Molecular characterization and evaluation of crude oil remediation potential of some rhizobia isolated from plant root nodules

Abstract

This study aimed to determine the molecular identities and genetic relatedness of rhizobia isolated from pigeon pea and pinto beans, and assess their remediation potential in the presence of 1 %, 3 % and 5 % (w/v) crude oil in minimal medium for 7 days incubation period. Standard microbiological and molecular methods which include amplification and purification of 16S rRNA, agarose gel electrophoresis, and sequencing. Results showed molecular identities of six rhizobia from pigeon peas as Bradyrhizobium diazoefficiens USDA122, Rhizobium leguminosarum WSM2304, Bradyrhizobium japonicum N61, Rhizobium leguminosarum N741, Rhizobium leguminosarum BIHIB1217, and Bradyrhizobium japonicum E109; and three rhizobia obtained from pinto beans were Rhizobium leguminosarum N871, Bradyrhizobium diazoefficiens USDA110 and Bradyrhizobium japonicum SEMIA5079. All tested rhizobia (9) showed petroleum degradation ability, as they all grew in the 1, 3 and 5 % (w/v) crude oil minimal medium under laboratory conditions. B. diazoefficiens USDA122 showed the highest optical density (OD) value of 1.184 ± 0.05 on 7th day at 1 % (w/v) crude oil contamination, while R. leguminosarum N741 has the lowest OD value of 0.372 ± 0.02 at 5 % (w/v) crude oil on 7^th day. For all the rhizobia, increase occurred throughout incubation period at 1, 3 and 5 % (w/v) except Rhizobium leguminosarum N741 and R. leguminosarum BIHIB1217. In conclusion, the association of R. leguminosarum BIHIB1217 and R. leguminosarum N871 from pigeon pea and pinto beans respectively, were found most effective in crude oil degradation and thus they are recommended as a promising association for remediation of crude oil spilled soils.