆科植物，提出了与rosR、pssA和pssY基因序列互补的引物集I-III。进一步设计了四组引物（iv ~ vii）用于豆黄菌生物品种的区分。利用从波兰不同地区的不同寄主植物结核中分离的广泛的豆科芽孢杆菌菌株，对该方法的有效性进行了检验。我们证明了引物组I-III具有很高的区分能力，可以区分豆科蒿和两个密切相关的物种，豆科蒿和大蒜。这种新方法适用于鉴定豆科芽孢杆菌菌株，起源于结核或土壤，其中许多其他密切相关的细菌预计存在。根据rosR和pssA基因的核苷酸序列，确定了所选豆科致病菌分离株的系统发育关系。我们的结果表明，rosR和pssA可能是区分和确定一组豆科发菌之间关系的有用标记。
Rhizobia, producing species-specific exopolysaccharides (EPSs), comprise a very diverse group of soil bacteria that are able to establish nitrogen-fixing symbioses with legumes. Based on the sequences of R. leguminosarum EPS synthesis genes, a sensitive and reliable PCR-based method for identification and subsequent discrimination between Rhizobium species has been developed and tested. For identification of R. leguminosarum, primer sets I-III complementary to sequences of rosR, pssA and pssY genes were proposed. Further sets of primers (IV-VII) were designed for discrimination between R. leguminosarum biovars. The usefulness of the method was examined using a wide range of R. leguminosarum strains isolated from different host plants nodules originating from different regions of Poland. We demonstrate a high discriminating power of primer sets I-III that allow distinguishing R. leguminosarum and two closely related species, R. etli and R. gallicum. This new approach is applicable to identification of R. leguminosarum strains, originating from nodules or soil, where many other closely related bacteria are expected to be present. Based on the nucleotide sequence of rosR and pssA genes, phylogenetic relationships of selected R. leguminosarum isolates were determined. Our results indicate that both rosR and pssA might be useful markers to differentiate and define relationships within a group of R. leguminosarum strains.