Supply of acetyl-CoA to N2-fixing bacteroids: insights from the mutational and proteomic analyses of Sinorhizobium meliloti

Authors

  • Olga Onishchuk All-Russia Research Institute for Agricultural Microbiology, Shosse Podbel'skogo, 3, Saint Petersburg, 196608, Russian Federation https://orcid.org/0000-0002-5378-7826
  • Oxana Kurchak All-Russia Research Institute for Agricultural Microbiology, Shosse Podbel'skogo, 3, Saint Petersburg, 196608, Russian Federation https://orcid.org/0000-0003-3555-7426
  • Elena Chizhevskaya All-Russia Research Institute for Agricultural Microbiology, Shosse Podbel'skogo, 3, Saint Petersburg, 196608, Russian Federation https://orcid.org/0000-0002-7715-8696
  • Boris Simarov All-Russia Research Institute for Agricultural Microbiology, Shosse Podbel'skogo, 3, Saint Petersburg, 196608, Russian Federation https://orcid.org/0000-0002-6893-557X
  • Nikolay Provorov All-Russia Research Institute for Agricultural Microbiology, Shosse Podbel'skogo, 3, Saint Petersburg, 196608, Russian Federation https://orcid.org/0000-0001-9091-9384

DOI:

https://doi.org/10.21638/spbu03.2021.203

Abstract

Rhizobia represent a diverse group of gram-negative bacteria capable of fixing atmospheric nitrogen in symbiosis with leguminous plants. Mechanisms of symbiotic efficiency are important to study not only to reveal the “fine tuning” of the host–symbiont supra-organismal genetic system emergence, but also to develop agriculture with minimal environmental risks. In this paper we demonstrate that among seven genes whose inactivation by Tn5 insertions results in an increased efficiency of rhizobia (Sinorhizobium meliloti) symbiosis with alfalfa (Eff++ phenotype), six genes are involved in the metabolism of small molecules. One of them (SMc04399) encodes for acetate-CoA transferase catalyzing the formation of acetyl-CoA from acyl-CoA. Since acetyl-CoA is required for operation of the Krebs cycle, providing ATP for symbiotic N2 fixation, we suggest that a significant portion of this coenzyme utilized by bacteroids is provided by the plant cell supporting the energy-consuming nitrogenase reaction. Proteomic data analysis allow us to reveal the lability of enzymatic pathways which are involved in bacteroids in the production and catabolism of acetyl-CoA and which should be modified to obtain the Eff++ phenotype. This phenotype was developed also after inactivation of NoeB protein which is involved in the host-specific nodulation and is characterized by an elevated production in wild type S. meliloti bacteroids, suggesting a multifunctional role of noeB in the symbiosis operation.

Keywords:

Sinorhizobium meliloti, symbiotic efficiency, metabolism of small molecules, acetyl CoA, acetate-CoA transferase, transposon (Tn5) mutagenesis, proteomic analysis

Downloads

Download data is not yet available.
 

References

Antonets, K. S., Onishchuk, O. P., Kurchak, O. N., Volkov, K. V., Lykholay, A. N., Andreeva, E. A., Andronov, E. E., Pinaev, A. G., Provorov, N. A., and Nizhnikov, A. A. 2018. Proteomic profile of the bacterium Sinorhizobium meliloti depends on its life form and host plant species. Molecular Biology 52(5):779–785. https://doi.org/10.1134/S0026898418050038

Ardourel, M., Lortet, G., Maillet, F., Roche, P., Truchet, G., Promé, J. C., and Rosenberg, C. 1995. In Rhizobium meliloti, the operon associated with the nod box n5 comprises nodL, noeA and noeB, three host-range genes specifically required for the nodulation of particular Medicago species. Molecular Microbiology 17(4):687–699. https://doi.org/10.1111/j.1365-2958.1995.mmi_17040687.x

Beringer, J. E. 1974. R factor transfer in Rhizobium leguminosarum. Microbiology 84(1):188–198. https://doi.org/10.1099/00221287-84-1-188

Cabanes, D., Boistard, P., and Batut, J. 2000. Symbiotic induction of pyruvate dehydrogenase genes from Sinorhizobium meliloti. Molecular Plant-Microbe Interactions 13(5):483–493. https://doi.org/10.1094/MPMI.2000.13.5.483

Chizhevskaya, E. P., Onishchuk, O. P., Andronov, E. E., and Simarov, B. V. 2011. Use of the site-directed mutagenesis method to study the functions of the SMb20332 gene in alfalfa nodule bacteria Sinorhizobium meliloti. Sel'skokhozyaistvennaya biologiya 3:55–61. (In Russian)

Contador, C. A., Lo, S. K., Chan, S. H. J., and Lam, H. M. 2020. Metabolic analyses of nitrogen fixation in the soybean microsymbiont Sinorhizobium fredii using constraint-based modeling. mSystems 5(1):e00516-19. https://doi.org/10.1128/mSystems.00516-19

Dunn, M. F. 1998. Tricarboxylic acid cycle and anaplerotic enzymes in rhizobia. FEMS Microbiology Reviews 22(2):105–123. https://doi.org/10.1111/j.1574-6976.1998.tb00363.x

Fedorov, S. N. and Simarov, B. V. 1987. Obtaining mutants with altered symbiotic properties in Rhizobium meliloti under the impact of UV rays. Sel'skokhozyaistvennaya biologiya 9:44–49. (In Russian)

Gao, M., Nguyen, H., González, I. S., and Teplitski, M. 2016. Regulation of fixLJ by Hfq controls symbiotically important genes in Sinorhizobium meliloti. Molecular Plant-Microbe Interactions 29(11):844–853. https://doi.org/10.1094/MPMI-09-16-0182-R

Green, R. T., East, A. K., Karunakaran, R., Downie, J. A., and Poole, P. S. 2019. Transcriptomic analysis of Rhizobium leguminosarum bacteroids in determinate and indeterminate nodules. Microbial Genomics 5(2):e000254. https://doi.org/10.1099/mgen.0.000254

Lakin, G. F. 1980. Biometrics. Moscow, Visshaya Shkola. 293 p. (In Russian)

Lerouge, P., Roche, P., Faucher, C., Maillet, F., Truchet, G., and Prome, J. C. 1990. Symbiotic host-specificity of Rhizobium meliloti is determined by a sulphated and acylated glucosamine oligosaccharide signal. Nature 344(6268):781–784. https://doi.org/10.1038/344781a0

Liu, A., Contador, C. A., Fan, K., and Lam, Y.-M. 2018. Interaction and regulation of carbon, nitrogen, and phosphorus metabolisms in root nodules of legumes. Frontiers in Plant Science 9:1860. https://doi.org/10.3389/fpls.2018.01860

Mulley, G., Lopez-Gomez, M., Zhang, Y., Terpolilli, J., Prell, J., Finan, T., and Poole, P. 2010. Pyruvate is synthesized by two pathways in pea bacteroids with different efficiencies for nitrogen fixation. Journal of Bacteriology 192(19):4944–4953. https://doi.org/10.1128/JB.00294-10

Novikova, A. T. and Simarov, B. V. 1979. Methodical recommendations for obtaining new strains of nodule bacteria and assessing their effectiveness. Leningrad. 50 p. (In Russian)

Onishchuk, O. P., Chizhevskaya, E. P., Kurchak, O. N., Andronov, E. E., and Simarov, B. V. 2014. Identification of new genes of the nodule bacteria Sinorhizobium meliloti involved in control of the efficiency of symbiosis with Medicago sativa. Ecological Genetics 12(1):39–47. https://doi.org/10.17816/ecogen12139-47 (In Russian)

Onishchuk, O. P., Sharypova, L. A., and Simarov, B. V. 1994. Isolation and characterization of the Rhizobium meliloti Tn5-mutants with impaired nodulation competitiveness. Plant and Soil 197(1):267–274. https://doi.org/10.1007/BF00007953

Onishchuk, O. P., Vorobyov, N. I., Provorov, N. A., and Simarov, B. V. 2009. Symbiotic activity of alfalfa rhizobia (Sinorhizobium meliloti) with genetic modifications in dicarboxylate transport. Ecological Genetics 7(2):3–10. https://doi.org/10.17816/ecogen72 (In Russian)

Provorov, N. A., Onishchuk, O. P., Yurgel, S. N., Kurchak, O. N., Chizhevskaya, E. P., Vorobyov, N. I., Zatovskaya, T. V., and Simarov, B. V. 2014. Construction of highly-effective symbiotic bacteria: evolutionary models and genetic approaches. Russian Journal of Genetics 50(11):1273–1285. https://doi.org/10.1134/S1022795414110118

Roche, P., Debelle, F., Maillet, F., Lerouge, P., Faucher, C., and Truchet, G. 1991. Molecular basis of symbiotic host specificity in Rhizobium meliloti: nodH and nodPQ genes encode the sulfation of lipo-oligosaccharide signals. Cell 67(6):1131–1143. https://doi.org/10.1016/0092-8674(91)90290-F

Sharypova, L. A., Onishchuk, O. P., Chesnokova, O. N., Fomina-Eshchenko, J. G., and Simarov, B. V. 1994. Isolation and characterization of Rhizobium meliloti Tn5 mutants showing enhanced symbiotic effectiveness. Microbiology 140(2):463–470. https://doi.org/10.1099/00221287-140-3-463

Simon, R., Priefer, U., and Pühler, A. 1983. A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in gram negative bacteria. Biotechnology 1(11):784–791. https://doi.org/10.1038/nbt1183-784

Werner, M., Semsey, S., Sneppen, K., and Krishna, S. 2009. Dynamics of uptake and metabolism of small molecules in cellular response systems. Plos ONE 4(3):e4923. https://doi.org/10.1371/journal.pone.0004923

Biological Communications

Downloads

Published

2021-06-30

How to Cite

Onishchuk, O., Kurchak, O., Chizhevskaya, E., Simarov, B., & Provorov, N. (2021). Supply of acetyl-CoA to N<sub>2</sub>-fixing bacteroids: insights from the mutational and proteomic analyses of <em>Sinorhizobium meliloti</em>. Biological Communications, 66(2), 124–128. https://doi.org/10.21638/spbu03.2021.203

Issue

Vol. 66 No. 2 (2021)

Section

Full communications

Categories

License

Articles of Biological Communications are open access distributed under the terms of the License Agreement with Saint Petersburg State University, which permits to the authors unrestricted distribution and self-archiving free of charge.