Artificial activation of nif gene expression in nodule bacteria Ex Planta

Abstract

Background. Rhizobia are the most effective nitrogen-fixing organisms that can fix nitrogen only in symbiosis with leguminous plants. The general transcriptional activator of nitrogen fixation genes in diazotrophic bacteria is NifA. In this work, the possibility of modifying the regulation of nitrogen fixation in the nodule bacteria Mesorhizobium, Ensifer and Rhizobium was studied by introducing an additional copy of the nifA gene into the bacterial genomes during the regulation of induced bacterial promoters.

Materials and methods. A series of expression genetic constructs with NifA genes of nodule bacteria strains under the control of an inducible promoter Pm were created. The resulting constructs were transformed into strains of nodule bacteria. The obtained recombinant strains were investigated for the appearance of their nitrogen-fixing activity in the free-living state.

Results. It was shown that the expression of nifA in recombinant cells of all three genera of bacteria leads to the appearance of insignificant nitrogenase activity. At the same time, the level of nitrogenase activity does not have a correlation with the level of expression of the introduced nifA gene, which, most likely, is a consequence of the multilevel regulation of nitrogen fixation.

Conclusion. The possibility of artificial activation of nitrogenase activity in nodule bacteria in the free-living state by introducing the NifA regulatory protein gene into bacteria was shown.

Keywords

nitrogen fixation / nodule bacteria / nifA / recombinant bacteria / plasmid vector / promoter

Funding

CCU “Biomika” of the Institute of Biochemistry and Genetics of UFRC RASЦКП «Биомика» Института биохимии и генетики УФИЦ РАН()

Russian Foundation for Basic ResearchРоссийский фонд фундаментальных исследований()

Russian Foundation for Basic ResearchРоссийский фонд фундаментальных исследований()

Innovation Research GrantГрант по инновационным исследованиям «У.М.Н.И.К.»()

Russian Science FoundationРоссийский научный фонд()

RIGHTS & PERMISSIONS

Baymiev An.K., Gumenko R.S., Vladimirova A.A., Akimova E.S., Vershinina Z.R., Baymiev Al.K.