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Fungal toxins can have various adverse health effects, including carcinogenic, teratogenic or hepatotoxic impacts. In addition, fungal alteration has also a negative impact on agricultural plant production. The use of chemical fungicides to control mycotoxin contamination is increasingly controversial and regulated. More environmentally friendly methods are therefore being explored. Essential oils, as compounds extracted from plants, are liquids whose specific aromatic compounds give each essential oil its own unique characteristics. Due to their rich chemical composition, essential oils (EOs) have many interesting properties, including antifungal activities. The objective of the present study was to analyze volatile chemical composition of EOs (Cymbopogon schoenanthus, Cymbopogon nardus and Eucalyptus camaldulensis) by GC/MS and to investigate their effects on the growth, sporulation and mycotoxin production of Aspergillus flavus, Aspergillus carbonarius and Fusarium verticillioides (aflatoxin B1, ochratoxin A and fumonisin B1, respectively). In addition, EOs influence on aflatoxin B1 (AFB1) and fumonisin B1 (FB1) biosynthesis pathways was explored using real-time qRT-PCR. The results obtained in vitro, by direct contact with the EOs and by diffusion of their volatile compounds, showed that the essential oils had inhibitory effects on the growth and the production of mycotoxins of the 3 fungal strains and modified the expression of some toxin synthesis genes. We conclude that the recorded effects were dependent on the combined effects of the EOs type, the fungal strains and the doses studied.

International audience; The Vfm quorum sensing (QS) system is preponderant for the virulence of different species of the bacterial genus Dickeya. The vfm gene cluster encodes 26 genes involved in the production, sensing or transduction of the QS signal. To date, the Vfm QS signal has escaped detection by analytical chemistry methods. However, we report here a strain-specific polymorphism in the biosynthesis genes vfmO and vfmP, which is predicted to be related to the production of different analogues of the QS signal. Consequently, the Vfm communication could be impossible between strains possessing different variants of the genes vfmO/P. We constructed three Vfm QS biosensor strains possessing different vfmO/P variants and compared these biosensors for their responses to samples prepared from 34 Dickeya strains possessing different vfmO/P variants. A pattern of specificity was demonstrated, providing evidence that the polymorphism in the genes vfmO/P determines the biosynthesis of different analogues of the QS signal. Unexpectedly, this vfmO/P-dependent pattern of specificity is linked to a polymorphism in the ABC transporter gene vfmG, suggesting an adaptation of the putative permease VfmG to specifically bind different analogues of the QS signal. Accordingly, we discuss the possible involvement of VfmG as co-sensor of the Vfm two-component regulatory system

International audience; Soil is a major receptor of manufactured nanomaterials (NMs) following unintentional releases or intentional uses. Ceria NMs have been shown to undergo biotransformation in plant and soil organisms with a partial Ce(IV) reduction into Ce(III), but the influence of environmentally widespread soil bacteria is poorly understood. We used high-energy resolution fluorescencedetected X-ray absorption spectroscopy (HERFD-XAS) with an unprecedented detection limit to assess Ce speciation in a model soil bacterium (Pseudomonas brassicacearum) exposed to CeO2 NMs of different sizes and shapes. The findings revealed that the CeO2 NM’s size drives the biotransformation process. No biotransformation was observed for the 31 nm CeO2 NMs, contrary to 7 and 4 nm CeO2 NMs, with a Ce reduction of 64 ± 14% and 70 ± 15%, respectively. This major reduction appeared quickly, from the early exponential bacterial growth phase. Environmentally relevant organic acid metabolites secreted by Pseudomonas, especially in the rhizosphere, were investigated. The 2- keto-gluconic and citric acid metabolites alone were able to induce a significant reduction in 4 nm CeO2 NMs. The high biotransformation measured for

International audience; Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) are the biological vectors of arboviruses of global importance in animal health. We characterized the physicochemical parameters that determine the density and composition of the main Culicoides species of veterinary interest in larval habitats of the Niayes region of Senegal. For this purpose, we combined larval and substrate sampling in the field in different habitat types with adult emergence and physicochemical analyses in the laboratory. Three major habitat types were identified, conditioning the predominant species of Culicoides and pH and the amount of organic matter were positively correlated with the abundance of larvae and emerging Culicoides, as opposed to salinity. The diversity of emerging Culicoides was positively correlated with pH while it was negatively correlated with salinity. Culicoides distinctipennis was the predominant species in the larval habitat group of freshwater lake edges. In the larval habitat group of pond and puddle edges, C. oxystoma and C. nivosus were predominant; both species were again most abundant in the larval habitat group of saltwater lake edges. These variabilities in physicochemical parameters support the distribution of different Culicoides species in different habitat groups. These results make it possible to implement effective, selective and environmental-friendly control measures but also to improve current models for estimating the abundance of adult vector populations at a local scale.

International audience; Numerous differentin vitroassays, labeled as "antioxidant assays," are used intensively to predict the antioxidant capacity of phenolic compounds. Most of these methods give valuable information in terms of chemical reactivities but also present some weaknesses. It may be difficult to use them to predict antioxidant capacity in real conditions. Indeed, lipid oxidation is a complex reaction, with numerous paths and components, and antioxidant action can occur via a multitude of mechanisms, especially when different phases coexist in the lipid-based formulation. Yet, correctly combining selectedin vitromethods to extract complementary information with respect to antioxidant behaviors would be much better and will help reducing the gap between prediction and efficacy in the finished product. Thus, we hereby propose a methodology to evaluate the antioxidant properties of 12 selected natural polyphenolic extracts based on the appropriate combination ofin vitroassays. The scores obtained with the DPPH test, the measure of the chelation capacity, the evaluation of antioxidants efficiency in emulsions (CAT and VESICAT assays), or in bulk oils were submitted to a statistical treatment. This analysis allowed a ranking on their global antioxidant capacities and the creation of clusters depending on their mechanisms of action and the type of media where the tests were performed.

Malaria, babesiosis, trypanosomosis, and leishmaniasis are some of the most life-threatening parasites, but the range of drugs to treat them is limited. An effective, safe, and low-cost drug with a large activity spectrum is urgently needed. For this purpose, an aryl amino alcohol derivative called Alsinol was resynthesized, screened in silico, and tested against Plasmodium, Babesia, Trypanosoma, and Leishmania. In silico Alsinol follows the Lipinski and Ghose rules. In vitro it had schizontocidal activity against Plasmodium falciparum and was able to inhibit gametocytogenesis; it was particularly active against late gametocytes. In malaria-infected mice, it showed a dose-dependent activity similar to chloroquine. It demonstrated a similar level of activity to reference compounds against Babesia divergens, and against promastigotes, and amastigotes stages of Leishmania in vitro. It inhibited the in vitro growth of two African animal strains of Trypanosoma but was ineffective in vivo in our experimental conditions. It showed moderate toxicity in J774A1 and Vero cell models. The study demonstrated that Alsinol has a large spectrum of activity and is potentially affordable to produce. Nevertheless, challenges remain in the process of scaling up synthesis, creating a suitable clinical formulation, and determining the safety margin in preclinical models. Departamento Administrativo de Ciencia, Tecnología e Innovación (COLCIENCIAS) Revisión por pares

The discovery and deployment of new broad-spectrum resistance (R) genes from cultivated rice and its wild relatives is a strategy to broaden the genetic basis of modern rice cultivars to combat rice blast disease. Oryza glaberrima possessing many valuable traits for tolerance to biotic and abiotic stresses, is an elite gene pool for improvement of Asian cultivated rice. An introgression line IL106 derived from O. glaberrima (Acc. IRGC100137) confers complete resistance to Magnaporthe oryzae in blast nursery. Genetic analysis using 2185 BC6F2 progenies derived from a cross between IL106 and the recurrent parent Dianjingyou 1 showed that IL106 harbors a single dominant resistance gene against M. oryzae strain 09BSH-10-5A. This gene was preliminarily mapped on the long arm of chromosome 6 of rice in a region of ca. 0.9 cM delimited by two SSR markers (RM20650 and RM20701). In order to finely map this gene, 17,100 additional progenies were further analyzed. As a result, this gene was further narrowed down to a region flanked by two molecular markers STS69-15 and STS69-7, and co-segregated with 3 molecular markers, RM20676, STS69-21 and STS69-22 on the long arm of chromosome 6. Based on reference genome sequences, this R gene was mapped in silico in 76.1-Kb and 67.7-Kb physical intervals, and containing 4 and 3 NBS-LRR candidate genes in O. sativa cultivar Nipponbare and O. glaberrima cultivar CG14, respectively. Because no blast resistance gene was finely mapped in this physical interval before, this R gene was considered as not described yet and designated as Pi69(t), which is the first identified and finely mapped blast R gene from O. glaberrima, as far as we know. Evaluation of IL106 with 151 blast strains collected from 6 countries in Asia showed that 148 strains are avirulent on IL106, suggesting that Pi69(t) is a broad-spectrum blast R gene, and a promising resistant resource for improvement of Asian cultivated rice.

Plant bioactive extracts represent a major resource for identifying drugs and adjuvant therapy for type 2 diabetes. To promote early screening of plants’ antidiabetic potential, we designed a four in vitro tests strategy to anticipate in vivo bioactivity. Two antidiabetic plants were studied: Ocimum gratissimum L. (Oc) leaf extract and Musanga cecropoides R. Br. ex Tedlie (Mu) stem bark extract. Chemical compositions were analyzed by LCMS and HPLC. Antidiabetic properties were measured based on (1) INS-1 cells for insulin secretion, (2) L6 myoblast cells for insulin sensitization (Glut-4 translocation), (3) L6 myoblast cells for protection against hydrogen peroxide (H2O2) oxidative stress (cell mortality), and (4) liver microsomial fraction for glucose-6-phosphastase activity (G6P). Oc extract increased insulin secretion and insulin sensitivity, whereas it decreased oxidative stress-induced cell mortality and G6P activity. Mu extract decreased insulin secretion and had no effect on insulin sensitivity or G6P activity, but it increased oxidative stress-induced cell mortality. Results were compared with NCRAE, an antidiabetic plant extract used as reference, previously characterized and reported with increased insulin secretion and insulin sensitivity, protection against oxidative stress, and decreased G6P activity. The proposed set of four in vitro tests combined with chemical analysis provided insight into the interest in rapid early screening of plant extract antidiabetic potential to anticipate pharmaco-toxicological in vivo effects.

Black coffee twig borer is a new but rapidly spreading insect pest of coffee in Uganda. Female beetles bore into primary branches/twigs and cultivate an ambrosia fungus for feeding their larvae. Thus, controlling the fungus means depriving the brood a source of food. Three fungicides, chlorothalonil (Glider), tebuconazole (Orius 25EW) and dimethomorph + mancozeb (Volar) were evaluated in vitro for their effectiveness in inhibiting mycelial growth of ambrosia fungus associated with the beetle. The pathogen was exposed to four concentrations (1.5x, 1.25x, 1.0x and 0.5x times the manufacturer recommended rate) incorporated into potato dextrose agar using inhibition and food poisoning techniques. The three fungicides inhibited fungal growth to some extent, even at the lowest concentration (0.5x) and percentage inhibition was significantly different (P≤0.05) from each other. Tebucozanole caused 100% growth inhibition irrespective of concentration and technique used while chlorothalonil and dimethomorph + mancozeb caused less than 40% inhibition for both techniques. Therefore, research should determine effectiveness of tebucozanole for suppressing fungal growth under field conditions for diminishing beetle incidence and fungal pathogenic effects in infested branches. This will pave way for integration of use of tebucozanole into overall Integrated Pest Management package (IPM) for the beetle in Uganda. Key words: Ambrosia-fungus, black-coffee-twig-borer, chlorothalonil, dimethomorph + mancozeb, fungicides, tebuconazole, Xylosandrus-compactus.

International audience; Among the most common plants used by fish farmers in Java, 18 were selected for deeper ethnobotanical and laboratory investigation into their antimicrobial activity. These plants are mainly used against fish diseases, but with no precise therapeutic indication, leading to low specificity of use. Leaves, which were the most commonly used part of plants, were generally placed directly in water or less frequently added to the feed. Disk diffusion tests showed that 15 plants of the 18 (83%) displayed varying degrees of antibacterial activity and that Streptococcus agalactiae was more sensitive than Aeromonas hydrophila. Crude ethanolic extracts (EE) were more active than water macerates (WM), and minimum inhibitory concentrations (MICs) ranged from 12.5 to 25 mg/mL. Only the EE of Piper betle had an inhibitory effect against A. hydrophila. No MIC was found for any of the WM studied here. Furthermore, both the EE and, to a lesser extent, the WM of active plants were able to reduce bacterial growth by more than 99%, even at doses below their MICs. These results suggest that these plant extracts have a potential for eco‐friendly prevention of bacterial fish diseases.

International audience; Bacterial wilt caused by Ralstonia solanacearum is one of the world’s most important soil-borne plant diseases. In Martinique, French West Indies, a highly virulent new pathogenic variant of this bacterium (phylotype IIB/4NPB) severely impacts tomato production. Here we report on the effect of R. solanacearum CFBP 6783, classified in phytotype IIB/4NPB, on tomato plantlets grown under strict in vitro culture conditions in the presence or absence of the arbuscular mycorrhizal fungus Rhizophagus irregularis MUCL 41833. A mycelium donor plant (i.e. Crotalaria spectabilis) was used for rapid, uniform mycorrhization of tomato plantlets that were subsequently infected by the bacterium. Bacterial wilt was significantly delayed and the incidence of the disease consequently reduced in the mycorrhizal tomato plantlets. Conversely, R. solanacearum did not affect root colonization by the AMF within the 16 days of the experiment. These results suggested that the mycorrhizal fungus was able to reduce bacterial wilt symptoms, probably by eliciting defence mechanisms in the plant.

Introduction Dans le cadre de la reconstruction du ligament croise anterieur au demi-tendineux quatre brins (DT4) se pose la question de la technique utilisee pour le creer. Nous proposons de comparer mecaniquement des greffes quatre brins obtenues avec les tendons des muscles ischio-jambiers, droit interne (DI) et demi-tendineux (DT), faufilees (F) et non faufilees (NF). Materiel et methodes Les tendons DI et DT sont preleves sur six sujets cadaveriques embaumes. Des greffes a quatre brins sont realisees : 12 DI et 12 DT. Les greffes du cote droit sont NF, celles du cote gauche sont F. Chaque greffe est testee selon le meme protocole experimental. La greffe est placee sur une machine de traction. Une pre-charge de 1 N lui est appliquee puis l’effort est amene a 50 N et maintenu pendant 30 s. Mille cycles de chargement-dechargement d’amplitude 200 N sont ensuite appliques a la frequence de 1 Hz. Enfin, la greffe est allongee jusqu’a rupture a la vitesse de 20 mm/min. L’evolution de l’allongement de la greffe au cours des cycles, la raideur avant rupture et la charge a rupture sont etudiees et comparees avec une etude statistique. Resultats Au 1000e cycle, l’allongement des greffes NF (moyenne : 4,20 mm ; DTNF : 3,51 mm) est superieur aux greffes F (moyenne : 3,44 mm ; DTF : 3,17 mm), mais ces differences ne sont pas statistiquement significatives. La raideur avant rupture pour les greffes NF (moyenne : 307 N/mm ; DTNF : 336 N/mm) est inferieure a celle des greffes F (moyenne : 322 N/mm ; DTF : 343 N/mm), mais cette difference n’est pas significative. La charge a rupture est statistiquement differente entre les greffes NF (moyenne : 843 N ; DTNF : 924 N) et F (moyenne : 1133 N ; DTF : 1263 N) (p > 0,01). Discussion Les tendons etant preleves sur sujets embaumes, leurs proprietes mecaniques sont alterees, neanmoins, cette limitation n’empeche pas la comparaison NF/F. L’absence de difference significative peut-etre expliquee par le nombre limite de sujets malgre l’utilisation des deux ischio-jambiers. Conclusion Au regard des resultats, la greffe faufilee semble preferable.

Deberdt, P., Perrin, B., Coranson-Beaudu, R., Duyck, P.-F., and Wicker, E. 2012. Effect of Allium fistulosum extract on Ralstonia solanacearum populations and tomato bacterial wilt. Plant Dis. 96:687-692. To control bacterial wilt (Ralstonia solanacearum, phylotype IIB/4NPB), the antimicrobial effect of Allium fistulosum aqueous extract was assessed as a preplant soil treatment. Three concentrations of extract (100, 50, and 25%, 1:1 [wt/vol]) were evaluated by in vitro inhibition assay and in vivo experiments in a growth chamber. In vitro, A. fistulosum (100 and 50%) suppressed growth of R. solanacearum. Preplant treatment of the soil with A. fistulosum extract significantly reduced the R. solanacearum populations. No pathogen was detected in the soil after treatment with 100% concentrated extract from the third day after application until the end of the experiment. A. fistulosum also significantly reduced the incidence of tomato bacterial wilt. In the untreated control, the disease affected 61% of the plants whereas, with 100 and 50% extracts, only 6 and 14% of the plants, respectively, were affected. These results suggest that A. fistulosum extracts could be used in biocontrol-based management strategies for bacterial wilt of tomato. Bacterial wilt caused by Ralstonia solanacearum occurs worldwide, affects hundreds of plant species, and is among the most widespread plant diseases in tropical and subtropical regions (17), including the West Indies. It affects both cash and subsistence

Decoction of Strychnopsis thouarsii is used in the Malagasy traditional medicine to combat malaria. We have shown that this traditional remedy prevents malaria infection by targeting Plasmodium at its early liver stage. Bioassay-guided fractionation of S. thouarsii stem barks extracts, using a rodent Plasmodium yoelii liver stage parasites inhibition assay, led to isolate the new morphinan alkaloid tazopsine (1) together with sinococuline (2) and two other new related morphinan analogs, 10-epi-tazopsine (3) and 10-epi-tazoside (4). Structures were characterized by 2D NMR, MS, and CD spectral analysis. Compounds 1-3 were found to fully inhibit the rodent P. yoelii liver stage parasites in vitro.

In this study, three strains of Trypanosoma cruzi were isolated at the same time and in the same endemic region in Mexico from a human patient with chronic chagasic cardiomyopathy (RyC-H); vector (Triatoma barberi) (RyC-V); and rodent reservoir (Peromyscus peromyscus) (RyC-R). The three strains were characterized by multilocus enzyme electrophoresis, random amplified polymorphic DNA, and by pathological profiles in experimental animals (biodemes). Based on the analysis of genetic markers the three parasite strains were typed as belonging to T. cruzi I major group, discrete typing unit 1. The pathological profile of RyC-H and RyC-V strains indicated medium virulence and low mortality and, accordingly, the strains should be considered as belonging to biodeme Type III. On the other hand, the parasites from RyC-R strain induced more severe inflammatory processes and high mortality (> 40%) and were considered as belonging to biodeme Type II. The relationship between genotypes and biological characteristics in T. cruzi strains is still debated and not clearly understood. An expert committee recommended in 1999 that Biodeme Type III would correspond to T. cruzi I group, whereas Biodeme Type II, to T. cruzi II group. Our findings suggest that, at least for Mexican isolates, this correlation does not stand and that biological characteristics such as pathogenicity and virulence could be determined by factors different from those identified in the genotypic characterization.

International audience; Here we characterized the effect of the ectomycorrhizal symbiosis on the genotypic and functional diversity of soil Pseudomonas fluorescens populations and analysed its possible consequences in terms of plant nutrition, development and health. - Sixty strains of P. fluorescens were isolated from the bulk soil of a forest nursery, the ectomycorrhizosphere and the ectomycorrhizas of the Douglas fir ( Pseudostuga menziesii ) seedlings- Laccaria bicolor S238N. They were characterized in vitro with the following criteria: ARDRA, phosphate solubilization, siderophore, HCN and AIA production, genes of N 2 -fixation and antibiotic synthesis, in vitro confrontation with a range of phytopathogenic and ectomycorrhizal fungi, effect on the Douglas fir– L. bicolor symbiosis. - For most of these criteria, we demonstrated that the ectomycorrhizosphere significantly structures the P. fluorescens populations and selects strains potentially beneficial to the symbiosis and to the plant. - This prompts us to propose the ectomycorrhizal symbiosis as a true microbial complex where multitrophic interactions take place. Moreover it underlines the fact that this symbiosis has an indirect positive effect on plant growth, via its selective pressure on bacterial communities, in addition to its known direct positive effect.

Objectives: To assess the relationship between the presence of DHFR and DHPS mutations in Plasmodium falciparum, parasite in vitro resistance, and in vivo efficacy of sulfadoxine-pyrimethamine (SP) treatment. Patients and methods: Measurement of SP treatment efficacy in malaria-infected children in Gabon was combined with in vitro tests of susceptibility to pyrimethamine and cycloguanil, and molecular genotyping at several DHFR and DHPS loci of parasites isolated before treatment. DHFR was studied at codons 108, 51, and 59, whereas DHPS gene was typed at positions 436, 437, 540 and 581. Results: SP treatment was effective in 86% of children by day 28. Seventy-five percent of isolates were in vitro resistant to pyrimethamine and 65.5% to cycloguanil. No mutation was detected at codons 540 and 581 of the DHPS gene. Most isolates (71.8%) presented with the triple mutant DHFR genotype, whereas 64.3% combined at least three DHFR and one DHPS mutations. The increase in the number of DHFR mutations was associated with an increase in in vitro resistance to pyrimethamine and cycloguanil; three DHFR mutations conferred pyrimethamine and to a lesser extent cycloguanil resistance. Treatment failures only occurred with isolates presenting at least two DHFR mutations (S108N and C59R) and one DHPS mutation (S436A or A437G), but SP treatment of infections with such parasites gave treatment success in 82.0% of children. Conclusions: DHFR mutations that lead to high-level in vitro resistance to pyrimethamine plus 1-2 DHPS mutations are not sufficient to induce in vivo failure of SP treatment in young children from Gabon.

PSA (Promastigote Surface Antigen) belongs to a family of membrane-bound and secreted proteins present in several Leishmania (L.) species. PSA is recognized by human Th1 cells and provides a high degree of protection in vaccinated mice. We evaluated humoral and cellular immune responses induced by a L. amazonensis PSA protein (LaPSA-38S) produced in a L. tarentolae expression system. This was done in individuals cured of cutaneous leishmaniasis due to L. major (CCLm) or L. braziliensis (CCLb) or visceral leishmaniasis due to L. donovani (CVLd) and in healthy individuals. Healthy individuals were subdivided into immune (HHR-Lm and HHR-Li: Healthy High Responders living in an endemic area for L. major or L. infantum infection) or non immune/naive individuals (HLR: Healthy Low Responders), depending on whether they produce high or low levels of IFN-γ in response to Leishmania soluble antigen. Low levels of total IgG antibodies to LaPSA-38S were detected in sera from the studied groups. Interestingly, LaPSA-38S induced specific and significant levels of IFN-γ, granzyme B and IL-10 in CCLm, HHR-Lm and HHR-Li groups, with HHR-Li group producing TNF-α in more. No significant cytokine response was observed in individuals immune to L. braziliensis or L. donovani infection. Phenotypic analysis showed a significant increase in CD4+ T cells producing IFN-γ after LaPSA-38S stimulation, in CCLm. A high positive correlation was observed between the percentage of IFN-γ-producing CD4+ T cells and the released IFN-γ. We showed that the LaPSA-38S protein was able to induce a mixed Th1 and Th2/Treg cytokine response in individuals with immunity to L. major or L. infantum infection indicating that it may be exploited as a vaccine candidate. We also showed, to our knowledge for the first time, the capacity of Leishmania PSA protein to induce granzyme B production in humans with immunity to L. major and L. infantum infection. This study received financial support from EU's Seventh Framework Programme (FP7) (RAPSODI project, grant agreement number 223341). Sí

AnaEE-France research infrastructure services for experimental studies on soil biodiversity and associated ecological functions. 1. Global Soil Biodiversity Conference : Assessing Soil Biodiversity and its Role for Ecosystem Services

Nickel (Ni)-tolerant ectomycorrhizal Pisolithus albus was isolated from extreme ultramafic soils that are naturally rich in heavy metals. This study aimed to identify the specific molecular mechanisms associated with the response of P. albus to nickel. In presence of high concentration of nickel, P. albus Ni-tolerant isolate showed a low basal accumulation of nickel in its fungal tissues and was able to perform a metal efflux mechanism. Three genes putatively involved in metal efflux were identified from the P. albus transcriptome, and their overexpression was confirmed in the mycelium that was cultivated in vitro in the presence of nickel and in fungal tissues that were sampled in situ. Cloning these genes in yeast provided significant advantages in terms of nickel tolerance (+ 31% Ni EC50) and growth (+ 83% u) compared with controls. Furthermore, nickel efflux was also detected in the transformed yeast cells. Protein sequence analysis indicated that the genes encoded a P-type-ATPase, an ABC transporter and a major facilitator superfamily permease (MFS). This study sheds light on a global mechanism of metal efflux by P. albus cells that supports nickel tolerance. These specific responses to nickel might contribute to the fungal adaptation in ultramafic soil.

La szaibélyite, hydroxypyroborate de formule Mg2B2O5 H2O, possède des propriétés analogues à celles des minéraux fibreux regroupés sous le terme d’amiante (caractéristiques structurales, physicochimiques et thermiques, en particulier sa possible décomposition avec libération d’eau par élévation de température). Elle pourrait donc servir, comme l’amiante, de retardateur de flamme. La question qui se pose, avant de l’envisager comme substitut de l’amiante (on connaît les problèmes de santé publique posés), est de conclure à sa non nocivité. La réelle preuve demande une étude de grande ampleur in vivo avec des rats. Cela dépasse nos capacités et, dans le présent travail, nous nous sommes limités à des essais préliminaires de solubilité in vitro dans un milieu pulmonaire analogue. Les fibres proviennent d’échantillons de skarns (Roumanie) à calcite + szaibélyite (+ d’autres minéraux d’abondance relative mineure), après ou non lixiviation à l’acide. Les observations aux MEB montrent que les fibres ont une longueur de quelques à plus d’une dizaine de µ (maximum 200 µ) ; leur diamètre va de 0,2 à quelques µ (maximum 15 µ) ; surface spécifique : 2 m2 par g (BET). Les caractéristiques thermiques ont été évaluées à l’aide des techniques DTA, DTG et TGA ; on note en particulier la présence d’un pic marqué de décomposition autour de T = 650°C. Le milieu pulmonaire analogue est défini par les composés suivants : CaCl2, NH4Cl, NaCl, NaHCO3, Na2HPO4, Na2SO4 10H2O, (Na3Citrate)5 5H2O, glycocolle. 0,5 g de fibres sont mises dans un litre de solution, en système fermé, ou ouvert avec balayage par un mélange gazeux N-CO2 à la température de 37°. Durée des expériences : quelques heures à quelques jours. On surveille les teneurs en Magnésium et Bore dans la solution au cours du temps. Résultats : en quelques jours on observe une diminution de quelques 10 à 30 % du volume de fibres initial ; on peut s’attendre à une dissolution complète en quelques semaines. Ce résultat est bien meilleur que pour les amiantes s.l. Les indices KNB et KI montrent a priori que la szaibélyite est très biosoluble et non biopersistante. Les résultats n’ont pas été interprétés quantitativement en termes de constante cinétique de dissolution, ni discutés pour leur pertinence (le ratio entre fibres et solution pleurale en milieu pulmonaire est-il comparable ?). Résumé de travaux inédits conduits à l’Ecole des Mines de Saint-Etienne (années 1997-2000) sous la direction de B.G. par les étudiants en stage : Nathalie Debouzy, Loreta Dumitrescu, Rémi Girodon, Florence Grillet, Caroline Horber, Moussa Manaa et Claire Rambeau. Remerciements à Blandine Albert, Essaïd Bilal, Clare Collier, Stefan Marincea, Jacques Moutte, Ciprian Teleman et Olivier Valfort. Soutien de l’ANVAR, de l’Ecole des Mines et du Geological Survey of Romania.

Nous avons étudié une collection de 76 nouveaux isolats bactériens de nodules obtenus par piégeage sur plantes d'Acacia tortilis subsp. raddiana in vitro, inoculées avec des suspensions de sols prélevés dans divers sites au nord (Tunisie) et au sud (Sénégal, Mauritanie) du Sahara, en comparaison avec 54 souches de références. L'analyse des nouveaux isolats par RAPD et SDS-PAGE a montré une grande diversité taxonomique parmi les isolats d'A. tortilis subsp. raddiana. En effet, bien que toutes les souches puissent être qualifiées de « rhizobium à croissance rapide », elles se répartissent en au moins 9 groupes différents, dont plusieurs ne correspondent à aucun des groupes de rhizobia déjà décrits. Aucune relation entre la position taxonomique des souches, leur origine géographique, les conditions édaphiques ou la profondeur d'isolement n'a pu être établie. La caractérisation symbiotique des souches d'A. tortilis subsp. raddiana a, au contraire, montré une grande homogénéité des caractéristiques symbiotiques majeures. Toutes les souches nodulent également A. Senegal, Prosopis julifora et Leucaena leucocephala, qui constituent avec A. tortilis subsp. raddiana un groupe d'inoculation. L'analyse des profils chromatographiques des facteurs Nod (FN) d'une quinzaine de souches réparties dans les différents groupes taxonomiques montre qu'ils sont très similaires. L'homogénéité structurale des FN des souches d'A. tortilis subsp. raddiana a été confirmée par la détermination de la structure des FN de quelques souches représentatives. Cette étroite relation entre FN et plante d'isolement/spectre d'hôte indépendamment de la taxonomie des souches suggère l'existence d'une forte pression de sélection de la part de la plante hôte et confirme nos observations précédentes, à savoir que les facteurs Nod sont de bons indicateurs du potentiel de nodulation d'un rhizobium donné. We studied a collection of 76 new bacterial isolates from nodules developed on Acacia tortilis subsp. raddiana plants inoculated in vitro with soil suspensions harvested in different sites in North (Tunisia) and South (Senegal, Mauritania) parts of Sahara. RAPD and SDS-PAGE analyses in comparison with 54 reference strains from diverse origins showed great taxonomie diversity among the new isolates. All of them are fast-growing and form at least 9 different groups, several of which could correspond to undescribed groups. No relationship could be established between taxonomie groups, geographical origin (North/South of Sahara) of the strains, edaphic conditions, or depth of soil sampling (down to - 32 m). Opposite to their taxonomie diversity, the new isolates share identical main symbiotic properties. All strains nodulating A. tortilis subsp. raddiana can also nodulate Prosopis juliflora, Acacia Senegal and Leucaena leucocephala, these four plants then constituting a cross-inoculation group. By thin-layer chromatography, we showed that 20 A. tortilis subsp. raddiana strains, representative of the different taxonomie groups, share very similar Nod factor (NF) profiles, independently from their geographical origins. NF structure homogeneity was confirmed by NF structure determination of some representative strains. The consistency between NF and plant of isolation/nodulation spectrum, independently of the taxonomie position of the strains suggests a high selection pressure from the plant, and confirms our previous observations that NF are good markers for predicting the nodulation potential of a given rhizobial strain.

The in vitro activities of dihydroartemisinin (the biologically active metabolite of artemisinin derivatives), chloroquine, monodesethylamodiaquine (the biologically active metabolite of amodiaquine), quinine, mefloquine, halofantrine, and pyrimethamine were assessed in 65 African isolates of #Plasmodium falciparum$ from Yaoundé, Cameroon using an isotopic microtest. The 50% inhibitory concentration (IC50) values for dihydroartemisinin were within a narrow range from 0.25 to 4.56 nM, with a geometric mean of 1.11 nM (95% confidence interval = 0.96-1.28 nM). Dihydroartemisinin was equally active (P is greater than 0.05) against the chloroquine-sensitive isolates (geometric mean IC50 = 1.25 nM, 95% confidence interval = 0.99-1.57 nM) and the chloroquine-resistant isolates (geometric mean IC50 = 0.979 nM, 95% confidence interval = 0.816-1.18 nM). A significant positive correlation was observed between the responses to dihydroartemisinin and mefloquine (r = 0.662) or halofantrine (r = 0.284), suggesting in vitro cross-resistance. There was no correlation between the responses to dihydroartemisinin and other antimalarial drugs. (Résumé d'auteur)

Etude de trois souches d'Arthrobotrys oligospora : Caractérisation biologique et effets sur Meloidogyne mayaguensis parasite de la tomate au Sénégal. Three strains (ORS 18690 S2, ORS 18691 S6 and ORS 18693 S5) of the nematophagous fungus Arthrobotrys oligospora have been isolated in Senegal for the first time. In vitro, two strains (ORS 18690 S2 and ORS 18693 S5) of them trapped 100 % and the other (ORS 18691 S5) 80 %> of 7-day-old juvenile Meloidogyne mayaguensis within 48h. Optimal growth occured at 25-30°C and at a pH 5.6, but salinity inhibited development. In order to test the ability of fungi to control M. mayaguensis in pots on tomato, the fungus was incorporated into compost blocks or in vermiculite before sowing or subsequent transplanting. In pot experiments both strains reduced nematode populations ans stimulated seedling growth. However, these effects were higher in compost blocks than in vermiculite.

Trente-cinq agents antimicrobiens différents ont été testés in vitro sur 42 souches de Mycobacterium farcinogenes, 13 de Mycobacterium senegalense et 17 de Nocardia farcinica sur un milieu solide avec extrait de levure glucosé. La plupart des souches étaient sensibles à l'amikacine (2 g/ml), à la doxycycline (8 g/ml), à l'érythromycine (64 g/ml), au sulfate de lividomycine (16 g/ml), à la minocycline (8 g/ml), à l'acide nalidixique (64 g/ml), au phosphate d'oléandomycine (64 g/ml), à l'oxytétracycline (64 g/ml), à la rifampicine (64 g/ml), au sulfate de streptomycine (64 g/ml) et à la vancomycine (64 g/ml). Une nette résistance aux médicaments antituberculeux a été observée dans la plupart des souches testées, bien que M. senegalense et N. farcinica aient montré beaucoup plus de résistance que M. farcinogenes. Les différentes réponses des médicaments aux agents antimicrobiens se sont avérées utiles pour différencier M. farcinogenes et M. Senegalense, ainsi que ces deux derniers et N. farcinica. Ceci pourrait avoir des conséquences thérapeutiques significatives dans le traitement du farcin.

Cowpea (Vigna unguiculata (L.) Walp.) is one of the most important grain legumes in sub-Saharian regions. It contributes to man food security by providing a protein-rich diet. However, its production is limited by abiotic stresses such as salinity. This study aims to evaluate the salt tolerance of 15 cowpea cultivars, at germination stage. The seed germination process consisted of sowing them in agarified water (8 g·L−1) supplemented with 6 different concentrations of NaCl (0, 10, 50, 100, 150, and 200 mM). Results highlighted that high salt concentrations drastically reduced germination and significantly delayed the process for all varieties. A cowpea varietal effect towards the salt tolerance was noticed. Genotypes Diongoma, 58-78, and 58-191 were more salt-tolerant cultivars while Mougne and Yacine were more salt-sensitive ones as confirmed in the three groups of the dendrogram. NaCl effects on the early vegetative growth of seedlings were assessed with a tolerant (58-191) and a susceptible (Yacine) cultivar. Morphological (length and dry biomass) and physiological (chlorophyll and proline contents) parameter measurements revealed a negative effect of high (NaCl). However, 58-191 was much more salt tolerant, and the chlorophyll and proline contents were higher than those of Yacine genotype at increasing salt concentrations.

In this study, we successfully cryopreserved cotyledonary somatic embryos of diploid and triploid Arachis pintoi cytotypes using the encapsulation-dehydration technique. The highest survival rates were obtained when somatic embryos were encapsulated in calcium alginate beads and precultured in agitated (80 rpm) liquid establishment medium (EM) with daily increasing sucrose concentration (0.50, 0.75, and 1.0 M). The encapsulated somatic embryos were then dehydrated with silica gel for 5 h to 20% moisture content (fresh weight basis) and cooled either rapidly (direct immersion in liquid nitrogen, LN) or slowly (1ºC min-1 from 25ºC to -30ºC followed by immersion in LN). Beads were kept in LN for a minimum of 1 h and then were rapidly rewarmed in a 30ºC water-bath for 2 min. Finally, encapsulated somatic embryos were post-cultured in agitated (80 rpm) liquid EM with daily decreasing sucrose concentration (0.75 and 0.5 M) and transferred to solidified EM. Using this protocol, we obtained 26% and 30% plant regeneration from cryopreserved somatic embryos of diploid and triploid cytotypes. No morphological abnormalities were observed in any of the plants regenerated from cryopreserved embryos and their genetic stability was confirmed with 10 isozyme systems and nine RAPD profiles. Fil: Rey, Hebe Yolanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste. Instituto de Botánica del Nordeste (i); Argentina; Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina; Fil: Faloci, Mirta Mabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste. Instituto de Botánica del Nordeste (i); Argentina; Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina; Fil: Medina, Ricardo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste. Instituto de Botánica del Nordeste (i); Argentina; Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina; Fil: Dolce, Natalia Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste. Instituto de Botánica del Nordeste (i); Argentina; Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina; Fil: Engelmann, Florent. Centre National de la Recherche Scientifique. Institut de Recherche Pour Le Developpement; Francia; Fil: Mroginski, Luis Amado. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste. Instituto de Botánica del Nordeste (i); Argentina; Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina

Ce mémoire a reçu un "Crelan Awards" dans la catégorie "masters bioingénieurs en sciences agronomiques" en langue française (cf. URL).; Master; La bactérie tellurique Ralstonia solanacearum, responsable du flétrissement bactérien de la tomate, constitue une contrainte biotique majeure dans les systèmes de cultures maraîchères en Martinique comme dans toute la ceinture tropicale et subtropicale humide. Dans le contexte actuel de réduction de l’usage des produits phytosanitaires, le développement de stratégies alternatives de gestion des bioagresseurs est indispensable pour permettre aux agriculteurs de continuer à produire en quantité et en qualité satisfaisantes. Dans cette optique, des études sur l’introduction de plantes dites de services en association ou en rotation avec la tomate pour la bioprotéger contre le flétrissement bactérien mobilisent un effort de recherche considérable. Un des objectifs majeur est de stimuler les communautés microbiennes de la rhizosphère et en particulier les champignons mycorhiziens à arbuscules (CMA) pour la bioprotection des plants de tomate contre le flétrissement bactérien. Les CMA, en symbiose mutualiste avec la majorité des plantes terrestres, favorisent la nutrition minérale et hydrique et augmentent la résistance aux stress abiotiques et biotiques des végétaux. Ils induisent un biocontrôle caractérisé par la réduction des symptômes des maladies provoquées principalement par les champignons, les nématodes et les bactéries phytopathogènes du sol. L’objectif de ce travail est de déterminer comment associer des plantes potentiellement mycorhizotrophes et biorégulatrices à la tomate pour la bioprotéger contre le flétrissement bactérien. Parmi les espèces candidates, celles du genre Allium spp et Crotalaria spp ont été étudiées comme plantes de services pour leurs potentiels biorégulateur et mycorhizotrophe. La stratégie expérimentale mise en oeuvre a consisté à évaluer dans des systèmes de culture in vitro associant une plantule de tomate et un CMA, le potentiel bioprotecteur des CMA contre R. solanacearum. Cette expérience principale repose sur deux expériences préliminaires qui ont permis d’évaluer d’une part le potentiel de C. spectabilis et A. fistulosum comme plantes-donneuses de mycorhizes pour la tomate et d’évaluer d’autre part la dynamique de colonisation des plants de tomate par un CMA. L’incidence du flétrissement au cours du temps a été mesurée selon deux échelles différentes (le pourcentage de plantes flétries (PWP) et l’échelle de He (DI)). Ces expériences ont été complétées par une expérience exploratoire, visant à étudier le comportement in vitro de C. spectabilis et A. fistulosum mycorhizés en présence de R. solanacearum. L’analyse in vitro, de la dynamique de colonisation des plants de tomates par un CMA, a permis de mettre en évidence une colonisation maximum des racines (20 ± 3%) par le CMA en 13 jours. De plus, les résultats obtenus in vitro, montre que la pré-mycorhization des plants de tomates par R. irregularis MUCL 41833 diminue l’incidence du flétrissement causé par R. solanacearum. Ces résultats s’expliquent par différents mécanismes qui peuvent être synergiques. Ainsi les hypothèses de compétition pour les sites de colonisations, d’induction de mécanismes de défense et de modifications de la composition biochimique des exsudats, ont été discutées, comme facteurs de bioprotection, induits par les CMA. Ces recherches s’insèrent comme une piste originale et porteuse quant à la manière d’aborder l’insertion des plantes de services mycorhizotrophes et à propriétés allélopathiques dans une rotation culturale. Les résultats obtenus permettent de prétendre à une maitrise plus fine du potentiel bioprotecteur des CMA contre le flétrissement bactérien de la tomate.

Two cryopreservation procedures using aluminium cryo-plates, termed V-Cryo-plate and D-Cryo-plate, were successfully developed for in vitro mat rush (Juncus decipiens Nakai) basal stem buds. Multiple stems induced in liquid MS medium containing 8.9 μM BA by roller culture were cut into small clumps, plated on solid MS medium and cultured for 1 week at 25°C. Clumps that had produced many buds were cold-hardened at 5°C for 1-2 months. The buds with basal stems were dissected from small clumps and precultured overnight at 25°C on solid MS medium containing 0.3 M sucrose. Precultured buds were placed on aluminium cryo-plates and embedded in calcium alginate gel. Osmoprotection was performed by immersing the cryo-plates for 30 min at 25°C in loading solution (2 M glycerol + 1.0 M sucrose). In the D-Cryo-plate procedure, the buds were dehydrated to 27-25% moisture content (fresh weight) by placing the cryo-plates in the air current of a laminar flow cabinet for 2 to 3 h. In the V-Cryo-plate procedure, buds were dehydrated by immersing the cryoplates in PVS2 vitrification solution for 40 min at 25°C. In both procedures, cooling was performed by placing the cryo-plates in uncapped cryotubes, which were immersed in liquid nitrogen. For rewarming, cryo-plates were immersed in medium with 1.0 M sucrose for 20 min at room temperature. Regrowth of cryopreserved buds of line Kitakei 2' using D-Cryoplate and V-Cryo-plate procedures, was 90% and 80%, respectively. The two procedures were applied to 20 additional mat rush lines. Using the V-Cryo-plate procedure resulted in regrowth ranging between 13.3 and 86.7%, with an average of 52.5%. The D-Cryo-plate led to regrowth ranging between 73.3 and 96.7%, with an average of 86.3%. The D-Cryo-plate procedure will facilitate cryostorage of mat rush germplasm.

Selon la FAO, deux milliards de personnes souffrent de la « faim cachée » qui correspond aux carences en micronutriments incluant la carence en vitamine A, principale cause de cécité dans le monde. Par ailleurs, une alimentation excédentaire est source de stress oxydant pour le corps, facteur de risque de nombreuses maladies non transmissibles comme le diabète et les maladies cardiovasculaires. Contenus dans les produits végétaux, certains caroténoïdes représentent une source indirecte de vitamine A et pourraient participer aux effets bénéfiques de la consommation de fruits et légumes via des propriétés antioxydantes. Dans cette thèse, le comportement in vitro des caroténoïdes et rétinoïdes pendant la digestion a été étudié, depuis leur libération de la matrice alimentaire jusqu'à leurs interactions avec les cellules intestinales. Mes recherches s'articulent autour de la micellisation : processus clé de l'absorption des caroténoïdes et rétinoïdes. Dans un premier temps, la stabilité et la bioaccessibilité en digestion in vitro (DIV) de ces composés purs ont été comparées à celles de leurs homologues issus d'un jus de carotte, d'épinards crus et cuits et d'une farine fortifiée. Dans un deuxième temps, un nouveau modèle d'oxydation de nano-émulsions intestinales à base de sels biliaires (SB) a été développé. Les relations entre structure des micelles mixtes, réactivité à l'oxydation et protection par des antioxydants ont été étudiées grâce notamment au suivi de la dégradation des acides gras. Enfin, des expériences préliminaires ont été menées sur la réactivité enzymatique de cellules coliques (Caco-2) à divers oxydants et aux micelles mixtes. Pendant la DIV, le β-carotène et palmitate de rétinyl purs étaient particulièrement instables contrairement à la lutéine et à l'acétate de rétinyl. De plus, la matrice alimentaire protège ces composés et favorise leur bioaccessibilité surtout lorsqu'elle est broyée ou cuite. En effet, les prétraitements ont un impact conséquent sur la libération des caroténoïdes et doivent être considérés comme des moyens sérieux pour optimiser les apports. Les nano-émulsions intestinales mises au point se composaient de micelles mixtes sphériques ou cylindriques, ces dernières étant les moins résistantes à l'oxydation. En effet, la distribution des molécules de SB doit être différente selon la morphologie de la micelle, impactant ainsi leur réactivité. Dans nos conditions expérimentales, l'AAPH fut le seul oxydant efficace. L'α-tocophérol et la lutéine ont significativement ralenti la dégradation des acides gras contrairement au β-carotène. Leur place au sein de la micelle (coeur ou bordure) pourrait expliquer ces observations. Enfin, aucun des oxydants testés n'a significativement modifié l'activité catalase des cellules Caco-2. En revanche, la mise en contact avec les micelles à base de SB ont significativement diminué l'activité des 3 enzymes suivies. L'effet était positivement corrélé à la concentration en SB dont la conjugaison fut un élément déterminant. Finalement, le β-carotène, la lutéine, et l'acétate de rétinyl présents dans ces micelles ont en partie rétabli l'activité de la catalase contrairement au rétinol, palmitate de rétinyl et à l'α-tocophérol.

The antimalarial action of methylene blue (MB) was first noted by Paul Ehrlich in the late 19th century. Although it has only sporadically been adopted as a serviceable drug, the resolution of its antimalarial action seems warranted, as it is currently used for the treatment of various methemoglobinemias. In this work we have used MB, and its analogues Azures A (AZA), B (AZB), C (AZC), and thionin (TH), as well as the oxazine Celestine blue (CB) and azine Phenosaphranin (PS). All MB analogues inhibit the growth of various strains of Plasmodium falciparum in culture with IC50s in the 2 x 10(-9)-1 x 10(-7) M range, with the rank order MB approximately AZAAZBAZCTHPSCB. The IC50s for a mammalian cell line were in the 3 x 10(-6)-4 x 10(-5) M range, and the rank order was TH approximately AZBAZA approximately PSAZC approximately CBMB. As MB could affect cell growth through the oxidation of NADPH, we tested the action of the various compounds on the hexose-monophosphate shunt activity. Appreciable activation of the shunt was observed at 1 x 10(-5) M in both cell types, thus accounting for inhibition of growth of mammalian cells but not of parasites. All compounds were found to complex with heme in a rank order similar to their antimalarial effect. It is therefore suggested that MB and its congeners act by preventing the polymerization of heme, which is produced during the digestion of host cell cytosol in the parasite food vacuole, into hemozoin. In this respect, these compounds seem to act similarly to the 4-aminoquinoline antimalarials. All compounds effectively suppressed the growth of P. vinckei petteri in vivo with IC50 in the 1.2-5.2 mg/kg range, and MB and AZB suppressed P. yoelii nigeriensis in the 9-11 mg/kg range (i.e. at doses similar to those of chloroquine). The potential toxicity of these compounds may restrict their clinical use, but their impressive antimalarial activities suggest that the phenothiazine structure could serve as a lead compound for further drug development.

Two strains (ORS 18692 S7 and ORS 18692 S5) of the nematophagous fungus Arthrobotrys oligospora have been isolated for the first time in Senegal. In vitro, both strains trapped 98% of groups of 7-day-old Meloidogyne mayaguensis juveniles within 48 h. Their optimal growth occurred at 25-30 C and at a pH of 5.6, but salinity inhibited their development. In order to test the fungi on M. mayaguensis in pots and in mixed populations of Meloidogyne spp. on tomato in the field, the fungus was incorporated into compost blocks before sowing and subsequent transplanting. In pot experiments during the cool season (28 C), both strains reduced nematode populations and stimulated seedling growth. In field trials during the hot season (35 C), both strains were efficient in reducing Meloidogyne populations. The feasibility of combining the use of compost blocks with the introduction of A. oligospora is discussed. Pour la premie re fois, deux souches du champignon ne matophage Arthrobotrys oligospora (souches ORS 18692 S5...

Dans le cadre de la promotion de systèmes de culture alternatifs à moindres intrants chimiques, des études sur l’introduction de plantes de service ayant vocation à réduire l’usage des pesticides et engrais azotés mobilisent un effort de recherche considérable. Dans le cadre de la production bananière pour laquelle la nuisibilité causée par les nématodes phytoparasites constitue l’une des principales contraintes, une des principales alternatives à l’utilisation de nématicides chimiques repose sur l’usage raisonné en rotation ou en association culturale de plantes de service à propriétés nématorégulatrices. Parmi les espèces candidates, celles du genre Crotalaria spp. ont largement été étudiées en raison de leur aptitude à contrôler les nématodes phytoparasites. Une méta-analyse révèle cependant que même si de nombreux auteurs s’accordent à reconnaître les propriétés allélopathiques des plantes de cette espèce, ils sont également enclins à reconnaître la forte variabilité d’expression de ces propriétés vis-àvis du contrôle des nématodes selon les situations expérimentées. L’objectif de ce travail repose sur la compréhension du déterminisme biochimique des exsudats.[br/] La stratégie expérimentale mise en oeuvre a consisté à tester en conditions contrôlées le comportement biochimique des exsudats racinaires de deux espèces de crotalaires (C. retusa, C. spectabilis) et d’une espèce sensible, le bananier dessert (Musa spp.) en fonction i) de conditions biotiques variées - i.e. présence ou non de nématodes (Radopholus similis et Meloidogyne arenaria), et présence ou non de mycorhizes-, ii) de conditions abiotiques contrastées – i.e. utilisation d’un sol naturel vs. substrat neutre, ajout ou non d’engrais chimique. La méthodologie innovante des « plantes à traire » (Brevet INRA) a été requise pour pouvoir travailler sur les exsudats racinaires des plantes conditionnées plutôt que sur des lixiviats, comme réalisé dans des expérimentations antérieures. [br/] Les résultats obtenus en spectrophométrie visible montrent que la présence ou l’absence de nématodes affecte de manière importante les profils biochimiques des plantes étudiées. En présence du nématode M. arenaria les profils biochimiques des deux crotalaires sont également modifiés. Des biotests réalisés à l’aide des exsudats racinaires conditionnés au préalable par les modifications biotiques ont permis de démontrer l’effet nématostatique des crotalaires. Des analyses biochimiques en GC-MS mettent en évidence de manière plus précise des molécules actives mises en jeu sous différentes modalités biotiques et abiotiques. [br/] Ces recherches, de portée générique, permettront de mieux comprendre les conditions de l’environnement qui régulent les profils biochimiques des exsudats racinaires des crotalaires et par extension, seront applicables aux recherches menées sur d’autres plantes de service candidates. Ceci aidera, à terme, à définir les conditions environnementales à réunir, pour orienter de manière optimale les effets allélopathiques dans une perspective de nématorégulation naturelle., As part of the promotion of alternative cropping systems at lower chemical inputs, studies on the introduction of cover crops designed to reduce the use of pesticides and nitrogen fertilizers mobilize a considerable research effort. Under banana production for which the harmfulness caused by plant-parasitic-nematodes is one of the main constraints, one of the main alternatives to the use of chemical nematicides is based on the rational use in rotation or intercropping of cover crops to protect crops from plant-parasitic-nematodes. [br/] Among the candidate species, the species from the genus of Crotalaria spp. have been widely studied because of their ability to control plant parasitic-nematodes. A meta-analysis reveals that although many authors agree allelopathic properties of these species, they are also likely to recognize the high variability of expression of these properties vis-à-vis the control of nematodes according to the situations experienced. The objective of this work is focused on the understanding of the biochemical determinism of root exudates under environment biotic and abiotic variations. [br/] The experimental strategy was to test under controlled conditions the biochemical behavior of root exudates of two species of crotalarias (C. retusa, C. spectabilis) and a susceptible species, the dessert banana crop (Musa spp.) with i) modification of biotic conditions - ie presence or absence of nematodes (Meloidogyne arenaria and Radopholus similis), and presence or absence of mycorrhiza- ii) contrasting abiotic conditions - ie using a natural soil vs. neutral substrate, whether to add fertilizer. The innovative methodology of "milking plants" (Patent INRA) was required in order to work on the root exudates of plants packed rather than leachate, as done in previous experiments.[br/] The results obtained in visible spectrophotometry show that the presence or absence of nematodes affect significantly the biochemical profiles of the studied plants. In the presence of nematode M. arenaria biochemical profiles of both crotalarias are also changed. Bioassays conducted using root exudates conditioned in advance by biotic changes have demonstrated the effect of nématostatique crotalarias. Biochemical GC-MS demonstrate more clearly active molecules that are involved in various biotic and abiotic conditions. [br/] This research, generic scope, to better understand the environmental conditions that regulate biochemical profiles of root exudates crotalarias and by extension, will apply to research on other plants service candidates. This will ultimately define the environmental conditions to meet, to guide optimal allelopathic effects from the perspective of natural nématorégulation.

Background Sexual reproduction is common in eukaryotic microorganisms, with few species reproducing exclusively asexually. However, in some organisms, such as fungi, asexual reproduction alternates with episodic sexual reproduction events. Fungi are thus appropriate organisms for studies of the reasons for the selection of sexuality or clonality and of the mechanisms underlying this selection. Magnaporthe oryzae, an Ascomycete causing blast disease on rice, reproduces mostly asexually in natura. Sexual reproduction is possible in vitro and requires (i) two strains of opposite mating types including (ii) at least one female-fertile strain (i.e. a strain able to produce perithecia, the female organs in which meiosis occurs). Female-fertile strains are found only in limited areas of Asia, in which evidence for contemporary recombination has recently been obtained. We induced the forced evolution of four Chinese female-fertile strains in vitro by the weekly transfer of asexual spores (conidia) between Petri dishes. We aimed to determine whether female fertility was rapidly lost in the absence of sexual reproduction and whether this loss was controlled genetically or epigenetically. Results All the strains became female-sterile after 10 to 19 rounds of selection under asexual conditions. As no single-spore isolation was carried out, the observed decrease in the production of perithecia reflected the emergence and the invasion of female-sterile mutants. The female-sterile phenotype segregated in the offspring of crosses between female-sterile evolved strains and female-fertile wild-type strains. This segregation was maintained in the second generation in backcrosses. Female-sterile evolved strains were subjected to several stresses, but none induced the restoration of female fertility. This loss of fertility was therefore probably due to genetic rather than epigenetic mechanisms. In competition experiments, female-sterile mutants produced similar numbers of viable conidia to wild-type strains, but released them more efficiently. This advantage may account for the invasion of our populations by female-sterile mutants. Conclusions We show for the first time that, in the absence of sexual reproduction, female-sterile mutants of M. oryzae rice strains can arise and increase in abundance in asexual generations. This change in phenotype was frequent and probably caused by mutation. These results suggest that female fertility may have been lost rapidly during the dispersion of the fungus from Asia to the rest of the world.

The semi-automated erythrocyte cellular antioxidant activity (ERYCA) method and Hydrophilic Oxygen Radical Absorbance Capacity (H-ORAC) assay were used to assess the ability of aqueous acetone extracts from 34 common tropical fruits to delay hemolysis of human erythrocytes and loss of fluorescein fluorescence respectively, when exposed to free peroxyl radicals induced by 2,2'-azobis (2-amidinopropane hydrochloride) (AAPH). The ERYCA assay quantifies both direct scavenging of free radicals and cell-mediated antioxidant protection (Cell-MAP). Because the H-ORAC method only analyzes the direct scavenging of free radicals in the reaction medium, Cell-MAP was deduced from the results of both assays. The results showed that Cell-MAP of fruit extracts on erythrocytes is, on average 3 to 4 times more than protection from direct scavenging of free radicals in the surrounding medium. The Cell-MAP of fruit extracts ranged from 0 to almost 4500 ?mol quercetin equivalents (QE) per 100 g on fresh weight basis, with tropical highland berries, cashew apple, mamey and 3 guavas species scoring the highest. Correlation coefficients among ERYCA, H-ORAC, Cell-MAP and total polyphenolic (TPP) content (assessed using a modified method excluding non-phenolics interference) were very high (r2>0.88 for all correlations), even though numerous discrepancies existed between the results of different methods. The analysis of Cell-MAP sheds new light on antioxidant rich food. (Resume d'auteur)

Cryopreservation (liquid nitrogen, -196°C) is the only safe and cost-effective option for long-term -conservation of genetic resources of non-orthodox seed species. Cryopreservation protocols have been developed for various materials including seeds, dormant buds, cell suspensions, calli, apices, zygotic, and somatic embryos of numerous plant species. Zygotic embryos or embryonic axes of almost 100 different species and somatic embryos of almost 40 different species from both temperate and tropical climates, comprising crops, fruit, and forest trees as well as wild species, whose seeds displayed orthodox, intermediate, and recalcitrant storage characteristics, have been successfully cryopreserved. With zygotic embryos and embryonic axes, the desiccation technique has been used with the majority of the species tested, leading to highly variable survival and recovery after freezing, especially during earlier experiments. More recently, new cryopreservation techniques viz. encapsulation-dehydration and vitrification have been employed, leading to generally improved results. With somatic embryos, different cryopreservation methods have been used viz. desiccation, pre-growth-desiccation, encapsulation-dehydration, vitrification, encapsulation-vitrification, and droplet-vitrification. There are also a few examples of the utilisation of slow controlled freezing, which correspond to the earlier experiments performed with somatic embryos. The development and application of cryopreservation is significantly more advanced for somatic embryos, in comparison with zygotic embryos, mainly because of the different origin and characteristics of the species treated. In most cases, zygotic embryos originate from tropical, wild species, for which knowledge and techniques relevant to the development of cryopreservation protocols are limited, or even non-existent. By contrast, somatic embryos are generally produced from cultivated species, which have already been studied extensively and for which propagation techniques are already operational. A number of technical possibilities to explore exist in order to improve the development of cryopreservation protocols for zygotic embryos and embryonic axes. For both categories of materials, the utilisation of analytical techniques has proved to be extremely useful to assist in the development of cryopreservation protocols.