scholarly journals A Virtual Nodule Environment (ViNE) for modelling the inter-kingdom metabolic integration during symbiotic nitrogen fixation

2019 ◽  
Author(s):  
George C diCenzo ◽  
Michelangelo Tesi ◽  
Thomas Pfau ◽  
Alessio Mengoni ◽  
Marco Fondi
Keyword(s):  
Nitrogen Fixation ◽  
Plant Growth ◽  
Sinorhizobium Meliloti ◽  
Symbiotic Nitrogen Fixation ◽  
Nitrogenase Activity ◽  
Plant Mitochondria ◽  
Experimental Studies ◽  
Free Oxygen ◽  
Balance Analysis ◽  
Peribacteroid Space

ABSTRACTBiological associations are often premised upon metabolic cross-talk between the organisms, with the N2-fixing endosymbiotic relationship between rhizobia and leguminous plants being a prime example. Here, we report the in silico reconstruction of a metabolic network of a Medicago truncatula plant nodulated by the bacterium Sinorhizobium meliloti. The nodule tissue of the model contains five spatially distinct developmental zones and encompasses the metabolism of both the plant and the bacterium. Flux balance analysis (FBA) suggested that the majority of the metabolic costs associated with symbiotic nitrogen fixation are directly related to supporting nitrogenase activity, while a minority is related to the formation and maintenance of nodule and bacteroid tissue. Interestingly, FBA simulations suggested there was a non-linear relationship between the rate of N2-fixation per gram of nodule and the rate of plant growth; increasing the N2-fixation efficiency was associated with diminishing returns in terms of plant growth. Evaluating the metabolic exchange between the symbiotic partners provided support for: i) differentiating bacteroids having access to sugars (e.g., sucrose) as a major carbon source, ii) ammonium being the major nitrogen export product of N2-fixing bacteria, and iii) N2-fixation being dependent on the transfer of protons from the plant cytoplasm to the bacteria through acidification of the peribacteroid space. Our simulations further suggested that the use of C4-dicarboxylates by N2-fixing bacteroids may be, in part, a consequence of the low concentration of free oxygen in the nodule limiting the activity of the plant mitochondria. These results demonstrate the power of this integrated model to advance our understanding of the functioning of legume nodules, and its potential for hypothesis generation to guide experimental studies and engineering of symbiotic nitrogen fixation.

scholarly journals Increase in Alfalfa Nodulation, Nitrogen Fixation, and Plant Growth by Specific DNA Amplification in Sinorhizobium meliloti

1999 ◽  
Vol 65 (6) ◽  
pp. 2716-2722 ◽  
Author(s):  
Marcela Castillo ◽  
Margarita Flores ◽  
Patrick Mavingui ◽  
Esperanza Martínez-Romero ◽  
Rafael Palacios ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Plant Growth ◽  
Copy Number ◽  
Sinorhizobium Meliloti ◽  
Nitrogenase Activity ◽  
Regulatory Gene ◽  
Dna Amplification ◽  
Moderate Increase ◽  
Multicopy Plasmid ◽  
Plant Nitrogen

ABSTRACT To improve symbiotic nitrogen fixation on alfalfa plants,Sinorhizobium meliloti strains containing different average copy numbers of a symbiotic DNA region were constructed by specific DNA amplification (SDA). A DNA fragment containing a regulatory gene (nodD1), the common nodulation genes (nodABC), and an operon essential for nitrogen fixation (nifN) from the nod regulon region of the symbiotic plasmid pSyma ofS. meliloti was cloned into a plasmid unable to replicate in this organism. The plasmid then was integrated into the homologous DNA region of S. meliloti strains 41 and 1021, which resulted in a duplication of the symbiotic region.Sinorhizobium derivatives carrying further amplification were selected by growing the bacteria in increased concentrations of an antibiotic marker present in the integrated vector. Derivatives of strain 41 containing averages of 3 and 6 copies and a derivative of strain 1021 containing an average of 2.5 copies of the symbiotic region were obtained. In addition, the same region was introduced into both strains as a multicopy plasmid, yielding derivatives with an average of seven copies per cell. Nodulation, nitrogenase activity, plant nitrogen content, and plant growth were analyzed in alfalfa plants inoculated with the different strains. The copy number of the symbiotic region was critical in determining the plant phenotype. In the case of the strains with a moderate increase in copy number, symbiotic properties were improved significantly. The inoculation of alfalfa with these strains resulted in an enhancement of plant growth.

scholarly journals Succinate Transport Is Not Essential for Symbiotic Nitrogen Fixation by Sinorhizobium meliloti or Rhizobium leguminosarum

2017 ◽  
Vol 84 (1) ◽  
Author(s):  
Michael J. Mitsch ◽  
George C. diCenzo ◽  
Alison Cowie ◽  
Turlough M. Finan
Keyword(s):  
Nitrogen Fixation ◽  
Carbon Source ◽  
Sinorhizobium Meliloti ◽  
Rhizobium Leguminosarum ◽  
Symbiotic Nitrogen Fixation ◽  
Sequencing Analysis ◽  
Wild Type ◽  
Content Type ◽  
Transcriptional Changes ◽  
Symbiotic Properties

ABSTRACTSymbiotic nitrogen fixation (SNF) is an energetically expensive process performed by bacteria during endosymbiotic relationships with plants. The bacteria require the plant to provide a carbon source for the generation of reductant to power SNF. While C4-dicarboxylates (succinate, fumarate, and malate) appear to be the primary, if not sole, carbon source provided to the bacteria, the contribution of each C4-dicarboxylate is not known. We address this issue using genetic and systems-level analyses. Expression of a malate-specific transporter (MaeP) inSinorhizobium melilotiRm1021dctmutants unable to transport C4-dicarboxylates resulted in malate import rates of up to 30% that of the wild type. This was sufficient to support SNF withMedicago sativa, with acetylene reduction rates of up to 50% those of plants inoculated with wild-typeS. meliloti.Rhizobium leguminosarumbv. viciae 3841dctmutants unable to transport C4-dicarboxylates but expressing themaePtransporter had strong symbiotic properties, withPisum sativumplants inoculated with these strains appearing similar to plants inoculated with wild-typeR. leguminosarum. This was despite malate transport rates by the mutant bacteroids being 10% those of the wild type. An RNA-sequencing analysis of the combinedP. sativum-R. leguminosarumnodule transcriptome was performed to identify systems-level adaptations in response to the inability of the bacteria to import succinate or fumarate. Few transcriptional changes, with no obvious pattern, were detected. Overall, these data illustrated that succinate and fumarate are not essential for SNF and that, at least in specific symbioses,l-malate is likely the primary C4-dicarboxylate provided to the bacterium.IMPORTANCESymbiotic nitrogen fixation (SNF) is an economically and ecologically important biological process that allows plants to grow in nitrogen-poor soils without the need to apply nitrogen-based fertilizers. Much research has been dedicated to this topic to understand this process and to eventually manipulate it for agricultural gains. The work presented in this article provides new insights into the metabolic integration of the plant and bacterial partners. It is shown that malate is the only carbon source that needs to be available to the bacterium to support SNF and that, at least in some symbioses, malate, and not other C4-dicarboxylates, is likely the primary carbon provided to the bacterium. This work extends our knowledge of the minimal metabolic capabilities the bacterium requires to successfully perform SNF and may be useful in further studies aiming to optimize this process through synthetic biology approaches. The work describes an engineering approach to investigate a metabolic process that occurs between a eukaryotic host and its prokaryotic endosymbiont.

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.

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.

scholarly journals In Silico Insights into the Symbiotic Nitrogen Fixation in Sinorhizobium meliloti via Metabolic Reconstruction

PLoS ONE ◽  
2012 ◽  
Vol 7 (2) ◽  
pp. e31287 ◽  
Author(s):  
Hansheng Zhao ◽  
Mao Li ◽  
Kechi Fang ◽  
Wenfeng Chen ◽  
Jing Wang
Keyword(s):  
Nitrogen Fixation ◽  
In Silico ◽  
Sinorhizobium Meliloti ◽  
Symbiotic Nitrogen Fixation ◽  
Metabolic Reconstruction

scholarly journals About the work of Vinnytsia regional branch of the Ukrainian Society of Plant Physiologists

2020 ◽  
Vol 52 (5) ◽  
pp. 422-433
Author(s):  
V.G. Kuryata ◽  
Keyword(s):  
Nitrogen Fixation ◽  
Abscisic Acid ◽  
Plant Development ◽  
Acid Content ◽  
Symbiotic Nitrogen Fixation ◽  
Nitrogenase Activity ◽  
Free Form ◽  
Linear Growth Rate ◽  
Wide Range ◽  
Source Sink

The work and the main scientific results of the Vinnytsia branch of the USPP from the moment of its formation to the present time are analyzed. The main direction of research is exogenous regulation of plants source-sink relations in the systems «photosynthesis—growth», «depot of assimilates—growth» in the heterotrophic phase of plant development, and «macrosymbiont—microsymbiont» in symbiotic nitrogen fixation systems. Gibberellin and antigibberellin preparations (retardants) were used to create different tension of source-sink relations. It was found that under the influence of retardants, the decrease in the linear growth rate was accompanied by an increase in the cytokinins content with a decrease in the indoleacetic acid content in stems and leaves tissues of a wide range of plants. The free gibberellins activity was lower compared to control. At the same time, the content of these phytohormones antagonist, abscisic acid, increased. The retardants increased the stem branching, leaves number, weight and leaf surface area, optimized the leaves mesostructural organization, resulting in an increase in the photosynthetic net productivity. These changes lead to increased crop productivity. Under the conditions of artificial combination of external (light/dark) and hormonal factors (gibberellic acid and retardants) during seed germination, changes in the functioning of the source-sink system in the heterotrophic stage of plant development from seeds with different reserve substances types were studied. Gibberellin stimulated the starch breakdown in both light and dark, but under conditions of germination in the dark, the rate of reserve seed starch use was higher. Gibberellin also stimulated the seed reserve proteins hydrolysis, but the process began after intensive starch hydrolysis. Peculiarities of regulation of source-sink relations in the system «macro—microsymbiont» during the processes of symbiotic nitrogen fixation under the action of antigibberellin preparations — retardants are revealed. It was found that the typical reaction of soybean plants to the paclobutrazol application was a decrease in the free form gibberellins activity and an increase in the abscisic acid content, which led to changes in morphogenesis. Under the retardants action, the processes of formation of the symbiotic complex «soybean—Bradyrhizobium japonicum» intensify, nitrogenase activity increased, crop yield rised.

Bidirectional Selection for Nitrogenase Activity and Shoot Dry Weight Among Late Generation Progenies of a Virginia x Spanish Peanut Cross1

1986 ◽  
Vol 13 (2) ◽  
pp. 86-89 ◽  
Author(s):  
S. Arrendell ◽  
J. C. Wynne ◽  
G. H. Elkan ◽  
T. J. Schneeweis
Keyword(s):  
Nitrogen Fixation ◽  
Plant Growth ◽  
Growth Habit ◽  
Nitrogenase Activity ◽  
Dry Weight ◽  
Fruit Weight ◽  
Shoot Dry Weight ◽  
Plant Growth Habit ◽  
The Mean ◽  
Selection For

Abstract Improvement of the host contribution to nitrogen fixation has been proposed as a method of increasing nitrogen fixation. Significant variability and generally high broad-sense heritability estimates (.60 ± .27 to .82 ± .26 for nitrogenase activity and .53 ± .29 to .85 ± .26 for shoot dry weight) have been reported for F2-derived families from a cross between the Virginia (Arachis hypogaea L. ssp. hypogaea var. hypogaea) cultivar NC 6 and the Spanish (ssp. fastigiata Waldron vulgaris Harz.) breeding line 922, indicating selection for increased nigtogen fixation should be effective in this population. Lines from this population were chosen randomly from F2-derived families selected for high and low nitrogenase activity and high and low shoot dry weight after evaluation at three dates and two locations in each of 2 years (F5 and F6 generations). This study's objectives were to evaluate the N2-fixing ability of the selected lines and to evaluate the association between plant growth habit and N2 fixation. Twenty-four lines in each of the four selection groups and the parents, NC 6 and 922, were evaluated at two sampling dates and two locations. Mean nitrogenase activity of lines selected for increased nitrogenase activity was significantly greater than the mean of the lines selected for low nitrogenase activity. Improved nitrogenase activity was associated with increased fruit weight. The fruit weight mean of the group selected for increased fruit weight. The fruit weight mean of the group selected for increased nitrogenase activity was 39% greater than the mean of the group selected for low nitrogenase activity. Mean shoot dry weight of lines selected for increased shoot dry weight was significantly greater than the mean of the lines selected for low shoot dry weight; however, the fruit weight means of these two groups did not differ. It was hypothesized that selection for increased N2 fixation in a population derived from a cross between Virginia and Spanish types would eliminate genotypes with Spanish growth habit. Groups selected for high nitrogenase activity and for high shoot dry weight had longer and wider leaflets, longer cotyledonary laterals and greater main stem height than did their respective low selection groups. However, these traits chosen to characterize plant growth habit were inadequate in discriminating parental growth habits. Consequently, the data neither substantiated nor refuted the hypothesis.

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 Growth, photosynthesis, nodule nitrogen and carbon fixation in the chickpea cultivars under salt stress

2004 ◽  
Vol 16 (3) ◽  
pp. 137-146 ◽  
Author(s):  
Neera Garg ◽  
Ranju Singla
Keyword(s):  
Nitrogen Fixation ◽  
Salt Stress ◽  
Carbon Fixation ◽  
Symbiotic Nitrogen Fixation ◽  
Nitrogenase Activity ◽  
Saline Stress ◽  
Negative Effects ◽  
Leaf Chlorophyll ◽  
Mediterranean Origin ◽  
Salt Affected Soils

Four cultivars of chickpea, two of them of Mediterranean origin (kabuli), CSG 9651, BG 267 and two Indian (desi) types, CSG 8962, DCP 92-3, differing in their salt sensitivities were identified after screening ten genotypes in saline soils. The cultivars CSG 9651 and CSG 8962 were salt tolerant while BG 267 and DCP 92-3 were salt sensitive, respectively. The seeds of different cultivars were inoculated with Mesorhizobium ciceri, strain F: 75 and the plants were grown in the greenhouse. After the establishment of symbiosis, 15-day-old seedlings were administered doses of saline at varying concentrations (0, 4, 6, 8 dSm-1 NaCl, Na2SO4, CaCl2). Plants were harvested at 40, 70 and 100 days after sowing, for analyses. The main aim was to compare the relative salt tolerance of both desi and kabuli cultivars in terms of nitrogen fixation and carbon metabolism, as well as to ascertain whether the negative effects of saline stress on nitrogen fixation were due to a limitation of photosynthate supply to the nodule or to a limitation on the nodular metabolism that sustains nitrogenase activity. Plant growth, nodulation and nitrogenase activity was more severely affected in BG 267 and DCP 92-3 under salinity treatments (6 and 8 dSm-1) compared with CSG 9651 and CSG 8962. Nodule number as well as nodule mass increased under salt stress in CSG 9651 and CSG 8962 which might be responsible for their higher nitrogen fixation. Salinity reduced leaf chlorophyll and Rubisco activities in all cultivars. However, tolerant cultivars CSG 9651 and CSG 8962 showed smaller declines than the sensitive ones. Phosphoenolpyruvate carboxylase (PEPCase) activity increased significantly in the nodules of tolerant cultivars under salt stress at all harvests, and this was clearly related to salt concentrations. Our results suggest that in salt-affected soils tolerant cultivars have more efficient nodulation and support higher rates of symbiotic nitrogen fixation than the sensitive cultivars.

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.

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