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Pea has a high potential in agroecology, however, it may suffer from a susceptibility to iron deficiency in calcareous soils. This susceptibility varies depending on cultivars. Pseudomonad siderophores, pyoverdines (pvd), were shown to promote iron nutrition in Arabidopsis and tobacco. The hypothesis was therefore raised that variation in iron susceptibility between pea cultivars could be related to differences in their ability to recruit fluorescent pseudomonads (fp) contributing to siderophore-mediated iron nutrition. Our aims were to compare i) the impact of a susceptible (S) and a tolerant (T) cultivar of pea on fp biodiversity with a focus on their pvd, and in return ii) the impact of the corresponding bacteria and pvd on pea iron growth and nutrition. fp associated with roots of S & T and corresponding pvd were compared. In return, representative pvd were tested for their impact on plant growth, iron content and ionome (HRICP-MS). Differences recorded between fp associated with T and S roots were related to i) their low susceptibility to Fe deficiency and ii) their pvd types as characterized by isoelectrofocalisation. Representatives of major pvd types impacted differently S & T iron content, plant ionome and growth. The results obtained support the principle of an extended phenotype in which associated bacteria contribute to plant nutrition, showing the importance to consider root microbiota to develop strategies aiming at overcoming iron susceptibility in pea.

Soil samples were collected from different rhizosphere plants grown in SEKEM farm, Bilbis, El-Sharkyia governorate, Egypt. Four fluorescent pseudomonads isolates, out of seventy one, were selected according to their efficiency to produce IAA, cyanide, antagonistic effect and high ability to solubilize potassium and phosphorus. These isolates were identified using 16S rRNA gene sequencing technique to be different strains of Pseudomonas otitidis. Growth curve and growth kinetics were determined on king’s broth medium. Ps. otitidis SE8 gave the highest specific growth rate, multiplication rate and number of generation being 0.287 h−1, 0.4147 h−1 and 4.98 h−1, respectively. This strain also showed the lowest of doubling time. Ps. otitidis SE8 and OL2 had higher activity to solubilize phosphorus than other tested strains. Bacterial strain SE8 also gave a considerable amount of soluble potassium as compared with other strains being 80.7 ppm. The maximum IAA and gibberellic acid production was also recorded by Pseudomonas SE8 and OL2. A remarkable quantity of siderophores was detected in the case of Ps. otitidis SE8 being 28.20 mM DFOM. Application of these isolates as inoculants for tomato plants in green house was performed. The results showed that inoculation of tomato seedling with Pseudomonas strains led to suppress the soil-borne pathogen, increased of NPK uptake and supported tomato plant growth.

In agricultural ecosystems, pest insects, pathogens, and reduced soil fertility pose major challenges to crop productivity and are responsible for significant yield losses worldwide. Management of belowground pests and diseases remains particularly challenging due to the complex nature of the soil and the limited reach of conventional agrochemicals. Boosting the presence of beneficial rhizosphere organisms is a potentially sustainable alternative and may help to optimize crop health and productivity. Field application of single beneficial soil organisms has shown satisfactory results under optimal conditions. This might be further enhanced by combining multiple beneficial soil organisms, but this remains poorly investigated. Here, we inoculated wheat plots with combinations of three beneficial soil organisms that have different rhizosphere functions and studied their effects on crop performance. Plant beneficial Pseudomonas bacteria, arbuscular mycorrhizal fungi (AMF), and entomopathogenic nematodes (EPN), were inoculated individually or in combinations at seeding, and their effects on plant performance were evaluated throughout the season. We used traditional and molecular identification tools to monitor their persistence over the cropping season in augmented and control treatments, and to estimate the possible displacement of native populations. In three separate trials, beneficial soil organisms were successfully introduced into the native populations and readily survived the field conditions. Various Pseudomonas, mycorrhiza, and nematode treatments improved plant health and productivity, while their combinations provided no significant additive or synergistic benefits compared to when applied alone. EPN application temporarily displaced some of the native EPN, but had no significant long-term effect on the associated food web. The strongest positive effect on wheat survival was observed for Pseudomonas and AMF during a season with heavy natural infestation by the frit fly, Oscinella frit, a major pest of cereals. Hence, beneficial impacts differed between the beneficial soil organisms and were most evident for plants under biotic stress. Overall, our findings indicate that in wheat production under the test conditions the three beneficial soil organisms can establish nicely and are compatible, but their combined application provides no additional benefits. Further studies are required, also in other cropping systems, to fine-tune the functional interactions among beneficial soil organisms, crops, and the environment., The project was funded by grants of the Swiss National Research Program 68 on the “Sustainable Use of Soil as a Resource” from the Swiss National Science Foundation (406840_143065 awarded to TT and FM, 406840_143141 awarded to CK and MM, 406840_143097 to MvdH, OF, and WC). GJ was supported by an assistantship from the University of Neuchâtel, and a Swiss Government Excellence Scholarships supported XC for Foreign Scholars. The Government of Portugal supports RCH with an Investigator Program award (IF/00552/2014).

Many species or strains of the genus Pseudomonas have been characterized as plant growth promoting rhizobacteria (PGPR). We used a combination of phenotypic and genotypic techniques to analyze the community of fluorescent Pseudomonas strains in the rhizosphere of commercially grown Mentha piperita (peppermint). Biochemical techniques, Amplified rDNA Restriction Analysis (ARDRA), and 16S rRNA gene sequence analysis revealed that the majority of the isolated native fluorescent strains were P. putida. Use of two Repetitive Sequence-based PCR (rep-PCR) techniques, BOX-PCR and ERIC-PCR, allowed us to evaluate diversity among the native strains and to more effectively distinguish among them. PGPR activity was tested for the native strains and reference strain P. fluorescens WCS417r. Micropropagated M. piperita plantlets were exposed to microbial volatile organic compounds (mVOCs) emitted by the bacterial strains, and plant biomass parameters and production of essential oils (EOs) were measured. mVOCs from 11 of the native strains caused an increase in shoot fresh weight. mVOCs from three native strains (SJ04, SJ25, SJ48) induced changes in M. pierita EO composition. The mVOCs caused a reduction of metabolites in the monoterpene pathway, for example menthofuran, and an increase in menthol production. Menthol production is the primary indicator of EO quality. The mVOCs produced by native strains SJ04, SJ25, SJ48, and strain WCS417r were analyzed. The obtained mVOC chromatographic profiles were unique for each of the three native strains analyzed, containing varying hydrocarbon, aromatic, and alogenic compounds. The differential effects of the strains were most likely due to the specific mixtures of mVOCs emitted by each strain, suggesting a synergistic effect occurs among the compounds present.

Organic amendments influence chemical and microbial compositions in soils and also susceptibility to plant diseases. The purpose of this study was to establish different parameters that interfere with pathogen growth in soil. Four different organic-amendment regimes, i.e. slurry, compost, slurry-dung and compost-slurry-dung, were applied to two fields proximate to each other on the same farm and covered with the same grass-clover ley in the last two years preceding sampling. Before that period, there were differences in cropping and management practices between both fields. Chemical analyses of the soils revealed no differences between organic amendments, whereas significant differences were present between fields in all C, N and pH values. Growth of R. solani AG2-2IIIB in soil, measured by damping-off in sugar beet plants, was influenced by the interaction of organic amendment with field type. Diversity and evenness values of the microbial communities, studied by PCR-DGGEs specific for bacteria, fungi, Pseudomonas and ammonia-oxidizing beta-proteobacteria, revealed different patterns, i.e. no differences between organic amendments, but clear differences between fields. Multivariate analyses done on individual species of the four groups, as represented by band location and size in PCR-DGGE fingerprints, and by inclusion of chemical and R. solani AG2-2IIIB growth parameters as `environmental' variables, revealed strong and occasionally significant effects of organic matter content, water-dissolvable organic carbon and pH on microbial communities. It was therefore concluded that different organic amendments had different effects on pathogen growth in soils of both fields and that organic matter content and pH influenced soil microbial compositions most.

Aims: To determine the role of phenazines (PHZ) and lipopeptide surfactants (LPs) produced by Pseudomonas in suppression of stem rot disease of groundnut, caused by the fungal pathogen Sclerotium rolfsii. Methods and Results: In vitro assays showed that PHZ-producing Pseudomonas chlororaphis strain Phz24 significantly inhibited hyphal growth of S. rolfsii and suppressed stem rot disease of groundnut under field conditions. Biosynthesis and regulatory mutants of Phz24 deficient in PHZ production were less effective in pathogen suppression. Pseudomonas strains SS101, SBW25, and 267, producing viscosin or putisolvin-like LPs, only marginally inhibited hyphal growth of S. rolfsii and did not suppress stem rot disease. In contrast, Pseudomonas strain SH-C52, producing the chlorinated LP thanamycin, inhibited hyphal growth of S. rolfsii and significantly reduced stem rot disease of groundnut in nethouse and field experiments, whereas its thanamycin-deficient mutant was less effective. Conclusions: Phenazines and specific lipopeptides play an important role in suppression of stem rot disease of groundnut by root-colonizing Pseudomonas strains. Significance and Impact of the Study: Pseudomonas strains Phz24 and SH-C52 showed significant control of stem rot disease. Treatment of seeds or soil with these strains provides a promising supplementary strategy to control stem rot disease of groundnut

This paper briefly reviews research on the causative agents of blackleg and soft rot diseases of potato, namely Pectobacterium and Dickeya species, and the disease syndrome, including epidemiological and aetiological aspects. It critically evaluates control methods used in practice based on the avoidance of the contamination of plants, in particular the use of seed testing programmes and the application of hygienic procedures during crop production. It considers the perspective of breeding and genetic modification to introduce resistance. It also evaluates the application of physical and chemical tuber treatments to reduce inoculum load and examines the possibility of biocontrol using antagonistic bacteria and bacteriophages.

The denitrabon of 2,4,6-trinitrotoluene (TNT) can produce mono- or dinkro aromatic compounds susceptible to microbial mineralization. In the present study, denitration of TNT and other nitro aromatic compounds was investigated with a solid-phase extract obtained from the culture supernatant of Pseudomonas aeruginosa ESA-5 grown on a chemically defined aerobic medium. When the C-18 solid-phase extract containing extracellular catalysts (EC) was incubated with TNT and NAD(P)H, we observed a significant release of nitrite. The concentration of nitrite released in the reaction medium was strongly dependent on the concentration of NAD(P)H and EC. Denitration also occurred with two TNT-related molecules, 2,4,6-trinkrobenzaldehyde, and 2,4,6-trinftrobenzyl alcohol. The release of nitrite was coupled with the formation of two polar metabolites, and mass spectrometry analyses indicated that each of these compounds had lost two nitro groups from the trinitro aromatic parent molecule. During this process, the productionof toxic reduced TNT metabolites was minimal. The incubation of EC with TNT, NAD(P)H, and specific scavengers of reactive oxygen species suggested the involvement of superoxide radicals (O-2(center dot-)) and hydrogen peroxide in the denitration process. Results obtained in this study reveal for the first time that extracellular small-molecular-weight substance(s) of bacterial origin can serve as green catalyst) to initiate TNT denitration. In addition, this study gives clear evidence for the production of a TNT metabolite bearing a single nitro group following a denitration reaction with catalyst(s) of biotic origin.

In previous work, several bacterial groups that show a response to fruiting bodies (the mycosphere) of the ectomycorrhizal fungus Laccaria proxima were identified. We here extend this work to a broader range of fungal fruiting bodies sampled at two occasions. PCR-DGGE analyses showed clear effects of the mycosphere of diverse fungi on the total bacterial and Pseudomonas communities in comparison with those in the corresponding bulk soil. The diversities of the Pseudomonas communities increased dramatically in most of the mycospheres tested, which contrasted with a decrease of the diversity of the total bacterial communities in these habitats. The data also indicated the existence of universal (i.e. Pseudomonas poae, P. lini, P. umsongensis, P. corrugata, P. antarctica and Rahnella aquatilis) as well as specific (i.e. P. viridiflava and candidatus Xiphinematobacter americani) fungiphiles, defined as bacteria adapted to the mycospheres of, respectively, three or more or just one fungal species. The selection of such fungiphiles was shown to be strongly related to their capacities to use particular carbonaceous compounds, as evidenced using principal components analyses of BIOLOG-based substrate utilization tests. The differentiating compounds, i.e. l-arabinose, l-leucine, m-inositol, m-arabitol, d-mannitol and d-trehalose, were tentatively linked to compounds known to occur in mycosphere exudates.

International audience; The mineral weathering ability of 45 bacterial strains belonging to the genus Collimonas and coming from various terrestrial environments was compared to that of 5 representatives from the closely related genera Herbaspirillum and Janthinobacterium. Using glucose as the sole carbon source in a microplate assay for quantifying the release of iron and protons from biotite, all Collimonas strains proved to be very efficient weathering agents, in contrast to the Herbaspirillum and Janthinobacterium strains. The weathering phenotype was also evident during growth of collimonads on mannitol and trehalose, but not on gluconic acid. All Collimonas strains were able to solubilize inorganic phosphorus and produce gluconic acid from glucose, suggesting that acidification is one of the main mechanisms used by these bacteria for mineral weathering. The production of siderophores may also be involved, but this trait, measured as the ability of collimonads to mobilize iron, was shared with Herbaspirillum and Janthinobacterium strains. These findings are discussed in an ecological context that recognizes collimonads as mycophagous (fungal-eating) and efficient mineral weathering bacteria and suggests that this ability has evolved as an adaptation to nutrient-poor conditions, possibly as part of a mutualistic relationship with mycorrhizal fungi.

Bacterial endophytes live inside plants for at least part of their life cycle. Studies of the interaction of endophytes with their host plants and their function within their hosts are important to address the ecological relevance of endophytes. The modulation of ethylene levels in plants by bacterially produced 1-aminocyclopropane-1-carboxylate deaminase is a key trait that enables interference with the physiology of the host plant. Endophytes with this capacity might profit from association with the plant, because colonization is enhanced. In turn, host plants benefit by stress reduction and increased root growth. This mechanism leads to the concept of 'competent' endophytes, defined as endophytes that are equipped with genes important for maintenance of plant-endophyte associations. The ecological role of these endophytes and their relevance for plant growth are discussed here.

The bacterial communities in the Laccaria proxima mycosphere (soil from beneath fruiting bodies) and the corresponding bulk soil were compared by cultivation-dependent and cultivation-independent methods. To assess the distribution of type III secretion systems (TTSS), a PCR-based system for the broad detection of a highly conserved gene involved in TTSS, that is hrcR, was developed and used to assay the cultured bacteria from the L. proxima mycosphere and surrounding bulk soil. PCR-DGGE based on the 16S ribosomal RNA gene showed the selection of presumably mycosphere-specific bacterial groups in the mycosphere of L. proxima compared to the bulk soil in 3 sampling years. Moreover, plate counts revealed that the numbers of culturable heterotrophic bacteria were increased in the mycosphere as compared to the bulk soil. Strikingly, the percentage of randomly picked isolates that carried the hrcR gene showed a significant increase, from 2.8 in the bulk to 13.4 in the mycosphere soil. The increase could be mainly attributed to the emergence of a hrcR positive Pseudomonas fluorescens, denoted BS053, which constituted the most dominant species in the culturable mycosphere communities. This organism was, together with a hrcR-positive Burkholderia terrae BS110, exclusively found in mycosphere soil. Direct detection of hrcR genes using a cultivation-independent approach showed the selection of several hrcR gene types uniquely in the mycosphere, indicating the selection of several TTSS-harboring bacterial species. Thus, different bacteria were found to be enriched in the L. proxima mycosphere and TTSS can be involved in some of the interactions with the fungal host.

Aims: Phytophthora capsici is a major pathogen of black pepper and zoospores play an important role in the infection process. Fluorescent pseudomonads that produce biosurfactants with zoosporicidal activities were isolated from the black pepper rhizosphere in Vietnam, and their genotypic diversity and potential to control Phy. capsici root rot was determined. Methods: Biosurfactant-producing pseudomonads were genotypically and biochemically characterized by BOX-polymerase chain reaction (PCR), 16S-rDNA sequencing, reverse-phase-high-performance liquid chromatography and liquid chromatography-masss spectrometry analyses. Results: Biosurfactant-producing fluorescent pseudomonads make up c. 1.3% of the culturable Pseudomonas population in the rhizosphere of black pepper. Although BOX-PCR revealed substantial genotypic diversity, the isolates were shown to produce the same biosurfactants and were all identified as Pseudomonas putida. When applied to black pepper stem cuttings, several of the biosurfactant-producing strains provided significant disease control. In absence of the disease, several of the bacterial strains promoted shoot and root growth of black pepper stem cuttings. Conclusions: Biosurfactant-producing pseudomonads indigenous to the rhizosphere of black pepper plants are genotypically diverse and provide a novel resource for the control of Phy. capsici root rot and growth promotion of black pepper stem cuttings. Significance and Impact of the Study: The results of this study provide a strong basis for further development of supplementary strategies with antagonistic bacteria to control foot and root rot of black pepper and to promote plant growth.

Chili ( Capsicum annuum L. ) is one of the vegetable commodities with high economic value. Until now, the necessity rate of chili is still high but the production is still low. One of the obstacles, which highly influence chili production, is wilt disease due to Ralstonia solanacearum and Fusarium oxysporum . This study aims at identifying the best fluorescent pseudomonads isolate in suppressing the development of the primary wilt disease due to Ralstonia solanacearum and F. oxysprorum in the field. This study employs Completely Randomized Design that comprises 3 treatments. The three treatments are the type of fluorescent pseudomonads which is Pf-122 isolate, Pf-160 isolate, and Pf-B isolate. The observation parameter on suppressing the disease development is perceived from incubation period of the initial appearance of symptoms, disease index and the change of stem tissue color. The result of the study shows that the longest incubation period is treatment with fluorescent pseudomonads of Pf-122 isolate and Pf-160 isolate. The chili is treated with fluorescent pseudomonads of Pf-122 isolate and Pf-160 isolate, the initial appearance of symptoms at the 8th day after inoculation. In the treatment with fluorescent pseudomonads of Pf-122 isolate and Pf-160 isolate compared to control, is able to delay the appearance of symptom for 3 days. The disease development starts from 10th to 30th day, the treatment with Pf-122 isolate shows the slowest development compared to other treatment and control. In the last observation, the disease index with treatment of Pf-122 isolate is the lowest which is 37%, then it is followed by Pf-B and Pf-160 which are 39% and 40% respectively, whereas in the control it reaches 67%. For the color change on stem tissue, there is no real difference of the three treatments of fluorescent pseudomonads application but it differs and smaller from the control.

Under conditions of iron limitation Pseudomonas fluorescens ATCC 17400 produces two siderophores, pyoverdine, and a second siderophore quinolobactin, which itself results from the hydrolysis of the unstable molecule 8-hydroxy-4-methoxy-2-quinoline thiocarboxylic acid (thioquinolobactin). Pseudomonas fluorescens ATCC 17400 also displays a strong in vitro antagonism against the Oomycete Pythium, which is repressed by iron, suggesting the involvement of a siderophore(s). While a pyoverdine-negative mutant retains most of its antagonism, a thioquinolobactin-negative mutant only slowed-down Pythium growth, and a double pyoverdine-, thioquinolobactin-negative mutant, which does not produce any siderophore, totally lost its antagonism against Pythium. The siderophore thioquinolobactin could be purified and identified from spent medium and showed anti-Pythium activity, but it was quickly hydrolysed to quinolobactin, which we showed has no antimicrobial activity. Analysis of antagonism-affected transposon mutants revealed that genes involved in haem biosynthesis and sulfur assimilation are important for the production of thioquinolobactin and the expression of antagonism.

Analysis of microbial genome sequences have revealed numerous genes involved in antibiotic biosynthesis. In Pseudomonads, several gene clusters encoding non-ribosomal peptide synthetases (NRPSs) were predicted to be involved in the synthesis of cyclic lipopeptide (CLP) antibiotics. Most of these predictions, however, are untested and the association between genome sequence and biological function of the predicted metabolite is lacking. Here we report the genome-based identification of previously unknown CLP gene clusters in plant pathogenic Pseudomonas syringae strains B728a and DC3000 and in plant beneficial Pseudomonas fluorescens Pf0-1 and SBW25. For P. fluorescens SBW25, a model strain in studying bacterial evolution and adaptation, the structure of the CLP with a predicted 9-amino acid peptide moiety was confirmed by chemical analyses. Mutagenesis confirmed that the three identified NRPS genes are essential for CLP synthesis in strain SBW25. CLP production was shown to play a key role in motility, biofilm formation and in activity of SBW25 against zoospores of Phytophthora infestans. This is the first time that an antimicrobial metabolite is identified from strain SBW25. The results indicate that genome mining may enable the discovery of unknown gene clusters and traits that are highly relevant in the lifestyle of plant beneficial and plant pathogenic bacteria

Environmental concerns have led to the need of sustainable use of natural resources. The conventional agriculture caused considerable impacts on soils and waters. It is important to change certain agricultural managements to environmental cleaner techniques. The sustainable agriculture has pointed many approaches and techniques to reduce environmental impact. One of those strategies is the utilization of the soil microbiota to induce plant growth, control of plant diseases and biodegradation of xenobiotic compounds. Studies on the relationship between roots and microbiota are essential to achieve viable agricultural applications. These studies indicate that one of the most abundant microorganisms in the rhizosphere (area around the roots) is fluorescent Pseudomonas spp. They have been considered as an alternative to agrochemicals for controlling plant diseases and increasing plant development. This review addresses the main findings on fluorescent Pseudomonads. It summarizes and discusses significant aspects of this general topic, including (i) rhizosphere as a microhabitat; (ii) taxonomic, genetic and ecological aspects of fluorescent Pseudomonads in the rhizosphere; (iii) mechanisms of Plant Growth Promoting and Biological Control and (iv) commercial use of PGPR in agriculture.As preocupações com o meio ambiente têm levado à necessidade do uso sustentável dos recursos naturais. A agricultura convencional tem causado impactos consideráveis no solo e nas águas. É importante mudar certas técnicas agrícolas por outras consideradas mais limpas para o meio ambiente. Uma estratégia é a utilização da microbiota do solo para induzir o crescimento de plantas, controlar fitomoléstias e para fazer a biodegradação de compostos xenobióticos. Estudos sobre a relação entre as raízes e a microbiota são essenciais para conseguir aplicações viáveis na agricultura. Estes estudos têm indicado que um dos mais abundantes microrganismos na rizosfera (região ao redor da raiz) é Pseudomonas spp. do grupo fluorescente. Estes têm sido considerados como alternativa tanto aos agrotóxicos no controle de doença de plantas como na indução do seu desenvolvimento. Esta revisão mostra os principais conhecimentos sobre Pseudomonas spp. do grupo fluorescente. Esta resume e discute aspectos significantes deste tópico, incluindo, (i) rizosfera como microhabitat; (ii) aspectos taxonômicos, genéticos e ecológicos de Pseudomonas spp. do grupo das fluorescentes na rizosfera; (iii) mecanismos de Promoção de Crescimento de Plantas e Controle Biológico e (iv) uso comercial de PGPR na agricultura.

The phaC1 gene from Pseudomonas oleovorans, coding for the Pha-C1 polymerase, was introduced into the potato genome. Transgenic callus and plant lines which transcribed and translated the transgene were selected and cell suspension cultures from the wild type and transgenic lines were established. The substrate for the Pha-C1 polymerase, 3-(R)-hydroxyoctanoate, was provided to the growth medium. In the transgenic lines, but not in the wild type or in transgenic cell suspension cultures without Pha-C1 expression, evidence of medium-chain-length polyhydroxyalkanoate accumulation ranging from 0.02 to 9.7 mg of polymer per gram of dry weight was observed after feeding in the growth medium the substrate. (C) 2003 Elsevier Science B.V. All rights reserved.

affiche, résumé publié; Iron is an essential element for plants and microbes. However, in most cultivated soils, the concentration of iron available for these living organisms is very low since its solubility is controlled by stable hydroxides, oxyhydroxides and oxides. The high demand for iron by plants and microorganisms in the rhizosphere together with its low availability in soils leads to a strong competition for this nutrient among living organisms. To face this competition, plants and microorganisms have developed active strategies of iron uptake. In non graminaceous plants (strategy I), iron uptake relies on acidification and reduction of Fe+++ in Fe++ which incorporated in the roots by iron transporters. Active iron uptake by microorganisms relies on siderophores showing high affinity for iron. We have previously shown that plants of Arabidopsis thaliana (strategy I) supplemented with Fe-pyoverdine had a higher iron content than those supplemented Fe- EDTA [1]. Iron from pyoverdine was not incorporated through the major iron transporter IRT1 as indicated by the similar iron content of the wild-type plant and IRT1 mutant knockout iron transporter IRT1. Furthermore, pyoverdine was shown to be incorporated as indicated by its presence in planta based on enzyme-linked immunosorbent assay measurement of pyoverdine and on 15N of 15N-pyoverdine. Taken together, these observations suggest that iron from Fe-pyoverdine was not incorporated in planta through the strategy I. In the present, we explored the possible incorporation of iron from pyoverdine at the cellular level. For that purpose, on Arabidopsis when cultivated in the presence of Fe-EDTA (50µM) or Fe-pyoverdine (50µM), we analyzed the immunolocalization of pyoverdine in roots by confocal microscopy and performed ultrastructural studies with transmission electron microscopy (TEM). Plants were cultivated in vitro with these chelates during seven days. Immunolabeling were performed with pyoverdine antibody as primary antibody and fluorescent secondary antibody for confocal microscopy and colloidal gold coupled to secondary antibody for immunogold labeling by TEM. Observations with confocal microscopy clearly indicated the presence of pyoverdine in fresh tissue sections. This immunolocalization revealed the presence of pyoverdine in root apoplasmic space. Observations with TEM showed the more abundant presence of vesicles in root apoplasm of plants when cultured with Fe-pyoverdine than with Fe-EDTA (Figure 1). Despite that pyoverdine immunogold labeling of roots sections did not allow to reveal the formal presence of pyoverdine in these vesicles, the present results suggest that the incorporation of Fe- pyoverdine might rely on endocytosis.

Bacteria are believed to play an important role in the fitness and biochemistry of sponges (Porifera). Pseudomonas species (Gammaproteobacteria, Pseudomonadales) are capable of colonizing a broad range of eukaryotic hosts, but knowledge of their diversity and function in freshwater invertebrates is rudimentary. We assessed the diversity, structure and antimicrobial activities of Pseudomonas spp. in the freshwater sponge Ephydatia fluviatilis. Polymerase Chain Reaction - Denaturing Gradient Gel Electrophoresis (PCR-DGGE) fingerprints of the global regulator gene gacA revealed distinct structures between sponge-associated and free-living Pseudomonas communities, unveiling previously unsuspected diversity of these assemblages in freshwater. Community structures varied across E. fluviatilis specimens, yet specific gacA phylotypes could be detected by PCR-DGGE in almost all sponge individuals sampled over two consecutive years. By means of whole-genome fingerprinting, 39 distinct genotypes were found within 90 fluorescent Pseudomonas isolates retrieved from E. fluviatilis. High frequency of in vitro antibacterial (49%), antiprotozoan (35%) and anti-oomycetal (32%) activities was found among these isolates, contrasting less-pronounced basidiomycetal (17%) and ascomycetal (8%) antagonism. Culture extracts of highly predation-resistant isolates rapidly caused complete immobility or lysis of cells of the protozoan Colpoda steinii. Isolates tentatively identified as P. jessenii, P. protegens and P. oryzihabitans showed conspicuous inhibitory traits and correspondence with dominant sponge-associated phylotypes registered by cultivation-independent analysis. Our findings suggest that E. fluviatilis hosts both transient and persistent Pseudomonas symbionts displaying antimicrobial activities of potential ecological and biotechnological value. European Regional Development Fund (ERDF) through the COMPETE (Operational Competitiveness Programme); national funds through FCT (Foundation for Science and Technology) [PEst-C/MAR/LA0015/2011]; FCT-funded project [PTDC/BIA-MIC/3865/2012]; Federation of European Microbiological Societies (FEMS) info:eu-repo/semantics/publishedVersion

International audience; Several reports have highlighted that forest soil samples are more phylum-rich than agricultural soil samples. However, little is known about the structure and richness of the bacterial communities in forest soil. Using high-throughput next generation 454 pyrosequencing, we deeply investigated the diversity of bacterial communities colonizing the oak rhizosphere niche and the surrounding soil. From three spatially independent soil samples, we obtained over 300 000 partial 16S rRNA gene sequences. The most abundant bacterial groups were the Acidobacteria, Proteobacteria and unclassified bacteria. Multifactorial analysis of the relative proportions of the different phyla revealed a net differentiation of the bacterial communities present in the rhizosphere and soil environments, suggesting an oak rhizosphere effect. Significantly more b-, g- and unclassified Proteobacteria inhabited the rhizosphere when compared with the surrounding soil. Conversely, significantly more unclassified bacteria were detected in the bulk soil than in the rhizosphere, demonstrating that the soil remains a challenging reservoir of complexity. This work increases our understanding of the niche effect on bacterial diversity and on the rare phylogenetic groups inhabiting the soil.

International audience; Increasing interest has been given so far to the role of mycorrhizal symbiosis on plant diversity and ecosystem productivity. However much remains unknown about the effect of ectomycorrhizal fungal diversity on plant growth and rhizobial symbiosis. The purpose of this study is to investigate the influence of ectomycorrhizal diversity on root nodulation and plant nutrient uptake during plant growth. A gradient of ectomycorrhizal diversity was created (ranged from one to six fungal isolates) on Australian Acacia (Acacia mangium) seedlings grown on a phosphate-deficient soil in controlled conditions. The results dearly show (i) an enhancement of A. mangium growth due to ectomycorrhizal fungal inoculation, (ii) as well as significant effects of increasing ectomycorhizal diversity and soil ectomycorrhizal mycelium density on whole plant biomass and plant nutrient uptake. They also reveal an effect of ectomycorrhizal diversity effect on the functioning of the nitrogen-fixing symbiosis. These results emphasize the need to manage soil ectomycorrhizal potential (abundance and diversity of ectomycorrhizal mycelium) in forest management practices in order to optimize N-2 fixation and seedling growth, leading thus to dramatic improvements in outplanting performances with fast-growing tree legumes on various reafforestation sites, more particularly in tropical and Mediterranean areas. (C) 2012 Elsevier Ltd. All rights reserved.

Analysis of the 16S rRNA gene sequences generated from Xerocomus pruinatus and Scleroderma citrinum ectomycorrhizospheres revealed that similar bacterial communities inhabited the two ectomycorrhizospheres in terms of phyla and genera, with an enrichment of the Burkholderia genus. Compared to the bulk soil habitat, ectomycorrhizospheres hosted significantly more Alpha -, Beta -, and Gammaproteobacteria .

Aims: To determine the role of phenazines (PHZ) and lipopeptide surfactants (LPs) produced by Pseudomonas in suppression of stem rot disease of groundnut, caused by the fungal pathogen Sclerotium rolfsii. Methods and Results: In vitro assays showed that PHZ-producing Pseudomonas chlororaphis strain Phz24 significantly inhibited hyphal growth of S. rolfsii and suppressed stem rot disease of groundnut under field conditions. Biosynthesis and regulatory mutants of Phz24 deficient in PHZ production were less effective in pathogen suppression. Pseudomonas strains SS101, SBW25, and 267, producing viscosin or putisolvin-like LPs, only marginally inhibited hyphal growth of S. rolfsii and did not suppress stem rot disease. In contrast, Pseudomonas strain SH-C52, producing the chlorinated LP thanamycin, inhibited hyphal growth of S. rolfsii and significantly reduced stem rot disease of groundnut in nethouse and field experiments, whereas its thanamycin-deficient mutant was less effective. Conclusions: Phenazines and specific lipopeptides play an important role in suppression of stem rot disease of groundnut by root-colonizing Pseudomonas strains. Significance and Impact of the Study: Pseudomonas strains Phz24 and SH-C52 showed significant control of stem rot disease. Treatment of seeds or soil with these strains provides a promising supplementary strategy to control stem rot disease of groundnut

International audience; Mycorrhizal symbiosis is a key component of a sustainable soil-plant system, governing the cycles of major plant nutrients and vegetation cover. The mycorrhizosphere includes plants roots, the mycorrhizal fungi, and a complex microbial compartment. A large number of methods have been used to characterize the genetic and functional diversity of these soil microbial communities. We present here a review of the multivariate data analysis methods that have been used in 16 research articles published in the 2005-2009 period. "Descriptive" multivariate data analysis methods have been priviledged over classical "predictive" methods and univariate statistical tests. Data sets, multivariate data analysis methods, graphical outputs and interpretation results are presented and explained in details on several examples coming from some of the 16 articles. These multivariate and graphical methods are available in the ade4 package for the R statistical software. The discussion underlines the importance of using appropriate statistical methods to obtain a good description of soil microbiological and biochemical indicators and a good evaluation of soil quality.

SPE EA Ecoldur CT3 Enjeu2 S. Ghirardi : ancienne thèsarde de l'UMR 1229 MSE, n'a jmas intégré l'UMR 1347 Agroécologie (mais sur PDF 1347 !) T. Corberand : ancien agent de l'UMR 1229 MSE, n'a jmas intégré l'UMR 1347 Agroécologie (mais sur PDF 1347 !); Rhizosphere competence of fluorescent pseudomonads is a prerequisite for the expression of their beneficial effects on plant growth and health. To date, knowledge on bacterial traits involved in rhizosphere competence is fragmented and derived mostly from studies with model strains. Here, a population approach was taken by investigating a representative collection of 23 Pseudomonas species and strains from different origins for their ability to colonize the rhizosphere of tomato plants grown in natural soil. Rhizosphere competence of these strains was related to phenotypic traits including: (1) their carbon and energetic metabolism represented by the ability to use a wide range of organic compounds, as electron donors, and iron and nitrogen oxides, as electron acceptors, and (2) their ability to produce antibiotic compounds and N-acylhomoserine lactones (N-AHSL). All these data including origin of the strains (soil/rhizosphere), taxonomic identification, phenotypic cluster based on catabolic profiles, nitrogen dissimilating ability, siderovars, susceptibility to iron starvation, antibiotic and N-AHSL production, and rhizosphere competence were submitted to multiple correspondence analyses. Colonization assays revealed a significant diversity in rhizosphere competence with survival rates ranging from approximately 0.1 % to 61 %. Multiple correspondence analyses indicated that rhizosphere competence was associated with siderophore-mediated iron acquisition, substrate utilization, and denitrification. However, the catabolic profile of one rhizosphere-competent strain differed from the others and its competence was associated with its ability to produce antibiotics phenazines and N-AHSL. Taken together, these data suggest that competitive strains have developed two types of strategies to survive in the rhizosphere.

This paper briefly reviews research on the causative agents of blackleg and soft rot diseases of potato, namely Pectobacterium and Dickeya species, and the disease syndrome, including epidemiological and aetiological aspects. It critically evaluates control methods used in practice based on the avoidance of the contamination of plants, in particular the use of seed testing programmes and the application of hygienic procedures during crop production. It considers the perspective of breeding and genetic modification to introduce resistance. It also evaluates the application of physical and chemical tuber treatments to reduce inoculum load and examines the possibility of biocontrol using antagonistic bacteria and bacteriophages.

Many agricultural soils of Iran have high pH, resulting in low availability of Fe and Zn. The potentials of nonsymbiotic plant growth-promoting rhizbacteria (PGPR) for stimulating plant growth have been extensively used during recent decades. This experiment was carried out in growth chamber to evaluate the effects of siderophoreproducing Pseudomonads on the growth as well as Fe and Zn uptake of wheat. A randomized complete block design experiments was conducted using with Alborz genotype (an efficient phytosiderophore-producing bread wheat) treated with either 7NSK2 strain as a siderophore positive (sid+) or with MPFM1 mutant strain of the same isolate as a siderophore negative (sid-) treatments with three replications. The potentials of these strains for auxin production and phosphate solubilizing activity were evaluated by standard methods. The results showed that inoculation with sid+ strain increased dry matter production in shoots as compared with the control (sterile condition) or with sid - strain. Likewise, the concentration of chlorophyll a in leaves of sid+ and sid - treatments were 1.27 and 0.41 g mg-1 of fresh weight, respectively, and the concentration of chlorophyll b were measured to be 1.09 and 0.35 g mg-1 of fresh weight, respectively, indicating significantly more chlorophyll formation due to inoculation with sid+ as compared with sid -. The uptake of Fe by roots and its rate of translocation to the shoots were greater for the sid+ treated plants as compared with the sid - treated ones, indicating that siderophores increased the rate of Fe uptake by wheat. The effect of microbial inoculation on shoot Zn was not significant, but increased the concentration of Zn on roots compared with control. The results suggested that the siderophores of Pseudomonads may involve on increasing bioavailability of iron.

We examined the effects of an ectomycorrhizal (EM) fungus, Pisolithus sp., on of the growth of Pinus halepensis (Allepo pine) seedlings, soil microbial functions and rock phosphate solubilization in a un-disinfected soil amended or not with a Moroccan rock phosphate. Allepo pine seedlings were inoculated with an EM fungus (Pisolithus sp. strain PH4) isolated from a P. halepensis plantation and selected for its high ability to mobilize P from an inorganic form of phosphate. After 4 month's culture in a disinfected substrate, plants were transferred in 10 l-containers filled with a natural forest soil and amended or not with rock phosphate powder. After 12 month's culturing, the growth, needle nutrient concentrations of P. halepensis plants were measured. Soil microbial catabolic diversity was assessed by measuring CO2 production of substrate induced respiration responses. Fluorescent pseudomonads were isolated from each soil treatment and tested in axenic conditions for their ability to solubilize a source of inorganic phosphate. The results clearly showed that (i) P. halepensis growth was greatly promoted by the presence of the ectomycorrhizal fungus Pisolithus strain PH4 in a disinfected soil/vermiculite mixture and in a non disinfected soil, (ii) ectomycorrhizal inoculation induced significant changes in the functions of soil microbial communities and selected microorganisms potentially beneficial to the plant growth (i.e. phosphate-solubilizing fluorescent pseudomonad) and (iii) rock phosphate solubilisation was mainly dependent on EM inoculation and mycorrhizosphere microorganisms. These results were in accordance with previous studies where it was demonstrated that EM symbiosis has a beneficial effect on plant growth through a direct effect on the host plant but also an indirect effect via a selective pressure on soil microbiota that favours microorganisms potentially beneficial to plant growth.

Natural disease-suppressive soils provide an untapped resource for the discovery of novel beneficial microorganisms and traits. For most suppressive soils, however, the consortia of microorganisms and mechanisms involved in pathogen control are unknown. To date, soil suppressiveness to Fusarium wilt disease has been ascribed to carbon and iron competition between pathogenic Fusarium oxysporum and resident non-pathogenic F. oxysporum and fluorescent pseudomonads. In this study, the role of bacterial antibiosis in Fusarium wilt suppressiveness was assessed by comparing the densities, diversity and activity of fluorescent Pseudomonas species producing 2,4-diacetylphloroglucinol (DAPG) (phlD+) or phenazine (phzC+) antibiotics. The frequencies of phlD+ populations were similar in the suppressive and conducive soils but their genotypic diversity differed significantly. However, phlD genotypes from the two soils were equally effective in suppressing Fusarium wilt, either alone or in combination with non-pathogenic F. oxysporum strain Fo47. A mutant deficient in DAPG production provided a similar level of control as its parental strain, suggesting that this antibiotic does not play a major role. In contrast, phzC+ pseudomonads were only detected in the suppressive soil. Representative phzC+ isolates of five distinct genotypes did not suppress Fusarium wilt on their own, but acted synergistically in combination with strain Fo47. This increased level of disease suppression was ascribed to phenazine production as the phenazine-deficient mutant was not effective. These results suggest, for the first time, that redox-active phenazines produced by fluorescent pseudomonads contribute to the natural soil suppressiveness to Fusarium wilt disease and may act in synergy with carbon competition by resident non-pathogenic F. oxysporum.