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Discrimination of Rhizobium tropici and R. leguminosarum strains by PCR‐specific amplification of 16S–23S rDNA spacer region fragments and denaturing gradient gel electrophoresis (DGGE)
Author(s) -
De Oliveira V. M.,
Coutinho H. L. C.,
Sobral B. W. S.,
Guimarães C. T.,
Van Elsas J. D.,
Manfio G. P.
Publication year - 1999
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1046/j.1365-2672.1999.00480.x
Subject(s) - temperature gradient gel electrophoresis , biology , 16s ribosomal rna , microbiology and biotechnology , gel electrophoresis , polymerase chain reaction , ribosomal dna , bacteria , gene , genetics , phylogenetics
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.