ISOLATION AND SCREENING OF ANTI-MICROBIAL COMPOUNDS PRODUCED BY FLUORESCENT PSEUDOMONADS AGAINST PV.  XANTHOMONAS AXONOPODIS  PUNICAE

Madhu S  Giri; Chidanand A Rabinal; Raju J; Jayalakshmi  K

doi:10.48165/

Authors

Madhu S Giri Central Coffee Research Institute, Coffee Research Station Post, Chikkamagaluru Disitrict - 577 117, Karnataka
Chidanand A Rabinal College of Agricultural Science and Applied Research, Bharatiya Engineering, Science and Technology 2 Innovation University, Gownivarpalli, Gorantla Mandal, Andhra Pradesh – 515 231
Raju J ICAR-Directorate of Onion Garlic Research, Rajgurunagar, Pune, Maharashtra- 410 505
Jayalakshmi K ICAR-Directorate of Onion Garlic Research, Rajgurunagar, Pune, Maharashtra- 410 505

DOI:

https://doi.org/10.48165/

Keywords:

Anti-microbial compounds, Bacterial blight,, Fluorescent, Pseudomonads

Abstract

Xanthomonas axonopodis pv. punicae is the most prevalent pathogen in majority of the pomegranate cultivating areas of India which causes bacterial blight. There is a need to develop ecologically friendly methods to prevent pomegranate production loss caused by this disease. The present study aimed to identify the most efficient native strains of antagonistic fluorescent pseudomonads against X. axonopodis pv. punicae, as well as the anti-microbial compounds produced by such efficient strains. Out of 99 colonies of fluorescent pseudomonads collected from different rhizosphere soils of pomegranate, the dual culture assay against X. axonopodis pv. punicae revealed that 83 colonies showed inhibitory effect while nine strains were found highly potent antagonists. In the present study, the bio-assay of crude cell free extracts of nine efficient strains, only five strains showed inhibition ofX. axonopodis pv. punicae culture. The crude cell free extract of these five strains were separated on a TLC plate which yielded four to six metabolites with different Rf values ranged from 0.23 to 0.86. Eluted portion of all the metabolites failed to show the inhibition activity exceptonly one metabolite produced by FP-64 with Rf value of 0.67 whichshowed an inhibition of 1.50 cmin diameter.

References

Chin-A-Woeng, T.F.C., G.V. Bloemberg and B.J.J. Lugtenberg, 2003: Phenazines and their role in biocontrol by Pseudomonas bacteria. New Phytol., 157: 503-23.

Dahiya, J.S., D.L. Woods and J.P. Jewari, 1988: Control of Rhizoctonia solani, causal agent of brown girdling root rot of rapeseed, by Pseudomonas fluorescens. Bot. Bull. Acad. Sin., 29: 135-142.

Dowling, D.N. and F. O’ Gara, 1994: Metabolites of Pseudomonas involved in biocontrol of plant disease. Trends Biotechnol., 12: 133-141.

Haas, D. and G. Defago, 2005: Biological control of soil borne pathogens by fluorescent pseudomonads. Nature Rev. Microbiol., AOP, Published online 10th March 2005, pp. 1-13.

Han, J.S., J.H. Cheng, T.M. Yoon, J. Song, A. Rajkarnikar, W.G. Kim, I.D. Yoo, Y.Y. Yang and J.W. Suh, 2005: Biological control agent of common scab disease by antagonistic strain Bacillus sp. sunhua. J. Appl. Microbiol., 99: 213-221.

Hingorani, M.K. and N.J. Singh, 1959: Xanthomonas punicae sp. Nov. on pomegranate (Punica granatum L.). Indian J. Agric. Sci., 29: 45-48.

Howell, C.R. and R.D. Stipanovic, 1979: Control of Rhizoctonia solani on cotton seedlings with Pseudomonasfluorescens and with an antibiotic produced by the bacterium. Phytopathology, 69: 480-482.

Howell, C.R. and R.D. Stipanovic, 1980: Suppression of Pythium ultimum – induced dampi ng - off of cotton seedlings by Pseudomonas fluorescens and its antibiotic pyoluteorin. Phytopathology, 70: 712-715. Notz, R., M. Maurhofer, U. Schnider-Keel, B. Duffy, D. Haas, G. Defagom, 2001: Biotic factors affecting expression of the 2,4-diacetylphloroglucinol biosynthesis gene phlA in Pseudomonas fluorescens strains CHA0 in the rhizosphere. Phytopathology, 91: 876-881.

Raaijmakers, J. M., R.F. Bonsall and D.M. Weller, 1999: Effect of population density of Pseudomonas fluorescens on production of 2,4- diacetyl-phloroglucinol in the rhizosphere of wheat. Phytopathology, 89: 470-475

Raaijmakers, J. M., M. Vlami and J.T. De Souza, 2002: Antibiotic production by bacterial biocontrol agents. Anton. Van Leeuw, 81: 537–47.

Shanahan, P., D.J. O’ Sullivan, P. Simpson, J.D. Glennon and F. O’Gara, 1992: Isolation of 2,4- diacetylphloroglucinol from a fluorescent p s eu do mo na d a n d i nve st i ga t i on of physiological parameters influencing its production. Appl. Environ. Microbiol., 58: 353-

Sharifi-Tehrani, A., M. Zala, A. Natsch, Y. Moenne- Loccoz and G. Defago. 1998: Biocontrol of soil- borne fungal plant diseases by 2,4- diacetylphloroglucinol producing fluorescent pseudomonads with different restriction profiles of amplified 16S rDNA. Eur. J. Plant Pathol., 104: 631-343.

Thamashow, L. S. and D.M. Weller, 1988: Role of phenazine antibiotic from Pseudomonas f luorescens i n biologi cal control of Gaeumanomyces graminis var. tritici. J. Bacteriol., 170: 3499-3508.

Thmashow, L. S. and D.M. Weller, 1995: Current concepts in the use of introduced bacteria for biological disease control: mechanisms and antifungal metabolites. In: Plant – Microbe Interactions, Vol.1, Edited by G. Stacy and N. Keen. Chapmanand Hall, New York, 187-235.

Vauterin, L., B. Haste, K. Kersters, and J. Swings, 1995: Reclassification of Xanthomonas. Int. J. Syst. Bacteriol., 45: 475-489.

Ve l u sa my, P. , J . E. I mm a n u e l a n d S . S . Gnanamanickam, 2006: Biological control of rice bacterial blight by plant-associated bacteria producing 2,4-diacetylphlrglucinol. Can. J. Microbiol., 52: 56-65.

Weller, D.M. and R.J. Cook, 1983: Suppression of take all of wheat by seed treatments with fluorescent Pseudomonds, Phytopathology, 73: 403.

Hu, H. B., Y.Q. Xu, F. Cheng, X.H. Zhang, and B. Hur, 2005: Isolation and characterization of a new Pseudomonas strain produced both phenazine 1-carboxylic acid and pyoluteorin. J. Microbiol. Biotechnol., 15: 86-90.

Huang, J., Y. Xu, H. Zhang, Y. Li, X. Huang and B. Re, 2009: Temperature-dependent expression of phzM and its regulatory genes lasI and ptsP in rhizosphere isolate Pseudomonas sp. Strain M185. Appl. Environ. Microbio., 75: 6568-80.

Jagdeesh, K.S., 2000 : Selection of rhizobactria antagonistic to Ralstonia solanacerum EF Smith causing bacterial wilt in tomato and their control mechanisms. Ph.D. Thesis, Univ. Agric.

Sci., Dharwad, Karnataka (India).

Keel, C., P.H. Wirthner, T.H. Oberhansli, C. Voisard,

D. Berger, D. Haas and G. Defago, 1990: Pseudomonads as antagonists of plant pathogens in the rhizosphere: role of the antibiotic 2,4-diacetylphloroglucinol in the suppression of black root rot of tobacco. Symbiosis, 9: 327-341.

Keel, C., Weller, D.M., A. Natsch, G. Defago, R.J. Cook, and L.S. Thomashow, 1996: Conservation of the 2,4-diacetylphloroglucinol biosynthesis locus among fluorescent Pseudomonas strains from diverse geographic locations. Appl. Environ. Microbiol., 62: 552-536.

Kraus, J. and J.E. Loper, 1992: Lack of evidence for a role of antifungal metabolite production by P. fluorescens Pf-5 in biological control of Pythium damping off of cucumber. Phytopathology, 82: 264-271.

Lindeberg, G.D., 1981: An antibiotic lethal to fungi.

Plant Dis., 65: 680-683.

Mallesh, S.B. 2008: Plant growth promoting rhizobacteria, their characterization and mechanisms in the suppression of soil borne pathogens of Coleus and Ashwagandha. Ph.D. Thesis, Univ. Agric. Sci., Dharwad, Karnataka (India).

Morrissey, J. P., U.F. Walsh, A. O’Donnell, Y. Moenne- Loccoz and F. O’Gara, 2002: Exploitation of genetically modified inoculants for industrial ecology applications. Anton. Van Leeuw, 81: 599-606.