scholarly journals Rhizobium-Induced Calcium Spiking in Lotus japonicus

2003 ◽  
Vol 16 (4) ◽  
pp. 335-341 ◽  
Author(s):  
Jeanne M. Harris ◽  
Rebecca Wais ◽  
Sharon R. Long

Legumes and rhizobium bacteria form a symbiosis that results in the development of nitrogen-fixing nodules on the root of the host plant. The earliest plant developmental changes are triggered by bacterially produced nodulation (Nod) factors. Within minutes of exposure to Nod factors, sharp oscillations in cytoplasmic calcium levels (calcium spiking) occur in epidermal cells of several closely related legumes. We found that Lotus japonicus, a legume that follows an alternate developmental pathway, responds to both its bacterial partner and to the purified bacterial signal with calcium spiking. Thus, calcium spiking is not restricted to a particular pathway of nodule development and may be a general component of the response of host legumes to their bacterial partner. Using Nod factor-induced calcium spiking as a tool to identify mutants blocked early in the response to Nod factor, we show that the L. japonicus Ljsym22-1 mutant but not the Ljsym30 mutant fails to respond to Nod factor with calcium spiking.

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Eiichi Murakami ◽  
Jeryl Cheng ◽  
Kira Gysel ◽  
Zoltan Bozsoki ◽  
Yasuyuki Kawaharada ◽  
...  

Recognition of Nod factors by LysM receptors is crucial for nitrogen-fixing symbiosis in most legumes. The large families of LysM receptors in legumes suggest concerted functions, yet only NFR1 and NFR5 and their closest homologs are known to be required. Here we show that an epidermal LysM receptor (NFRe), ensures robust signalling in L. japonicus. Mutants of Nfre react to Nod factors with increased calcium spiking interval, reduced transcriptional response and fewer nodules in the presence of rhizobia. NFRe has an active kinase capable of phosphorylating NFR5, which in turn, controls NFRe downstream signalling. Our findings provide evidence for a more complex Nod factor signalling mechanism than previously anticipated. The spatio-temporal interplay between Nfre and Nfr1, and their divergent signalling through distinct kinases suggests the presence of an NFRe-mediated idling state keeping the epidermal cells of the expanding root system attuned to rhizobia.


eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Anna Malolepszy ◽  
Simon Kelly ◽  
Kasper Kildegaard Sørensen ◽  
Euan Kevin James ◽  
Christina Kalisch ◽  
...  

Morphogens provide positional information and their concentration is key to the organized development of multicellular organisms. Nitrogen-fixing root nodules are unique organs induced by Nod factor-producing bacteria. Localized production of Nod factors establishes a developmental field within the root where plant cells are reprogrammed to form infection threads and primordia. We found that regulation of Nod factor levels by Lotus japonicus is required for the formation of nitrogen-fixing organs, determining the fate of this induced developmental program. Our analysis of plant and bacterial mutants shows that a host chitinase modulates Nod factor levels possibly in a structure-dependent manner. In Lotus, this is required for maintaining Nod factor signalling in parallel with the elongation of infection threads within the nodule cortex, while root hair infection and primordia formation are not influenced. Our study shows that infected nodules require balanced levels of Nod factors for completing their transition to functional, nitrogen-fixing organs.


2008 ◽  
Vol 21 (8) ◽  
pp. 1118-1127 ◽  
Author(s):  
Jean-Philippe Combier ◽  
Helge Küster ◽  
Etienne-Pascal Journet ◽  
Natalija Hohnjec ◽  
Pascal Gamas ◽  
...  

Nod factors are key bacterial signaling molecules regulating the symbiotic interaction between bacteria known as rhizobia and leguminous plants. Studying plant host genes whose expression is affected by Nod factors has given insights into early symbiotic signaling and development. Here, we used a double supernodulating mutant line that shows increased sensitivity to Nod factors to study the Nod factor-regulated transcriptome. Using microarrays containing more than 16,000 70-mer oligonucleotide probes, we identified 643 Nod-factor-regulated genes, including 225 new Nod-factor-upregulated genes encoding many potential regulators. Among the genes found to be Nod factor upregulated, we identified and characterized MtRALFL1 and MtDVL1, which code for two small putative peptide regulators of 135 and 53 amino acids, respectively. Expression analysis confirmed that these genes are upregulated during initial phases of nodulation. Overexpression of MtRALFL1 and MtDVL1 in Medicago truncatula roots resulted in a marked reduction in the number of nodules formed and in a strong increase in the number of aborted infection threads. In addition, abnormal nodule development was observed when MtRALFL1 was overexpressed. This work provides evidence for the involvement of new putative small-peptide regulators during nodulation.


1996 ◽  
Vol 42 (2) ◽  
pp. 187-190 ◽  
Author(s):  
Kodjo Tomekpe ◽  
Marcelle Holsters ◽  
Bernard Dreyfus

Azorhizobium caulinodans ORS571 and Sinorhizobium teranga ORS51 and ORS52 are symbionts of the same host plant Sesbania rostrata. In nature, A. caulinodans nodulates more competitively the stem-located infection sites of Sesbania rostrata. Sinorhizobium strains, although frequently present in root nodules, are seldom found in stem nodules. One probable explanation for this phenomenon is the more abundant presence of Azorhizobium on the leaf and stem surfaces of the host plant. Work presented here hints at other plausible factors that determine the greater "stem specificity" of Azorhizobium. We found that under experimental conditions in which roots are not inoculated, all strains nodulated stems very well. However, ORS51 and ORS52 were much more sensitive than ORS571 to suppression of stem nodulation by previous root inoculation. The introduction of the regulatory nodD gene from A. caulinodans diminished the sensitivity to this suppression, probably by enhanced nod gene expression and subsequent Nod factor production. Our hypothesis is that the greater infectivity of ORS571 is due to a more efficient production of mitogenic Nod factors at stem-located infection sites, thereby more readily overcoming systemic suppression caused by previous root inoculations.Key words: autoregulation, nitrogen fixation, rhizobial ecology, systemic suppression of nodulation.


2010 ◽  
Vol 23 (1) ◽  
pp. 58-66 ◽  
Author(s):  
Anita S. Bek ◽  
Jørgen Sauer ◽  
Mikkel B. Thygesen ◽  
Jens Ø. Duus ◽  
Bent O. Petersen ◽  
...  

Formation of functional nodules is a complex process depending on host–microsymbiont compatibility in all developmental stages. This report uses the contrasting symbiotic phenotypes of Lotus japonicus and L. pedunculatus, inoculated with Mesorhizobium loti or the Bradyrhizobium sp. (Lotus), to investigate the role of Nod factor structure and Nod factor receptors (NFR) for rhizobial recognition, infection thread progression, and bacterial persistence within nodule cells. A key contribution was the use of 800 MHz nuclear magnetic resonance spectroscopy and ultrahigh-performance liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry for Nod factor analysis. The Nod factor decorations at the nonreducing end differ between Bradyrhizobium sp. (Lotus) and M. loti, and the NFR1/NFR5 extracellular regions of L. pedunculatus and L. japonicus were found to vary in amino acid composition. Genetic transformation experiments using chimeric and wild-type receptors showed that both receptor variants recognize the structurally different Nod factors but the later symbiotic phenotype remained unchanged. These results highlight the importance of additional checkpoints during nitrogen-fixing symbiosis and define several amino acids in the LysM domains as expendable for perception of the two differentially carbamoylated Nod factors.


2006 ◽  
Vol 19 (8) ◽  
pp. 914-923 ◽  
Author(s):  
Hiroki Miwa ◽  
Jongho Sun ◽  
Giles E. D. Oldroyd ◽  
J. Allan Downie

Nodulation (Nod)-factor signaling molecules are essential for rhizobia to initiate the nitrogen-fixing symbiotic interaction with legumes. Using a dual dye ratiometric calcium imaging technique, we have shown that 10 nM Nod factor added to roots of Lotus japonicus seedlings induces an intra-cellular calcium increase (calcium flux) that precedes oscillations in intracellular calcium (calcium spiking). The calcium flux was not observed with 1 or 0.1 nM Nod factor, which did induce calcium spiking. The calcium flux was variable in timing of initiation and duration and was observed in approximately half of the root hairs examined. Representatives from 11 complementation groups of symbiotically defective mutants were analyzed for the calcium flux. Mutants from four groups (sym6, ccamk, sym35, and nin) which retained calcium spiking all showed a normal calcium flux. Two classes of mutants (nfr1 and nfr5) lacked both calcium influx and calcium spiking, whereas five classes of mutants (symRK, castor, pollux, nup133, and sym24) defective for calcium spiking retained a calcium flux. There was no correlation between calcium spiking and induction of root hair deformation by Nod factor. We propose that increased bacterial numbers within infection foci in root hairs leads to accumulation of Nod factor to sufficient levels to activate the calcium flux, and this may drive infection thread growth.


Author(s):  
Bikash Raul ◽  
Oindrila Bhattacharjee ◽  
Amit Ghosh ◽  
Priya Upadhyay ◽  
Kunal Tembhare ◽  
...  

Root nodule symbiosis (RNS) is the pillar behind sustainable agriculture and plays a pivotal role in the environmental nitrogen cycle. Most of the genetic, molecular, and cell-biological knowledge on RNS come from model legumes that exhibit a root-hair mode of bacterial infection in contrast to the Dalbergoid legumes exhibiting crack-entry of rhizobia. As a step towards understanding this important group of legumes, we have combined microscopic analysis and temporal transcriptome to obtain a dynamic view of plant gene expression during Arachis hypogaea (peanut) nodule development. We generated a comprehensive transcriptome data by mapping the reads to A. hypogaea, and two diploid progenitor genomes. Additionally, we performed BLAST searches to identify nodule-induced yet-to-be annotated peanut genes. Comparison between peanut, Medicago truncatula, Lotus japonicus, and Glycine max showed upregulation of 61 peanut orthologs among 111 tested known RNS-related genes, indicating conservation in mechanisms of nodule development among members of the Papilionoid family. Unlike model legumes, recruitment of class 1 phytoglobin derived symbiotic hemoglobin (SymH) in peanut indicates diversification of oxygen scavenging mechanisms in the Papilionoid family. Finally, absence of cysteine-rich motif-1 containing-NCRs, but the recruitment of defensin like NCRs suggest a diverse molecular mechanism of terminal bacteroid differentiation. In summary, our work describes genetic conservation and diversification in legume-rhizobial symbiosis in the Papilionoid family, as well as among members of the Dalbergoid legumes.


1999 ◽  
Vol 12 (9) ◽  
pp. 820-824 ◽  
Author(s):  
T. Laeremans ◽  
C. Snoeck ◽  
J. Mariën ◽  
C. Verreth ◽  
E. Martínez-Romero ◽  
...  

Phaseolus vulgaris is a promiscuous host plant that can be nodulated by many different rhizobia representing a wide spectrum of Nod factors. In this study, we introduced the Rhizobium tropici CFN299 Nod factor sulfation genes nodHPQ into Azorhizobium caulinodans. The A. caulinodans transconjugants produce Nod factors that are mostly if not all sulfated and often with an arabinosyl residue as the reducing end glycosylation. Using A. caulinodans mutant strains, affected in reducing end decorations, and their respective transconjugants in a bean nodulation assay, we demonstrated that bean nodule induction efficiency, in decreasing order, is modulated by the Nod factor reducing end decorations fucose, arabinose or sulfate, and hydrogen.


2002 ◽  
Vol 80 (9) ◽  
pp. 907-915 ◽  
Author(s):  
Walter F Giordano ◽  
Michelle R Lum ◽  
Ann M Hirsch

We have initiated studies on the molecular biology and genetics of white sweetclover (Melilotus alba Desr.) and its responses to inoculation with the nitrogen-fixing symbiont Sinorhizobium meliloti. Early nodulin genes such as ENOD40 serve as markers for the transition from root to nodule development even before visible stages of nodule formation are evident. Using Northern blot analysis, we found that the ENOD40 gene was expressed within 6 h after inoculation with two different strains of S. meliloti, one of which overproduces symbiotic Nod factors. Inoculation with this strain resulted in an additional increase in ENOD40 gene expression over a typical wild-type S. meliloti strain. Moreover, the increase in mRNA brought about by the Nod-factor-overproducing strain 24 h after inoculation was correlated with lateral root formation by using whole-mount in situ hybridization to localize ENOD40 transcripts in lateral root meristems and by counting lateral root initiation sites. Cortical cell divisions were not detected. We also found that nodulation occurred more rapidly on white sweetclover in response to the Nod-factor-overproducing strain, but ultimately there was no difference in nodulation efficiency in terms of nodule number or the number of roots nodulated by the two strains. Also, the two strains could effectively co-colonize the host when inoculated together, although a few host cells were occupied by both strains.Key words: ENOD40, Nod factor, Melilotus, Sinorhizobium, symbiosis.


2018 ◽  
Author(s):  
Daniela Tsikou ◽  
Estrella E. Ramirez ◽  
Ioanna S. Psarrakou ◽  
Jaslyn E. Wong ◽  
Dorthe B. Jensen ◽  
...  

SUMMARYPost-translational modification of receptor proteins is involved in activation and de-activation of signaling systems in plants. Both ubiquitination and deubiquitination have been implicated in plant interactions with pathogens and symbionts. Here we presentLjPUB13, a PUB-ARMADILLO repeat E3 ligase that specifically ubiquitinates the kinase domain of the Nod Factor receptor NFR5 and has a direct role in nodule organogenesis events inLotus japonicus. Phenotypic analyses of three LORE1 retroelement insertion plant lines revealed thatpub13plants display delayed and reduced nodulation capacity and retarded growth.LjPUB13expression is spatially regulated during symbiosis withMesorhizobium loti, with increased levels in young developing nodules. Thus,LjPUB13 is an E3 ligase with a positive regulatory role during the initial stages of nodulation inL. japonicus.


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