Production and Optimization of Pseudomonas fluorescens Biomass and Metabolites for Biocontrol of Strawberry Grey Mould

Wafaa M. Haggag, Mostafa Abo El Soud

Biotechnology and Genetic Engineering Unit, National Research Center, Dokki, Cairo, Egypt..
Department of Plant Pathology, National Research Centre, Dokki, Cairo, Egypt;.

DOI: 10.4236/ajps.2012.37101                

Abstract

Pseudomonas species have been widely studied as biological agents (BCAs) and it is alternative to the application of chemical fungicides. Our objective was to optimize nutritional and environmental conditions of the isolated Pseudomonas fluorescens fp-5 for biomass and metabolites production and to evaluate itsagainst the grey mould disease caused by Botrytis cinerea on strawberry plants under field conditions. Pseudomonas fluorescens, showed antagonistic properties, in vitro, against thepathogen Botrytiscinerea. Effect of the separated secondary metabolites on the fungal growth by broth dilution technique and antifungal activity by agar well diffusion technique was studied. Response surface methodology was used to investigate the effects of four fermentation parameters (pH, incubation time, carbon and nitrogen concentrations) on biomass and bioactivemetabolites [antibiotic phenazin and siderophore] production. Glycerol was found to be the best carbon source for improved biomass and metabolites production. Meanwhile, peptone and yeast extract were found to be the best nitrogen source. Analysis of each formulation revealed that glycerol oil at 0.01% the best oil used for protect P. fluorescens for 3 months Under natural condition, P. fluorescens formulation was effective in reducing B. cinerea disease in strawberry leaves and fruits. Pre-harvest treatment protected fruits from Botrytis post-harvest disease in comparing of fungicide. In addition, the obtained results showed that bacterial treatment significantly increased thegrowth parameters as well as dry weights and yield.

Keywords

Botrytis Bunch Rot; Pseudomonas fluorescens; Strawberry Blight

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Haggag, Wafaa, M. (2008). Isolation of bioactive antibiotic peptides from Bacillus brevis and Bacillus polymyxa against Botrytis grey mould in strawberry. Archives Journal of Phyto-pathology and Plant Protection(German) . 2008;41(7): 477 – 491
[2]	Zhang, H., L. Wang, Y. Dong, S. Jiang, J. Cao, and R.Meng (2007). Postharvest biological control ofgray mold decay of strawberry with Rhodotorulaglutinis. Biological Control 40: 287-292.
[3]	Williamson, B., B. Tudzynski, P. Tud-zynski, and J. A. L. van Kan (2007).Botrytis cinerea: the cause of grey mould disease. Molecular Plant Pathology 8: 561-580.
[4]	Sutton, J.C. (1995). Evaluation of micro-organisms for biocontrol: Botrytis cinereaand strawberry, a case study. In: Advances in Plant Pathology, eds. J. H. Andrews, and I. C. Tommerup, Academic
[5]	Press, San Diego, USA. [3]vanKan, J.A.L. (2006). Licensed to kill: the lifestyle of a necrotrophic plant pathogen. Trends in Plant Science11, 247-253.
[6]	Haggag, Wafaa, M., and Mohamed, H.A.A. (2007). Biotechnological Aspects of Microorganisms used in Plant Biological Control (Article). American-Eurasian Journal of Sustainable Agriculture, 1(1):7-12
[7]	Meyer, J. M. andAbdallah, M.A. (1978 ). The florescent pigment of Pseudo-monas fluorescens biosynthesis, purification and physical-chemical properties. J Gen Microbiol, vol. 107, pp 319-328.
[8]	Rachid, D. and Ahmed, B. (2005). Effect of iron and growth inhibitors on siderophores production by Pseudo-monas fluorescens. African J. Biotechnol, vol. 4, pp 697-702.
[9]	Voisard, C, Keel, C., Hass, D. andDefago, G. (1989). Cyanide production by Pseudomonas fluorescenssuppress helps black root rot of tobacco under gnotobiotic conditions, EMBO J,vol. 8, pp 351–358.
[10]	María, E. and D.M.E. de Villegas, (2007). “Biotechnological production of sidero-phores,” in Microbial Siderophores, vol.12, A. Varma, S. Chincholkar, Eds. Berlin, Heidelberg: Springer Verlag, pp. 219-231
[11]	Mohamed N., Lherminier J., Farmer M.J., Fromentin J., BenoN., Houot V., Milat M.L., Blein J.P., 2007 Defense responses in grapevine leaves against Botrytis cinerea induced by application of a Pythiumoligandrum strain or its elicitin, oligandrin, to roots. Phytopathology 97: 611–620.
[12]	Silva , G. and de Almeida, E . (2006). Production of Yellow-Green Fluorescent Pigment by Pseudomonas fluorescens.Brazilian Archives ofBiology and Technology. Vol.49, n. 3 : pp. 411-419.
[13]	Cohen, J.I.; Falconi, C. and Komen, J. (1998). Strategic decisions for agricultural biotechnology.Synthesis of four policy seminars. 38:1-11.
[14]	SEUK, C., PAULITA, T., BAKER, R. (1988). Attributes associate with increased bio-control activity of fluorescent Pseudomonads. J. Plant Pathol. 4(3): 218-225.