Characterization of rhizobial bacteria isolated from arsenic-contaminated site in south-eastern Kurdistan province and their influence on plant growth

Symbiotic bacteria associated with the root of legume plants were isolated from soil and the root nodules of alfalfa (Medicago sativaL.), chickpea (Cicer arietinum L.) and yellows alfalfa (Melilotus officinalis L.) grown in arsenic-contaminated areas in south eastern part of Kurdistan province, Iran. According to physiological and biochemical properties, isolates were identified as the members of Rhizobium and Ensifer genus. The presence of arsenic resistant system known as ars system was confirmed by amplification using primers corresponding to arsC gene. Growth rate of strains in different concentrations of arsenate was investigated in YEMB medium supplemented with 100-400 mM arsenate. Alfalfa,s isolates (AB1, AB3), chickpea,s isolates (PA2, PC2), yellow alfalfa,s isolate (YA1) and standard strain (Sinorhizobium meliloti SM117) were tolerant to 350 mM arsenate and (AB1, PA2 and YA1) isolates were moderately capable to grow at 400 mM arsenate. Among the identified strains, AB1 and PA2, were selected for greenhouse experiments. In order to evaluate the effect of arsenic contamination on legume-rhizobia symbiosis and biomass production of alfalfa and chickpea plants, an experiment was carried out based on a completely randomized design with a factorial arrangement in three replications. Experiment factors consisted of, three levels of rhizobia inoculation (with and without inoculation of AB1 and PA2) and five levels of arsenic concentrations (0. 10, 50, 75 and 100 mgkg-1 soil) under greenhouse conditions for 8 weeks. Results obtained in this study indicated that the fresh weight and dry weight of alfalfa and chickpea (shoot and root) were decreased as the arsenic concentration of the soil was increased. The results also showed that fresh and dry shoot and root weight of alfalfa and chickpea were significantly higher in rhizobia-inoculated treatments compared to non-inoculated plants.