Development and Partial Characterization of Nearly Isogenic Pea Lines (<i>Pisum sativum</i> L.) that Alter Uptake Hydrogenase Activity in Symbiotic <i>Rhizobium</i>

Phillips, Donald A.; Kapulnik, Yoram; Bedmar, Eulogio J.; Joseph, Cecillia M.

doi:10.1104/pp.92.4.983

Cited by 9 publications

(8 citation statements)

References 29 publications

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“…Similarly, high variability of effectiveness of rhizobia was reported in chickpea, mungbean, Vicia, Lathyrus, pea, soybean, lima bean, cowpea, bush bean, peanut, Leucaena leucocephala, tinga pea, alfalfa and clover [6,17,19,[31][32][33].…”

Section: Discussionmentioning

confidence: 98%

Molecular diversity of native rhizobia trapped by five field pea genotypes in Indian soils

Wadhwa

Dudeja

Yadav

2010

J. Basic Microbiol.

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Five pea cultivars; HFP 4, HVP 3-5, HFP 9426, Jayanti and Hariyal, being grown in CCS Haryana Agricultural University farm were used to isolate native rhizobia. Selected 54 rhizobia, from all cultivars, were authenticated as rhizobia by plant infectivity test. Along with nodulation, symbiotic effectiveness in terms of symbiotic ratios showed wide range of effectiveness of pea rhizobia from 1.11 to 5.0. DNA of all the 54 rhizobia was extracted and amplified by PCR, using ERIC and 16S rDNA primers. Dendrogram based on ERIC profiles of these 54 rhizobia showed the formation of 13 subclusters at 80% level of similarity. Dendrogram based on RFLP of 16S rDNA by three restriction endonucleases; Msp I, Csp 6I and Rsa I; also formed 13 subclusters at 80% level of similarity. However, positioning of subclusters was different from that of ERIC based dendrogram. Majority of the isolates i.e. 64.8% by ERIC profiles and 44.4% by RFLP of 16S rDNA formed one cluster. Isolates from same nodule were not 100% similar. Considering each cluster representing a rhizobial genotype, both techniques used to assess molecular diversity indicated the presence of 13 genotypes of field pea rhizobia in CCS Haryana Agricultural University farm soil. Two pea rhizobial genotypes were able to nodulate all the five pea cultivars. Furthermore, high strain richness index (0.43-0.5) of field pea rhizobia was observed by both the techniques.

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Section: Discussionmentioning

confidence: 98%

Molecular diversity of native rhizobia trapped by five field pea genotypes in Indian soils

Wadhwa

Dudeja

Yadav

2010

J. Basic Microbiol.

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“…An area of which we know little about genetically but that deserves keen research attention is the host-mediated controlled expression of the bacterial symbiont's hydrogenase (Nautiyal et al, 1988;Neves and Hungria, 1987;Phillips et al, 1990). This interesting phenomenon has led to the proposal of a third hydrogenase phenotype, in addition to the Hup' and Hup-phenotypes, called the host-regulated (Hup-hr) types (Van Berkum, 1990).…”

Section: Host-regulated Hup Expressionmentioning

confidence: 96%

Toward More Productive, Efficient, and Competitive Nitrogen-Fixing Symbiotic Bacteria

Maier

Triplett

1996

Critical Reviews in Plant Sciences

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The prospects of developing strains of legume nodule bacteria that provide higher productivity of leguminous plants are described. The generic, biochemical, physiological, regulatory, and economic constraints that govern the ability of private and public efforts to construct better inoculants for legume nodulation are discussed. Success in constructing better inoculants requires a two-pronged approach. First, strains need to be improved in order to compete successfully with indigenous strains for root nodulation of legumes. Several loci have been identified to date that affect competitiveness for strain nodule occupancy. Usually mutations in these loci affect the ability of a strain to form nodules rapidly and efficiently. Other loci, such as those that confer antibiotic production, can be added to strains to enhance nodulation competitiveness when co-inoculated with antibiotic-sensitive strains. Second, the inoculum strains must be improved with respect to symbiotic nitrogen fixation. Efforts to enhance the symbiotic productivity of legume nodule bacteria either by mutation or genetic engineering are also described. The best characterized example of these is the hydrogenase system. Due to nitrogenase-dependent catalysis of proton reduction, diazotrophs evolve large amounts of H,. An approach to maximize the efficiency of symbiotic N, fixation, and therefore of legume productivity, is to construct strains of Rhizobium with the ability to oxidize this otherwise wasted H,. The electrons produced by H, oxidation are funneled through energy-conserving electron transport chains. Our knowledge of the genetics and biochemistry of H, oxidation in Bradyrhizobium japonicum and Rhizobium leguminosarum has developed rapidly in recent years. At least 20 genes are needed for these bacteria to manufacture and efficiently express a nickel-containing H,-uptake hydrogenase. These genes include those encoding regulatory elements, posttranslational processing enzymes, nickel-sensing and nickel-metabolism proteins, and electron transport components for integrating the electrons from H, oxidation into the respiratory chain. Some of the components for oxidizing H, in the symbiotic N, fixing bacteria are distinct from the analogous components in (nonsymbiotic) H, oxidizing bacteria.

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“…viciae (25,36), indicating that the potential for hydrogenase activity of this species has been underestimated in the past. Nickel-dependent stimulation of the activity of the nickel-containing enzymes urease and hydrogenase has been reported previously for soybeans grown in low-nickel soil (2).…”

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Nickel availability to pea (Pisum sativum L.) plants limits hydrogenase activity of Rhizobium leguminosarum bv. viciae bacteroids by affecting the processing of the hydrogenase structural subunits

et al. 1994

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Rhizobium leguminosarum bv. viciae UPM791 induces the synthesis of an [NiFe] hydrogenase in pea (Pisum sativum L.) bacteroids which oxidizes the H2 generated by the nitrogenase complex-inside the root nodules. The synthesis of this hydrogenase requires the genes for the small and large hydrogenase subunits (hupS and hupL, respectively) and 15 accessory genes clustered in a complex locus in the symbiotic plasmid. We show here that the bacteroid hydrogenase activity is limited by the availability of nickel to pea plants. Addition of Ni2+ to plant nutrient solutions (up to 10 mg/liter) resulted in sharp increases (up to 15-fold) in hydrogenase activity. This effect was not detected when other divalent cations (Zn2+, Co2+, Fe2+, and Mn2+) were added at the same concentrations. Determinations of the steady-state levels of hupSL-specific mRNA indicated that this increase in hydrogenase activity was not due to stimulation of transcription of structural genes. Immunoblot analysis with antibodies raised against the large and small subunits of the hydrogenase enzyme demonstrated that in the low-nickel situation, both subunits are mainly present in slow-migrating, unprocessed forms. Supplementation of the plant nutrient solution with increasing nickel concentrations caused the conversion of the slow-migrating forms of both subunits into fast-moving, mature forms. This nickel-dependent maturation process of the hydrogenase subunits is mediated by accessory gene products, since bacteroids from H2 uptake-deficient mutants carrying TnS insertions in hupG and hupK and in hypB and hypE accumulated the immature forms of both hydrogenase subunits even in the presence of high nickel levels.Most hydrogen uptake hydrogenases are membrane-bound, heterodimeric iron-sulfur proteins containing nickel ([NiFe] hydrogenases) (for reviews, see references 9, 37, and 46). Bacteria forming nodules on legume roots synthesize uptake hydrogenases which recycle the H2 evolved by nitrogenase in the nodules (5, 27) and contribute to the overall efficiency of the N2 fixation process (6). These bacteria include Rhizobium leguminosarum bv. viciae and Bradyrhizobium japonicum, the microsymbionts of peas and soybeans, respectively. The genetic determinants for H2 uptake (hup genes) in R. leguminosarum bv. viciae UPM791 are clustered in a 20-kb DNA region of the symbiotic plasmid and have been isolated in cosmid pAL618 (24). This cosmid has the capacity to confer H2 uptake activity on Hup-strains of R. leguminosarum bv. viciae and Rhizobium etli in symbiosis with peas and beans, respectively (25). The DNA region spanning the H2 uptake gene cluster has been sequenced, and 17 genes, closely linked and oriented in the same direction, were identified (15,38,40). The first two genes, hupS and hupL, encode the hydrogenase structural polypeptides (14). The predicted proteins for the small (360 amino acid residues, including a leader peptide of 45 residues) and the large (596 amino acid residues) subunits are homologous to the corresponding hydrogenase structur...

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Development and Partial Characterization of Nearly Isogenic Pea Lines (Pisum sativum L.) that Alter Uptake Hydrogenase Activity in Symbiotic Rhizobium

Cited by 9 publications

References 29 publications

Molecular diversity of native rhizobia trapped by five field pea genotypes in Indian soils

Molecular diversity of native rhizobia trapped by five field pea genotypes in Indian soils

Toward More Productive, Efficient, and Competitive Nitrogen-Fixing Symbiotic Bacteria

Nickel availability to pea (Pisum sativum L.) plants limits hydrogenase activity of Rhizobium leguminosarum bv. viciae bacteroids by affecting the processing of the hydrogenase structural subunits

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