scholarly journals Transcription of ENOD8 in Medicago truncatula Nodules Directs ENOD8 Esterase to Developing and Mature Symbiosomes

2008 ◽  
Vol 21 (4) ◽  
pp. 404-410 ◽  
Author(s):  
Laurent Coque ◽  
Purnima Neogi ◽  
Catalina Pislariu ◽  
Kimberly A. Wilson ◽  
Christina Catalano ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Medicago Truncatula ◽  
Sinorhizobium Meliloti ◽  
Immunoblot Analysis ◽  
Nodule Formation ◽  
Nitrogenous Compounds ◽  
Symbiosome Membrane ◽  
The Matrix ◽  
And Function ◽  
Infected Nodule

In Medicago truncatula nodules, the soil bacterium Sinorhizobium meliloti reduces atmospheric dinitrogen into nitrogenous compounds that the legume uses for its own growth. In nitrogen-fixing nodules, each infected cell contains symbiosomes, which include the rhizobial cell, the symbiosome membrane surrounding it, and the matrix between the bacterium and the symbiosome membrane, termed the symbiosome space. Here, we describe the localization of ENOD8, a nodule-specific esterase. The onset of ENOD8 expression occurs at 4 to 5 days postinoculation, before the genes that support the nitrogen fixation capabilities of the nodule. Expression of an ENOD8 promoter–gusA fusion in nodulated hairy roots of composite transformed M. truncatula plants indicated that ENOD8 is expressed from the proximal end of interzone II to III to the proximal end of the nodules. Confocal immunomicroscopy using an ENOD8-specific antibody showed that the ENOD8 protein was detected in the same zones. ENOD8 protein was localized in the symbiosome membrane or symbiosome space around the bacteroids in the infected nodule cells. Immunoblot analysis of fractionated symbiosomes strongly suggested that ENOD8 protein was found in the symbiosome membrane and symbiosome space, but not in the bacteroid. Determining the localization of ENOD8 protein in the symbiosome is a first step in understanding its role in symbiosome membrane and space during nodule formation and function.

scholarly journals Expression of Medicago truncatula Genes Responsive to Nitric Oxide in Pathogenic and Symbiotic Conditions

2008 ◽  
Vol 21 (6) ◽  
pp. 781-790 ◽  
Author(s):  
Alberto Ferrarini ◽  
Matteo De Stefano ◽  
Emmanuel Baudouin ◽  
Chiara Pucciariello ◽  
Annalisa Polverari ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Medicago Truncatula ◽  
Regulatory Networks ◽  
Sinorhizobium Meliloti ◽  
Biotic Interactions ◽  
Hypersensitive Reaction ◽  
Nodule Formation ◽  
Polymorphism Analysis ◽  
Symbiotic Interaction ◽  
Physiological Relevance

Nitric oxide (NO) is involved in diverse physiological processes in plants, including growth, development, response to pathogens, and interactions with beneficial microorganisms. In this work, a dedicated microarray representing the widest database available of NO-related transcripts in plants has been produced with 999 genes identified by a cDNA amplified fragment length polymorphism analysis as modulated in Medicago truncatula roots treated with two NO donors. The microarray then was used to monitor the expression of NO-responsive genes in M. truncatula during the incompatible interaction with the foliar pathogen Colletotrichum trifolii race 1 and during the symbiotic interaction with Sinorhizobium meliloti 1021. A wide modulation of NO-related genes has been detected during the hypersensitive reaction or during nodule formation and is discussed with special emphasis on the physiological relevance of these genes in the context of the two biotic interactions. This work clearly shows that NO-responsive genes behave differently depending on the plant organ and on the type of interaction, strengthening the need to consider regulatory networks, including different signaling molecules.

scholarly journals Structure and antimicrobial activity of NCR169, a nodule-specific cysteine-rich peptide of Medicago truncatula

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Noriyoshi Isozumi ◽  
Yuya Masubuchi ◽  
Tomohiro Imamura ◽  
Masashi Mori ◽  
Hironori Koga ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Medicago Truncatula ◽  
Mechanism Of Action ◽  
Disulfide Bonds ◽  
Sinorhizobium Meliloti ◽  
Bound State ◽  
Disulfide Linkage ◽  
Model Legume ◽  
Nmr Structures ◽  
And Function

AbstractA model legume, Medicago truncatula, has over 600 nodule-specific cysteine-rich (NCR) peptides required for symbiosis with rhizobia. Among them, NCR169, an essential factor for establishing symbiosis, has four cysteine residues that are indispensable for its function. However, knowledge of NCR169 structure and mechanism of action is still lacking. In this study, we solved two NMR structures of NCR169 caused by different disulfide linkage patterns. We show that both structures have a consensus C-terminal β-sheet attached to an extended N-terminal region with dissimilar features; one moves widely, whereas the other is relatively stapled. We further revealed that the disulfide bonds of NCR169 contribute to its structural stability and solubility. Regarding the function, one of the NCR169 oxidized forms could bind to negatively charged bacterial phospholipids. Furthermore, the positively charged lysine-rich region of NCR169 may be responsible for its antimicrobial activity against Escherichia coli and Sinorhizobium meliloti. This active region was disordered even in the phospholipid bound state, suggesting that the disordered conformation of this region is key to its function. Morphological observations suggested the mechanism of action of NCR169 on bacteria. The present study on NCR169 provides new insights into the structure and function of NCR peptides.

scholarly journals Aphid infestation differently affects the defences of nitrate-fed and nitrogen-fixing Medicago truncatula and alters symbiotic nitrogen fixation

2020 ◽  
Vol 287 (1934) ◽  
pp. 20201493
Author(s):  
Gaurav Pandharikar ◽  
Jean-Luc Gatti ◽  
Jean-Christophe Simon ◽  
Pierre Frendo ◽  
Marylène Poirié
Keyword(s):  
Nitrogen Fixation ◽  
Medicago Truncatula ◽  
Sinorhizobium Meliloti ◽  
Acyrthosiphon Pisum ◽  
Symbiotic Nitrogen Fixation ◽  
Pea Aphid ◽  
Systemic Resistance ◽  
Aphid Infestation ◽  
Plant Nitrogen ◽  
Pathogenesis Related Protein

Legumes can meet their nitrogen requirements through root nodule symbiosis, which could also trigger plant systemic resistance against pests. The pea aphid Acyrthosiphon pisum , a legume pest, can harbour different facultative symbionts (FS) influencing various traits of their hosts. It is therefore worth determining if and how the symbionts of the plant and the aphid modulate their interaction. We used different pea aphid lines without FS or with a single one ( Hamiltonella defensa , Regiella insecticola, Serratia symbiotica ) to infest Medicago truncatula plants inoculated with Sinorhizobium meliloti (symbiotic nitrogen fixation, SNF) or supplemented with nitrate (non-inoculated, NI). The growth of SNF and NI plants was reduced by aphid infestation, while aphid weight (but not survival) was lowered on SNF compared to NI plants. Aphids strongly affected the plant nitrogen fixation depending on their symbiotic status, suggesting indirect relationships between aphid- and plant-associated microbes. Finally, all aphid lines triggered expression of Pathogenesis-Related Protein 1 ( PR1 ) and Proteinase Inhibitor (PI) , respective markers for salicylic and jasmonic pathways, in SNF plants, compared to only PR1 in NI plants. We demonstrate that the plant symbiotic status influences plant–aphid interactions while that of the aphid can modulate the amplitude of the plant's defence response.

scholarly journals Sinorhizobium meliloti Controls Nitric Oxide–Mediated Post-Translational Modification of a Medicago truncatula Nodule Protein

2015 ◽  
Vol 28 (12) ◽  
pp. 1353-1363 ◽  
Author(s):  
Pauline Blanquet ◽  
Liliana Silva ◽  
Olivier Catrice ◽  
Claude Bruand ◽  
Helena Carvalho ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nitrogen Fixation ◽  
Medicago Truncatula ◽  
Sinorhizobium Meliloti ◽  
Molecular Mechanisms ◽  
Root Nodules ◽  
Plant Protein ◽  
Post Translational Modification ◽  
Model Legume ◽  
Bacterial Mutants

Nitric oxide (NO) is involved in various plant-microbe interactions. In the symbiosis between soil bacterium Sinorhizobium meliloti and model legume Medicago truncatula, NO is required for an optimal establishment of the interaction but is also a signal for nodule senescence. Little is known about the molecular mechanisms responsible for NO effects in the legume-rhizobium interaction. Here, we investigate the contribution of the bacterial NO response to the modulation of a plant protein post-translational modification in nitrogen-fixing nodules. We made use of different bacterial mutants to finely modulate NO levels inside M. truncatula root nodules and to examine the consequence on tyrosine nitration of the plant glutamine synthetase, a protein responsible for assimilation of the ammonia released by nitrogen fixation. Our results reveal that S. meliloti possesses several proteins that limit inactivation of plant enzyme activity via NO-mediated post-translational modifications. This is the first demonstration that rhizobia can impact the course of nitrogen fixation by modulating the activity of a plant protein.

scholarly journals EFFECT OF PHOSPHATE ON NODULE PRIMORDIA OF SOYBEAN (Glycine max Merrill) IN ACID SOILS IN RHIZOTRON EXPERIMENTS

2016 ◽  
Vol 5 (2) ◽  
pp. 37 ◽  
Author(s):  
Setiyo Hadi Waluyo ◽  
Tek An Lie ◽  
Leendert ’t Mannetje
Keyword(s):  
Nitrogen Fixation ◽  
Root Growth ◽  
Acid Soils ◽  
Nodule Formation ◽  
Soybean Plant ◽  
Limiting Nutrients ◽  
West Sumatra ◽  
And Function ◽  
Biological Nitrogen ◽  
Positive Effect

To clarify whether P had a direct or indirect effect on the nodulation  process of soybean grown in acid soils from Sitiung, West Sumatra, Indonesia, a series of rhizotron experiments, with special attention given to formation of nodule primordia, was conducted at Laboratory of  Microbiology, Wageningen University in 1998-2000. It was shown that Ca and P were essential nutrients for root growth, nodule formation, and growth of soybean in the acid soils (Oxisols). Ca increased root growth, number of nodule primordia, nodules, and growth of the soybean plant. This positive effect of Ca was increased considerably by the application of P. Ca and P have a synergistic effect on biological nitrogen fixation (BNF) of soybean in acid soils. Ca is important for the establishment of nodules, whilst P is essential for the development and function of the formed nodules. P increased number of nodule primordia, thus it also has an important role in the initiation of nodule formation. From this study, it can be concluded that Ca and P are the most limiting nutrients for BNF of soybean in the acid soils of Sitiung, West Sumatra, Indonesia.

scholarly journals Specific Host-Responsive Associations Between Medicago truncatula Accessions and Sinorhizobium Strains

2017 ◽  
Vol 30 (5) ◽  
pp. 399-409 ◽  
Author(s):  
Théophile Kazmierczak ◽  
Marianna Nagymihály ◽  
Florian Lamouche ◽  
Quentin Barrière ◽  
Ibtissem Guefrachi ◽  
...  
Keyword(s):  
Medicago Truncatula ◽  
Sinorhizobium Meliloti ◽  
Root Nodules ◽  
Nodule Formation ◽  
Bacterial Strains ◽  
Symbiotic Efficiency ◽  
Model Legume ◽  
Functional Studies ◽  
Multiple Parameters ◽  
Nitrogen Fixation Activity

Legume plants interact with rhizobia to form nitrogen-fixing root nodules. Legume-rhizobium interactions are specific and only compatible rhizobia and plant species will lead to nodule formation. Even within compatible interactions, the genotype of both the plant and the bacterial symbiont will impact on the efficiency of nodule functioning and nitrogen-fixation activity. The model legume Medicago truncatula forms nodules with several species of the Sinorhizobium genus. However, the efficiency of these bacterial strains is highly variable. In this study, we compared the symbiotic efficiency of Sinorhizobium meliloti strains Sm1021, 102F34, and FSM-MA, and Sinorhizobium medicae strain WSM419 on the two widely used M. truncatula accessions A17 and R108. The efficiency of the interactions was determined by multiple parameters. We found a high effectiveness of the FSM-MA strain with both M. truncatula accessions. In contrast, specific highly efficient interactions were obtained for the A17-WSM419 and R108-102F34 combinations. Remarkably, the widely used Sm1021 strain performed weakly on both hosts. We showed that Sm1021 efficiently induced nodule organogenesis but cannot fully activate the differentiation of the symbiotic nodule cells, explaining its weaker performance. These results will be informative for the selection of appropriate rhizobium strains in functional studies on symbiosis using these M. truncatula accessions, particularly for research focusing on late stages of the nodulation process.

scholarly journals Transcriptomic Analysis of Sinorhizobium meliloti and Medicago truncatula Symbiosis Using Nitrogen Fixation–Deficient Nodules

2015 ◽  
Vol 28 (8) ◽  
pp. 856-868 ◽  
Author(s):  
Claus Lang ◽  
Sharon R. Long
Keyword(s):  
Gene Expression ◽  
Nitrogen Fixation ◽  
Bacterial Infection ◽  
Medicago Truncatula ◽  
Sinorhizobium Meliloti ◽  
Cell Envelope ◽  
Root Nodules ◽  
Bacterial Cells ◽  
Nitrogen Fixing ◽  
Bacterial Genes

The bacterium Sinorhizobium meliloti interacts symbiotically with legume plant hosts such as Medicago truncatula to form nitrogen-fixing root nodules. During symbiosis, plant and bacterial cells differentiate in a coordinated manner, resulting in specialized plant cells that contain nitrogen-fixing bacteroids. Both plant and bacterial genes are required at each developmental stage of symbiosis. We analyzed gene expression in nodules formed by wild-type bacteria on six plant mutants with defects in nitrogen fixation. We observed differential expression of 482 S. meliloti genes with functions in cell envelope homeostasis, cell division, stress response, energy metabolism, and nitrogen fixation. We simultaneously analyzed gene expression in M. truncatula and observed differential regulation of host processes that may trigger bacteroid differentiation and control bacterial infection. Our analyses of developmentally arrested plant mutants indicate that plants use distinct means to control bacterial infection during early and late symbiotic stages.

scholarly journals EFFECT OF PHOSPHATE ON NODULE PRIMORDIA OF SOYBEAN (Glycine max Merrill) IN ACID SOILS IN RHIZOTRON EXPERIMENTS

2016 ◽  
Vol 5 (2) ◽  
pp. 37 ◽  
Author(s):  
Setiyo Hadi Waluyo ◽  
Tek An Lie ◽  
Leendert ’t Mannetje
Keyword(s):  
Nitrogen Fixation ◽  
Root Growth ◽  
Acid Soils ◽  
Nodule Formation ◽  
Soybean Plant ◽  
Limiting Nutrients ◽  
West Sumatra ◽  
And Function ◽  
Biological Nitrogen ◽  
Positive Effect

To clarify whether P had a direct or indirect effect on the nodulation  process of soybean grown in acid soils from Sitiung, West Sumatra, Indonesia, a series of rhizotron experiments, with special attention given to formation of nodule primordia, was conducted at Laboratory of  Microbiology, Wageningen University in 1998-2000. It was shown that Ca and P were essential nutrients for root growth, nodule formation, and growth of soybean in the acid soils (Oxisols). Ca increased root growth, number of nodule primordia, nodules, and growth of the soybean plant. This positive effect of Ca was increased considerably by the application of P. Ca and P have a synergistic effect on biological nitrogen fixation (BNF) of soybean in acid soils. Ca is important for the establishment of nodules, whilst P is essential for the development and function of the formed nodules. P increased number of nodule primordia, thus it also has an important role in the initiation of nodule formation. From this study, it can be concluded that Ca and P are the most limiting nutrients for BNF of soybean in the acid soils of Sitiung, West Sumatra, Indonesia.

scholarly journals Spatiotemporal cytokinin signaling imaging reveals IPT3 function in nodule development in Medicago truncatula

2021 ◽  
Author(s):  
Paolo M. Triozzi ◽  
Thomas B. Irving ◽  
Henry W. Schmidt ◽  
Zachary P. Keyser ◽  
Sanhita Chakraborty ◽  
...  
Keyword(s):  
High Resolution ◽  
Medicago Truncatula ◽  
Sinorhizobium Meliloti ◽  
Root Nodules ◽  
Nodule Development ◽  
Nodule Formation ◽  
Symbiotic Association ◽  
Loss Of Function ◽  
Cytokinin Signaling ◽  
Tissue Specific

ABSTRACTMost legumes can establish a symbiotic association with soil rhizobia that triggers the development of root nodules. These nodules host the rhizobia and allow them to fix nitrogen efficiently. The perception of bacterial lipo-chitooligosaccharide (LCO) signal in the epidermis initiates a signaling cascade that allows rhizobial intracellular infection in the root and de-differentiation and activation of cell division that gives rise to the nodule. Nodule organogenesis and rhizobial infection need to be coupled in space and time for successful nodulation. The plant hormone cytokinin (CK) acts as an essential positive regulator of nodule organogenesis, and specific CK receptors are required for nodule formation. Temporal regulation of tissue-specific CK signaling and biosynthesis in response to LCOs or Sinorhizobium meliloti inoculation in Medicago truncatula remains poorly understood. In the present study, using a fluorescence-based CK sensor (TCSn::nls:tGFP), we performed a high-resolution tissue-specific temporal characterization of the CK response’s sequential activation during root infection and nodule development in M. truncatula after inoculation with S. meliloti. Loss-of-function mutants of the CK-biosynthetic gene ISOPENTENYL TRANSFERASE 3 (IPT3) showed impairment of nodulation, suggesting that IPT3 is required for nodule development in M. truncatula. Simultaneous live imaging of pIPT3::tdTOMATO and the CK sensor showed that IPT3 induction in the root stele at the base of nodule primordium contributes to CK biosynthesis, which in turn promotes expression of positive regulators of nodule organogenesis in M. truncatula.One-sentence summaryHigh-resolution spatiotemporal imaging of cytokinin signaling reveals IPT3 function during indeterminate nodule development in Medicago truncatula

scholarly journals Sinorhizobium medicae WSM419 genes that improve symbiosis between Sinorhizobium meliloti Rm1021 and Medicago truncatula Jemalong A17 and in other symbiotic systems

2021 ◽  
Author(s):  
Prithwi Ghosh ◽  
Katie N. Adolphsen ◽  
Svetlana N. Yurgel ◽  
Michael L. Kahn
Keyword(s):  
Plant Growth ◽  
Medicago Truncatula ◽  
Pathogenic Bacteria ◽  
Sinorhizobium Meliloti ◽  
Rhizobium Leguminosarum ◽  
Nodule Formation ◽  
New Genes ◽  
Sinorhizobium Medicae ◽  
Different Strains ◽  
Vigorous Plant

Some soil bacteria called rhizobia can interact symbiotically with legumes in which they form nodules on the plant roots where they can reduce atmospheric dinitrogen to ammonia, a form of nitrogen that can be used by growing plants. Rhizobia/plant combinations can differ in how successful this symbiosis is—Sinorhizobium meliloti Rm1021 forms a relatively ineffective symbiosis with Medicago truncatula Jemalong A17 but Sinorhizobium medicae WSM419 is able to support more vigorous plant growth. Using proteomic data from free-living and symbiotic S. medicae WSM419, we previously identified a subset of proteins that were not closely related to any S. meliloti Rm1021 proteins and speculated that adding one or more of these proteins to S. meliloti Rm1021 would increase its effectiveness on M. truncatula A17. Three genes, Smed_3503, Smed_5985, and Smed_6456, were cloned into S. meliloti Rm1021 downstream of the E. coli lacZ promoter. Strains with these genes increased nodulation and improved plant growth, individually and in combination with one another. Smed_3503, renamed iseA (increased symbiotic effectiveness) had the largest impact, increasing M. truncatula biomass by 61%. iseA homologs were present in all currently sequenced S. medicae strains but were infrequent in other Sinorhizobium isolates. Rhizobium leguminosarum bv. viciae 3841 containing iseA led to more nodules on pea and lentil. Split root experiments with M. truncatula A17 indicated that S. meliloti Rm1021 carrying the S. medicae iseA is less sensitive to plant induced resistance to rhizobial infection, suggesting an interaction with the plant’s regulation of nodule formation. IMPORTANCE The legume symbiosis with rhizobia is highly specific. Rhizobia that can nodulate and fix nitrogen on one legume species are often unable to associate with a different species. The interaction can be more subtle—symbiotically enhanced growth of the host plant can differ substantially when nodules are formed by different rhizobial isolates of a species, much like disease severity can differ when conspecific isolates of pathogenic bacteria infect different cultivars. Much is known about bacterial genes essential for a productive symbiosis, but less is understood about genes that marginally improve performance. We used a proteomic strategy to identify Sinorhizobium genes that contribute to plant growth differences that are seen when two different strains nodulate M. truncatula A17. These genes could also alter the symbiosis between R. leguminosarum bv. viciae 3841 and pea or lentil, suggesting that this approach may identify new genes that may more generally contribute to symbiotic productivity.

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