Charaterization of non-ribosomial lipopeptides as biopesticides

Le premier objectif de ce travail a été d’étudier l’implication des lipopeptides de Bacillus spp. dans la colonisation de la rhizosphère de tomates. Alors que seules les souches produisant de la surfactine sont capables de coloniser un milieu synthétique, toutes les souches testées, colonisent la rhizosphère de tomates avec une plus ou moins bonne efficacité quelque soit leur propriété de production de lipopeptide(s). L’efficacité de la colonisation des racines de tomates est principalement espèce-dépendante. Ce n’est que quand une souche est déjà une bonne colonisatrice que la surfactine semble améliorer cette propriété. Le deuxième objectif a été de tester l’effet des lipopeptides surfactine et mycosubtiline contre le phytopathogène obligatoire de la laitue : Bremia lactucae. À l’échelle du laboratoire, la mycosubtiline à 100 mg/L réduit le pourcentage de plantes infestées de 70 %. La surfactine ne montre aucun effet contre le champignon. Un mélange de mycosubtiline et de surfactine à 50 mg/L chacun diminue le pourcentage de plantes infestées de 65 %. Il semble diminuer le nombre de spores par plante infestée alors que cette propriété n’est pas remarquée avec les autres traitements. L’utilisation de la mycosubtiline dans une serre de culture limite la maladie aux symptômes les moins sévères et protège les plantes saines d’une contamination croisée. L’action des lipopeptides dans la colonisation des racines par Bacillus spp. n’avait jamais été validée in situ. De même, c’est la première fois que l’activité des lipopeptides est testée contre un phytopathogène obligatoire.The first aim of this work was to study the role played by the lipopeptides of Bacillus spp. in the colonization of the tomato rhizosphere. While only the strains producing surfactin are able to colonize a synthetic agar medium, all the strains are able to colonize the rhizosphere of tomatoes with a more or less good efficiency, whatever the lipopeptide(s) they have the capability to produce. The efficiency of the colonization of the tomato rhizosphere is species-dependant. However, surfactine seems to improve the efficiency of only the good colonizing strains. The second aim of this thesis was to test the effect of surfactin and mycosubtilin against a biotrophic parasite of lettuce: Bremia lactucae. Used at 100 mg/L, mycosubtilin reduces the percentage of infested plants of 70 %. Surfactin does not have effect against the fungy. A mixture of mycosubtilin and surfactin at both 50 mg/L decreases the percentage of infested plants of 65 %. This mixture seems to reduce the number of spore per infested plant while this property was not found for the other treatments. The use of mycosubtilin in a greenhouse confines the disease to the lowest classes of severity and protects the healthy plants from a cross contamination.The efficiency of lipopeptides of Bacillus spp. in root colonization by these bacteria was never tested in situ before. Furthermore, this is the first time that the activity of lipopeptides is validated against an obligate phytopathogen

Caractérisation des lipopeptides d'origine non ribosomique comme biopesticides

Le premier objectif de ce travail a été d étudier l implication des lipopeptides de Bacillus spp. dans la colonisation de la rhizosphère de tomates. Alors que seules les souches produisant de la surfactine sont capables de coloniser un milieu synthétique, toutes les souches testées, colonisent la rhizosphère de tomates avec une plus ou moins bonne efficacité quelque soit leur propriété de production de lipopeptide(s). L efficacité de la colonisation des racines de tomates est principalement espèce-dépendante. Ce n est que quand une souche est déjà une bonne colonisatrice que la surfactine semble améliorer cette propriété. Le deuxième objectif a été de tester l effet des lipopeptides surfactine et mycosubtiline contre le phytopathogène obligatoire de la laitue : Bremia lactucae. À l échelle du laboratoire, la mycosubtiline à 100 mg/L réduit le pourcentage de plantes infestées de 70 %. La surfactine ne montre aucun effet contre le champignon. Un mélange de mycosubtiline et de surfactine à 50 mg/L chacun diminue le pourcentage de plantes infestées de 65 %. Il semble diminuer le nombre de spores par plante infestée alors que cette propriété n est pas remarquée avec les autres traitements. L utilisation de la mycosubtiline dans une serre de culture limite la maladie aux symptômes les moins sévères et protège les plantes saines d une contamination croisée. L action des lipopeptides dans la colonisation des racines par Bacillus spp. n avait jamais été validée in situ. De même, c est la première fois que l activité des lipopeptides est testée contre un phytopathogène obligatoire.The first aim of this work was to study the role played by the lipopeptides of Bacillus spp. in the colonization of the tomato rhizosphere. While only the strains producing surfactin are able to colonize a synthetic agar medium, all the strains are able to colonize the rhizosphere of tomatoes with a more or less good efficiency, whatever the lipopeptide(s) they have the capability to produce. The efficiency of the colonization of the tomato rhizosphere is species-dependant. However, surfactine seems to improve the efficiency of only the good colonizing strains. The second aim of this thesis was to test the effect of surfactin and mycosubtilin against a biotrophic parasite of lettuce: Bremia lactucae. Used at 100 mg/L, mycosubtilin reduces the percentage of infested plants of 70 %. Surfactin does not have effect against the fungy. A mixture of mycosubtilin and surfactin at both 50 mg/L decreases the percentage of infested plants of 65 %. This mixture seems to reduce the number of spore per infested plant while this property was not found for the other treatments. The use of mycosubtilin in a greenhouse confines the disease to the lowest classes of severity and protects the healthy plants from a cross contamination.The efficiency of lipopeptides of Bacillus spp. in root colonization by these bacteria was never tested in situ before. Furthermore, this is the first time that the activity of lipopeptides is validated against an obligate phytopathogen.LILLE1-Bib. Electronique (590099901) / SudocSudocFranceF

Biopesticides, a complementary and alternative approach to the use of agrochemicals. A review

Peer reviewe

Biofilm formation is determinant in tomato rhizosphere colonization by Bacillus velezensis FZB42

National audienceIn this work, the behavior in tomato rhizosphere of Bacillus velezensis FZB42 was analyzed taking into account the surfactin production, the use of tomato roots exudate as substrates, and the biofilm formation. B. velezensis FZB42 and B. amyloliquefaciens S499 have a similar capability to colonize tomato rhizosphere. Little difference in this colonization was observed with surfactin non producing B. velezensis FZB42 mutant strains. B. velezensis is able to grow in the presence of root exudate and used preferentially sucrose, maltose, glutamic, and malic acids as carbon sources. A mutant enable to produce exopolysaccharide (EPS-) was constructed to demonstrate the main importance of biofilm formation on rhizosphere colonization. This mutant had completely lost its ability to form biofilm whatever the substrate present in the culture medium and was unable to efficiently colonize tomato rhizosphere

Les lipopeptides de Bacillus sp. : des molécules à haut potentiel pour la lutte biologique contre les champignons phytopathogènes

International audienc

Mycosubtilin and surfactin are efficient, low ecotoxicity molecules for the biocontrol of lettuce downy mildew.

The use of surfactin and mycosubtilin as an eco-friendly alternative to control lettuce downy mildew caused by the obligate pathogen Bremia lactucae was investigated. Preliminary ecotoxicity evaluations obtained from three different tests revealed the rather low toxicity of these lipopeptides separately or in combination. The EC50 (concentration estimated to cause a 50 % response by the exposed test organisms) was about 100 mg L(-1) in Microtox assays and 6 mg L(-1) in Daphnia magna immobilization tests for mycosubtilin and 125 mg L(-1) and 25 mg L(-1) for surfactin, respectively. The toxicity of the mixture mycosubtilin/surfactin (1:1, w/w) was close to that obtained with mycosubtilin alone. In addition, the very low phytotoxic effect of these lipopeptides has been observed on germination and root growth of garden cress Lepidium sativum L. While a surfactin treatment did not influence the development of B. lactucae on lettuce plantlets, treatment with 100 mg L(-1) of mycosubtilin produced about seven times more healthy plantlets than the control samples, indicating that mycosubtilin strongly reduced the development of B. lactucae. The mixture mycosubtilin/surfactin (50:50 mg L(-1)) gave the same result on B. lactucae development as 100 mg L(-1) of mycosubtilin. The results of ecotoxicity as well as those obtained in biocontrol experiments indicated that the presence of surfactin enhances the biological activities of mycosubtilin. Mycosubtilin and surfactin were thus found to be efficient compounds against lettuce downy mildew, with low toxicity compared to the toxicity values of chemical pesticides. This is the first time that Bacillus lipopeptides have been tested in vivo against an obligate pathogen and that ecotoxic values have been given for surfactin and mycosubtilin

Novel biosurfactants with biocontrol activity and low ecotoxicity.

International audienc