Symbiotic Nitrogen Fixation at the Late Stage of Nodule Formation in Lotus japonicus and Other Legume Plants

2000 ◽  
Vol 113 (4) ◽  
pp. 467-473 ◽  
Author(s):  
Shigeyuki Tajima ◽  
Kenichi Takane ◽  
Mika Nomura ◽  
Hiroshi Kouchi
Keyword(s):  
Nitrogen Fixation ◽  
Late Stage ◽  
Symbiotic Nitrogen Fixation ◽  
Lotus Japonicus ◽  
Nodule Formation ◽  
Legume Plants

scholarly journals Αναπρογραμματισμός του μεταβολισμού του Lotus japonicus σε μεταγραφικό, βιοχημικό και μεταβολομικό επίπεδο κατά τη συμβιωτική αζωτοδέσμευση

2016 ◽  
Author(s):  
Χρυσάνθη Καλλονιάτη
Keyword(s):  
Nitrogen Fixation ◽  
Metabolite Profiling ◽  
Symbiotic Nitrogen Fixation ◽  
Sulfur Metabolism ◽  
Lotus Japonicus ◽  
Nitrogen Fixing ◽  
Model Legume ◽  
Whole Plant ◽  
Plant Level ◽  
Molecular And Biochemical Mechanisms

Symbiotic nitrogen fixation in legumes takes place in specialized organs called nodules,which become the main source of assimilated nitrogen for the whole plant. Symbiotic nitro‐gen fixation requires exquisite integration of plant and bacterial metabolism and involvesglobal changes in gene expression and metabolite accumulation in both rhizobia and thehost plant. In order to study the metabolic changes mediated by symbiotic nitrogen fixationon a whole‐plant level, metabolite levels were profiled by gas chromatography–mass spec‐trometry in nodules and non‐symbiotic organs of Lotus japonicus plants uninoculated or in‐oculated with M. loti wt,  ΔnifA or  ΔnifH fix‐ strains. Furthermore, transcriptomic andbiochemical approaches were combined to study sulfur metabolism in nodules, its link tosymbiotic nitrogen fixation, and the effect of nodules on whole‐plant sulfur partitioning andmetabolism. It is well established that nitrogen and sulfur (S) metabolism are tightly en‐twined and sulfur is required for symbiotic nitrogen fixation, however, little is known aboutthe molecular and biochemical mechanisms governing sulfur uptake and assimilation duringsymbiotic nitrogen fixation. Transcript profiling in Lotus japonicus was combined with quan‐tification of S‐metabolite contents and APR activity in nodules and in non‐symbiotic organsof plants uninoculated or inoculated with M. loti wt, ΔnifA or ΔnifH fix‐ strains. Moreover,sulfate uptake and its distribution into different plant organs were analyzed and 35S‐flux intodifferent S‐pools was monitored. Metabolite profiling revealed that symbiotic nitrogen fixa‐tion results in dramatic changes of many aspects of primary and secondary metabolism innodules which leads to global reprogramming of metabolism of the model legume on awhole‐plant level. Moreover, our data revealed that nitrogen fixing nodules represent athiol‐rich organ. Their high APR activity and 35S‐flux into cysteine and its metabolites in com‐bination with the transcriptional up‐regulation of several genes involved in sulfur assimila‐tion highlight the function of nodules as a new site of sulfur assimilation. The higher thiolcontent observed in non‐symbiotic organs of nitrogen fixing plants in comparison touninoculated plants cannot be attributed to local biosynthesis, indicating that nodules couldserve as a novel source of reduced sulfur for the plant, which triggers whole‐plant repro‐gramming of sulfur metabolism. Interestingly, the changes in metabolite profiling and theenhanced thiol biosynthesis in nodules and their impact on the whole‐plant sulfur, carbonand nitrogen economy are dampened in fix‐ plants, which in most respects metabolically re‐sembled uninoculated plants, indicating a strong interaction between nitrogen fixation andsulfur and carbon metabolism.

scholarly journals The rhizobial autotransporter determines the symbiotic nitrogen fixation activity of Lotus japonicus in a host-specific manner

2020 ◽  
Vol 117 (3) ◽  
pp. 1806-1815 ◽  
Author(s):  
Yoshikazu Shimoda ◽  
Yuki Nishigaya ◽  
Hiroko Yamaya-Ito ◽  
Noritoshi Inagaki ◽  
Yosuke Umehara ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Host Plant ◽  
Symbiotic Nitrogen Fixation ◽  
Root Nodules ◽  
Lotus Japonicus ◽  
Dependent Manner ◽  
Strain Specificity ◽  
Passenger Domain ◽  
Aspartic Peptidase ◽  
Specific Manner

Leguminous plants establish endosymbiotic associations with rhizobia and form root nodules in which the rhizobia fix atmospheric nitrogen. The host plant and intracellular rhizobia strictly control this symbiotic nitrogen fixation. We recently reported a Lotus japonicus Fix− mutant, apn1 (aspartic peptidase nodule-induced 1), that impairs symbiotic nitrogen fixation. APN1 encodes a nodule-specific aspartic peptidase involved in the Fix− phenotype in a rhizobial strain-specific manner. This host-strain specificity implies that some molecular interactions between host plant APN1 and rhizobial factors are required, although the biological function of APN1 in nodules and the mechanisms governing the interactions are unknown. To clarify how rhizobial factors are involved in strain-specific nitrogen fixation, we explored transposon mutants of Mesorhizobium loti strain TONO, which normally form Fix− nodules on apn1 roots, and identified TONO mutants that formed Fix+ nodules on apn1. The identified causal gene encodes an autotransporter, part of a protein secretion system of Gram-negative bacteria. Expression of the autotransporter gene in M. loti strain MAFF3030399, which normally forms Fix+ nodules on apn1 roots, resulted in Fix− nodules. The autotransporter of TONO functions to secrete a part of its own protein (a passenger domain) into extracellular spaces, and the recombinant APN1 protein cleaved the passenger protein in vitro. The M. loti autotransporter showed the activity to induce the genes involved in nodule senescence in a dose-dependent manner. Therefore, we conclude that the nodule-specific aspartic peptidase, APN1, suppresses negative effects of the rhizobial autotransporter in order to maintain effective symbiotic nitrogen fixation in root nodules.

scholarly journals CRISPR/Cas9 knockout of leghemoglobin genes in Lotus japonicus uncovers their synergistic roles in symbiotic nitrogen fixation

2019 ◽  
Vol 224 (2) ◽  
pp. 818-832 ◽  
Author(s):  
Longlong Wang ◽  
Maria Carmen Rubio ◽  
Xian Xin ◽  
Baoli Zhang ◽  
Qiuling Fan ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Symbiotic Nitrogen Fixation ◽  
Lotus Japonicus

scholarly journals The SNARE Protein SYP71 Expressed in Vascular Tissues Is Involved in Symbiotic Nitrogen Fixation in Lotus japonicus Nodules

2012 ◽  
Vol 160 (2) ◽  
pp. 897-905 ◽  
Author(s):  
Tsuneo Hakoyama ◽  
Ryo Oi ◽  
Kazuya Hazuma ◽  
Eri Suga ◽  
Yuka Adachi ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Symbiotic Nitrogen Fixation ◽  
Lotus Japonicus ◽  
Snare Protein ◽  
Vascular Tissues

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 Genome Editing in Cowpea Vigna unguiculata Using CRISPR-Cas9

2019 ◽  
Vol 20 (10) ◽  
pp. 2471 ◽  
Author(s):  
Jie Ji ◽  
Chunyang Zhang ◽  
Zhongfeng Sun ◽  
Longlong Wang ◽  
Deqiang Duanmu ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Genome Editing ◽  
Vigna Unguiculata ◽  
Symbiotic Nitrogen Fixation ◽  
Nodule Formation ◽  
Guide Rnas ◽  
Legume Crop ◽  
Receptor Like Kinase ◽  
Agronomical Traits

Cowpea (Vigna unguiculata) is widely cultivated across the world. Due to its symbiotic nitrogen fixation capability and many agronomically important traits, such as tolerance to low rainfall and low fertilization requirements, as well as its high nutrition and health benefits, cowpea is an important legume crop, especially in many semi-arid countries. However, research in Vigna unguiculata is dramatically hampered by the lack of mutant resources and efficient tools for gene inactivation in vivo. In this study, we used clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (Cas9). We applied the CRISPR/Cas9-mediated genome editing technology to efficiently disrupt the representative symbiotic nitrogen fixation (SNF) gene in Vigna unguiculata. Our customized guide RNAs (gRNAs) targeting symbiosis receptor-like kinase (SYMRK) achieved ~67% mutagenic efficiency in hairy-root-transformed plants, and nodule formation was completely blocked in the mutants with both alleles disrupted. Various types of mutations were observed near the PAM region of the respective gRNA. These results demonstrate the applicability of the CRISPR/Cas9 system in Vigna unguiculata, and therefore should significantly stimulate functional genomics analyses of many important agronomical traits in this unique crop legume.

Influence of chlorsulfuron on rhizobial growth, nodule formation, and nitrogen fixation with chickpea

2004 ◽  
Vol 55 (10) ◽  
pp. 1059 ◽  
Author(s):  
A. Anderson ◽  
J. A. Baldock ◽  
S. L. Rogers ◽  
W. Bellotti ◽  
G. Gill
Keyword(s):  
Nitrogen Fixation ◽  
Symbiotic Nitrogen Fixation ◽  
Defined Medium ◽  
Application Rate ◽  
Field Application ◽  
Nodule Formation ◽  
Negative Effects ◽  
Legume Crops ◽  
Cell Inoculum

Sulfonylurea residues have been found to inhibit the growth of some legume crops and pastures in seasons following application. Negative effects of these herbicides on symbiotic nitrogen fixation by legume crops and pastures have been demonstrated. Reductions in nitrogen fixation may result from a direct effect of the herbicide on rhizobial growth and/or an indirect effect on plant growth. In this study the influence of chlorsulfuron on the growth of chickpea rhizobia [Mesorhizobium ciceri (CC1192)], the growth of chickpea plants, and the extent of nodulation and nitrogen fixation by the chickpea/rhizobia symbiosis were examined. In vitro studies (in yeast mannitol broth and a defined medium) showed that chlorsulfuron applied at double the recommended field application rate did not influence the growth of chickpea rhizobia. An experiment using 14C-labelled chlorsulfuron was conducted to determine if rhizobial cells exposed to chlorsulfuron could deliver the herbicide to the point of root infection and nodule formation. Approximately 1% of the herbicide present in the rhizobial growth medium remained with the cell/inoculum material after rinsing with 1/4 strength Ringer’s solution. This was considered unlikely to affect chickpea growth, nodulation, or nitrogen fixation. A pot experiment was used to define the influence of chlorsulfuron on the growth, nodulation, and nitrogen fixation of chickpeas. The presence of chlorsulfuron in the soil reduced the nodulation and nitrogen fixation of the chickpea plants. Pre-exposing rhizobia to chlorsulfuron before inoculating them into pots with germinating chickpea seeds, reduced the number of nodules formed by 51%. Exposure of chickpeas and chickpea rhizobia to chlorsulfuron can adversely affect the formation and activity of symbiotic nitrogen-fixing nodules, even when only the rhizobial inoculant is exposed briefly to the herbicide.

INFLUENCE OF ROOT TEMPERATURE ON THE EARLY GROWTH AND SYMBIOTIC NITROGEN FIXATION OF NODULATED LOTUS CORNICULATUS PLANTS

1970 ◽  
Vol 50 (5) ◽  
pp. 569-575 ◽  
Author(s):  
H. T. KUNELIUS ◽  
K. W. CLARK
Keyword(s):  
Nitrogen Fixation ◽  
Ammonium Nitrate ◽  
Environmental Conditions ◽  
Symbiotic Nitrogen Fixation ◽  
Lotus Corniculatus ◽  
Early Growth ◽  
Birdsfoot Trefoil ◽  
Nodule Formation ◽  
Root Temperature ◽  
Nitrogen Yields

Three birdsfoot trefoil (Lotus corniculatus L.) cultivars, inoculated with one of six Lotus rhizobia strains or dependent on ammonium nitrate, were grown in diSPo growth pouches under controlled environmental conditions at five root temperatures (9–30 C) for 35 days after nodule formation. When the plants were dependent on symbiotic nitrogen fixation, the highest dry weights and nitrogen yields per plant were obtained at 18 or 24 C depending on symbiotic combination. At 9 and 12 C, nitrogen fixation was depressed and the growth was poor. The dry weights of plants at 9 C were 19 to 45% of those at 24 C. At 30 C the growth and nitrogen fixation were generally depressed. At all root temperatures the growth of plants dependent on symbiotic nitrogen fixation was inferior to that of plants receiving combined nitrogen (NH4NO3). Significant interactions indicate that the nitrogen fixing ability of cultivars was dependent on both root temperature and the strain of Lotus rhizobia.

scholarly journals Nitrate inhibits nodule organogenesis through inhibition of cytokinin biosynthesis in Lotus japonicus

2020 ◽  
Author(s):  
Jieshun Lin ◽  
Yuda Purwana Roswanjaya ◽  
Wouter Kohlen ◽  
Jens Stougaard ◽  
Dugald Reid
Keyword(s):  
Nitrogen Fixation ◽  
Developmental Process ◽  
Root Zone ◽  
Lotus Japonicus ◽  
Nodule Development ◽  
Nodule Formation ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Cytokinin Biosynthesis ◽  
Nitrate Inhibition

AbstractLegumes balance nitrogen acquisition from soil nitrate with symbiotic nitrogen fixation. Nitrogen fixation requires establishment of a new organ, which is a cytokinin dependent developmental process in the root. We found cytokinin biosynthesis is a central integrator, balancing nitrate signalling with symbiotic acquired nitrogen. Low nitrate conditions provide a permissive state for induction of cytokinin by symbiotic signalling and thus nodule development. In contrast, high nitrate is inhibitory to cytokinin accumulation and nodule establishment in the root zone susceptible to nodule formation. This reduction of symbiotic cytokinin accumulation was further exacerbated in cytokinin biosynthesis mutants, which display hypersensitivity to nitrate inhibition of nodule development, maturation and nitrogen fixation. Consistent with this, cytokinin application can rescue nodulation and nitrogen fixation of biosynthesis mutants in a concentration dependent manner. These inhibitory impacts of nitrate on symbiosis occur in a Nlp1 and Nlp4 dependent manner and contrast with the positive influence of nitrate on cytokinin biosynthesis that occurs in non-symbiotic species. Altogether this shows that legumes, as exemplified by Lotus japonicus, have evolved a different cytokinin response to nitrate compared to non-legumes.One sentence summaryCytokinin biosynthesis is suppressed by nitrate in Lotus japonicus, providing a mechanism for nitrate inhibition of symbiotic nodule organogenesis.

scholarly journals A Dicarboxylate Transporter, LjALMT4, Mainly Expressed in Nodules of Lotus japonicus

2016 ◽  
Vol 29 (7) ◽  
pp. 584-592 ◽  
Author(s):  
Kojiro Takanashi ◽  
Takayuki Sasaki ◽  
Tomohiro Kan ◽  
Yuka Saida ◽  
Akifumi Sugiyama ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Symbiotic Nitrogen Fixation ◽  
Lotus Japonicus ◽  
Specific Gene ◽  
Vascular Bundles ◽  
Transport Activity ◽  
Organ Specific ◽  
Infected Cells ◽  
Photosynthetic Products ◽  
Strong Candidate

Legume plants can establish symbiosis with soil bacteria called rhizobia to obtain nitrogen as a nutrient directly from atmospheric N2 via symbiotic nitrogen fixation. Legumes and rhizobia form nodules, symbiotic organs in which fixed-nitrogen and photosynthetic products are exchanged between rhizobia and plant cells. The photosynthetic products supplied to rhizobia are thought to be dicarboxylates but little is known about the movement of dicarboxylates in the nodules. In terms of dicarboxylate transporters, an aluminum-activated malate transporter (ALMT) family is a strong candidate responsible for the membrane transport of carboxylates in nodules. Among the seven ALMT genes in the Lotus japonicus genome, only one, LjALMT4, shows a high expression in the nodules. LjALMT4 showed transport activity in a Xenopus oocyte system, with LjALMT4 mediating the efflux of dicarboxylates including malate, succinate, and fumarate, but not tricarboxylates such as citrate. LjALMT4 also mediated the influx of several inorganic anions. Organ-specific gene expression analysis showed LjALMT4 mRNA mainly in the parenchyma cells of nodule vascular bundles. These results suggest that LjALMT4 may not be involved in the direct supply of dicarboxylates to rhizobia in infected cells but is responsible for supplying malate as well as several anions necessary for symbiotic nitrogen fixation, via nodule vasculatures.

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