Phylogenetic relationship and genetic diversity of Agrobacterium spp. isolated in Poland based on gyrB gene sequence analysis and RAPD
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The genetic diversity of 47 strains of Agrobacterium originating from different host plants and geographical locations in Poland, together with 12 strains from other countries was investigated. It was analyzed using RFLP of DNA fragment amplified with primers UP-1 and UP-2r flanking part of gyrB and parE genes, gyrB sequencing and randomly amplified polymorphic DNA (RAPD) technique. On the basis of obtained results, we found the majority of agrobacteria isolated in Poland belong to biovar 2. However, among others, three strains distinct from type strains of all the known Agrobacterium species, were discovered. All three methods showed no correlation between genetic diversity and geographical origin or the host plant of all studied strains but they revealed high diversity of the tested agrobacteria. The highest diversity was observed within strains of biovar 1, whereas those of biovar 2 were found to be the more homogenous group. The topology of the constructed gyrB tree corresponds to topologies of 16S and 23S rDNA trees obtained in this and other studies, but the gyrB tree had deeper branching. In the case of RAPD, it was possible to find a unique DNA fingerprint for almost each strain tested. The gyrB gene appeared to be a good phylogenetic marker with high discrimination power allowing better differentiation between species and strains, whereas the RAPD technique can serve as a tool for single strain typing.
KeywordsCrown gall Fruit trees Hairy-roots
Authors wishes to thank: Noëlle Amarger (INRA, France), Xavier Nesme (University of Lyon, France), Esperanza Martinez Romero (CCG, Mexico), Xinhua Sui (ChAU, China), Marta Zawadzka (IH, Poland) for providing bacterial strains. This work was partly supported by the Polish Scientific Committee (KBN) Grant 580/E-177/SPB/COST/P-06/DZ 447/2002-2005.
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