Authors: Malgorzata Waleron Krzysztof Waleron Joanna Kamasa Wlodzimierz Przewodowski Ewa Lojkowska
Publish Date: 2011/05/31
Volume: 131, Issue: 2, Pages: 341-
Abstract
The utility of polymorphism analysis was determined for differentiation of the following subspecies of the Grampositive plant pathogenic bacterium Clavibacter michiganensis C m subsp michiganensis C m subsp sepedonicus C m subsp insidiosus C m subsp nebraskensis and C m subsp tessellarius Specific primers designed for amplification of the housekeeping genes recA rpoB and rpoD generated 827 1037 and 862bp DNA fragments respectively PCR products obtained from 40 C michiganensis strains were analysed using RFLP with four restriction endonucleases and those PCR products with specific RFLP patterns were sequenced The genotypes discriminated after PCR–RFLP were specific for each subspecies and also allowed for differentiation of C m subsp michiganensis strains Sequence analysis of the recA rpoB and rpoD gene fragments also distinguished C michiganensis subspecies and was useful for phylogenetic analysis of all subspecies For rapid inexpensive and effective differentiation of the five subspecies in this research we recommend the amplification of recA and/or rpoD gene fragments and digestion of the PCR products with the restriction endonuclease FnuDIIThe authors thank Bogumila Tyszkiewicz from the State Plant Health and Seed Inspection Service in Gdansk and Dr Joanna Pulawska from the Research Institute of Pomology and Floriculture in Skierniewice for providing Cm strains This work was supported by the Ministry of Science and Higher Education Project No N 303 086 32/2778
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