de Oliveira V M, Coutinho H L, Sobral B W, Guimarães C T, van Elsas J D, Manfio G P
Fundação André Tosello, Campinas, SP, Brazil.
Lett Appl Microbiol. 1999 Feb;28(2):137-41. doi: 10.1046/j.1365-2672.1999.00480.x.
With the aim of detecting Rhizobium species directly in the environment, specific PCR primers for Rh. tropici and Rh. leguminosarum were designed on the basis of sequence analysis of 16S-23S rDNA spacer regions of several Rh. tropici, Rh. leguminosarum and Agrobacterium rhizogenes strains. Primer specificity was checked by comparison with available rDNA spacer sequences in databases, and by PCR using DNA from target and reference strains. Sequence polymorphisms of rDNA spacer fragments among strains of the same species were detected by denaturing gradient gel electrophoresis (DGGE). The specific PCR primers designed in this study could be applied to evaluate the diversity of Rh. tropici and Rh. leguminosarum by analysing the polymorphisms of 16S-23S spacer rDNA amplified from either whole-cell or soil-extracted DNA.
为了直接在环境中检测根瘤菌属物种，基于几种热带根瘤菌、豌豆根瘤菌和发根农杆菌菌株的16S - 23S rDNA间隔区序列分析，设计了针对热带根瘤菌和豌豆根瘤菌的特异性PCR引物。通过与数据库中可用的rDNA间隔区序列进行比较，以及使用来自目标菌株和参考菌株的DNA进行PCR来检查引物特异性。通过变性梯度凝胶电泳（DGGE）检测同一物种菌株间rDNA间隔区片段的序列多态性。本研究设计的特异性PCR引物可通过分析从全细胞或土壤提取的DNA中扩增的16S - 23S间隔rDNA的多态性，用于评估热带根瘤菌和豌豆根瘤菌的多样性。
