Carbon Dioxide Requirement for Growth of Legume Nodule Bacteria

Lowe, R. H.; Evans, Harold J.

doi:10.1097/00010694-196212000-00001

Cited by 64 publications

(37 citation statements)

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“…Lowe and Evans (9) and Johnson et al (5) found C02-fixing enzymes and malate synthetase in R. japonicum bacteroids, but did not determine the role or contribution of these enzymes to the metabolism of intact bacteroids. Our data suggest that carboxylases and malate synthetase are functional in intact bacteroids and that a substantial fraction of acetate carbon is utilized for direct malate formation.…”

Section: Discussionmentioning

confidence: 96%

Organic Acid Metabolism by Isolated Rhizobium japonicum Bacteroids

Stovall¹,

Cole²

1978

Plant Physiol.

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Rhizobium japonicum bacteroids isolated from soybean (Glycine max L.) nodules oxidized "C-labeled succinate, pyruvate, and acetate in a manner consistent with operation of the tricarboxylic acid cycle and a partial glyoxylate cycle. Substrate carbon was incorporated into all major cellular components (cell wall + membrane, nucleic acids, and protein).Much research has been done on select aspects of nitrogen fixation by leguminous plants, such as nodule formation and the biochemistry and regulation of nitrogen fixation. However, relatively little attention has been given to carbon metabolism in nodules. Since nodular metabolism ofcarbon compounds provides both the reductant and energy source to support nitrogen fixation, as well as furnishing carbon skeletons from which amino acids are synthesized and transported from the nodules to aerial portions of the plant, we thought that an overview of nodule carbon metabolism represented a logical extension of existing knowledge of nodule function.There is good evidence that cultured Rhizobiumjaponicum cells possess a functional TCA' cycle (6,7). Although several researchers have assumed that bacteroids also possess a TCA cycle (1,2,12, 16) (4,11,14). Fresh wt of nodules was determined and the nodules were surface-sterilized for 5 min in 10 ml of 4% (w/v) NaCIO and rinsed several times in a total volume of 200 ml of sterile distilied H20. All subsequent steps were performed under aseptic conditions. Nodules were crushed in 10 ml of sterile 0.1 M Na-phosphate buffer (pH 7.4), and filtered through four layers of cheesecloth. The filtrate was centrifuged at l,OOOg for 5 min. The pellet of bacteroids was washed once with P04 buffer. Bacteroid preparations contained approximately 2 mg of protein/ml for field-grown plants and 0.8 mg of protein/ml for greenhouse-grown plants. Protein was determined by the Lowry method (10), modified by boiling the bacteroids in 0.1 M NaOH for 10 min to solubilize the protein prior to adding the reagents used in the Lowry assay.Radiorespirometry

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Section: Discussionmentioning

confidence: 96%

Organic Acid Metabolism by Isolated Rhizobium japonicum Bacteroids

Stovall¹,

Cole²

1978

Plant Physiol.

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“…H2-uptake-positive (Hup+) strains of R. japonicum cultured on usual media failed to show hydrogenase activity. Strains of R. japonicum capable of synthesizing the hydrogenase system, however, expressed activity when supplied with limited 02, a source of H2, and a low concentration of carbon substrates in the medium (13).The essentiality of CO2 for growth of five major species of Rhizobium, including R. japonicum, was demonstrated in this laboratory (14). The CO2 requirement could not be replaced by any one of a series of organic metabolites or by yeast extract.…”

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confidence: 87%

Autotrophic growth of H ₂ -uptake-positive strains of Rhizobium japonicum in an atmosphere supplied with hydrogen gas

Hanus

Maier

Evans

1979

Proc. Natl. Acad. Sci. U.S.A.

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123

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Previous research from this laboratory has demonstrated C02-fixing and H2-uptake capacities of certain strains of Rhizobium japonicum. In this report we have shown that SR, a H2euptake-positive (Hup+) strain of R. japonicum, is capable of autotrophic growth with H2 as the energy source. Growth occurred on mineral salts/vitamins/Noble agar, mineral salts/vitamins liquid medium (0.27 jig of C as vitamins per ml), and in mineral salts liquid medium with no added vitamins when cultures were provided with NH4CI and incubated in an atmosphere containing H2, CO2, 02, and N2. Little or no growth occurred when either H2 or CO2 was omitted from the atmosphere or when the culture was inoculated with SR3, a Hupmutant of SR. Growth was measured by protein synthesis, fixed organic carbon, and increase in cell number in liquid cultures. The organism that grew autotrophically was verified as B.japonicum by (i) apparent purity on streak plates; (ii) retention of the double antibiotic resistance markers; and (iii) its capability to nodulate soybeans. H2-and CO2-supported growth was demonstrated for three additional Hup+ wild type L japonicum strains (USDA 136, 311b 6, and 3I1b 143), while three Hupwild-type strains (USDA 120, 3Ilb 144, and USDA 117) were incapable of growth on the Noble agar medium containing mineral salts/vitamins in the H2/CO2/O2/N2 atmosphere. This demonstrated capability of Hup B. japonicum strains to grow autotrophically requires revision of current concepts regarding conditions for survival and competition of these bacteria in the soil and their relationships to other microorganisms. Beijerinck in 1888 (1) isolated nodule-forming bacteria from the root nodules of several different leguminous plants and referred to them as "Bacillus radicola." Since this pioneering discovery the microorganisms that nodulate roots of legumes have been classified into several species of Rhizobium primarily on the basis of their capacities to form N2-fixing nodules on the roots of certain groups of leguminous hosts (2). All rhizobia have been considered as aerobic chemoorganotrophs that grow best on complex media (3). Pentoses and hexoses are usually supplied as sources of carbon and energy for culture of the slow-growing species of which R. japonicum is an example (3).A hydrogenase system involved in H2 oxidation in root nodules of Pisum sativum was identified by Phelps and Wilson (4) and rediscovered by Dixon (5, 6). Nodules of P. sativum formed by R. leguminosarum strains ONA 311 and 314 consumed H2, but nodules produced by other strains of this species lacked this capability (7). H2-uptake activity of nodules formed by R. japonicum 3Ilb 110 was observed by Schubert et al. (8,9); however, only 7 of 32 R. japonicum strains surveyed by Carter et al. (10) synthesized sufficient hydrogenase in nodule bacteroids to recycle the major portion of the H2 produced during N2 fixation. The H2-uptake reaction in soybean nodule bacteroids was characterized by McCrae et al. (11). Recently, Emerich et al. (12) showed that H2 oxidat...

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“…In addition to directly or indirectly regulating specific genes involved in a variety of metabolic processes (Encarnació n et al, 2003;Heinz & Streit, 2003;Streit et al, 1996), the B-group vitamin biotin has profound effects on the physiology of rhizobia by acting as the prosthetic group in reactions catalysed by the biotin-dependent carboxylases (Encarnació n et al, 1995;Lowe & Evans, 1962;Watson et al, 2001;West & Wilson, 1940). The function of biotin in these reactions is the activation and transfer of CO 2 to an acceptor molecule such as pyruvate or an acyl-CoA, and organisms may contain one to several different biotin-dependent carboxylases.…”

Section: Introductionmentioning

confidence: 99%

“…ACC and/or PCC activities have been reported in several rhizobia (DeHertogh et al, 1964;Dunn et al, 2002;Encarnació n et al, 1995;Lowe & Evans, 1962) and could be catalysed in each species by distinct ACCs or PCCs and/or by a single acyl-CoA carboxylase. Western blot analysis of Rhizobium etli extracts revealed two biotin-containing proteins in addition to PYC (Dunn et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

Biochemical characterization of a Rhizobium etli monovalent cation-stimulated acyl-coenzyme A carboxylase with a high substrate specificity constant for propionyl-coenzyme A

2004

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Biotin has a profound effect on the metabolism of rhizobia. It is reported here that the activities of the biotin-dependent enzymes acetyl-coenzyme A carboxylase (ACC; EC 6.4.1.2) and propionyl-coenzyme A carboxylase (PCC; EC 6.4.1.3) are present in all species of the five genera comprising the Rhizobiaceae which were examined. Evidence is presented that the ACC and PCC activities detectable in Rhizobium etli extracts are catalysed by a single acyl-coenzyme A carboxylase. The enzyme from R. etli strain 12-53 was purified 478-fold and displayed its highest activity with propionyl-CoA as substrate, with apparent K m and V max values of 0?064 mM and 2885 nmol min or Cs + markedly activated the enzyme, which was essentially inactive in their absence. Electrophoretic analysis indicated that the acyl-CoA carboxylase was composed of a 74 kDa biotin-containing a subunit and a 45 kDa biotin-free b subunit, and gel chromatography indicated a total molecular mass of 620 000 Da. The strong kinetic preference of the enzyme for propionyl-CoA is consistent with its participation in an anaplerotic pathway utilizing this substrate.

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Carbon Dioxide Requirement for Growth of Legume Nodule Bacteria

Cited by 64 publications

References 0 publications

Organic Acid Metabolism by Isolated Rhizobium japonicum Bacteroids

Organic Acid Metabolism by Isolated Rhizobium japonicum Bacteroids

Autotrophic growth of H ₂ -uptake-positive strains of Rhizobium japonicum in an atmosphere supplied with hydrogen gas

Biochemical characterization of a Rhizobium etli monovalent cation-stimulated acyl-coenzyme A carboxylase with a high substrate specificity constant for propionyl-coenzyme A

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Carbon Dioxide Requirement for Growth of Legume Nodule Bacteria

Cited by 64 publications

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Organic Acid Metabolism by Isolated Rhizobium japonicum Bacteroids

Organic Acid Metabolism by Isolated Rhizobium japonicum Bacteroids

Autotrophic growth of H 2 -uptake-positive strains of Rhizobium japonicum in an atmosphere supplied with hydrogen gas

Biochemical characterization of a Rhizobium etli monovalent cation-stimulated acyl-coenzyme A carboxylase with a high substrate specificity constant for propionyl-coenzyme A

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Autotrophic growth of H ₂ -uptake-positive strains of Rhizobium japonicum in an atmosphere supplied with hydrogen gas