scholarly journals Paraburkholderia phymatum Homocitrate Synthase NifV Plays a Key Role for Nitrogenase Activity during Symbiosis with Papilionoids and in Free-Living Growth Conditions

Cells ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 952
Author(s):  
Paula Bellés-Sancho ◽  
Martina Lardi ◽  
Yilei Liu ◽  
Sebastian Hug ◽  
Marta Adriana Pinto-Carbó ◽  
...  
Keyword(s):  
Nitrogenase Activity ◽  
Root Nodules ◽  
Growth Conditions ◽  
Lysine Biosynthesis ◽  
Metabolome Analysis ◽  
Plant Host ◽  
Free Living ◽  
Bacterial Enzyme ◽  
Homocitrate Synthase ◽  
Iron Molybdenum Cofactor

Homocitrate is an essential component of the iron-molybdenum cofactor of nitrogenase, the bacterial enzyme that catalyzes the reduction of dinitrogen (N2) to ammonia. In nitrogen-fixing and nodulating alpha-rhizobia, homocitrate is usually provided to bacteroids in root nodules by their plant host. In contrast, non-nodulating free-living diazotrophs encode the homocitrate synthase (NifV) and reduce N2 in nitrogen-limiting free-living conditions. Paraburkholderia phymatum STM815 is a beta-rhizobial strain, which can enter symbiosis with a broad range of legumes, including papilionoids and mimosoids. In contrast to most alpha-rhizobia, which lack nifV, P. phymatum harbors a copy of nifV on its symbiotic plasmid. We show here that P. phymatum nifV is essential for nitrogenase activity both in root nodules of papilionoid plants and in free-living growth conditions. Notably, nifV was dispensable in nodules of Mimosa pudica despite the fact that the gene was highly expressed during symbiosis with all tested papilionoid and mimosoid plants. A metabolome analysis of papilionoid and mimosoid root nodules infected with the P. phymatum wild-type strain revealed that among the approximately 400 measured metabolites, homocitrate and other metabolites involved in lysine biosynthesis and degradation have accumulated in all plant nodules compared to uninfected roots, suggesting an important role of these metabolites during symbiosis.

scholarly journals Effects of Engineered Sinorhizobium meliloti on Cytokinin Synthesis and Tolerance of Alfalfa to Extreme Drought Stress

2012 ◽  
Vol 78 (22) ◽  
pp. 8056-8061 ◽  
Author(s):  
Ji Xu ◽  
Xiao-Lin Li ◽  
Li Luo
Keyword(s):  
Nitrogen Fixation ◽  
Drought Stress ◽  
Sinorhizobium Meliloti ◽  
Nitrogenase Activity ◽  
Root Nodules ◽  
Severe Drought ◽  
Control Strain ◽  
Free Living ◽  
Content Type ◽  
Host Tolerance

ABSTRACTCytokinin is required for the initiation of leguminous nitrogen fixation nodules elicited by rhizobia and the delay of the leaf senescence induced by drought stress. A few free-living rhizobia have been found to produce cytokinin. However, the effects of engineered rhizobia capable of synthesizing cytokinin on host tolerance to abiotic stresses have not yet been described. In this study, two engineeredSinorhizobiumstrains overproducing cytokinin were constructed. The tolerance of inoculated alfalfa plants to severe drought stress was assessed. The engineered strains, which expressed theAgrobacterium iptgene under the control of different promoters, synthesized more zeatins than the control strain under free-living conditions, but their own growth was not affected. After a 4-week inoculation period, the effects of engineered strains on alfalfa growth and nitrogen fixation were similar to those of the control strain under nondrought conditions. After being subjected to severe drought stress, most of the alfalfa plants inoculated with engineered strains survived, and the nitrogenase activity in their root nodules showed no apparent change. A small elevation in zeatin concentration was observed in the leaves of these plants. The expression of antioxidant enzymes increased, and the level of reactive oxygen species decreased correspondingly. Although theiptgene was transcribed in the bacteroids of engineered strains, the level of cytokinin in alfalfa nodules was identical to that of the control. These findings suggest that engineeredSinorhizobiumstrains synthesizing more cytokinin could improve the tolerance of alfalfa to severe drought stress without affecting alfalfa nodulation or nitrogen fixation.

scholarly journals MtMOT1.2 is responsible for molybdate supply toMedicago truncatulanodules

2018 ◽  
Author(s):  
Patricia Gil-Díez ◽  
Manuel Tejada-Jiménez ◽  
Javier León-Mediavilla ◽  
Jiangqi Wen ◽  
Kirankumar S. Mysore ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Symbiotic Nitrogen Fixation ◽  
Nitrogenase Activity ◽  
Reductase Activity ◽  
Root Nodules ◽  
Loss Of Function ◽  
Intracellular Membrane ◽  
Wild Type ◽  
Endodermal Cells ◽  
Iron Molybdenum Cofactor

ABSTRACTSymbiotic nitrogen fixation in legume root nodules requires a steady supply of molybdenum for synthesis of the iron-molybdenum cofactor of nitrogenase. This nutrient has to be provided by the host plant from the soil, crossing several symplastically disconnected compartments through molybdate transporters, including members of the MOT1 family. MtMOT1.2 is aMedicago truncatulaMOT1 family member located in the endodermal cells in roots and nodules. Immunolocalization of a tagged MtMOT1.2 indicates that it is associated to the plasma membrane and to intracellular membrane systems, where it would be transporting molybdate towards the cytosol, as indicated in yeast transport assays. A loss-of-functionmot1.2-1mutant showed reduced growth compared to wild-type plants when nitrogen fixation was required, but not when nitrogen was provided as nitrate. While no effect on molybdenum-dependent nitrate reductase activity was observed, nitrogenase activity was severely affected, explaining the observed difference of growth depending on nitrogen source. This phenotype was the result of molybdate not reaching the nitrogen-fixing nodules, since genetic complementation with a wild-typeMtMOT1.2gene or molybdate-fortification of the nutrient solution, both restored wild-type levels of growth and nitrogenase activity. These results support a model in which MtMOT1.2 would mediate molybdate delivery by the vasculature into the nodules.

scholarly journals Medicago truncatulaMOT1.3 is a plasma membrane molybdenum transporter required for nitrogenase activity in root nodules

2017 ◽  
Author(s):  
Manuel Tejada-Jiménez ◽  
Patricia Gil-Díez ◽  
Javier León-Mediavilla ◽  
Jiangqi Wen ◽  
Kirankumar S. Mysore ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Nitrogenase Activity ◽  
Root Nodules ◽  
List Type ◽  
Knockout Mutant ◽  
Model Legume ◽  
Molybdate Transport ◽  
The One ◽  
Iron Molybdenum Cofactor

SummaryMolybdenum, as a component of the iron-molybdenum cofactor of nitrogenase, is essential for symbiotic nitrogen fixation. This nutrient has to be provided by the host plant through molybdate transporters.Members of the molybdate transporters family MOT1 were identified in the model legumeMedicago truncatulaand their expression in nodules determined. Yeast toxicity assays, confocal microscopy, and phenotypical characterization of aTnt1insertional mutant line were carried out in the oneM. truncatulaMOT1 family member expressed specifically in nodules.Among the five MOT1 members present inM. truncatulagenome,MtMOT1.3is the only one uniquely expressed in nodules. MtMOT1.3 shows molybdate transport capabilities when expressed in yeast. Immunolocalization studies revealed that MtMOT1.3 is located in the plasma membrane of nodule cells. Amot1.3-1knockout mutant showed an impaired growth concomitant with a reduction in nitrogenase activity. This phenotype was rescued by increasing molybdate concentrations in the nutritive solution, or upon addition of an assimilable nitrogen source. Furthermore,mot1.3-1plants transformed with a functional copy ofMtMOT1.3showed a wild type-like phenotype.These data are consistent with a model in which MtMOT1.3 would be responsible for introducing molybdate into nodule cells, which will be later used to synthesize functional nitrogenase.

Carbon Costs of Nitrogenase Activity in Legume Root Nodules determined using Acetylene and Oxygen

1983 ◽  
Vol 34 (8) ◽  
pp. 951-963 ◽  
Author(s):  
J. F. WITTY ◽  
F. R. MINCHIN ◽  
J. E. SHEEHY
Keyword(s):  
Nitrogenase Activity ◽  
Root Nodules ◽  
Carbon Costs

Seasonal pattern of energy use, respiration, and nitrogenase activity in root nodules of Myrica gale

1983 ◽  
Vol 61 (11) ◽  
pp. 2937-2942 ◽  
Author(s):  
Christa R. Schwintzer ◽  
John D. Tjepkema
Keyword(s):  
Energy Cost ◽  
Energy Use ◽  
Nitrogenase Activity ◽  
Root Nodules ◽  
Seasonal Pattern ◽  
Uptake Ratio ◽  
Experimental Conditions ◽  
Myrica Gale ◽  
Net Production ◽  
N Requirement

Annual CO2 evolution, H2 evolution, and C2H2 reduction were measured in root nodules from a vigorous Myrica gale stand in a Massachusetts peatland at 3-week intervals in 1980. Nodule activity was approximately the same under the experimental conditions (excised nodules reducing C2H2) as in nature (attached nodules reducing N2) and the CO2 evolution to O2 uptake ratio averaged 1.07. Nitrogenase activity was first detectable in late May, reached its maximum [Formula: see text] in mid-July, and disappeared in late October. The seasonal pattern of CO2 evolution was similar except that it continued at low rates when nitrogenase activity was absent. Hydrogen evolution was barely detectable. The energy cost of nitrogen fixation, expressed as the molar CO2:C2H4 ratio, was relatively low [Formula: see text] throughout the period of substantial nitrogenase activity and had a mean annual value of 4.9. Annual N2 fixation was estimated to be 2.8 g N m−2year−1, contributing about 33% of the annual N requirement measured in 1979. Annual C use by nodules was about 21.0 g C m−2 year−1. If this C were available for additional net production, it would increase it by about 5.5%.

Use of products of straw decomposition by N2-fixing (C2H2-reducing) populations of bacteria in three soils from wheat-growing areas

1986 ◽  
Vol 37 (1) ◽  
pp. 1 ◽  
Author(s):  
MM Roper ◽  
DM Halsall
Keyword(s):  
Nitrogenase Activity ◽  
New South ◽  
Carbon Sources ◽  
Clay Content ◽  
Free Living ◽  
Straw Decomposition ◽  
South Wales ◽  
History Of ◽  
Straw Incorporation ◽  
The Difference

The potential for N2 fixation by free-living bacteria using straw as a source of energy was evaluated in three soils (one from Gunnedah and two from Cowra) representative of the wheat belt in New South Wales. All three soils had a history of straw incorporation. The abilities of the respective microbial populations to use a range of carbon sources, including potential products of decomposition of straw, was determined and compared with the size and composition of each population. Neutral to alkaline (pH 7.4) soil of high (51%) clay content from Gunnedah produced higher rates of nitrogenase activity with straw than more acid (pH 5.6) lower (17%) clay containing soil from Cowra (site B). Gunnedah soil also contained a larger population of N2-fixing bacteria which used a broader range of energy sources than soil from either Cowra site B or Cowra site W (pH 5.8, clay content 34%). There was little difference in the composition of the N2-fixing populations in each of the soils except that Azotobacter spp. were absent from the acid Cowra soils. It was concluded that the difference in behaviour of the respective N2-fixing populations was primarily due to the physical characteristics of the soil affecting the numbers and activities of diazotrophic microorganisms. In addition some soil environments failed to support specific organisms.

scholarly journals The landscape of cytokinin binding by a plant nodulin

2013 ◽  
Vol 69 (12) ◽  
pp. 2365-2380 ◽  
Author(s):  
M. Ruszkowski ◽  
K. Szpotkowski ◽  
M. Sikorski ◽  
M. Jaskolski
Keyword(s):  
Root Nodules ◽  
Outer Cortex ◽  
Plant Host ◽  
Symbiotic Interaction ◽  
Model Legume ◽  
Hydrophobic Cavity ◽  
Density Maps ◽  
Loop Element ◽  
Α Helix ◽  
Related Proteins

Nodulation is an extraordinary symbiotic interaction between leguminous plants and nitrogen-fixing bacteria (rhizobia) that assimilate atmospheric nitrogen (in root nodules) and convert it into compounds suitable for the plant host. A class of plant hormones called cytokinins are involved in the nodulation process. In the model legumeMedicago truncatula, nodulin 13 (MtN13), which belongs to the pathogenesis-related proteins of class 10 (PR-10), is expressed in the outer cortex of the nodules. In general, PR-10 proteins are small and monomeric and have a characteristic fold with an internal hydrophobic cavity formed between a seven-stranded antiparallel β-sheet and a C-terminal α-helix. Previously, some PR-10 proteins not related to nodulation were found to bind cytokinins such astrans-zeatin. Here, four crystal structures of the MtN13 protein are reported in complexes with several cytokinins, namelytrans-zeatin,N6-isopentenyladenine, kinetin andN6-benzyladenine. All four phytohormones are bound in the hydrophobic cavity in the same manner and have excellent definition in the electron-density maps. The binding of the cytokinins appears to be strong and specific and is reinforced by several hydrogen bonds. Although the binding stoichiometry is 1:1, the complex is actually dimeric, with a cytokinin molecule bound in each subunit. The ligand-binding site in each cavity is formed with the participation of a loop element from the other subunit, which plugs the only entrance to the cavity. Interestingly, a homodimer of MtN13 is also formed in solution, as confirmed by small-angle X-ray scattering (SAXS).

scholarly journals The Novel Genes emmABC Are Associated with Exopolysaccharide Production, Motility, Stress Adaptation, and Symbiosis in Sinorhizobium meliloti

2009 ◽  
Vol 191 (19) ◽  
pp. 5890-5900 ◽  
Author(s):  
Jennifer Morris ◽  
Juan E. González
Keyword(s):  
Sinorhizobium Meliloti ◽  
Genetic Factors ◽  
Leguminous Plant ◽  
The Novel ◽  
Two Component System ◽  
Cell Component ◽  
Plant Host ◽  
Free Living ◽  
Novel Genes ◽  
Exopolysaccharide Production

ABSTRACT The nitrogen-fixing symbiont Sinorhizobium meliloti senses and responds to constantly changing environmental conditions as it makes its way through the soil in search of its leguminous plant host, Medicago sativa (alfalfa). As a result, this bacterium regulates various aspects of its physiology in order to respond appropriately to stress, starvation, and competition. For example, exopolysaccharide production, which has been shown to play an important role in the ability of S. meliloti to successfully invade its host, also helps the bacterium withstand osmotic changes and other environmental stresses. In an effort to further elucidate the intricate regulation of this important cell component, we set out to identify genetic factors that may affect its production. Here we characterize novel genes that encode a small protein (EmmA) and a putative two-component system (EmmB-EmmC). A mutation in any of these genes leads to increased production of the symbiotically important exopolysaccharide succinoglycan. In addition, emm mutants display membrane-associated defects, are nonmotile, and are unable to form an optimal symbiosis with alfalfa, suggesting that these novel genes may play a greater role in the overall fitness of S. meliloti both during the free-living stage and in its association with its host.

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