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How does l-glutamate transport relate to selection of mixed nitrogen sources in Rhizobium leguminosarum biovar trifolii MNF1000 and cowpea Rhizobium MNF2030?

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Abstract

When growing on a mixture of ammonia and l-glutamate as nitrogen sources, Rhizobium leguminosarum biovar trifolii MNF1000 utilizes ammonia exclusively, while cowpea Rhizobium MNF2030 utilizes both compounds at similar rates. l-Glutamate transport in both strain MNF1000 and MNF2030 is active, giving rise to a 60-fold concentration gradient across the membrane of cells of strain MNF2030. Both strains produce two kinetically distinguishable glutamate transport systems under all conditions of growth — a high affinity system with an apparent K m of 0.06–0.17 μM but of relatively low V max, and a low affinity system with a K m of 1.2–6.7\ μM, but of higher overall capacity. l-Glutamate transport activity in cells of MNF2030 was relatively insensitive to the presence of ammonia in the growth medium. By contrast, ammonia in the growth medium resulted in low activities of glutamate transport in cells of MNF1000 which were provided with a carbon source, offering one explanation for the failure of this strain to use glutamate in the presence of ammonia. However, in cells of MNF1000 growing on glutamate as sole source of carbon and nitrogen, the glutamate transport system is synthesized, even in the presence of accumulated or added ammonia. This suggests that the regulation of the glutamate permease also depends on availability of carbon source.

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Abbreviations

CCCP:

carbonyl cyanide m-chlorophenyl hydrazone

HEPES:

N-hydroxyethylpiperazine-N′-2-ethanesulphonic acid

References

  • Bernt E, Bergmeyer HU (1974) l-Glutamate UV-assay with glutamate dehydrogenase and NAD. In: Bergmeyer HU (ed) Methods of enzymatic analysis, 2nd edn, vol 4. Academic Press, New York, pp 1704–1708

    Google Scholar 

  • Botsford JL (1984) Osmoregulation in Rhizobium meliloti: inhibition of growth by salts. Arch Microbiol 137:124–127

    Google Scholar 

  • Brown CM, Dilworth MJ (1975) Ammonia assimilation by Rhizobium cultures and bacteroids. J Gen Microbiol 86:39–48

    Google Scholar 

  • Brown CM, Macdonald-Brown DS, Meers JL (1974) Physiological aspects of microbial inorganic nitrogen metabolism. Adv Microb Physiol 11:1–52

    Google Scholar 

  • D'Aniello A, D'Onofrio G, Pischetola M, Strazzullo L (1985) Effect of various substances on the colorimetric amino acid-ninhydrin reaction. Anal Biochem 144:610–611

    Google Scholar 

  • Dilworth MJ, Thorneley RNF (1981) Nitrogenase of Klebsiella pneumoniae. Hydrazine is a product of azide reduction. Biochem J 193:971–983

    Google Scholar 

  • Doi E, Shibata D, Matoba T (1981) Modified colorimetric ninhydrin methods for peptidase assay. Anal Biochem 118:173–184

    Google Scholar 

  • Fawcett JK, Scott JE (1960) A rapid and precise method for the determination of urea. J Clin Pathol 13:156–159

    Google Scholar 

  • Fottrell PF, Mooney P (1969) The regulation of some enzymes involved in ammonia assimilation by Rhizobium japonicum. J Gen Microbiol 59:211–214

    Google Scholar 

  • Glenn AR, Arwas R, McKay IA, Dilworth MJ (1984) Fructose metabolism in wild-type, fructokinase-negative and revertant strains of Rhizobium leguminosarum. J Gen Microbiol 130: 231–237

    Google Scholar 

  • Gross W, Ring K (1971) Active transport of glutamate in Streptomyces hydrogenans. I. Studies of uptake and pool size, and their interrelationship. Biochim Biophys Acta 233:652–665

    Google Scholar 

  • Halpern YS, Even-Shoshan A (1967) Properties of the l-glutamate transport system in Escherichia coli. J Bacteriol 93:1009–1016

    Google Scholar 

  • halpern YS, Umbarger HE (1961) Utilization of l-glutamate and 2-oxoglutaric acid as sole source of carbon by Escherichia coli. J Gen Microbiol 26:175–183

    Google Scholar 

  • Howitt SM, Gresshoff PM (1985) Ammonia regulation of glutamine synthetase in Rhizobium sp. ANU289. J Gen Microbiol 131: 1433–1440

    Google Scholar 

  • Jin HN, Glenn AR, Dilworth MJ (1988) Ammonium uptake by cowpea Rhizobium strain MNF2030 and Rhizobium trifolii MNF1001. Arch Microbiol 149:308–311

    Google Scholar 

  • Kahn ML, Kraus J, Somerville JE (1985) A model of nutrient exchange in the Rhizobium-legume symbiosis. In: Evans HJ, Bottomley PJ, Newton WE (eds) Nitrogen fixation research progress. Martinus Nijhoff, Dordrecht, The Netherlands, pp 193–199

    Google Scholar 

  • Kustu SG, McFarland NC, Esmon SPH, Ames GF (1979) Nitrogen control of glutamine synthetase and amino acid transport system. J Bacteriol 138:218–224

    Google Scholar 

  • Lowe RH, Evans HJ (1964) Preparation and some properties of a soluble nitrate reductase from Rhizobium japonicum. Biochim Biophys Acta 85:377–389

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    Google Scholar 

  • Ludwig RA (1978) Control of ammonia assimilation in Rhizobium 32H1. J Bacteriol 135:114–123

    Google Scholar 

  • Magasanik B (1978) Regulation in the hut system. In: Miller JH, Reznikoff WS (eds) The operon. Cold Spring Harbor Laboratory, Cold Spring Harbor, pp 373–387

    Google Scholar 

  • Magasanik B (1982) Genetic control of nitrogen assimilation in bacteria. Annu Rev Gen 16:135–168

    Google Scholar 

  • Mellor HY, Glenn AR, Dilworth MJ (1987) Symbiotic and competitive properties of motility mutants of Rhizobium trifolii TA1. Arch Microbiol 148:34–39

    Google Scholar 

  • Mohapatra SS, Gresshoff PM (1984) Carbon-nitrogen requirements for the expression of nitrogenase activity in cultured Parasponia-Rhizobium strain ANU289. Arch Microbiol 137:58–62

    Google Scholar 

  • O'Gara F, Shamugam KT (1976a) Regulation of nitrogen fixation by Rhizobium: export of fixed N2 as NH +4 . Biochim Biophys Acta 437:313–321

    Google Scholar 

  • O'Gara F, Shanmugam KT (1976b) Control of symbiotic nitrogen fixation in rhizobia: regulation of NH +4 assimilation. Biochim Biophys Acta 451:342–352

    Google Scholar 

  • Poole PS, Dilworth MJ, Glenn AR (1984) Acquisition of aspartase activity in Rhizobium leguminosarum WU235. J Gen Microbiol 130:881–886

    Google Scholar 

  • Poole PS, Franklin M, Glenn AR, Dilworth MJ (1985) The transport of l-glutamate by Rhizobium leguminosarum involves a common amino acid carrier. J Gen Microbiol 131:1441–1448

    Google Scholar 

  • Poole PS, Dilworth MJ, Glenn AR (1987) Ammonia is the preferred nitrogen source in several rhizobia. J Gen Microbiol 133:1707–1712

    Google Scholar 

  • Ratcliffe HD, Drozd JW, Bull AT (1983) The utilization of 5-oxoproline, ammonia and glutamate by Rhizobium leguminosarum in chemostat culture. J Gen Microbiol 129:1707–1712

    Google Scholar 

  • Reid KG, Utech NM, Holden JT (1970) Multiple transport components for dicarboxylic amino acids in Streptococcus faecalis. J Biol Chem 245:5261–5277

    Google Scholar 

  • Saroso S, Glenn AR, Dilworth MJ (1984) Carbon utilization by free-living and bacteroid forms of cowpea Rhizobium strain NGR234. J Gen Microbiol 130:1809–1814

    Google Scholar 

  • Smart JB, Dilworth MJ, Robson AD (1984) Effect of phosphate supply on phosphate uptake and alkaline phosphatase activity in rhizobia. Arch Microbiol 140:281–286

    Google Scholar 

  • Smith I (1960) Chromatographic techniques. Heinemann, London, UK

    Google Scholar 

  • Trinick MJ (1968) Nodulation of tropical legumes. I. Specificity in the Rhizobium symbiosis of Leucatna leucocephala. Exp Agric 4:243–253

    Google Scholar 

  • Tubbs RS (1976) Regulation of nitrogen fixation in Rhizobium spp. Appl Env Microbiol 32:483–488

    Google Scholar 

  • Udvardi MK, Salcom CL, Day DA (1988) Transport of l-glutamate across the bacteroid membrane but not the peribacteroid membrane from soybean root nodules. Mol Plant-Microb Interactions 1:250–254

    Google Scholar 

  • Vincent JM (1970) A manual for the practical study of root nodule bacteria. Blackwell Scientific Publications, Oxford

    Google Scholar 

Download references

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Jin, H.N., Glenn, A.R. & Dilworth, M.J. How does l-glutamate transport relate to selection of mixed nitrogen sources in Rhizobium leguminosarum biovar trifolii MNF1000 and cowpea Rhizobium MNF2030?. Arch. Microbiol. 153, 448–454 (1990). https://doi.org/10.1007/BF00248426

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