Molecular Methods for Assessement the Bacterial Communities from Different Type of Soils in Romania

Abstract

Rhizobia are soil bacteria that are capable to form nitrogen-fixing symbiosis with leguminous plants. This ability, as well as the diversity of microbial populations in the soil, and in the rhizosphere of host plants and non-host plants is influenced by several factors, including crop management. The aim of this work was the examination of the influence of some factors on indigenous populations of rhizobia in soils under different crop managements. The genetic diversity of rhizobial strains isolated directly from soil (free-living state) or from root nodules of three herbaceous perennial legumes was examined. The study was conducted in the experimental fields located in Moara Domnească area (South of Romania) and in the Brașov County. The characteristics of brown reddish soil were determined (nitrogen content, organic carbon content and pH). Counting of the rhizobia populations was done by most probable number estimation and by viable plate counts. Bacterial strains were isolated directly from soil samples or from root nodules of different plant species (Trifolium repens, T. pratense and Lotus corniculatus). The characterization of rhizobia was performed by DNA fingerprinting (ERIC PCR and BOX PCR) and the bacterial diversity of soils was examined by DGGE technique. The results revealed that the rhizobial diversity was significantly lower in soils under increased fertilization with N. A reduced intraspecific polymorphism was observed in the strains recovered from the same plant species (Trifolium spp.), whatever the origin of the plant (Moara Domnească or Brașov) but clear differences appeared to be related to the origin of nodules (red or white clover) as revealed by DNA fingerprints. However, various amplicon profiles were observed by DGGE when total DNA isolated from soils was examined, the differences being associated with the fertilization level.