Abstract
We observed that 2,4-diacetylphloroglucinol (DAPG), a major antimicrobial metabolite produced by a rhizoplane bacterium Pseudomonas fluorescens ECO-001 inhibited mycelial growth of a damping-off phytopathogen Aphanomyces cochlioides AC-5 through inducing excessive branching and curling in the hyphae. This study aimed to unravel the mode of action of DAPG caused excessive branching, curling and growth inhibition of AC-5 hyphae by detecting localized changes in the cortical filamentous actin (F-actin) organization by rhodamine-conjugated phalloidin. Confocal laser scanning microscopic observations revealed that both living bacteria and DAPG severely disrupted the organization of F-actin in the A. cochlioides hyphae in a similar manner. Furthermore, an inhibitor of F-actin polymerization, latrunculin B also induced similar growth inhibition, excessive branching and caused disruption of F-actin in the AC-5 hyphae. Our results suggested that growth inhibition and excessive branching induced in A. cochlioides by DAPG is likely to be linked to the disruption of F-actin cytoskeleton in the affected hyphae. This is the first report on disruption of cytoskeleton of a eukaryotic A. cochlioides by a well-known biocontrol metabolite DAPG secreted from a prokaryotic bacterium ECO-001.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ayyadurai N, Naik PR, Rao MS, Kumar RS, Samrat SK, Manohar M, Sakthivel N (2006) Isolation and characterization of a novel banana rhizosphere bacterium as fungal antagonist and microbial adjuvant in micropropagation of banana. J Appl Microbiol 100:926–937
Bachewich C, Heath IB (1998) Radial F-actin arrays precede new hypha formation in Saprolegnia: implications for establishing polar growth and regulating tip morphogenesis. J Cell Sci 111:2005–2016
Bangera MG, Thomashow LS (1999) Characterization of a gene cluster for synthesis of the polyketide antibiotic 2, 4-diacetylphloroglucinol from Pseudomonas fluorescens Q2–87. J Bacteriol 181:3155–3163
Bender CL, Rangaswamy V (1999) Polyketide production by plant-associated Pseudomonads. Annu Rev Phytopathol 37:175–196
Brawley SH, Robinson KR (1985) Cytochalasin treatment disrupts the endogenous currents associated with cell polarization in fucoid zygotes: studies of the role of F-actin in embryogenesis. J Cell Biol 100:1173–1184
Brazelton JN, Pfeufer EE, Sweat TA, Brian B, Gardener M, Coenen C (2008) 2, 4-diacetylphloroglucinol alters plant root development. Mol Plant-Microbe Interact 21:1349–1358
Chin-A-Woeng TFC, Priester WD, Bij AJVD, Lugten-burg BJJ (1997) Description of the colonization of a gnotobiotic tomato rhizosphere by Pseudomonas fluorescens biocontrol strain WCS365, using scanning electron microscopy. Mol Plant-Microbe Interact 10:79–86
de Souza JT, Arnould C, Deulvot C, Lemanceau P, Gianinazzi-Pearson V, Raaijmakers JM (2003) Effect of 2, 4-diacetylphloroglucinol on Pythium: cellular responses and variation in sensitivity among propagules and species. Phytopathology 93:966–975
Dick MW (2001) The Peronosporomycetes. In: McLaughlin DJ, McLaughlin EG, Lemke PA (eds) The mycota VII, Part A. Springer-Verlag, Systematics and Evolution, Berlin, pp 39–72
Fiorentini C, Matarrese P, Fattorossi A, Donelli G (1996) Okadaic acid induces changes in the organization of F-actin in intestinal cells. Toxicon 34:937–945
Heath IB, Gupta G, Bai S (2000) Plasma membrane-adjacent actin filaments, but not microtubules, are essential for both polarization and hyphal tip morphogenesis in Saprolegnia ferax and Neurospora crassa. Fungal Genet Biol 30:45–62
Hogan DA, Kolter R (2002) Pseudomonas-Candida interactions: an ecological role for virulence factors. Science 296:2229–2232
Hülskamp M (2004) Plant trichomes: a model for cell differentiation. Nat Mol Cell Biol 5:471–480
Islam MT (2008a) Disruption of ultrastructure and cytoskeleton network is involved with biocontrol of damping-off pathogen Aphanomyces cochlioides by Lysobacter sp. SB-K88. Biol Control 46:312–321
Islam MT (2008b) Dynamic rearrangement of F-actin organization triggered by host-specific plant signal is linked to morphogenesis of Aphanomyces cochlioides zoospores. Cell Motil Cytoskeleton 65:553–562
Islam MT, Tahara S (2001) Chemotaxis of fungal zoospores, with special reference to Aphanomyces cochlioides. Biosci Biotechnol Biochem 65:1933–1948
Isnansetyo A, Cui L, Hiramatsu K, Kamei Y (2003) Antibacterial activity of 2, 4-diacetylphloroglucinol produced by Pseudomonas sp. AMSN isolated from a marine alga, against vancomycin-resistant Staphylococcus aureus. Int J Antimicrob Agents 22:545–547
Kamei Y, Isnansetyo A (2003) Lysis of methicillin-resistant Staphylococcus aureus by 2, 4-diacetylphloroglucinol produced by Pseudomonas sp. AMSN isolated from a marine alga. Int J Antimicrob Agents 21:71–74
Leira F, Cabado AG, Vieytes MR, Roman Y, Alfonso A, Botanas LM, Yasamoto T, Malaguti C, Rossini GP (2002) Characterization of F-actin depolymerization as a major toxic event induced by pectenotoxin-6 in neuroblastoma cells. Biochem Pharmacol 63:1979–1988
Marchand PA, Weller DM, Bonsall RF (2000) Convenient synthesis of 2, 4-diacetylphloroglucinol, a natural antibiotic involved in the control of take-all disease of wheat. J Agric Food Chem 48:1882–1887
Miller DD, de Rajuter NCA, Bisseling T, Emons AMC (1999) The role of actin in root hair morphogenesis: studies with lipochito-oligisaccharide as a growth stimulator and cytochalasin as an actin perturbing drug. Plant J 17:141–154
Peleg AY, Tampakakis E, Fuchs BB, Eliopoulos GM, Moellering RCJ, Mylonakis E (2008) Prokaryotic-eukaryotic interactions identified by using Caenorhabditis elegans. Proc Natl Acad Sci USA 105:14585–14590
Revenu C, Athman R, Robine S, Louvard D (2004) The co-workers of actin filaments: from cell structures to signals. Nat Rev Mol Cell Biol 5:635–646
Szymanski DB, Marks MD, Wick SM (1999) Organized F-actin is essential for normal trichome morphogenesis in Arabidopsis. Plant Cell 11:2331–2347
Takemoto D, Hardham AR (2004) The cytoskeleton as a regulator and target of biotic interactions in plants. Plant Physiol 136:3864–3878
Acknowledgments
We are thankful to Professor R. Yokosawa (Health Science University of Hokkaido, Hokkaido, Japan) for his kind gift of A. cochlioides (AC-5). Yuriko Aoyama, Abhinandan Deora and Toshiaki Ito of Hokkaido University, Japan deserve special thanks for their supports during CLSM, light microscopy and resizing micrographs, respectively. The financial support and a Postdoctoral Fellowship (to M.T.I) from the Japan Society for the Promotion of Science (JSPS), Japan is very much appreciated.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Islam, M.T., Fukushi, Y. Growth inhibition and excessive branching in Aphanomyces cochlioides induced by 2,4-diacetylphloroglucinol is linked to disruption of filamentous actin cytoskeleton in the hyphae. World J Microbiol Biotechnol 26, 1163–1170 (2010). https://doi.org/10.1007/s11274-009-0284-z
Received:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1007/s11274-009-0284-z