Abstract
Developmentally specific expression of Rhizobium spp. genes involved in symbiotic N2 fixation is known to operate through cascade regulation of various nif and fix operons. Fusion constructs of lacZ under symbiotic promoters P1 (for nifHDK operon) and P2 (for fixABCX operon) of Rhizobium meliloti were mobilized into Rhizobium spp. (Cicer) strains Rcd301 and RCR13. The assays for β-galactosidase activity to monitor the expression of lacZ under these promoters was performed in host backgrounds of Escherichia coli, R. meliloti, and Rhizobium spp. (Cicer). The enzyme assays indicated significant levels of expression from P1 and P2 promoters in chickpea rhizobia, specifically in symbiotic cells from nodules. However, as in R. meliloti, these promoters did not induce strong expression in free-living cells of Rhizobium spp. (Cicer). This indicates functional homology of R. meliloti promoters in rhizobium spp. (Cicer). Functional cross-reactivity of trans regulatory factors like NtrA, NtrC, and NifA between these rhizobia seems evident from the nodule-specific expression of P1 and P2 cis elements.
Similar content being viewed by others
References
Albright LM, Huala E, Ausubel FM (1989) Prokaryotic signal transduction mediated by sensor and regulator protein pairs. Annu Rev Genet 23:311–336
Ayanaba A, Haugland RA, Sadowsky MJ, Upchurch RG, Weiland KD, Zablotowicz RM (1986) Rapid coloured nodule assay for assessing root exudate-enhanced competitiveness of Bradyrhizobium japonicum. Appl Environ Microbiol 52:847–851
Bahl PN (1988) Chickpea. In: Baldev B, Ramanujam S, Jain HK (eds) Pulse crops. Oxford and IBH Publ, New Delhi, pp 95–131
Better M, Lewis B, Corbin D, Ditta G, Helinski DR (1983) Structural relationships among Rhizobium meliloti symbiotic promoters. Cell 35:479–485
Better M, Lewis B, Corbin D, Ditta G, Helinski DR (1985) Deletion analysis of Rhizobium meliloti symbiotic promoters. EMBO J 4:2419–2424
Boyer HS, Roulland-Dussoix D (1969) A complementation analysis of restriction and modification of DNA in Escherichia coli. J Mol Biol 41:459–472
David M, Daveran ML, Batut J, Dedieu A, Domergue O, Ghai J, Hertig C, Boistard P, Kahn D (1988) Cascade regulation of nif gene expression in Rhizobium meliloti. Cell 54:671–683
Ditta G (1986) Tn5 mapping of Rhizobium nitrogen fixation genes. Methods Enzymol 118:519–529
Ditta G, Stanfield S, Corbin D, Helinski DR (1980) Broad host range DNA cloning system for Gram-negative bacteria: Construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci USA 77:7347–7351
Ditta G, Schmidhauser T, Yakobson E, Lu P, Liang-X-W, Finlay DR, Guiney D, Helinski DR (1985) Plasmids related to the broad host range vector, pRK290, useful for gene cloning and for monitoring gene expression. Plasmid 13:149–153
Ditta G, Virts E, Palomares A, Kim CH (1987) The nifA gene of Rhizobium meliloti is oxygen regulated. J Bacteriol 169:3217–3223
Gaur YD, Sen AN (1979) Cross inoculation group specificity in Cicer-Rhizobium symbiosis. New Phytol 83:745–754
Gilles-Gonzalez MA, Ditta GS, Helinski DR (1991) A haemoprotein with kinase activity encoded by the oxygen sensor of Rhizobium meliloti. Nature (London) 350:170–172
Glazebrook J, Walker GC (1991) Genetic techniques in Rhizobium meliloti. Methods Enzymol 204:398–418
Gussin GN, Ronson CW, Ausubel FM (1986) Regulation of nitrogen fixtion genes. Annu Rev Genet 20:567–591
Khanuja SPS (1991) Construction of a gene bank of Cicer Rhizobium. Curr Sci 61:545–548
Khanuja SPS, Kumar S (1989) Symbiotic and galactose utilization properties of phage RMP 64-resistant mutants affecting three complementation groups in Rhizobium meliloti. J Genet 68:93–108
Long SR (1989) Rhizobium-legume nodulation: Life together in the underground. Cell 56:203–214
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor
McKnight T (1949) Efficiency of isolates of Rhizobium in cowpea group with proposed additions to this group. Queensl J Agric Sci 6:61–76
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, pp 354–358
Nambiar PTC, Ma SW, Iyer VN (1990) Limiting an insect infestation of nitrogen fixing root nodules of the pigeon pea (Cajanus cajan) by engineering the expression of an entomocidal gene in its root nodules. Appl Environ Microbiol 56:2866–2869
Rossen L, Davis EO, Johnston AWB (1987) Plant-induced expression of Rhizobium genes involved in host specificity and early stages of nodulation. Trends Biochem Sci 12:430–433
Singh A, Agarwala R, Kashyap LR, Kumar S (1988) Behaviour of expression of nifHDK and fixABCX promoters of Rhizobium meliloti in R. leguminosarum. Indian J Biochem Biophys 25:389–391
Skot L, Harrison SP, Nath A, Mytton LR, Clifford BC (1990) Expression of insecticidal activity in Rhizobium containing deltaendotoxin gene cloned from Bacillus thuringiensis subsp. tenebrionis. Plant and Soil 127:285–295
Sundaresan V, Ow DW, Ausubel FM (1983) Activation of Klebsiella pneumoniae and rhizobium meliloti nitrogenase promoters by gln regulatory proteins. Proc Natl Acad Sci USA 80:4030–4034
Vance CP, Egli MA, Griffith SM, Miller SS (1988) Plant regulated aspects of nodulation and N2 fixation. Plant Cell Environ 11:413–427
Vincent JM (1970) A manual for the practical study of the root nodule bacteria. Blackwell Scientific Publications, Oxford Edingburgh
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Khanuja, S.P.S., Suman, A., Singh, G. et al. Nodule-specific expression of Rhizobium meliloti symbiotic promoters P1 and P2 in chickpea-Rhizobium sp. symbiosis. Biol Fert Soils 18, 132–136 (1994). https://doi.org/10.1007/BF00336459
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00336459