Comparison of the genomic arrangement and the relative transcription of the nitrogenase genes in Rhizobium meliloti during symbiotic development in alfalfa root nodules

1982 ◽  
Vol 28 (12) ◽  
pp. 1330-1339 ◽  
Author(s):  
Alan S. Paau ◽  
Winston J. Brill
Keyword(s):  
Rhizobium Meliloti ◽  
Protein Pattern ◽  
Root Nodules ◽  
Symbiotic Association ◽  
Free Living ◽  
Genomic Arrangement ◽  
Infection Threads ◽  
Nitrogenase Genes ◽  
Specific Amplification ◽  
Alfalfa Root

In the course of the symbiotic development of free-living rhizobia to mature bacteroids in alfalfa nodules, mRNA transcripts of the nitrogenase structural genes are first detected when the rhizobia are inside the infection threads. These transcripts are most abundant in the mature bacteroids of young (6 to 8 weeks) root nodules and are present in a much lower level in bacteroids from senescent root nodules (13 weeks). Translation of these and related rhizobial genes essential for the symbiotic association apparently occurs very early in the symbiosis because the protein pattern of the "infection thread rhizobia" is very similar to that of the mature bacteroids and is different from that of the cultured vegetative rhizobia. Although bacteroids have a higher DNA content than the vegetative bacteria and are very different in the nucleoid organization, no specific amplification or rearrangement of the nitrogenase genes is detected in bacteroids.

scholarly journals The Rhizobium etli amtB Gene Coding for an NH4+ Transporter Is Down-Regulated Early During Bacteroid Differentiation

1998 ◽  
Vol 11 (3) ◽  
pp. 188-198 ◽  
Author(s):  
Rosarita Taté ◽  
Anna Riccio ◽  
Mike Merrick ◽  
Eduardo J. Patriarca
Keyword(s):  
Root Nodules ◽  
Ammonium Transport ◽  
Free Living ◽  
Gene Coding ◽  
Transcriptional Regulatory Mechanism ◽  
Transcriptional Regulatory ◽  
Infection Threads ◽  
Nitrogenase Genes ◽  
Uptake Activity ◽  
Free Living Conditions

During development of root nodules, Rhizobium bacteria differentiate inside the invaded plant cells into N2-fixing bacteroids. Terminally differentiated bacteroids are unable to grow using the ammonia (NH3 ) produced therein by the nitrogenase complex. Therefore, the nitrogen assimilation activities of bacteroids, including the ammonium (NH4 +) uptake activity, are expected to be repressed during symbiosis. By sequence homology the R. etli amtB (ammonium transport) gene was cloned and sequenced. As previously shown for its counterpart in other organisms, the R. etli amtB gene product mediates the transport of NH4 +. The amtB gene is cotranscribed with the glnK gene (coding for a PII-like protein) from a nitrogen-regulated σ54-dependent promoter, which requires the transcriptional activator NtrC. Expression of the glnKamtB operon was found to be activated under nitrogen-limiting, free-living conditions, but down-regulated just when bacteria are released from the infection threads and before transcription of the nitrogenase genes. Our data suggest that the uncoupling between N2-fixation and NH3 assimilation observed in symbiosomes is generated by a transcriptional regulatory mechanism(s) beginning with the inactivation of NtrC in younger bacteroids.

scholarly journals α-Galactoside Uptake in Rhizobium meliloti: Isolation and Characterization of agpA, a Gene Encoding a Periplasmic Binding Protein Required for Melibiose and Raffinose Utilization

1998 ◽  
Vol 180 (21) ◽  
pp. 5739-5748 ◽  
Author(s):  
Daniel J. Gage ◽  
Sharon R. Long
Keyword(s):  
Rhizobium Meliloti ◽  
Binding Protein ◽  
Root Nodules ◽  
Regulatory Protein ◽  
Ecological Niches ◽  
Periplasmic Binding Protein ◽  
Free Living ◽  
Gene Encoding ◽  
Intracellular Symbiont ◽  
Isolation And Characterization

ABSTRACT Rhizobium meliloti can occupy at least two distinct ecological niches; it is found in the soil as a free-living saprophyte, and it also lives as a nitrogen-fixing intracellular symbiont in root nodules of alfalfa and related legumes. One approach to understanding how R. meliloti alters its physiology in order to become an integral part of a developing nodule is to identify and characterize genes that are differentially expressed by bacteria living inside nodules. We used a screen to identify genes under the control of theR. meliloti regulatory protein NodD3, SyrM, or SyrA. These regulatory proteins are expressed by bacteria growing inside the root nodule. One gene isolated in this screen was mapped to pSymB and displayed complex regulation. The gene was downregulated by thesyrA gene product and also by glucose and succinate. This gene, referred to as agpA, encodes a periplasmic binding protein that is most similar to proteins from the periplasmic oligopeptide binding protein family. It is likely that AgpA binds α-galactosides, because α-galactosides induce the expression ofagpA, and agpA mutants cannot utilize or transport these sugars. Activity of anagpA::TnphoA fusion was downregulated by SyrA. Because syrA is known to be expressed at high levels in intracellular symbiotic R. meliloti and at low levels in the free-living bacteria, we propose that AgpA may belong to the class of gene products whose expression decreases when R. meliloti becomes an intracellular symbiont.

Formation and structure of root nodules induced on Macroptilium atropurpureum inoculated with various species of Rhizobium

1991 ◽  
Vol 69 (7) ◽  
pp. 1520-1532 ◽  
Author(s):  
Michael J. Trinick ◽  
Celia Miller ◽  
Paul A. Hadobas
Keyword(s):  
Rhizobium Leguminosarum ◽  
Rhizobium Meliloti ◽  
Root Nodules ◽  
Matrix Material ◽  
Immunogold Labelling ◽  
Infection Threads ◽  
Glass Tubes ◽  
Infected Cells ◽  
Macroptilium Atropurpureum ◽  
Nodule Tissue

Fifteen strains of Rhizobium leguminosarum biovar trifolii formed ineffective nodules and (or) nodule-like structures (rhizobia were re-isolated from both structures) on Macroptilium atropurpureum grown in enclosed glass tubes. Bacteria were observed among the parenchyma cells surrounding the nodule-like structures. One variant of R. leguminosarum biovar trifolii (NGR66/ST) isolated from M. atropurpureum formed nodules on this host that exhibited abnormal intercellular and intracellular infection. The bacteria (NGR66/ST) were contained within threadlike structures, surrounded by matrix material. The identities of the Rhizobium strains were confirmed serologically after reisolation and in sections of nodule tissue using immunogold labelling. Rhizobium leguminosarum biovar phaseoli strain NGR76 isolated from Phaseolus vulgaris formed nodules on M. atropurpureum resembling those formed by effective Bradyrhizobium strains. The association was partially effective in nitrogen fixation, and this was reflected in the nodule structure. The percentage of cells infected was lower than that in fully effective nodules. There was a high frequency of infected cells showing degeneration; these were located throughout the nodule tissue and were often adjacent to healthy infected cells. The rhizobia appeared to infect new nodule cells via infection threads, which were abundant both intercellularly and intracellularly in young, mature, and degenerating host nodule cells. Strains of R. leguminosarum biovar viceae and Rhizobium meliloti were unable to induce nodule-like structures on M. atropurpureum. Key words: Macroptilium, Bradyrhizobium, Rhizobium, microscopy, nodule, structure.

Effect of Low Nitrate Supply on Nitrogen Fixation in Alfalfa Root Nodules Induced by Rhizobium meliloti Strains with Varied Nitrate Reductase Activity

1989 ◽  
Vol 135 (2) ◽  
pp. 207-211 ◽  
Author(s):  
Cesáreo Arrese-Igor ◽  
José M. Estavillo ◽  
José I. Peña ◽  
Carmen Gonzalez-Murua ◽  
Pedro M. Aparicio-Tejo
Keyword(s):  
Nitrogen Fixation ◽  
Nitrate Reductase ◽  
Nitrate Reductase Activity ◽  
Rhizobium Meliloti ◽  
Reductase Activity ◽  
Root Nodules ◽  
Nitrate Supply ◽  
Alfalfa Root

scholarly journals Morphology of root nodules and nodule-like structures formed by Rhizobium and Agrobacterium strains containing a Rhizobium meliloti megaplasmid.

1983 ◽  
Vol 97 (3) ◽  
pp. 787-794 ◽  
Author(s):  
C H Wong ◽  
C E Pankhurst ◽  
A Kondorosi ◽  
W J Broughton
Keyword(s):  
Normal Development ◽  
Rhizobium Meliloti ◽  
Root Nodules ◽  
Root Hairs ◽  
Host Cells ◽  
Donor Strain ◽  
Intercellular Spaces ◽  
Infection Threads ◽  
Infected Cells ◽  
Rhizobium Sp

We examined expression of the megaplasmid pRme41b of Rhizobium meliloti in two different Rhizobium sp. Strains and in Agrobacterium tumefaciens. Transfer of pRme41b into these bacteria was facilitated by insertion of a recombinant plasmid coding for mobilization functions of RP4 into the nif region (Kondorosi, A., E. Kondorosi, C.E. Pankhurst, W. J. Broughton, and Z. Banfalvi, 1982, Mol. Gen. Genet., 188:433-439). In all cases, transconjugants formed nodule-like structures on the roots of Medicago sativa. These structures were largely composed of meristematic cells but they were not invaded by bacteria. Bacteria were found only within infection threads in root hairs, and within intercellular spaces of the outermost cells of the structures. The donor strain of R. meliloti containing pAK11 or pAK12 in pRme41b initially produced nodules on M. sativa that did not fix nitrogen (Fix-). In these nodules, bacteria were released from infection threads into the host cells but they did not multiply appreciably. Any bacteroids formed degenerated prematurely. In some cases, however, reversion to a Fix+ phenotype occurred after 4 to 6 wk. Bacteria released into newly infected cells in these nodules showed normal development into bacteriods.

scholarly journals Paraburkholderia phymatum Homocitrate Synthase NifV Plays a Key Role for Nitrogenase Activity during Symbiosis with Papilionoids and in Free-Living Growth Conditions

Cells ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 952
Author(s):  
Paula Bellés-Sancho ◽  
Martina Lardi ◽  
Yilei Liu ◽  
Sebastian Hug ◽  
Marta Adriana Pinto-Carbó ◽  
...  
Keyword(s):  
Nitrogenase Activity ◽  
Root Nodules ◽  
Growth Conditions ◽  
Lysine Biosynthesis ◽  
Metabolome Analysis ◽  
Plant Host ◽  
Free Living ◽  
Bacterial Enzyme ◽  
Homocitrate Synthase ◽  
Iron Molybdenum Cofactor

Homocitrate is an essential component of the iron-molybdenum cofactor of nitrogenase, the bacterial enzyme that catalyzes the reduction of dinitrogen (N2) to ammonia. In nitrogen-fixing and nodulating alpha-rhizobia, homocitrate is usually provided to bacteroids in root nodules by their plant host. In contrast, non-nodulating free-living diazotrophs encode the homocitrate synthase (NifV) and reduce N2 in nitrogen-limiting free-living conditions. Paraburkholderia phymatum STM815 is a beta-rhizobial strain, which can enter symbiosis with a broad range of legumes, including papilionoids and mimosoids. In contrast to most alpha-rhizobia, which lack nifV, P. phymatum harbors a copy of nifV on its symbiotic plasmid. We show here that P. phymatum nifV is essential for nitrogenase activity both in root nodules of papilionoid plants and in free-living growth conditions. Notably, nifV was dispensable in nodules of Mimosa pudica despite the fact that the gene was highly expressed during symbiosis with all tested papilionoid and mimosoid plants. A metabolome analysis of papilionoid and mimosoid root nodules infected with the P. phymatum wild-type strain revealed that among the approximately 400 measured metabolites, homocitrate and other metabolites involved in lysine biosynthesis and degradation have accumulated in all plant nodules compared to uninfected roots, suggesting an important role of these metabolites during symbiosis.

Expression of the nodulation and nitrogen fixation genes in Rhizobium meliloti during development

Genome ◽  
1989 ◽  
Vol 31 (1) ◽  
pp. 354-360 ◽  
Author(s):  
San Chiun Shen ◽  
Shui Ping Wang ◽  
Guan Qiao Yu ◽  
Jia Bi Zhu
Keyword(s):  
Nitrogen Fixation ◽  
Rhizobium Meliloti ◽  
Abnormal Development ◽  
Wild Type ◽  
Free Living ◽  
Nod Genes ◽  
Nodule Initiation ◽  
Fix Genes ◽  
Nitrogen Fixation Genes

Genes that specify nodulation (nod genes) are only active in the free-living rhizobia or in the nodule initiation state of rhizobia. As soon as the repression of nod genes occurs in the bacteroids of the nodule, nifA is induced, while ntrC is inactivated and thus the nifA-mediated nif/fix genes are turned on. Limitation of available oxygen brings about the induction of nifA, which reflects the actual status of nif/fix gene activities in symbiotic state of rhizobia. Oxygen thus appears to be a major symbiotic signal to the expression of bacteroid nif/fix genes. Mutation of nifA or shortage of nifA product in wild-type rhizobia caused by the inhibition of multicopy nifH/fixA promoters leads to an abnormal development of nodules and premature degradation of bacteroids in nodules.Key words: nitrogen fixation, nodulation, nif/fix regulation, nifA mutant.

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