[en] Phytopathogenic fungi affecting crop and post-harvested vegetables are a amajor threat to food production and food storage. To face these drawbacks, producers have become increasingly dependent on agrochemicals. However, intensive use of these compounds has led to the emergence of pathogen resistance and severe negative environmental impacts. There are also a number of plant diseases for which chemical solutions are ineffective or non-existent as well as an increasing demand by consumers for pesticide-free food. Thus, biological control through the use of natural antagonistic microorganisms has emerged as a promising alternative to chemical pesticides for more rational and safe crop management. RESULTS: The genome of the plant-associated B. amyloliquefaciens GA1 was sample sequenced. Several gene clusters involved in the synthesis of biocontrol agents were detected. Four gene clusters were shown to direct the synthesis of the cyclic lipopeptides surfactin, iturin A and fengycin as well as the iron-siderophore bacillibactin. Beside these non-ribosomaly synthetised peptides, three additional gene clusters directing the synthesis of the antibacterial polyketides macrolactin, bacillaene and difficidin were identified. Mass spectrometry analysis of culture supernatants led to the identification of these secondary metabolites, hence demonstrating that the corresponding biosynthetic gene clusters are functional in strain GA1. In addition, genes encoding enzymes involved in synthesis and export of the dipeptide antibiotic bacilysin were highlighted. However, only its chlorinated derivative, chlorotetaine, could be detected in culture supernatants. On the contrary, genes involved in ribosome-dependent synthesis of bacteriocin and other antibiotic peptides were not detected as compared to the reference strain B. amyloliquefaciens FZB42. CONCLUSION: The production of all of these antibiotic compounds highlights B. amyloliquefaciens GA1 as a good candidate for the development of biocontrol agents.
Ongena, Marc ; Université de Liège - ULiège > Gembloux Agro-Bio Tech > Gembloux Agro-Bio Tech
Halimi, Badre
Lara, Yannick ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Brans, Alain ; Université de Liège - ULiège > Centre d'ingénierie des protéines - GIGA-Management : Plate-forme production protéines
Joris, Bernard ; Université de Liège - ULiège > Département des sciences de la vie > Physiologie et génétique bactériennes - Centre d'ingénierie des protéines
Fickers, Patrick ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Language :
English
Title :
Bacillus amyloliquefaciens GA1 as a source of potent antibiotics and other secondary metabolites for biocontrol of plant pathogens.
Tournas, V., Spoilage of vegetable crops by bacteria and fungi and related health hazards (2005) Critical review in microbiology, 31, pp. 33-44
Lucy, M., Reed, E., Glick, B.R., Pplications of free living plant growth-promoting rhizobacteria (2004) Antonie Van Leeuwenhoek, 86, pp. A1-25
Somers, E., Vanderleyden, J., Srinivasan, M., Rhizosphere bacterial signalling: a love parade beneath our feet (2004) Critical review of microbiology, 30, pp. 205-240
Lugtenberg, B.J., Kamilova, F., Plant-Growth-Promoting Rhizobacteria (2009) Annual review of microbiology, 63, pp. 541-556. , 10.1146/annurev.micro.62.081307.162918, 19575558
Leclere, V., Bechet, M., Adam, A., Guez, J.S., Wathelet, B., Ongena, M., Thonart, P., Jacques, P., Mycosubtilin overproduction by Bacillus subtilis BBG100 enhances the organism's antagonistic and biocontrol activities (2005) Applied and environmental microbiology, 71, pp. 4577-4584. , 10.1128/AEM.71.8.4577-4584.2005, 1183317, 16085851
Chen, X., Scholz, R., Borriss, M., Junge, H., Mogel, G., Kunz, S., Borriss, R., Difficidin and bacilysin produced by plant-associated Bacillus amyloliquefaciens Dare efficient in controlling fire blight disease (2009) Journal of biotechnology, 140, pp. 38-44. , 10.1016/j.jbiotec.2008.10.015, 19061923
Senghor, A., Liang, W.L., Ho, W., Integrated control of Colletotrichum gloeosporioides on mango fruit in Taiwan by the combination of Bacillus subtilis and fruit bagging (2007) Biocontrol science and technology, 9, pp. 575-580
Yang, Z., Guo, H., Zhang, X., Study on the control of peach post-harvest diseases using Bacillus subtilis (2008) China Fruits, 23, pp. 35-38
Kotan, R., Dikbas, N., Bostan, H., Biological control of post harvest disease caused by Aspergillus flavus on stored lemon fruits (2009) African journal of biotechnology, 8, pp. 209-214
Favel, D., Commercialization and implementation of biocontrol (2005) Annual review of phytopathology, 43, pp. A337-359
Loper, J.E., Buyer, J.S., Siderophores in microbial interactions on plant-surfaces (1991) Molecular plant microbe interactions, 4, pp. 5-13
Stein, T., Bacillus subtilis antibiotics: structures, syntheses and specific functions (2005) Molecular microbiology, 56, pp. 845-857. , 10.1111/j.1365-2958.2005.04587.x, 15853875
Chen, X., Koumoutsi, A., Scholz, R., Eisenreich, A., Schneider, K., Heinemeyer, I., Morgenstern, B., Borriss, R., Comparative analysis of the complete genome sequence of the plant growth-promoting bacterium Bacillus amyloliquefaciens FZB42 (2007) Nature biotechnology, 25, pp. 1007-1014. , 10.1038/nbt1325, 17704766
Chen, X., Koumoutsi, A., Scholz, R., Schneider, K., Vater, J., Sussmuth, R., Piel, J., Borriss, R., Genome analysis of Bacillus amyloliquefaciens FZB42 reveals its potential for biocontrol of plant pathogens (2009) Journal of biotechnology, 140, pp. 27-37. , 10.1016/j.jbiotec.2008.10.011, 19041913
Toure, Y., Ongena, M., Jacques, P., Guiro, A., Thonart, P., Role of lipopeptides produced by Bacillus subtilis GA1 in the reduction of grey mould disease caused by Botrytis cinerea on apple (2004) Journal of applied microbiology, 96, pp. 1151-1160. , 10.1111/j.1365-2672.2004.02252.x, 15078533
Tsuge, K., Akiyama, T., Shoda, M., Cloning, sequencing, and characterization of the iturin A operon (2001) Journal of bacteriology, 183, pp. 6265-6273. , 10.1128/JB.183.21.6265-6273.2001, 100110, 11591669
Sutyak, K.E., Wirawan, R., Aroutcheva, A., Chikindas, M., Isolation of the Bacillus subtilis antimicrobial peptide subtilosin from the dairy product-derived Bacillus amyloliquefaciens (2008) Journal of applied microbiology, 104, pp. 1067-1074. , 10.1111/j.1365-2672.2007.03626.x, 2322860, 17976171
Chen, X., Vater, J., Piel, J., Franke, P., Scholz, R., Schneider, K., Koumoutsi, A., Borriss, R., Structural and functional characterization of three polyketide synthase gene clusters in Bacillus amyloliquefaciens FZB42 (2006) Journal of bacteriology, 188, pp. 4024-4036. , 10.1128/JB.00052-06, 1482889, 16707694
Schneider, K., Chen, X., Vater, J., Franke, P., Nicholson, G., Borriss, R., Sussmuth, R., Macrolactin is the polyketide biosynthesis product of the pks2 cluster of Bacillus amyloliquefaciens FZB42 (2007) Journal of natural product, 70, pp. 1417-1423
Rapp, C., Jung, G., Katzer, W., Loeffer, W., Chlorotetain from Bacillus subtilis, an antifungal dipeptide with an unusual chlorine containing amino acid (1988) Angewandte chemie interationnal edition, 12, pp. 1733-1734
Porwal, S., Lal, S., Cheema, S., Kalia, V.C., Phylogeny in aid of the present and novel microbial lineages: diversity in Bacillus (2009) PLoS One, 4, pp. e4438. , 10.1371/journal.pone.0004438, 2639701, 19212464
Zeigler, D., Gene sequences useful for predicting relatedness of whole genomes in bacteria (2003) International journal of systematic and evolutionary microbiology, 53, pp. 1893-1900. , 10.1099/ijs.0.02713-0, 14657120
Meier, J.L., Burkart, M.D., The chemical biology of modular biosynthetic enzymes (2009) Chemical society review, 38, pp. T2012-2045
Banat, I., Makkar, R., Cameotra, S., Potential commercial applications of microbial surfactants (2000) Applied microbiology and biotechnology, 53, pp. 495-508. , 10.1007/s002530051648, 10855707
Singh, P., Cameotra, S., Enhancement of metal bioremediation by use of microbial surfactants (2004) Biochemistry and Biophysical Research Communication, 319, pp. 291-297
Ron, E., Rosenberg, E., Natural roles of biosurfactants (2001) Environmental microbiology, 3, pp. 229-236. , 10.1046/j.1462-2920.2001.00190.x, 11359508
Mulligan, C., Environmental applications for biosurfactants (2005) Environmental pollution, 133, pp. 183-198. , 10.1016/j.envpol.2004.06.009, 15519450
Ongena, M., Jacques, P., Bacillus lipopeptides: versatile weapons for plant disease biocontrol (2008) Trends in microbiology, 16, pp. 115-125. , 10.1016/j.tim.2007.12.009, 18289856
Danhorn, T., Fuqua, C., Biofilm formation by plant-associated bacteria (2007) Annual review of microbiology, 61, pp. 401-422. , 10.1146/annurev.micro.61.080706.093316, 17506679
Kinsinger, R., Shirk, M., Fall, R., Rapid surface motility in Bacillus subtilis is dependent on extracellular surfactin and potassium ion (2003) Journal of bacteriology, 185, pp. 5627-5631. , 10.1128/JB.185.18.5627-5631.2003, 193742, 12949115
Bais, H., Fall, R., Vivanco, J.M., Biocontrol of Bacillus subtilis against infection of Arabidopsis roots by Pseudomonas syringae is facilitated by biofilm formation and surfactin production (2004) Plant physiology, 134, pp. 307-319. , 10.1104/pp.103.028712, 316310, 14684838
Hofemeister, J., Conrad, B., Adler, B., Hofemeister, B., Feesche, J., Kucheryava, N., Steinborn, G., Vater, J., Genetic analysis of the biosynthesis of non-ribosomal peptide- and polyketide-like antibiotics, iron uptake and biofilm formation by Bacillus subtilis A1/3 (2004) Molecular genetic and genomics, 272, pp. 363-378
Leclere, V., Marti, R., Bechet, M., Fickers, P., Jacques, P., The lipopeptides mycosubtilin and surfactin enhance spreading of Bacillus subtilis strains by their surface-active properties (2006) Archives of microbiology, 186, pp. 475-483. , 10.1007/s00203-006-0163-z, 16964493
Maget-Dana, R., Thimon, L., Peypoux, F., Ptak, M., Surfactin/iturin A interactions may explain the synergistic effect of surfactin on the biological properties of iturin A (1992) Biochimie, 74, pp. 1047-1051. , 10.1016/0300-9084(92)90002-V, 1292612
Asaka, O., Shoda, M., Biocontrol of Rhizoctonia solani damping-off of tomato with Bacillus subtilis RB14 (1996) Applied and environmental microbiology, 62, pp. 4081-4085. , 1388978, 16535440
Romero, D., de Vicente, A., Rakotoaly, R., Dufour, S., Veening, J., Arrebola, E., Cazorla, F., Perez-Garcia, A., The iturin and fengycin families of lipopeptides are key factors in antagonism of Bacillus subtilis toward Podosphaera fusca (2007) Molecular plant microbe interactions, 20, pp. 430-440. , 10.1094/MPMI-20-4-0430, 17427813
Ongena, M., Jourdan, E., Adam, A., Paquot, M., Brans, A., Joris, B., Arpigny, J.L., Thonart, P., Surfactin and fengycin lipopeptides of Bacillus subtilis as elicitors of induced systemic resistance in plants (2007) Environmental microbiology, 9, pp. 1084-1090. , 10.1111/j.1462-2920.2006.01202.x, 17359279
Conrath, U., Beckers, G., Flors, V., Garcia-Agustin, P., Jakab, G., Mauch, F., Newman, M., Mauch-Mani, B., Priming: getting ready for battle (2006) Molecular plant microbe interaction, 19, pp. 1062-1071
Jourdan, E., Henry, G., Duby, F., Dommes, J., Barthelemy, J.P., Thonart, P., Ongena, M., Insights into the defense-related events occurring in plant cells following perception of surfactin-type lipopeptide from Bacillus subtilis (2009) Molecular plant microbe interactions, 22, pp. 456-468. , 10.1094/MPMI-22-4-0456, 19271960
Kloepper, J., Leong, J., Teinzte, M., Schroth, M., Pseudomonas siderophore: a mechanism explaining disease-suppressive soil (1980) Current microbiology, 4, pp. 317-320
Wilson, K., Flor, J., Schwartz, R., Joshua, H., Smith, J., Pelak, B., Liesch, J.M., Hensens, O., Difficidin and oxydifficidin: novel broad spectrum antibacterial antibiotics produced by Bacillus subtilis. II. Isolation and physico-chemical characterization (1987) Journal of antibiotic (Tokyo), 40, pp. 1682-1691
Zimmerman, S., Schwartz, C., Monaghan, R., Pelak, B., Weissberger, B., Gilfillan, E., Mochales, S., Tejera, E., Difficidin and oxydifficidin: novel broad spectrum antibacterial antibiotics produced by Bacillus subtilis. I. Production, taxonomy and antibacterial activity (1987) Journal of antibiotic (Tokyo), 40, pp. 1677-1681
Zweerink, M., Edison, A., Difficidin and oxydifficidin: novel broad spectrum antibacterial antibiotics produced by Bacillus subtilis. III. Mode of action of difficidin (1987) Journal of antibiotic (Tokyo), 40, pp. 1692-1697
Gustafson, K., Roman, M., Fenical, W., The macrolactin, a novel class of antiviral and cytotoxic macrolides from a deep-sea marine bacterium (1989) Journal of the american chemical society, 111, pp. 7519-7524
Patel, P.S., Huang, S., Fisher, S., Pirnik, D., Aklonis, C., Dean, L., Meyers, E., Mayerl, F., Bacillaene, a novel inhibitor of procaryotic protein synthesis produced by Bacillus subtilis: production, taxonomy, isolation, physico-chemical characterization and biological activity (1995) Journal of antibiotic (Tokyo), 48, pp. 997-1003
Kenig, M., Abraham, E., Antimicrobial activities and antagonists of bacilysin and anticapsin (1976) Journal of general microbiology, 94, pp. 37-45
Chmara, H., Inhibition of glucosamine synthase by bacilysin and anticapsin (1985) Journal of general microbiology, 131, pp. 265-271
Righi, E., Giacomazzi, C.G., Bassetti, M., Bisio, F., Soro, O., McDermott, J.L., Varnier, O.E., Viscoli, C., Soft-tissue infection with Absidia corymbifera and kidney complications in an AIDS patient (2007) Medical Mycology, 45, pp. 637-640. , 10.1080/13693780701435358, 18033616
Phister, T.G., O'Sullivan, D.J., McKay, L., Identification of bacilysin, chlorotetaine, and iturin a produced by Bacillus sp. strain CS93 isolated from pozol, a Mexican fermented maize dough (2004) Applied and environmental microbiology, 70, pp. 631-634. , 10.1128/AEM.70.1.631-634.2004, 14711701, 321241
Yazgan, A., Ozcengiz, G., Marahiel, M., Tn10 insertional mutations of Bacillus subtilis that block the biosynthesis of bacilysin (2001) Biochimica biophysica Acta, 1518, pp. 87-94
Fickers, P., Leclere, V., Guez, J.S., Bechet, M., Coucheney, F., Joris, B., Jacques, P., Temperature dependence of mycosubtilin homologue production in Bacillus subtilis ATCC6633 (2008) Research in microbiology, 159, pp. 449-457. , 10.1016/j.resmic.2008.05.004, 18656330
Edgar, R., MUSCLE: multiple sequence alignment with high accuracy and high throughput (2004) Nucleic acids research, 32, pp. 1792-1797. , 10.1093/nar/gkh340, 390337, 15034147
Swofford, D., Waddell, P., Huelsenbeck, J., Foster, P., Lewis, P., Rogers, J., Bias in phylogenetic estimation and its relevance to the choice between parsimony and likelihood methods (2001) Systematic biology, 50, pp. 525-539. , 10.1080/106351501750435086, 12116651
Surzycki, S., A fast method to prepare random fragment sequencing libraries using a new procedure of DNA shearing by nebulization and electroporation (1990) Human molecular biology laboratory manual, 2003, pp. 100-106. , Oxford: Blackwell, Surzycki S
Landy, M., Warren, G.H., Rosenman, S.B., Colio, L.G., Bacillomycin: an antibiotic from Bacillus subtilis active against pathogenic fungi (1948) Proceedings of the Society for Experimental Biology and Medicine, 67, pp. 539-541
Fickers, P., Guez, J.S., Damblon, C., Leclere, V., Bechet, M., Jacques, P., Joris, B., High-level biosynthesis of the anteiso-C(17) isoform of the antibiotic mycosubtilin in Bacillus subtilis and characterization of its candidacidal activity (2009) Applied and environmrental microbiology, 75, pp. 4636-4640
Schwyn, B., Neiland, J.B., Universal chemical assay for the detection and determination of siderophores (1987) Analytical biochemistry, 160, pp. 47-56. , 10.1016/0003-2697(87)90612-9, 2952030
Miethke, M., Westers, H., Blom, E.J., Kuipers, O.P., Marahiel, M.A., Iron starvation triggers the stringent response and induces amino acid biosynthesis for bacillibactin production in Bacillus subtilis (2006) Journal of bacteriology, 188, pp. 8655-8657. , 10.1128/JB.01049-06, 1698241, 17012385
Phister, T., O'Sullivan, D., McKay, L., Identification of bacilysin, chlorotetain and iturin produced by Bacillus sp. Strain CS93 isolated from pozol, a Mexican fermented maize dough (2004) Applied and environmental microbiology, 70, pp. 631-634. , 10.1128/AEM.70.1.631-634.2004, 321241, 14711701