Creation of an inducible vector system based on the rhizobia nodA gene promoter
- Authors: Chubukova O.V.1, Vershinina Z.R.1, Matnyazov R.T.1, Baymiev A.K.1, Baymiev A.K.1
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Affiliations:
- Institute of Biochemistry and Genetics – Subdivision of the Ufa Federal Research Centre of the Russian Academy of Sciences
- Issue: Vol 19, No 1 (2021)
- Pages: 13-21
- Section: Genetic basis of ecosystems evolution
- URL: https://journals.rcsi.science/ecolgenet/article/view/48646
- DOI: https://doi.org/10.17816/ecogen48646
- ID: 48646
Cite item
Abstract
BACKGROUND: The possibility of changing the properties of rhizobial bacteria by giving them the ability to regulate the expression of additionally introduced genes into them is an urgent task both for fundamental science and for applied agrobiology, since this will make it possible to obtain microsymbionts with desired properties. An expression construct using the rhizobia regulatory system was created in this work. The rhizobia nodD gene encodes a regulatory protein that, in the presence of plant inducers, flavonoids, activates the transcription of nod-genes involved in the early stages of the formation of legume-rhizobium symbiosis.
MATERIALS AND METHODS: A vector construct containing the nodD gene from Rhizobium leguminosarum bv. trifoli under the regulation of its own promoter and the gfp gene under the regulation of the nodA gene promoter from the same rhizobia was obtained. Neorhizobium galegae CIAM 0702 were transformed with the vector construct.
RESULTS: It has been shown that in recombinant strains synthetic flavonoids are capable of inducing expression of gfp gene to varying degrees.
CONCLUSION: In the future, the results can be used to obtain rhizosphere microorganisms with a controlled synthesis of growth-stimulating and protective substances.
Keywords
Full Text
##article.viewOnOriginalSite##About the authors
Olga V. Chubukova
Institute of Biochemistry and Genetics – Subdivision of the Ufa Federal Research Centre of the Russian Academy of Sciences
Author for correspondence.
Email: chubukova@bk.ru
ORCID iD: 0000-0001-7553-9527
SPIN-code: 3297-4830
ResearcherId: A-3397-2014
PhD, Cand. Sci. (Biol.), Researcher
Russian Federation, 71 October Av., 450054 UfaZilya R. Vershinina
Institute of Biochemistry and Genetics – Subdivision of the Ufa Federal Research Centre of the Russian Academy of Sciences
Email: zilyaver@mail.ru
ORCID iD: 0000-0001-5480-5320
SPIN-code: 1866-7896
PhD, Cand. Sci. (Biol.), Researcher
Russian Federation, 71 October Av., 450054 UfaRustam T. Matnyazov
Institute of Biochemistry and Genetics – Subdivision of the Ufa Federal Research Centre of the Russian Academy of Sciences
Email: rmat@mail.ru
SPIN-code: 6798-7913
PhD, Cand. Sci. (Biol.)
Russian Federation, 71 October Av., 450054 UfaAndrey K. Baymiev
Institute of Biochemistry and Genetics – Subdivision of the Ufa Federal Research Centre of the Russian Academy of Sciences
Email: baymiev@anrb.ru
ORCID iD: 0000-0001-6637-9365
SPIN-code: 1919-5236
PhD, Cand. Sci. (Biol.)
Russian Federation, 71 October Av., 450054 UfaAleksey K. Baymiev
Institute of Biochemistry and Genetics – Subdivision of the Ufa Federal Research Centre of the Russian Academy of Sciences
Email: alex@anrb.ru
ORCID iD: 0000-0003-0606-6740
SPIN-code: 3771-4063
PhD, Cand. Sci. (Biol.), Head of Laboratory
Russian Federation, 71 October Av., 450054 UfaReferences
- Gopalakrishnan S, Sathya A, Vijayabharathi R, Varshney RK, et al. Plant growth promoting rhizobia: challenges and opportunities. 3 Biotech. 2015;5(4):355–377. doi: 10.1007/s13205-014-0241-x
- Bajmiev AnH, Gumenko RS, Vladimirova AA. et al. Artificial activation of nif gene expression in nodule bacteria ex planta. Ecological genetics. 2019;17(2):35–42. (In Russ.) doi: 10.17816/ecogen17235-42.
- Vershinina ZR, Khakimova LR, Lavina AM, et al. Effect of constitutive expression of the rapA1 gene on formation of bacterial biofilms and growth-stimulating activity of rhizobia. Microbiology. 2019;88(1):54–62. (In Russ.) doi: 10.1134/s0026365619010105
- Clua J, Roda C, Zaneti ME, Blanco FA. Compatibility between legumes and rhizobia for the establishment of a successful nitrogen fixing symbiosis. Genes (Basel). 2018;9(3):125. doi: 10.3390/genes9030125
- Andrews M, Andrews ME. Specificity in Legume-Rhizobia Symbioses. Int J Mol Sci. 2017;18(4):705. doi: 10.3390/ijms18040705
- Lindström K, Mousavi SA. Effectiveness of nitrogen fixation in rhizobia. Microb Biotechnol. 2020;13(5):1314–1335. doi: 10.1111/1751-7915.13517
- Liu C-W, Murray JD. The Role of Flavonoids in Nodulation Host-Range Specificity: An Update. Plants-Basel. 2016;5(3):33. doi: 10.3390/plants5030033
- Fauvart M, Michiels J. Rhizobial secreted proteins as determinants of host specificity in the rhizobium-legume symbiosis. FEMS Microbiol Lett. 2008;285(1):1–9. doi: 10.1111/j.1574-6968.2008.01254.x
- Jiménez-Guerrero I, Acosta-Jurado S, Del Cerro P, et al. Transcriptomic Studies of the Effect of nod Gene-Inducing Molecules in Rhizobia: Different Weapons, One Purpose. Genes (Basel). 2017;9(1):1. doi: 10.3390/genes9010001
- Hu H, Liu S, Yang Y, et al. In Rhizobium leguminosarum, NodD represses its own transcription by competing with RNA polymerase for binding sites. Nucleic Acids Res. 2000;28(14):2784–2793. doi: 10.1093/nar/28.14.2784
- Rossen L, Shearman CA, Johnston AWB, Downiel JA. The nodD gene of Rhizobium leguminosarum is autoregulatory and in the presence of plant exudate induces the nodA, B, C genes. EMBO J. 1985;4(13A):3369–3373. doi: 10.1002/j.1460-2075.1985.tb04092.x
- Spaink HP, Okker RJ, Wijffelman CA, et al. Promoters in the nodulation region of the Rhizobium leguminosarum Sym plasmid pRL1JI. Plant Mol Biol. 1987;9(1):27–39. doi: 10.1007/BF00017984
- Begum AA, Leibovitch S, Migner P, Zhang F. Specific flavonoids induced nod gene expression and pre-activated nod genes of Rhizobium leguminosarum increased pea (Pisum sativum L.) and lentil (Lens culinaris L.) nodulation in controlled growth chamber environments. J Exp Bot. 2001;52(360):1537–1543. doi: 10.1093/jexbot/52.360.1537
- Maj D, Wielbo J, Marek-Kozaczuk M, Skorupska A. Response to flavonoids as a factor influencing competitiveness and symbiotic activity of Rhizobium leguminosarum. Microbiol Res. 2010;165(1): 50–60. doi: 10.1016/j.micres.2008.06.002
- Spaink HP, Wijffelman CA, Pees E, et al. Rhizobium nodulation gene nodD as a determinant of host specificity. Nature. 1987;328:337–340. doi: 10.1038/328337a0
- Suominen L, Luukkainen R, Roos C, Lindström K. Activation of the nodA promoter by the nodD genes of Rhizobium galegae induced by synthetic flavonoids or Galega orientalis root exudate. FEMS Microbiol Lett. 2003;219(2):225–232. doi: 10.1016/s0378-1097(02)01206-5
- Hugria M, Jonston AWB, Phillips DA. Effects of flavonoids release naturally from bean (Phaseolus vulgaris) on nodD-regulated gene transcription in Rhizobium leguminosarum bv. phaseoli. Mol Plant Microbe Interact. 1992;5(3):199–203. doi: 10.1094/mpmi-5-199
- Dakora FD, Joseph CM, Phillips DA. Alfalfa (Medicago sativa L.) root exudates contain isoflavonoids in the presence of Rhizobium meliloti. Plant Physiol. 1993;101(3):819–824. doi: 10.1104/pp.101. 3.819
- Janczarek M, Urbanik-Sypniewska T, Skorupska А. Effect of authentic flavonoids and the exudate of clover roots on growth rate and inducing ability of nod genes of Rhizobium leguminosarum bv. trifolii. Microbiol Res. 1997;152(1):93–98. doi: 10.1016/s0944-5013(97)80028-6
- Feng J, Li Q, Hu HL, et al. Inactivation of the nod box distal half-site allows tetrameric NodD to activate nodA transcription in an inducer-independent manner. Nucleic Acids Res. 2003;31(12): 3143–3156. doi: 10.1093/nar/gkg411
- Stacey G. Bradyrhizobium japonicum nodulation genetics. FEMS Microbiol Lett. 1995;127(1-2):1–9. doi: 10.1111/j.1574-6968.1995.tb07441.x
- Suominen L, Roos C, Lortet G, et al. Identification and structure of the Rhizobium galegae common nodulation genes: evidence for horizontal gene transfer. Mol Biol Evol. 2001;18(6):907–916. doi: 10.1093/oxfordjournals.molbev.a003891
- Lin JJ. Electrotransformation of Agrobacterium. Methods Mol Biol. 1995;47:171–178. doi: 10.1385/0-89603-310-4:171.
- Mongiardini EJ, Ausmees N, Perez-Gimenez J, et al. The Rhizobial adhesion protein RapA1 is involved in adsorption of Rhizobia to plant roots but not in nodulation. FEMS Microbiol Ecol. 2008;65(2):279–288. doi: 10.1111/j.1574-6941.2008.00467.x
- Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA. 1979;76(9): 4350–4354. doi: 10.1073/pnas.76.9.4350
- Thompson JD, Higgins DG, Gibson TJ. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994;22(22):4673–4680. doi: 10.1093/nar/22.22.4673.
- Osterman J, Chizhevskaja EP, Andronov EE, et al. Galega orientalis is more diverse than Galega officinalis in Caucasus – whole-genome AFLP analysis and phylogenetics of symbiosis-related genes. Mol Ecol. 2011;20(22):4808–4821. doi: 10.1111/j.1365-294x.2011.05291.x.
- Machado D, Pueppke SG, Vinardell JM, et al. Expression of nodD1 and nodD2 in Sinorhizobium fredii, a Nitrogen-Fixing Symbiont of Soybean and Other Legumes. MPMI. 1998;11(5):375–382. doi: 10.1094/mpmi.1998.11.5.375.