The new taxonomic marker of nodulation bacteria of Rhizobium genus and its evolution

Abstract

The new taxonomic marker (hin-region) has been proposed, which gives possibility for Rhizobium bacteria study on “species — group of strains” level. Using this marker the groups of Rhizobium strains were determined, which could not be distinguished with other methods, and these results correlated with evolutionary similarity of the bacteria. The developed approach for creating marker systems allows to carry out effective inventory of inter- and intraspecies genetic diversity of nodulating bacteria and to evaluate perspectives of their use in agriculture. The proposed marker system was used for description of Rhizobium bacteria samples isolated from various ecological-geographical regions of Ukraine.

Genetic Diversity of Bacillus thuringiensis from Different Geo-Ecological Regions of Ukraine by Analyzing the 16S rRNA and gyrB Genes and by AP-PCR and saAFLP

“…We had modified the AFLP method developed and patented by M. Zabeau and P. Vos [36]; its suitability for the analysis of closely related Bt strains was assessed in this study. The phylogenetic relationships between closely related strains of various species belonging to the genus Rhizobium had been successfully analyzed using this modified saAFLP method [ 35 ]. The saAFLP procedure comprises three steps: (i) simultaneous treatment of the extracted bacterial DNA in the same tubes using one of the restriction endonucleases (XmaJI, XbaI, PstI) and ligation with a singlestranded adapter Ad.CT AG1; (ii ) PCR amplification with a single primer complementary to the Ad.CT AG1 sequence; (iii ) electrophoretic separation of the PCR products in agarose gel.…”

Root Nodule Bacteria: Perspectives of Monitoring Symbiotic Properties by Applying Genetic Markers

“…Наилучшим образом зарекомендовал себя метод анализа нуклеотидной последовательности гена 16S рРНК, который позволяет характеризовать штаммы не только на уровне вида, но и выделять геновиды (genospecies) ризобий (23,24). Также нередко прибегают к анализу межгенной последовательности rrn-rrl рибосомального оперона (ITS, intergenic sequence) или ее участков, например, hin-региона (25)(26)(27). Анализ указанных последовательностей позволяет определять не только видовую принадлежность штаммов, но и получать их штаммоспецифичные характеристики на уровне хромосомных маркеров (28), что, однако, не гарантирует наследования штаммом/штаммами генетических детерминант стрессоустойчивости и симбиотической активности.…”