Biologie des communautés dans la rhizosphère : analyse des interactions plantes - champignons mycorhizogènes à arbuscules - bactéries

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Methods for studying root colonization by introduced beneficial bacteria

e-Book; Some free-living rhizobacteria are considered as potential biocontrol and plant growth-promoting agents. Successful application of beneficial bacteria as microbial inoculants requires their presence and activity at the appropriate level, but even more, at the right time and place. Various markers are described in the literature to differentiate introduced bacteria from indigenous microflora and to visualize them. These markers are presented together with the methods currently applied to quantify bacterial densities and to characterize the distribution of introduced bacteria. The methods to quantify bacterial densities are either based on bacterial cultivation or not. Different types…

Strategies for analyzing interactions between arbuscular mycorrhiza and bacteria

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Medicago species affect the community composition of arbuscular mycorrhizal fungi associated with roots

National audience; The symbiosis between plants and arbuscular mycorrhizal fungi (AMF) is ancient and involves 80% of terrestrial plant families. The symbiotic association between AMF and plants was described to be non specific. However, AMF were reported to influence plant community diversity and productivity. On the other way, the effect of plant genotypes belonging to closely related species on AMF diversity has not been explored so far. The aim of this work was to assess the impact of four different Medicago species, M. laciniata, M. murex, M. polymorpha and M. truncatula cv. Jemalong J5, on the composition of AM fungal community, when cultivated in a silty-thin clay soil (Mas d’Imbert,…

Colonization pattern of primary tomato roots by Pseudomonas fluorescens A6RI characterized by dilution plating, flow cytometry, fluorescence, confocal and scanning electron microscopy

Early colonization of primary tomato roots, grown in vitro, by Pseudomonas fluorescens A6RI, introduced by seed bacterization, was monitored for 7 days in three different root zones (zone A, apex+elongation+young hairy zone; zone B, hairy zone; zone C, old hairy zone+collar). Bacterial quantification was assessed by enumeration of (i) colony forming units (cfu) after dilution plating and of (ii) total bacterial cells by flow cytometry. Bacterial distribution and organization in the root zones were analyzed by fluorescence, confocal and scanning electron microscopy. For all sampling dates and zones, the densities of total bacterial cells were significantly higher than those of the cfu. The k…

Sugar exchanges in arbuscular mycorrhiza: RiMST5 and RiMST6, two novel Rhizophagus irregularis monosaccharide transporters, are involved in both sugar uptake from the soil and from the plant partner

SPE IPM INRA UB CT1; International audience; Arbuscular mycorrhizal (AM) fungi are associated with about 80% of land plants. AM fungi provide inorganic nutrients to plants and in return up to 20% of the plant-fixed CO2 is transferred to the fungal symbionts. Since AM fungi are obligate biotrophs, unraveling how sugars are provided to the fungus partner is a key for understanding the functioning of the symbiosis. In this study, we identified two new monosaccharide transporters from Rhizophagus irregularis (RiMST5 and RiMST6) that we characterized as functional high affinity monosaccharide transporters. RiMST6 was characterized as a glucose specific, high affinity H(+) co-transporter. We prov…

Microdiversity of Burkholderiales associated with mycorrhizal and nonmycorrhizal roots of Medicago truncatula

The genetic diversity of bacterial communities associated with mycorrhizal and nonmycorrhizal roots of Medicago truncatula was characterized by two approaches. Firstly, phylogenetic analysis was performed on 164 partial 16S rRNA gene-intergenic spacer (IGS) sequences from operational taxonomic units previously shown to be preferentially associated with mycorrhizal roots. These sequences were distributed into three branches corresponding to Comamonadaceae, Oxalobacteraceae and Rubrivivax subgroups. Most sequences were obtained from mycorrhizal roots, indicating the preferential association of the corresponding families with mycorrhizal roots. A second phylogenetic analysis was performed on t…

Interactions between a mycorrhiza helper bacterium and arbuscular mycorrhizas

Pseudomonas fluorescens C7R12 was shown to promote arbuscular mycorrhization and to act as a ‘Mycorrhiza Helper Bacteria’ (MHB). The complex interactions between this model strain, Arbuscular Mycorrhizal (AM) fungi and host-plants were analysed by assessing the specificity of these interactions and by characterizing bacterial cell organization on mycorrhizal roots. Evaluation of the interaction specificity relied on the comparison of the bacterial effect on (i) the in vitro saprophytic growth of Glomus mosseae and Gigasopora rosea and (ii) the root colonization of two different plant species (Medicago truncatula and Lycopersicon esculentum) by the two AM fungal species. Characterization of …

Identification of sugar transporters in arbuscular mycorrhiza, from basic to applied science

Our study focuses on sugar transporters from both plant and fungal partners at the symbiotic interface to better understand biotrophic exchange systems. Thereby, a collection of putative contigs and ESTs of hexose transporters from Medicago truncatula and Fragaria x ananassa will be processed. The full length sequences will be cloned for functional complementation and uptake experiments in transport deficient yeast mutants. This work also investigates the influence of different mycorrhizal fungi on (1) the expression level of sugar transporters and (2) the economically relevant part of F. x ananassa by analyzing its impact on the plant and the fruit. To this aim, combination of phenological…

Effets de Pseudomonas fluorescens A6RI et de Glomus mosseae BEG12 sur la croissance et le développement racinaire de la tomate

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Colonization of Plant Roots by Pseudomonads and AM Fungi: A Dynamic Phenomenon, Affecting Plant Growth and Health

Because of their enormously large range of plant hosts and role in plant nutrition, arbuscular mycorrhizal (AM) fungi represent an extraordinarily fascinating field of study. Plant growth promotion effects by AM fungi were described as early as 1900 (Sthal 1900) and several data obtained in the second half of the last century support the idea that these microrganisms can act as biocontrol agents (BCA). The extent of root colonization is variable in different plants and under different environmental conditions (Giovannetti and Hepper 1985). Some effects of AM colonization on plants have been reported to be dependent on the degree of root colonization, while others have not. Root exudation an…

Colonization of adventitious roots ofMedicago truncatulabyPseudomonas fluorescensC7R12 as affected by arbuscular mycorrhiza

Pseudomonas fluorescens C7R12 was previously shown to promote colonization of Medicago truncatula roots by Glomus mosseae BEG12. To gain more insight into the interaction between C7R12 and BEG12, the cell organization of C7R12 was characterized on adventitious roots mycorrhized or not with BEG12 and on extraradical hyphae. Bacterial cell observations were made using the immuno-fluorescence technique and confocal laser scanning microscopy. Five types of cell organization, so-called organization types (OT), were identified: small or large single cells, cells by pair and cells in microcolonies or in strings. The frequencies of each OT on the roots were expressed as the percentage of observatio…

The symbiosis between Nicotiana tabacum and the endomycorrhizal fungus Funneliformis mosseae increases the plant glutathione level and decreases leaf cadmium and root arsenic contents

Over time, anthropogenic activities have led to severe cadmium (Cd) and arsenic (As) pollution in several environments. Plants inhabiting metal(loid)-contaminated areas should be able to sequester and detoxify these toxic elements as soon as they enter roots and leaves. We postulated here that an important role in protecting plants from excessive metal(loid) accumulation and toxicity might be played by arbuscular mycorrhizal (AM) fungi. In fact, human exploitation of plant material derived from Cd- and As-polluted environments may lead to a noxious intake of these toxic elements; in particular, a possible source of Cd and As for humans is given by cigarette and cigar smoke. We investigated …

Microbial ecology of the rhizosphere

Part II. Selected MethodsPlant-microbe Chapitre 9 : Interactions and Soil Quality; International audience

La protéomique comme moyen d'identification de protéines induites par le cadmium et différenciellement régulées dans les racines de pois colonisées par un champignon mycorhizogène à arbuscules

National audience

Identification of sugar transporters in arbuscular mycorrhiza

Our study focuses on sugar transporters from both plant and fungal partners at the symbiotic interface to better understand biotrophic exchange systems. Thereby, a collection of putative contigs and ESTs of hexose transporters from Medicago truncatula and Fragaria x ananassa will be processed. The full length sequences will be cloned for functional complementation and uptake experiments in transport deficient yeast mutants. This work also investigates the influence of different mycorrhizal fungi on (1) the expression level of sugar transporters and (2) the economically relevant part of F. x ananassa by analyzing its impact on the plant and the fruit. To this aim, combination of phenological…

Suppression of Rhizoctonia root-of tomato by Glomus mosseae BEG12 and Pseudomonas fluorescens A6RI is associated with their effect on the pathogen growth and on the root morphogenesis

Effect of arbuscular mycorrhiza establishment on the genetic structure and diversity of bacteria in the rhizosphere of Medicago truncatula

National audience; The rhizosphere is the volume of soil which is influenced by plant root activity. Plant roots release 20% of the photosynthates into the rhizosphere. This process sustains the development of an active free living microflora in the rhizosphere compared to the bulk soil. Plants establish also symbiotic associations with soil microorganisms. Arbuscular mycorrhizas (AM) are mutualistic symbioses between soil fungi and plant roots. AM contribute to plant adaptation to low fertility environments. AM are established by 80% of terrestrial plant families including major crop plants. AM are also ancient symbiosis (400 million years). AM are known to be non-specific associations. Ho…

Morphogenetic modifications induced by Pseudomonas fluorescens A6RI and Glomus mosseae BEG12 in the root system of tomato differ according to plant growth conditions

Summary • The ability of fluorescent pseudomonads and glomalean fungi to promote plant growth has been reported many times. However, little is known of their effects on root morphogenesis. Growth and root morphogenesis were compared in tomato ( Lycopersicon esculentum ) plants inoculated or not with a model strain of Pseudomonas fluorescens (A6RI) or with the arbuscular mycorrhizal fungus Glomus mosseae (BEG12). • Plants were cultivated in a sandy-loam soil mixed with sand at two different ratios (2 : 1 and 1 : 2), in gnotobiotic conditions. Plant growth was evaluated by measuring root and shoot fresh weight, and various morphometric parameters were compared. • Growth of control plants was …

Identification de transporteurs de sucres marqueurs de la mycorhize à arbuscules

Effect of Endomycorrhizal Infection on Root System Development in the Apple Rootstock (Malus domestica Borkh.) M26

A mycorrhizal fungus (Glomus intraradices) is able to promote shoot growth of the apple rootstock M26. We have shown that the root system is also modified by the presence of the endophyte. Mycorrhiza formation reduced the shoot to root ratio and increased the specific root length. We show that in apple modifications in root architecture are mainly due to a reduction in the elongation of the different root orders.

Targeted proteomics to identify cadmium-induced protein modifications in Glomus mosseae-inoculated pea roots

International audience