Identification of fast and slow growing rhizobia nodulating soybean ( Glycine max [L.] Merr) by a multiplex PCR reaction

Two DNA fragments, a 730-bp and a 900-bp fragment, one homologous to host cultivar specificity genes nolBT of Sinorhizobium fredii and the other one homologous to RSα, an insertion-like sequence present in Bradyrhizobium japonicum, were generated by polymerase chain reaction (PCR) with two pairs of...

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Bibliographic Details
Published in:FEMS microbiology letters 2003-12, Vol.229 (2), p.153-158
Main Authors: Pastorino, G.N, Martinez Alcántara, V, Balatti, P.A
Format: Article
Language:English
Subjects:
Bacteriology
Biological and medical sciences
Bradyrhizobium
Bradyrhizobium - genetics
Bradyrhizobium - growth & development
Bradyrhizobium japonicum
DNA, Bacterial - analysis
Fundamental and applied biological sciences. Psychology
Glycine max
Glycine max - microbiology
Microbiology
Miscellaneous
Molecular marker
Nitrogen Fixation
Polymerase chain reaction
Polymerase Chain Reaction - methods
Rhizobium
Sinorhizobium
Sinorhizobium fredii - genetics
Sinorhizobium fredii - growth & development
Sinorhizobium xinjiangensis
Soybean
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Summary:Two DNA fragments, a 730-bp and a 900-bp fragment, one homologous to host cultivar specificity genes nolBT of Sinorhizobium fredii and the other one homologous to RSα, an insertion-like sequence present in Bradyrhizobium japonicum, were generated by polymerase chain reaction (PCR) with two pairs of primers. The amount of each fragment generated by the multiplex PCR was proportional to the amount of template DNA present. The amplification of the 900-bp RSα fragment was more sensitive, since it was amplified from a smaller amount of template DNA than the 730-bp nolBT fragment. By running the multiplex reaction in the presence of template DNA isolated from different sources, we confirmed that the reaction can discriminate between S. fredii, Bradyrhizobium japonicum and Sinorhizobium xinjiangensis.
ISSN:0378-1097
1574-6968
DOI:10.1016/S0378-1097(03)00796-1