Isolation and identification of cucumber rhizospheric fluorescent pseudomonads and evaluation of their antagonistic potential as biocontrol agents

Document Type : Research Paper

Authors

1 Bu Ali Sina University

2 Department of Plant Protection, College of Agriculture, Bu-Ali Sina

Abstract

Fluorescent pseudomonads are important group of plant growth promoting bacteria that can improve the plant growth and restrict or suppress phytopathogens directly or indirectly. The aim of this study was to obtain fluorescent pseudomonads with efficient antagonistic activities. For performing this research, a total of 120 bacterial isolates belonging to Pseudomonas fluorescens and P. putida were isolated from different cucumber rhizospheric soil in Hamedan province. Then, the isolates were screened for their plant growth promoting traits and antibacterial activity. These isolates also were screened against Pseudomonas syringae pv. lachrymans. So, According to the results of in vitro studies, five antagonistic strains (BSU390, BSU630, BSU205, BSU161 and BSU391) with inhibition diameter >7 mm were selected. The range of inhibition diameter for these isolate was 7.66 to 22 mm. Also, during in vivo study, the strains were evaluated for their effect in suppressing disease development in terms of area under disease progress curve (AUDPC) and increasing biomass of cucumber. The results of greenhouse experiments revealed that strains BSU390, BSU205, and BSU630, showed high biocontrol efficacy. During the sole application, the strains significantly (P ≤ 0.05) increased plant height and dry matter compared to non-bacterized control. Hence, the study shows that these isolates have potential use in cucumber bioprotection, as PGPR or in an integrated disease management; However, their effectiveness under a variety of field conditions should be investigated.

 

Keywords


Ahemad, M. & Khan, M.S. 2012. Evaluation of plant growth promoting activities of rhizobacterium Pseudomonas putida under herbicide-stress. Annals Microbiology, 62: 1531-1540.
Alexander, D.B. & Zuberer, D.A. 1991. Use of chrome azurol S reagents to evaluate siderophore production by rhizosphere bacteria. Biology and Fertility of Soils, 2: 39-4.
Aliye, N., Fininsa, C. & Hiskias, Y. 2008. Evaluation of rhizosphere bacterial antagonists for their potential to bioprotect potato (Solanum tuberosum) against bacterial wilt (Ralstonia solanacearum). Biological Control. 47: 282-288.
Alstrom, S. & Burns, R.G. 1989. Cyanid production by rhizobacteria as a possible mechanism of plant growth inhibition. Biology and Fertility of Soils, 7: 232-238.
Arsenijevic, M., Obradovic, A., Stevanovic, D. & Ivanovic, M. 1998. Antagonistic effect of some saprophytic bacteria to Pseudomonas syringae pv. phaseolicola and Xanthomonas campestris pv. phaseoli. Biological Control of Fungal and Bacterial Plant Pathogens, IOBC Bulletin, 21 (9): 297-300.
Beneduzi, A., Ambrosini, A. & Passaglia, L.M.P. 2012. Plant growth-promoting rhizobacteria (PGPR): Their potential as antagonists and biocontrol agents. Genetics and Molecular Biology, 35, 4 (suppl): 1044-1051.
Bent, E. Tuzan, S. Chanway C.P. & Enebak, S. 2000. Alteration in plant growth and in root hormone levels of lodgeple pines inoculatied with rhizobacteria. Canadian Journal of Microbiology, 47: 793-800.
Bhattacharjee P.N. & Jha, D.K. 2012. Plant growth-promoting rhizobacteria (PGPR): emergence in agriculture. World Journal of Microbiology and Biotechnology, 28: 1327-1350.
Cappuccino, J.C. & Sherman, N. 1992. Microbiology: A Laboratory Manual. Third ed. Benjamin/Cummings Publishing Companies based in New York, pp. 125-179.
Carrim, A.J.I., Barbosa E.C. & Gonalves Vieira, J.D. 2006. Enzymatic activity of endophytic bacterial isolates of Jacaranda decurrens Cham. (Carobinha-docampo). Brazilian Archives of Biology and Technology. 49: 353-359.
Garrett, K.A. & Mundt, C.C. 2000. Host diversity can reduce potato late blight severity for focal and general patterns of primary inoculum. Phytopathology, 90: 1307-1312.
Glick, B.R. 2012. Plant Growth-Promoting Bacteria: Mechanisms and Applications. Hindawi Publishing Corporation, Scientifica.
Guo, J.H., Qi, H.Y., Guo, Y.H., Ge, H., Gong, L.Y., Zhang, L.X. & Sun, P.H. 2004. Biocontrol of tomato wilt by growth-promoting rhizobacteria. Biological Control, 29, 66-72.
Haas, D. & Défago, G. 2005. Biological control of soilborne pathogens by fluorescent pseudomonads. Nature Review of Microbiolology, 3: 307-319.
Hallmann, J., Quadt-Hallmann, A., Mahaffee, W.F. & Kloepper, J.W. 1997. Bacterial endophytes in agricultural crops. Canadian Journal of Microbiology, 43: 895-914.
Henok, K., Fassil, A. & Yaynu, H. 2007. Evaluation of of Pseudomonas fluorescens isolates as biocontrol agents against bacterial wilt caused by Ralstonia (Pseudomonas) solanacearum. Pest Management Journal of Ethiopia, 11: 9-18.
Illmer, P., Barbato, A. & Schinner, F. 1995. Solubilization of hardy soluble AlPO4 with P- solubilizing microorganisms. Soil Biology and Biochemistry, 27: 265-270.
King, E.O., Ward, M.K. & Raney, D.E. 1954. Two simple media for the demonstration of pyocyanin and fluorescein. Journal of Laboratory and Clinical Medicine. 44: 301-307.
Klement, Z., Rudolph, K. & Sand, D.C. 1990. Methods in phytobacteriology. Akademiai Kiado Budapest, 540pp.
Kopen, J., Hodrov, B. & Stewart, C.S. 1996. The isolation and characterization of a rumen chitinolytic bacterium. Letters in Applied Microbiology, 23: 195-198.
Lemessa, F. & Zeller, W. 2007. Screening rhizobacteria for biological control of Ralstonia solanacearum in Ethiopia. Biological Control, 42: 336-344.
Maurhofer, M., Reimmann, C., Schmidli-sacherer, P., Heeb, S., Haas, D., & Defago, G. 1998. Salicylic acid biosynthetic genes expressed in Pseudomonas fluorescens strain P3 improve the induction of system resistance in tobacco against tobacco necrosis virus. Phytopathology, 88: 678-684.
McInroy, J.A. & Kloepper, J.W. 1995. Population dynamics of endophytic bacteria in field-grown sweet corn and cotton. Canadian Journal of Microbiology, 41, 895-901.
Maleki, M., Mostafaee, S., Mokhtarnejad, L. & Farzaneh, M. 2010. Characterization of Pseudomonas fluorescens strain CV6 isolated from cucumber rhizosphere in Varamin as a potential biocontrol agent. Australian Journal of Crop Science, 4(9): 676-683.
Miller, J.H. 1974. Experiments in Molecular Genetics. 2nd ed. Cold Spring Harbor Lab., Cold Spring Harbor, NY.
Moreira, R.R., Nesi, C.N. & Mio, L.L.M.D. 2014. Bacillus spp. and Pseudomonas putida as inhibitors of the Colletotrichum acutatum group and potential to control Glomerella leaf spot. Biological Control, 72:30-37.
Pikovskaya, R.I. 1948. Mobilization of phosphorus in soil in connection with the vital activity of some microbial species. Mikrobiologiya, 17: 362-70.
Prabhat, N.J, Garima, G., Prameela J. & Mehrotra, R. 2013. Association of Rhizospheric/Endophytic Bacteria with Plants: A Potential Gateway to Sustainable Agriculture. Greener Journal of Agricultural Sciences, 3 (2): 073-084.
Ramesh, R. & Phadke, G.S. 2012. Rhizosphere and endophytic bacteria for the suppression of eggplant wilt caused by Ralstonia solanacearum. Crop Protection, 37: 35-41.
Ran, L.X., Liu, C.Y., Wu, G.J., van Loon, L.C. & Bakker, P.A.H.M., 2005. Suppression of bacterial wilt in Eucalyptus urophylla by fluorescent Pseudomonas spp. in China. Biological Control, 32: 111-120.
Ryan, A.D., Kinkel, L.L. & Schottel, J.L. 2004. Effect of pathogen isolate, potato cultivar and antagonist strain on potato scab severity and biological control. Biochemical Science and Technology, 14: 301-311.
Saharan, B.S. & Nehra, V. 2011. Plant Growth Promoting Rhizobacteria: A Critical Review. Life Sciences and Medicine Research, LSMR-21.
Schaad, N.W., Jones, J.B. & Chun, W. 2001. Laboratory Guide for Identification of Plant Pathogenic Bacteria. American Phytopathological Society. St. Paul, Minnesota, 373 pp.
Sierra, J.M. 1957. A simple method for the detection of lipolytic activity of micro-organisms and some observations on the influence of the contact between cells and fatty substances. Antonie van Leeuwenhock Ned. Tijdschr. Hyg. 23: 15-25.
Weller, D.M. & Cook, R.J. 1983. Suppression of take-all of wheat by seed treatments with fluorescent pseudomonads. Phytopathology, 78: 463-469.