Lack of the Flavin-Containing Monooxygenase FmoA Partially Impairs the Symbiotic Interaction of Mesorhizobium Huakuii with Astragalus Sinicus

2020 ◽  
Author(s):  
Hetao Wu ◽  
Qian Zou ◽  
Sha Luo ◽  
Donglan He ◽  
Xiaohua Li ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Target Genes ◽  
Symbiotic Nitrogen Fixation ◽  
Reductase Activity ◽  
Cumene Hydroperoxide ◽  
Nodule Development ◽  
Astragalus Sinicus ◽  
Symbiotic Interaction ◽  
Real Time Quantitative Pcr ◽  
Mesorhizobium Huakuii

Abstract Background: Flavin-containing monooxygenases (FMOs) catalyze the NADPH-dependent N- or S- oxygenation of numerous foreign chemicals, and thus may mediate interactions between microorganisms and their chemical environment. The aim of this study was to investigate the role of FMO in symbiotic nitrogen fixation of Mesorhizobium huakuii and its host plant Astragalus sinicus.Results: A mutation in M. huakuii fmoA gene was generated by homologous recombination. The fmoA mutant grew more slowly than its parental strain, and displayed decreased antioxidative capacity under higher concentration of H2O2 and cumene hydroperoxide (CUOOH), indicating that FmoA plays an important role in response to the peroxides. The fmoA mutant strain displayed no difference of peroxidase activity and glutathione reductase activity, but significantly lower level of glutathione and hydrogen peroxide content than the wild type. Real-time quantitative PCR showed that the fmoA gene expression is significantly up-regulated in three different stages of nodule development. The fmoA mutant was severely impaired in its rhizosphere colonization, and its symbiotic properties in Astragalus sinicus were drastically affected. Transcriptomes in root-nodule bacteroids were analyzed and compared. A total of 1233 genes were differentially expressed, of which 560 were up-regulated and 673 were down-regulated in HKfmoA bacteroids compared with that in 7653R bacteroids. The transcriptomic data allowed us to determine additional target genes, whose differential expression was able to explain the observed the changes of symbiotic phenotype in the mutant-infected nodules.Conclusions: The fmoA gene is essential for antioxidant capacity and symbiotic nitrogen fixation. Furthermore, RNA-Seq based global transcriptomic analysis provided a comprehensive view of M. huakuii fmoA gene involved in nodule senescence and symbiotic nitrogen fixation.

scholarly journals Antioxidation and symbiotic nitrogen fixation function of prxA gene in Mesorhizobium huakuii

2019 ◽  
Vol 8 (10) ◽  
Author(s):  
Sanjiao Wang ◽  
Tiantian Lu ◽  
Qiang Xue ◽  
Ke Xu ◽  
Guojun Cheng
Keyword(s):  
Nitrogen Fixation ◽  
Symbiotic Nitrogen Fixation ◽  
Mesorhizobium Huakuii

scholarly journals Digalactosyldiacylglycerol Synthase Gene MtDGD1 Plays an Essential Role in Nodule Development and Nitrogen Fixation

2019 ◽  
Vol 32 (9) ◽  
pp. 1196-1209
Author(s):  
Zaiyong Si ◽  
Qianqian Yang ◽  
Rongrong Liang ◽  
Ling Chen ◽  
Dasong Chen ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Essential Role ◽  
Symbiotic Nitrogen Fixation ◽  
Histochemical Staining ◽  
Nodule Development ◽  
Nodule Number ◽  
Nitrogen Fixation Activity ◽  
Fixation Activity ◽  
Infected Cells ◽  
Nodule Symbiosis

Little is known about the genes participating in digalactosyldiacylglycerol (DGDG) synthesis during nodule symbiosis. Here, we identified full-length MtDGD1, a synthase of DGDG, and characterized its effect on symbiotic nitrogen fixation in Medicago truncatula. Immunofluorescence and immunoelectron microscopy showed that MtDGD1 was located on the symbiosome membranes in the infected cells. β-Glucuronidase histochemical staining revealed that MtDGD1 was highly expressed in the infection zone of young nodules as well as in the whole mature nodules. Compared with the control, MtDGD1-RNA interference transgenic plants exhibited significant decreases in nodule number, symbiotic nitrogen fixation activity, and DGDG abundance in the nodules, as well as abnormal nodule and symbiosome development. Overexpression of MtDGD1 resulted in enhancement of nodule number and nitrogen fixation activity. In response to phosphorus starvation, the MtDGD1 expression level was substantially upregulated and the abundance of nonphospholipid DGDG was significantly increased in the roots and nodules, accompanied by corresponding decreases in the abundance of phospholipids such as phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol. Overall, our results indicate that DGD1 contributes to effective nodule organogenesis and nitrogen fixation by affecting the synthesis and content of DGDG during symbiosis.

Phenotypic reversion of nitrogenase in pleiotropic mutants of Rhizobium meliloti

1979 ◽  
Vol 25 (3) ◽  
pp. 298-301 ◽  
Author(s):  
Ilona Barabás ◽  
Tibor Sik
Keyword(s):  
Amino Acids ◽  
Nitrogen Fixation ◽  
Nitrate Reductase ◽  
Nitrate Reductase Activity ◽  
Symbiotic Nitrogen Fixation ◽  
Rhizobium Meliloti ◽  
Nitrogenase Activity ◽  
Reductase Activity ◽  
Amino Acid Utilization ◽  
Phenotypic Reversion

In two out of three pleiotropic mutants of Rhizobium meliloti, defective in nitrate reductase induced by amino acid utilization in vegetative bacteria and in symbiotic nitrogen fixation, nitrogenase activity could be restored completely by purines and partially by the amino acids L-glutamate, L-aspartate, L-glutamine, and L-asparagine. The compounds restoring effectiveness in nitrogen fixation did not restore nitrate reductase activity in vegetative bacteria. The restoration of effectiveness supports our earlier conclusion that the mutation is not in the structural gene for a suggested common subunit of nitrogenase and nitrate reductase.

Nodulation and symbiotic nitrogen fixation by genotypes of blackgram [Vigna mungo (L.) Hepper] as affected by fertiliser nitrogen

2002 ◽  
Vol 53 (4) ◽  
pp. 453
Author(s):  
B. Singh ◽  
K. Usha
Keyword(s):  
Nitrogen Fixation ◽  
Symbiotic Nitrogen Fixation ◽  
Unit Area ◽  
Reductase Activity ◽  
Farming Systems ◽  
Vigna Mungo ◽  
Atmospheric Nitrogen ◽  
Nitrogen Treatments ◽  
Nitrite Content ◽  
Applied Nitrogen

Intercropping with legumes and non-legumes is commonly practised in many parts of the world to maximise productivity per unit area of land. In India, blackgram or urd [Vigna mungo (L.) Hepper] is a popular pulse legume component of intercropping farming systems. Often, however, potential production is compromised, particularly in high fertiliser input systems, because blackgram competes with the non-legume component of the system for nitrogen in the soil. In order to identify lines of blackgram that could obtain the majority of their nitrogen requirements from symbiotic fixation of atmospheric nitrogen rather than from uptake of soil nitrogen, 50 genotypes were screened for tolerance to (applied) nitrogen in soil. The parameters used to appraise tolerance were extent of root nodulation, the amount of nitrogen fixed, nitrate reductase activity in roots and nodules, and nitrite content of roots and nodules. There were 2 nitrogen treatments applied as urea, viz. 40 (N40) and 120 (N120) kg N per ha. There were 3 genotypes whose nitrogen-fixing effectiveness was apparently unimpaired by applications of nitrogen to the soil. Genotype NC-59308 nodulated and fixed atmospheric nitrogen satisfactorily at both the lower and higher levels of applied nitrogen. At N40, genotypes EC-48215 and PLU-726 formed a great abundance of large nodules effective in nitrogen fixation; even at N120, both lines had better symbioses than the majority of the 50 blackgram lines originally screened. These 3 genotypes are deemed worthy of further examination for their suitability for intercropping systems. How this might be done is discussed.

scholarly journals Induction and Spatial Organization of Polyamine Biosynthesis During Nodule Development in Lotus japonicus

2004 ◽  
Vol 17 (12) ◽  
pp. 1283-1293 ◽  
Author(s):  
Emmanouil Flemetakis ◽  
Rodica C. Efrose ◽  
Guilhem Desbrosses ◽  
Maria Dimou ◽  
Costas Delis ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Symbiotic Nitrogen Fixation ◽  
Expression Patterns ◽  
Lotus Japonicus ◽  
Cyclin D3 ◽  
Nodule Development ◽  
Cdna Clones ◽  
Long Distance ◽  
Polyamine Biosynthesis ◽  
Transcript Levels

Putrescine and other polyamines are produced by two alternative pathways in plants. One pathway starts with the enzyme arginine decarboxylase (ADC; EC 4.1.1.19), the other with ornithine decarboxylase (ODC; EC 4.1.1.17). Metabolite profiling of nitrogen-fixing Lotus japonicus nodules, using gas chromatography coupled to mass spectrometry, revealed a two- to sixfold increase in putrescine levels in mature nodules compared with other organs. Genes involved in polyamine biosynthesis in L. japonicus nodules were identified by isolating cDNA clones encoding ADC (LjADC1) and ODC (LjODC) from a nodule library. Searches of the public expressed sequence tag databases revealed the presence of a second gene encoding ADC (LjADC2). Real-time reverse-transcription-polymerase chain reaction analysis showed that LjADC1 and LjADC2 were expressed throughout the plant, while LjODC transcripts were detected only in nodules and roots. Induction of LjODC and LjADC gene expression during nodule development preceded symbiotic nitrogen fixation. Transcripts accumulation was maximal at 10 days postinfection, when a 6.5-fold increase in the transcript levels of LjODC was observed in comparison with the uninfected roots, while a twofold increase in the transcript levels of LjADC1 and LjADC2 was detected. At later stages of nodule development, transcripts for ADC drastically declined, while in the case of ODC, transcript accumulation was higher than that in roots until after 21 days postinfection. The expression profile of genes involved in putrescine biosynthesis correlated well with the expression patterns of genes involved in cell division and expansion, including a L. japonicus Cyclin D3 and an α-expansin gene. Spatial localization of LjODC and LjADC1 gene transcripts in developing nodules revealed that both transcripts were expressed in nodule inner cortical cells and in the central tissue. High levels of LjADC1 transcripts were also observed in both nodule and connecting root vascular tissue, suggesting that putrescine and other polyamines may be subject to long-distance transport. Our results indicate that polyamines are primarily involved in physiological and cellular processes involved in nodule development, rather than in processes that support directly symbiotic nitrogen fixation and assimilation.

scholarly journals Lower nodule biomass with increased nitrogenase efficiency in Robinia pseudoacacia seedlings when grown under low soil phosphorus conditions

2020 ◽  
Vol 2 (11) ◽  
Author(s):  
Lindsay A. McCulloch ◽  
Stephen Porder
Keyword(s):  
Nitrogen Fixation ◽  
Plant Growth ◽  
Symbiotic Nitrogen Fixation ◽  
Soil Phosphorus ◽  
Robinia Pseudoacacia ◽  
Nodule Development ◽  
N Fixation ◽  
P Availability ◽  
Nodule Biomass ◽  
P Fertilizer

AbstractSymbiotic nitrogen (N) fixation is the largest non-anthropogenic N input to many terrestrial ecosystems. The energetic expense of symbiotic N fixation suggests soil phosphorus (P) availability may regulate symbiotic nitrogen fixation directly through nodule development and function, and/or indirectly through plant growth. Since P availability is heterogenous in the landscape, we sought to understand if symbiotic nitrogen fixation responds to both P availability and heterogeneity. To test how P availability affects symbiotic nitrogen fixation, we grew Robinia pseudoacacia seedlings under high (8.1 g P m−2) and low (0.2 g P m−2) conditions. Soil P heterogeneity was simulated by splitting roots into soil patches receiving P or no-P fertilizer. At the whole plant level, P availability limited seedling and nodule biomass. However, the low P treatment had higher nitrogenase efficiency (acetylene reduced (AR) g−1 nodule; a nodule efficiency proxy). High P seedlings had significantly more root and nodule biomass in the patches directly receiving P fertilizer, but patch proliferation was absent in the low P treatment. AR g−1 seedling did not differ between P treatments, suggesting P indirectly limited symbiotic nitrogen fixation through plant growth, rather than directly limiting symbiotic nitrogen fixation. This relatively consistent AR g−1 seedling across treatments demonstrates the ability of seedlings to respond to low P conditions with increased nitrogenase efficiency.

Nitrate modification of photosynthesis and photoassimilate export in young nodulated soybean plants

1982 ◽  
Vol 60 (12) ◽  
pp. 2665-2670 ◽  
Author(s):  
D. J. Ursino ◽  
D. M. Hunter ◽  
R. D. Laing ◽  
J. L. S. Keighley
Keyword(s):  
Nitrogen Fixation ◽  
Nitrate Reductase ◽  
Nutrient Solution ◽  
Nitrate Reductase Activity ◽  
Reductase Activity ◽  
Nodule Development ◽  
Nitrate Availability ◽  
Data Support ◽  
Nutrient Fertilization ◽  
Soybean Plants

Soybean plants (cv. Harosoy 63) inoculated with rhizobia were germinated from seed and beginning on day 7 after planting were subjected to one of four patterns of nutrient fertilization. One group received a nutrient solution containing 5 mM nitrate, a second group received nitrate-free nutrient solution, and two other groups received nitrate-containing solution either from days 7 to 13 or from days 14 to 20. On day 21 rates of leaf photosynthetic CO2 uptake and nitrate reductase activity were measured, as well as the capacities of the leaf to export recent photosynthate and of the nodules to reduce acetylene. The data support the hypothesis that sufficient nitrate availability in the leaves of young soybean plants can modify both photosynthetic CO2 uptake and light-mediated photoassimilate export to an extent that nodule development and the capacity for nitrogen fixation are reduced.

scholarly journals Characterization of a Mesorhizobium loti α-Type Carbonic Anhydrase and Its Role in Symbiotic Nitrogen Fixation

2009 ◽  
Vol 191 (8) ◽  
pp. 2593-2600 ◽  
Author(s):  
Chrysanthi Kalloniati ◽  
Daniela Tsikou ◽  
Vasiliki Lampiri ◽  
Mariangela N. Fotelli ◽  
Heinz Rennenberg ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Carbonic Anhydrase ◽  
Mutant Strain ◽  
Symbiotic Nitrogen Fixation ◽  
Plant Traits ◽  
Nitrogenase Activity ◽  
Nodule Development ◽  
Free Living ◽  
Model Legume ◽  
Mesorhizobium Loti

ABSTRACT Carbonic anhydrase (CA) (EC 4.2.1.1) is a widespread enzyme catalyzing the reversible hydration of CO2 to bicarbonate, a reaction that participates in many biochemical and physiological processes. Mesorhizobium loti, the microsymbiont of the model legume Lotus japonicus, possesses on the symbiosis island a gene (msi040) encoding an α-type CA homologue, annotated as CAA1. In the present work, the CAA1 open reading frame from M. loti strain R7A was cloned, expressed, and biochemically characterized, and it was proven to be an active α-CA. The biochemical and physiological roles of the CAA1 gene in free-living and symbiotic rhizobia were examined by using an M. loti R7A disruption mutant strain. Our analysis revealed that CAA1 is expressed in both nitrogen-fixing bacteroids and free-living bacteria during growth in batch cultures, where gene expression was induced by increased medium pH. L. japonicus plants inoculated with the CAA1 mutant strain showed no differences in top-plant traits and nutritional status but consistently formed a higher number of nodules exhibiting higher fresh weight, N content, nitrogenase activity, and δ13C abundance. Based on these results, we propose that although CAA1 is not essential for nodule development and symbiotic nitrogen fixation, it may participate in an auxiliary mechanism that buffers the bacteroid periplasm, creating an environment favorable for NH3 protonation, thus facilitating its diffusion and transport to the plant. In addition, changes in the nodule δ13C abundance suggest the recycling of at least part of the HCO3 − produced by CAA1.

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.

Sign in / Sign up

Export Citation Format

Share Document