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1.
Annu Rev Microbiol ; 76: 45-65, 2022 09 08.
Article in English | MEDLINE | ID: mdl-35395168

ABSTRACT

To suppress plant immunity and promote the intracellular infection required for fixing nitrogen for the benefit of their legume hosts, many rhizobia use type III secretion systems (T3SSs) that deliver effector proteins (T3Es) inside host cells. As reported for interactions between pathogens and host plants, the immune system of legume hosts and the cocktail of T3Es secreted by rhizobia determine the symbiotic outcome. If they remain undetected, T3Es may reduce plant immunity and thus promote infection of legumes by rhizobia. If one or more of the secreted T3Es are recognized by the cognate plant receptors, defense responses are triggered and rhizobial infection may abort. However, some rhizobial T3Es can also circumvent the need for nodulation (Nod) factors to trigger nodule formation. Here we review the multifaceted roles played by rhizobial T3Es during symbiotic interactions with legumes.


Subject(s)
Fabaceae , Rhizobium , Fabaceae/metabolism , Plant Immunity , Rhizobium/metabolism , Symbiosis/physiology , Type III Secretion Systems/metabolism
2.
Genes (Basel) ; 11(5)2020 05 07.
Article in English | MEDLINE | ID: mdl-32392829

ABSTRACT

On legume crops, formation of developmentally mature nodules is a prerequisite to efficient nitrogen fixation by populations of rhizobial bacteroids established inside nodule cells. Development of root nodules and concomitant microbial colonisation of plant cells are constrained by sets of recognition signals exchanged by infecting rhizobia and their legume hosts, with much of the specificity of symbiotic interactions being determined by the flavonoid cocktails released by legume roots and the strain-specific nodulation factors (NFs) secreted by rhizobia. Hence, much of Sinorhizobium fredii strain NGR234 symbiotic promiscuity was thought to stem from a family of >80 structurally diverse NFs and associated nodulation keys in the form of secreted effector proteins and rhamnose-rich surface polysaccharides. Here, we show instead that a mini-symbiotic plasmid (pMiniSym2) carrying only the nodABCIJ, nodS and nodD1 genes of NGR234 conferred promiscuous nodulation to ANU265, a derivative strain cured of the large symbiotic plasmid pNGR234a. The ANU265::pMiniSym2 transconjugant triggered nodulation responses on 12 of the 22 legumes we tested. On roots of Macroptilium atropurpureum, Leucaena leucocephala and Vigna unguiculata, ANU265::pMiniSym2 formed mature-like nodule and successfully infected nodule cells. While cowpea and siratro responded to nodule colonisation with defence responses that eventually eliminated bacteria, L. leucocephala formed leghemoglobin-containing mature-like nodules inside which the pMiniSym2 transconjugant established persistent intracellular colonies. This data shows seven nodulation genes of NGR234 suffice to trigger nodule formation on roots of many hosts and to establish chronic infections in Leucaena cells.


Subject(s)
Fabaceae/microbiology , Genes, Bacterial , Nitrogen Fixation/genetics , Plant Root Nodulation/genetics , Rhizobium/genetics , Root Nodules, Plant/microbiology , Symbiosis/genetics , Conjugation, Genetic , Fabaceae/immunology , Gene Dosage , Host Specificity , Plant Roots/metabolism , Plant Roots/microbiology , Plasmids/genetics , Replicon , Rhizobium/metabolism , Species Specificity
3.
Int J Syst Evol Microbiol ; 70(2): 1421-1430, 2020 Feb.
Article in English | MEDLINE | ID: mdl-32122457

ABSTRACT

For many smallholder farmers of Sub-Saharan Africa, pigeonpea (Cajanus cajan) is an important crop to make ends meet. To ascertain the taxonomic status of pigeonpea isolates of Côte d'Ivoire previously identified as bradyrhizobia, a polyphasic approach was applied to strains CI-1BT, CI-14A, CI-19D and CI-41S. Phylogeny of 16S ribosomal RNA (rRNA) genes placed these nodule isolates in a separate lineage from current species of the B. elkanii super clade. In phylogenetic analyses of single and concatenated partial dnaK, glnII, gyrB, recA and rpoB sequences, the C. cajan isolates again formed a separate lineage, with strain CI-1BT sharing the highest sequence similarity (95.2 %) with B. tropiciagri SEMIA 6148T. Comparative genomic analyses corroborated the novel species status, with 86 % ANIb and 89 % ANIm as the highest average nucleotide identity (ANI) values with B. elkanii USDA 76T. Although CI-1BT, CI-14A, CI-19D and CI-41S shared similar phenotypic and metabolic properties, growth of CI-41S was slower in/on various media. Symbiotic efficacy varied significantly between isolates, with CI-1BT and CI-41S scoring on the C. cajan 'Light-Brown' landrace as the most and least proficient bacteria, respectively. Also proficient on Vigna radiata (mung bean), Vigna unguiculata (cowpea, niébé) and additional C. cajan cultivars, CI-1BT represents a potential bioinoculant adapted to local soil conditions and capable of fostering the growth of diverse legume crops in Côte d'Ivoire. Given the data presented here, we propose the 19 C. cajan isolates to belong to a novel species called Bradyrhizobium ivorense sp. nov., with CI-1BT (=CCOS 1862T=CCMM B1296T) as a type strain.


Subject(s)
Bradyrhizobium/classification , Cajanus/microbiology , Phylogeny , Root Nodules, Plant/microbiology , Bacterial Typing Techniques , Base Composition , Bradyrhizobium/isolation & purification , Cote d'Ivoire , DNA, Bacterial/genetics , Fabaceae/microbiology , Genes, Bacterial , Nitrogen Fixation , Nucleic Acid Hybridization , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Symbiosis
4.
Front Microbiol ; 10: 154, 2019.
Article in English | MEDLINE | ID: mdl-30814981

ABSTRACT

During their lifecycle, from free-living soil bacteria to endosymbiotic nitrogen-fixing bacteroids of legumes, rhizobia must colonize, and cope with environments where nutrient concentrations and compositions vary greatly. Bacterial colonization of legume rhizospheres and of root surfaces is subject to a fierce competition for plant exudates. By contrast root nodules offer to rhizobia sheltered nutrient-rich environments within which the cells that successfully propagated via infection threads can rapidly multiply. To explore the effects on symbiosis of a slower rhizobia growth and metabolism, we deleted one or two copies of the three functional rRNA operons of the promiscuous Sinorhizobium fredii strain NGR234 and examined the impact of these mutations on free-living and symbiotic lifestyles. Strains with two functional rRNA operons (NGRΔrRNA1 and NGRΔrRNA3) grew almost as rapidly as NGR234, and NGRΔrRNA1 was as proficient as the parent strain on all of the five legume species tested. By contrast, the NGRΔrRNA1,3 double mutant, which carried a single rRNA operon and grew significantly slower than NGR234, had a reduced symbiotic proficiency on Cajanus cajan, Macroptilium atropurpureum, Tephrosia vogelii, and Vigna unguiculata. In addition, while NGRΔrRNA1 and NGR234 equally competed for nodulation of V. unguiculata, strain NGRΔrRNA1,3 was clearly outcompeted by wild-type. Surprisingly, on Leucaena leucocephala, NGRΔrRNA1,3 was the most proficient strain and competed equally NGR234 for nodule occupation. Together, these results indicate that for strains with otherwise identical repertoires of symbiotic genes, a faster growth on roots and/or inside plant tissues may contribute to secure access to nodules of some hosts. By contrast, other legumes such as L. leucocephala appear as less selective and capable of providing symbiotic environments susceptible to accommodate strains with a broader spectrum of competences.

5.
Mol Plant Microbe Interact ; 32(2): 208-216, 2019 Feb.
Article in English | MEDLINE | ID: mdl-30070615

ABSTRACT

Symbiotic nitrogen fixation between legumes and rhizobia involves a coordinated expression of many plant and bacterial genes as well as finely tuned metabolic activities of micro- and macrosymbionts. In spite of such complex interactions, symbiotic proficiency remains a resilient process, with host plants apparently capable of compensating for some deficiencies in rhizobia. What controls nodule homeostasis is still poorly understood and probably varies between plant species. In this respect, the promiscuous Sinorhizobium (Ensifer) fredii strain NGR234 has become a model to assess the relative contribution of single gene products to many symbioses. Here, we describe how a deletion in nifQ of NGR234 (strain NGRΔnifQ) makes nodules of Vigna unguiculata, V. radiata, and Macroptilium atropurpureum but not of the mimisoid tree Leucaena leucocephala, purple-red. This peculiar dark-nodule phenotype did not necessarily correlate with a decreased proficiency of NGRΔnifQ but coincided with a 20-fold or more accumulation of coproporphyrin III and uroporphyrin III in V. unguiculata nodules. Porphyrin accumulation was not restricted to plant cells infected with bacteroids but also extended to the nodule cortex. Nodule metal-homeostasis was altered but not sufficiently to prevent assembly and functioning of nitrogenase. Although the role of NifQ in donating molybdenum during assembly of nitrogenase cofactor FeMo-co makes it essential in free-living diazotrophs, our results highlight the dispensability of NifQ in many legume species.


Subject(s)
Bacterial Proteins , Metals , Nitrogen , Porphyrins , Symbiosis , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Homeostasis , Metals/metabolism , Nitrogen/metabolism , Nitrogen Fixation , Porphyrins/metabolism
6.
Front Microbiol ; 9: 2843, 2018.
Article in English | MEDLINE | ID: mdl-30519234

ABSTRACT

Sinorhizobium fredii indigenous populations are prevalent in provinces of Central China whereas Bradyrhizobium species (Bradyrhizobium japonicum, B. diazoefficiens, B. elkanii, and others) are more abundant in northern and southern provinces. The symbiotic properties of different soybean rhizobia have been investigated with 40 different wild soybean (Glycine soja) accessions from China, Japan, Russia, and South Korea. Bradyrhizobial strains nodulated all the wild soybeans tested, albeit efficiency of nitrogen fixation varied considerably among accessions. The symbiotic capacity of S. fredii HH103 with wild soybeans from Central China was clearly better than with the accessions found elsewhere. S. fredii NGR234, the rhizobial strain showing the broadest host range ever described, also formed nitrogen-fixing nodules with different G. soja accessions from Central China. To our knowledge, this is the first report describing an effective symbiosis between S. fredii NGR234 and G. soja. Mobilization of the S. fredii HH103 symbiotic plasmid to a NGR234 pSym-cured derivative (strain NGR234C) yielded transconjugants that formed ineffective nodules with G. max cv. Williams 82 and G. soja accession CH4. By contrast, transfer of the symbiotic plasmid pNGR234a to a pSym-cured derivative of S. fredii USDA193 generated transconjugants that effectively nodulated G. soja accession CH4 but failed to nodulate with G. max cv. Williams 82. These results indicate that intra-specific transference of the S. fredii symbiotic plasmids generates new strains with unpredictable symbiotic properties, probably due to the occurrence of new combinations of symbiotic signals.

7.
Front Microbiol ; 7: 1793, 2016.
Article in English | MEDLINE | ID: mdl-27891120

ABSTRACT

In smallholder farms of Côte d'Ivoire, particularly in the northeast of the country, Cajanus cajan (pigeonpea) has become an important crop because of its multiple beneficial facets. Pigeonpea seeds provide food to make ends meet, are sold on local markets, and aerial parts serve as forage for animals. Since it fixes atmospheric nitrogen in symbiosis with soil bacteria collectively known as rhizobia, C. cajan also improves soil fertility and reduces fallow time. Yet, seed yields remain low mostly because farmers cannot afford chemical fertilizers. To identify local rhizobial strains susceptible to be used as bio-inoculants to foster pigeonpea growth, root nodules were collected in six fields of three geographically distant regions of Côte d'Ivoire. Nodule bacteria were isolated and characterized using various molecular techniques including matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS) and DNA sequencing. These molecular analyses showed that 63 out of 85 nodule isolates belonged to two major clades of bradyrhizobia, one of which is known as the Bradyrhizobium elkanii super clade. Phylogenies of housekeeping (16S-ITS-23S, rpoB) and symbiotic (nifH) genes were not always congruent suggesting that lateral transfer of nitrogen fixation genes also contributed to define the genome of these bradyrhizobial isolates. Interestingly, no field-, plant-, or cultivar-specific effect was found to shape the profiles of symbiotic strains. In addition, nodule isolates CI-1B, CI-36E, and CI-41A that belong to distinct species, showed similar symbiotic efficiencies suggesting that any of these strains might serve as a proficient inoculant for C. cajan.

8.
Food Microbiol ; 55: 112-22, 2016 May.
Article in English | MEDLINE | ID: mdl-26742622

ABSTRACT

Acetic acid bacteria (AAB) are widespread microorganisms in nature, extensively used in food industry to transform alcohols and sugar alcohols into their corresponding organic acids. Specialized strains are used in the production of vinegar through the oxidative transformation of ethanol into acetic acid. The main AAB involved in the production of high-acid vinegars using the submerged fermentation method belong to the genus Komagataeibacter, characterized by their higher ADH stability and activity, and higher acetic acid resistance (15-20%), compared to other AAB. In this work, the bacteria involved in the production of high-acid spirit vinegar through a spontaneous acetic acid fermentation process was studied. The analysis using a culture-independent approach revealed a homogeneous bacterial population involved in the process, identified as Komagataeibacter spp. Differentially expressed proteins during acetic acid fermentation were investigated by using 2D-DIGE and mass spectrometry. Most of these proteins were functionally related to stress response, the TCA cycle and different metabolic processes. In addition, scanning and transmission electron microscopy and specific staining of polysaccharide SDS-PAGE gels confirmed that Komagataeibacter spp. lacked the characteristic polysaccharide layer surrounding the outer membrane that has been previously reported to have an important role in acetic acid resistance in the genus Acetobacter.


Subject(s)
Acetic Acid/metabolism , Alcoholic Beverages/microbiology , Alphaproteobacteria/metabolism , Bacterial Proteins/chemistry , Alphaproteobacteria/classification , Alphaproteobacteria/genetics , Alphaproteobacteria/ultrastructure , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Electrophoresis, Polyacrylamide Gel , Fermentation , Molecular Sequence Data , Phylogeny , Proteomics
9.
Appl Microbiol Biotechnol ; 99(13): 5547-62, 2015 Jul.
Article in English | MEDLINE | ID: mdl-25776061

ABSTRACT

Accurate identification of soil bacteria that form nitrogen-fixing associations with legume crops is challenging given the phylogenetic diversity of root nodule bacteria (RNB). The labor-intensive and time-consuming 16S ribosomal RNA (rRNA) sequencing and/or multilocus sequence analysis (MLSA) of conserved genes so far remain the favored molecular tools to characterize symbiotic bacteria. With the development of mass spectrometry (MS) as an alternative method to rapidly identify bacterial isolates, we recently showed that matrix-assisted laser desorption ionization (MALDI) time-of-flight (TOF) can accurately characterize RNB found inside plant nodules or grown in cultures. Here, we report on the development of a MALDI-TOF RNB-specific spectral database built on whole cell MS fingerprints of 116 strains representing the major rhizobial genera. In addition to this RNB-specific module, which was successfully tested on unknown field isolates, a subset of 13 ribosomal proteins extracted from genome data was found to be sufficient for the reliable identification of nodule isolates to rhizobial species as shown in the putatively ascribed ribosomal protein masses (PARPM) database. These results reveal that data gathered from genome sequences can be used to expand spectral libraries to aid the accurate identification of bacterial species by MALDI-TOF MS.


Subject(s)
Bacteria/chemistry , Bacteria/classification , Biodiversity , Ribosomal Proteins/analysis , Root Nodules, Plant/microbiology , Soil Microbiology , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Bacteria/genetics , Bacterial Proteins/genetics , Biomarkers/analysis , Cluster Analysis , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Molecular Sequence Data , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
10.
FEMS Microbiol Ecol ; 84(2): 421-32, 2013 May.
Article in English | MEDLINE | ID: mdl-23330958

ABSTRACT

Lake Cadagno is characterized by a compact chemocline that harbors high concentrations of various phototrophic sulfur bacteria. Four strains representing the numerically most abundant populations in the chemocline were tested in dialysis bags in situ for their ability to fix CO2. The purple sulfur bacterium Candidatus 'Thiodictyon syntrophicum' strain Cad16(T) had the highest CO2 assimilation rate in the light of the four strains tested and had a high CO2 assimilation rate even in the dark. The CO2 assimilation of the population represented by strain Cad16(T) was estimated to be up to 25% of the total primary production in the chemocline. Pure cultures of strain Cad16(T) exposed to cycles of 12 h of light and 12 h of darkness exhibited the highest CO2 assimilation during the first 4 h of light. The draft genome sequence of Cad16(T) showed the presence of cbbL and cbbM genes, which encode form I and form II of RuBisCO, respectively. Transcription analyses confirmed that, whereas cbbM remained poorly expressed throughout light and dark exposure, cbbL expression varied during the light-dark cycle and was affected by the available carbon sources. Interestingly, the peaks in cbbL expression did not correlate with the peaks in CO2 assimilation.


Subject(s)
Carbon Cycle , Carbon Dioxide/metabolism , Chromatiaceae/metabolism , Lakes/microbiology , Chromatiaceae/enzymology , Chromatiaceae/genetics , Lakes/chemistry , Photoperiod , Phototrophic Processes , Ribulose-Bisphosphate Carboxylase/genetics
11.
Appl Environ Microbiol ; 78(20): 7476-9, 2012 Oct.
Article in English | MEDLINE | ID: mdl-22865066

ABSTRACT

Cupriavidus taiwanensis forms proficient symbioses with a few Mimosa species. Inactivation of a type III protein secretion system (T3SS) had no effect on Mimosa pudica but allowed C. taiwanensis to establish chronic infections and fix nitrogen in Leucaena leucocephala. Unlike what was observed for other rhizobia, glutamate rather than plant flavonoids mediated transcriptional activation of this atypical T3SS.


Subject(s)
Bacterial Secretion Systems , Cupriavidus/physiology , Fabaceae/microbiology , Symbiosis , Cupriavidus/metabolism , Gene Knockout Techniques , Glutamates/metabolism , Proteins , Transcription, Genetic , Transcriptional Activation
12.
PLoS One ; 7(5): e37189, 2012.
Article in English | MEDLINE | ID: mdl-22615938

ABSTRACT

Rhizobia form a disparate collection of soil bacteria capable of reducing atmospheric nitrogen in symbiosis with legumes. The study of rhizobial populations in nature involves the collection of large numbers of nodules found on roots or stems of legumes, and the subsequent typing of nodule bacteria. To avoid the time-consuming steps of isolating and cultivating nodule bacteria prior to genotyping, a protocol of strain identification based on the comparison of MALDI-TOF MS spectra was established. In this procedure, plant nodules were considered as natural bioreactors that amplify clonal populations of nitrogen-fixing bacteroids. Following a simple isolation procedure, bacteroids were fingerprinted by analysing biomarker cellular proteins of 3 to 13 kDa using Matrix Assisted Laser Desorption/Ionization Time of Flight (MALDI-TOF) mass spectrometry. In total, bacteroids of more than 1,200 nodules collected from roots of three legumes of the Phaseoleae tribe (cowpea, soybean or siratro) were examined. Plants were inoculated with pure cultures of a slow-growing Bradyrhizobium japonicum strain G49, or either of two closely related and fast-growing Sinorhizobium fredii strains NGR234 and USDA257, or with mixed inoculants. In the fully automatic mode, correct identification of bacteroids was obtained for >97% of the nodules, and reached 100% with a minimal manual input in processing of spectra. These results showed that MALDI-TOF MS is a powerful tool for the identification of intracellular bacteria taken directly from plant tissues.


Subject(s)
Bacteria/isolation & purification , Root Nodules, Plant/microbiology , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Bacteriological Techniques/methods , Fabaceae/microbiology , Rhizome/microbiology , Sinorhizobium fredii/isolation & purification
13.
Syst Appl Microbiol ; 35(3): 139-44, 2012 May.
Article in English | MEDLINE | ID: mdl-22386960

ABSTRACT

Strain Cad16(T) is a small-celled purple sulfur bacterium (PSB) isolated from the chemocline of crenogenic meromictic Lake Cadagno, Switzerland. Long term in situ observations showed that Cad16(T) regularly grows in very compact clumps of cells in association with bacteria belonging to the genus Desulfocapsa in a cell-to-cell three dimensional structure. Previously assigned to the genus Lamprocystis, Cad16(T), was here reclassified and assigned to the genus Thiodictyon. Based on comparative 16S rRNA gene sequences analysis, isolate Cad16(T) was closely related to Thiodictyon bacillosum DSM234(T) and Thiodictyon elegans DSM232(T) with sequence similarities of 99.2% and 98.9%, respectively. Moreover, matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) analysis separated Cad16(T) from other PSB genera, Lamprocystis and Thiocystis. Major differences in cell morphology (oval-sphere compared to rod-shaped) and arrangement (no netlike cell aggregates), carotenoid group (presence of okenone instead of rhodopinal), chemolithotrophic growth as well as the ability to form syntrophic associations with a sulfate-reducing bacteria of the genus Desulfocapsa suggested a different species within the genus Thiodictyon. This isolate is therefore proposed and described as Candidatus "Thiodictyon syntrophicum" sp. nov., a provisionally novel species within the genus Thiodictyon.


Subject(s)
Bacterial Adhesion , Bacterial Physiological Phenomena , Chromatiaceae/classification , Chromatiaceae/isolation & purification , Deltaproteobacteria/physiology , Fresh Water/microbiology , Bacterial Typing Techniques , Chromatiaceae/chemistry , Chromatiaceae/physiology , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Microscopy , Molecular Sequence Data , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Switzerland
14.
J Proteomics ; 75(6): 1701-17, 2012 Mar 16.
Article in English | MEDLINE | ID: mdl-22155126

ABSTRACT

Acetic acid bacteria (AAB) are Gram-negative, strictly aerobic microorganisms that show a unique resistance to ethanol (EtOH) and acetic acid (AcH). Members of the Acetobacter and Gluconacetobacter genera are capable of transforming EtOH into AcH via the alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) enzymes and are used for the industrial production of vinegar. Several mechanisms have been proposed to explain how AAB resist high concentrations of AcH, such as the assimilation of acetate through the tricarboxylic acid (TCA) cycle, the export of acetate by various transporters and modifications of the outer membrane. However, except for a few acetate-specific proteins, little is known about the global proteome responses to AcH. In this study, we used 2D-DIGE to compare the proteome of Acetobacter pasteurianus LMG 1262(T) when growing in glucose or ethanol and in the presence of acetic acid. Interesting protein spots were selected using the ANOVA p-value of 0.05 as threshold and 1.5-fold as the minimal level of differential expression, and a total of 53 proteins were successfully identified. Additionally, the size of AAB was reduced by approximately 30% in length as a consequence of the acidity. A modification in the membrane polysaccharides was also revealed by PATAg specific staining.


Subject(s)
Acetic Acid/metabolism , Acetobacter/metabolism , Proteome/metabolism , Acetobacter/genetics , Acetobacter/ultrastructure , Culture Media/metabolism , Ethanol/metabolism , Fermentation , Glucose/metabolism , Microscopy, Electron, Scanning , Protein Biosynthesis , Protein Folding , Tandem Mass Spectrometry , Two-Dimensional Difference Gel Electrophoresis , Up-Regulation
15.
Microbiology (Reading) ; 157(Pt 10): 2745-2758, 2011 Oct.
Article in English | MEDLINE | ID: mdl-21719545

ABSTRACT

Rhizobia are a disparate collection of soil bacteria capable of reducing atmospheric nitrogen in symbiosis with legumes (Fix phenotype). Synthesis of the nitrogenase and its accessory components is under the transcriptional control of the key regulator NifA and is generally restricted to the endosymbiotic forms of rhizobia known as bacteroids. Amongst studied rhizobia, Sinorhizobium fredii strain NGR234 has the remarkable ability to fix nitrogen in association with more than 130 species in 73 legume genera that form either determinate, indeterminate or aeschynomenoid nodules. Hence, NGR234 is a model organism to study nitrogen fixation in association with a variety of legumes. The symbiotic plasmid pSfrNGR234a carries more than 50 genes that are under the transcriptional control of NifA. To facilitate the functional analysis of NifA-regulated genes a new transposable element, TnEKm-PwA, was constructed. This transposon combines the advantages of in vitro mutagenesis of cloned DNA fragments with a conditional read-out promoter from NGR234 (PwA) that reinitiates NifA-dependent transcription downstream of transposition sites. To test the characteristics of the new transposon, the nifQdctA1y4vGHIJ operon was mutated using either the Omega interposon or TnEKm-PwA. The symbiotic phenotypes on various hosts as well as the transcriptional characteristics of these mutants were analysed in detail and compared with the ineffective (Fix(-)) phenotype of strain NGRΔnifA, which lacks a functional copy of nifA. De novo transcription from inserted copies of TnEKm-PwA inside bacteroids was confirmed by qRT-PCR. Unexpectedly, polar mutants in dctA1 and nifQ were Fix(+) on all of the hosts tested, indicating that none of the six genes of the nifQ operon of NGR234 is essential for symbiotic nitrogen fixation on plants that form nodules of either determinate or indeterminate types.


Subject(s)
Bacterial Proteins/genetics , Dicarboxylic Acid Transporters/metabolism , Gene Expression Regulation, Bacterial , Operon , Promoter Regions, Genetic , Sinorhizobium fredii/genetics , Transcription Factors/metabolism , Bacterial Proteins/metabolism , Base Sequence , DNA Transposable Elements , Dicarboxylic Acid Transporters/genetics , Fabaceae/microbiology , Fabaceae/physiology , Molecular Sequence Data , Mutagenesis, Insertional , Sinorhizobium fredii/physiology , Symbiosis , Transcription Factors/genetics
16.
Arch Cardiovasc Dis ; 102(11): 743-8, 2009 Nov.
Article in English | MEDLINE | ID: mdl-19944389

ABSTRACT

BACKGROUND: Progress in techniques and equipment facilitates same-day-discharge percutaneous coronary intervention (PCI). AIM: To present initial experience of a same-day-discharge intention-to-treat ad hoc PCI strategy with a preferentially radial approach. METHODS: The first 102 same-day-discharge PCIs performed in our centre were analysed retrospectively. Subjects were stable or stabilised coronary patients, free of cardiac insufficiency, with a normal Allen test and residing within 60min of the centre. Discharge was authorized after six hours' event-free monitoring. The principal assessment criterion combined major adverse cardiovascular events, stroke, major haemorrhage and unscheduled medical consultation in the 30 days following PCI. Overall patient satisfaction and anxiety associated with same-day discharge were assessed by telephone questionnaire at some time after the intervention. RESULTS: Between January 2006 and March 2008, 95 day-hospital patients underwent 102 distinct PCIs (50 complex, 13 bifurcation lesions, nine intravascular ultrasound, 18 fractional flow reserve, two Rotablator procedures). Crossover to overnight admission was necessary for 5.9% (n=6) of interventions. Baseline clinical and angiographic characteristics were similar to those of an unselected Western population. At 30 days, four clinical events were observed (3.9% of interventions), which occurred within 6 hours (n=2) or after the 24th hour. Classical 24-hour admission would thus have provided no added benefit. On an analogue scale, overall satisfaction was high (8.9/10) and anxiety associated with same-day discharge was low (1.7/10). CONCLUSION: A same-day-discharge ad hoc PCI strategy proved reliable and safe for most patients selected a priori on simple clinical criteria, entailing no unexpected complications due to shorter medical monitoring.


Subject(s)
Angioplasty, Balloon, Coronary , Coronary Disease/therapy , Patient Discharge , Aged , Angioplasty, Balloon, Coronary/adverse effects , Angioplasty, Balloon, Coronary/methods , Angioplasty, Balloon, Coronary/psychology , Anxiety/etiology , Blood Coagulation Tests , Coronary Angiography , Coronary Disease/diagnostic imaging , Electrocardiography , Female , France , Humans , Length of Stay , Male , Middle Aged , Monitoring, Physiologic/methods , Patient Satisfaction , Pilot Projects , Radial Artery , Retrospective Studies , Risk Assessment , Severity of Illness Index , Surveys and Questionnaires , Time Factors , Treatment Outcome
17.
Trends Microbiol ; 17(10): 458-66, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19766492

ABSTRACT

Rhizobia are phylogenetically disparate alpha- and beta-proteobacteria that have achieved the environmentally essential function of fixing atmospheric nitrogen (N(2)) in symbiosis with legumes. All rhizobia elicit the formation of root - or occasionally stem - nodules, plant organs dedicated to the fixation and assimilation of nitrogen. Bacterial colonization of these nodules culminates in a remarkable case of sustained intracellular infection in plants. Rhizobial phylogenetic diversity raised the question of whether these soil bacteria shared a common core of symbiotic genes. In this article, we review the cumulative evidence from recent genomic and genetic analyses pointing toward an unexpected variety of mechanisms that lead to symbiosis with legumes.


Subject(s)
Alphaproteobacteria/physiology , Betaproteobacteria/physiology , Fabaceae/microbiology , Nitrogen Fixation , Plant Roots/microbiology , Root Nodules, Plant/microbiology , Symbiosis , Alphaproteobacteria/growth & development , Alphaproteobacteria/metabolism , Betaproteobacteria/growth & development , Betaproteobacteria/metabolism , Fabaceae/metabolism , Models, Biological , Plant Roots/metabolism
18.
Appl Environ Microbiol ; 75(12): 4035-45, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19376903

ABSTRACT

Rhizobium sp. strain NGR234 is a unique alphaproteobacterium (order Rhizobiales) that forms nitrogen-fixing nodules with more legumes than any other microsymbiont. We report here that the 3.93-Mbp chromosome (cNGR234) encodes most functions required for cellular growth. Few essential functions are encoded on the 2.43-Mbp megaplasmid (pNGR234b), and none are present on the second 0.54-Mbp symbiotic plasmid (pNGR234a). Among many striking features, the 6.9-Mbp genome encodes more different secretion systems than any other known rhizobia and probably most known bacteria. Altogether, 132 genes and proteins are linked to secretory processes. Secretion systems identified include general and export pathways, a twin arginine translocase secretion system, six type I transporter genes, one functional and one putative type III system, three type IV attachment systems, and two putative type IV conjugation pili. Type V and VI transporters were not identified, however. NGR234 also carries genes and regulatory networks linked to the metabolism of a wide range of aromatic and nonaromatic compounds. In this way, NGR234 can quickly adapt to changing environmental stimuli in soils, rhizospheres, and plants. Finally, NGR234 carries at least six loci linked to the quenching of quorum-sensing signals, as well as one gene (ngrI) that possibly encodes a novel type of autoinducer I molecule.


Subject(s)
Biological Transport , Membrane Transport Proteins/genetics , Membrane Transport Proteins/metabolism , Rhizobium/genetics , Rhizobium/metabolism , Sequence Analysis, DNA , DNA, Bacterial/genetics , Genes, Bacterial , Genome, Bacterial , Plasmids , Rhizobium/physiology
19.
Heart Vessels ; 23(2): 140-3, 2008 Mar.
Article in English | MEDLINE | ID: mdl-18389340

ABSTRACT

Aberrant coronary arteries are frequently observed in patients presenting with Fallot's tetralogy (TOF). Before the complete surgical repair of the TOF, the percutaneously performed pulmonary trunk (PT) angioplasty is often performed in order to temporarily increase the pulmonary circulation, thus increasing the pulmonary vessel size, finally improving surgical outcome. This case reports a 12-year-old boy with a TOF insufficiently improved by surgical correction, in whom a PT angioplasty with stent implantation was complicated by an extrinsic compression of an aberrant right coronary artery (RCA) causing a myocardial ischemia with severe hypotension. The RCA, originating from the left anterior descending coronary artery, passed through the aortic root and the PT and was thus compressed by the PT-stent. Finally the RCA was successfully treated with standard coronary balloon angioplasty and stenting, improving myocardial perfusion and the hemodynamics of the patient, who finally died several days thereafter due to septic shock and massive pulmonary embolism.


Subject(s)
Angioplasty, Balloon/adverse effects , Coronary Stenosis/etiology , Coronary Vessel Anomalies/complications , Pulmonary Valve Stenosis/therapy , Stents , Tetralogy of Fallot/therapy , Angioplasty, Balloon/instrumentation , Angioplasty, Balloon, Coronary/instrumentation , Cardiac Surgical Procedures , Child , Coronary Angiography , Coronary Circulation , Coronary Stenosis/diagnostic imaging , Coronary Stenosis/therapy , Coronary Vessel Anomalies/diagnostic imaging , Coronary Vessel Anomalies/physiopathology , Coronary Vessel Anomalies/therapy , Fatal Outcome , Humans , Hypotension/etiology , Male , Myocardial Ischemia/etiology , Pulmonary Circulation , Pulmonary Valve Stenosis/complications , Pulmonary Valve Stenosis/diagnostic imaging , Pulmonary Valve Stenosis/physiopathology , Tetralogy of Fallot/complications , Tetralogy of Fallot/diagnostic imaging , Tetralogy of Fallot/physiopathology
20.
Heart Vessels ; 23(1): 67-70, 2008 Jan.
Article in English | MEDLINE | ID: mdl-18273549

ABSTRACT

Myocardial bridge (MB) or tunneled coronary artery is an inborn abnormality, which implicates a systolic vessel compression with a persistent mid-late diastolic diameter reduction. Myocardial bridges are often observed during coronary angiography with an incidence of 0.5%-5.5%. The most involved coronary artery is the left anterior descending artery followed by the diagonal branches, the right coronary artery, and the left circumflex. The overall long-term prognosis is generally benign. However, several risk or precipitating factors (e.g., high heart rate, left ventricular hypertrophy, decreased peripheral vascular resistance) may trigger symptoms (most frequently angina). Herein, we describe two cases of symptomatic myocardial bridge, where medical treatment (i.e., inotropic negative drug) and coronary stenting were successfully utilized to treat this pathology. We also focus on the clinical presentation, and the diagnostic and therapeutic modalities to correctly manage this frequently observed congenital coronary abnormality, underlining the fact that in cases of typical angina symptoms without any significant coronary artery disease, MB should be considered as a possible differential diagnosis.


Subject(s)
Angina Pectoris/etiology , Coronary Vessel Anomalies/diagnostic imaging , Aged , Angina Pectoris/diagnostic imaging , Angina Pectoris/surgery , Blood Vessel Prosthesis Implantation/instrumentation , Coronary Angiography , Coronary Vessel Anomalies/complications , Coronary Vessel Anomalies/surgery , Diagnosis, Differential , Follow-Up Studies , Humans , Stents , Time Factors
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