Biological Control of Rhizocronia solani ( Pistachio Isolate) and Molecular Analysis of Chitinase Gene from the Best Bio-Control Agent

Document Type : Research Paper



Rhizoctonia solani, the causal agent of damping-off, root rot and stem rot, is one of the most important phytopathogenes with a wide host range which causes significant reduction in yield of pistachio (Pistacia vera). In search for finding chitinolytic biocontrol agents, actinomycetes producing chitinase enzyme were isolated from agricultural soils of Sirch, Kerman province . From tested isolates, Streptomyces isolate 410 significantly reduced the incidence of disease. Chitinase gene (603 bp) was cloned, sequenced and characterized from this isolate. The results showed that nucleotides encoding 200 amino acids and molecular weight of 21636.9 Da. This protein had the high homology with the family 19 chitinases glycosyl hydrolases which have the major role in the defense response against phytopathogens


Biological control, Streptomyces, Chitinase, Rhizoctonia solani, Pistachio.


Baharlouei A, Sharifi Sirchi GR, Shahidi Bonjar GH (2011). Biological control of Sclerotinia sclerotiorum (oilseed rape isolate) by an effective antagonist Streptomyces. African Journal of Biotechnology 10: 5785-5794.
Baharlouei A, Sharifi Sirchi GR, Shahidi Bonjar GH (2008). Investigation of Antagonistic Activity of Actinomycetes Against Sclerotinia sclerotiorum the Causal Agent of Rot of Oilseed Rape. The 2nd International Student Conference of Biotechnology and New Subjects, 15 Nov, Tehran, Iran (In Farsi).
Baharlouei A, Sharifi Sirchi GR, Shahidi Bonjar GH (2010). Isolation and Characterization of a Chitinase Gene Exhibiting Antifungal Activity Against Fusariumsolani f. sp. melongenae from an Iranian Streptomyces strain. Agricultural Biotechnology 9: 39-47.
Baharlouei A, Sharifi Sirchi GR, Shahidi Bonjar GH (2009). Isolation, Cloning and Sequencing of Chitinase Gene from Streptomyces plicatus. 13-15 August. The 6th National Biotechnologe Congress of I. R. Tehran. Iran (In Farsi).
Blaak H, Schnellmann J, Walter S, Henrissat B, Schrempf H (1993). Characteristics of an exochitinase from Streptomyces olivaceoviridis, its corresponding gene, putative protein domains and relationship to other chitinases. European Journal of Biochemistry 214: 659-669.
Chakraborty MR, Chatterjee NC (2008). Control of fusarium wilt of Solanum melongena by Trichoderma spp. Biological Plantarum 52: 582-586.
Chan YK, Wayne A, McCormick K, Seifert A (2003). Characterization of an Antifungal Soil Bacterium and its Antagonistic Activities Against Fusarium Species. Canadian Journal of Microbiology 49: 253–262.
Chi CC, Hanson EW (1965). In Vitro Effects of Streptomyces rimosus on Some Soil-Inhabiting Pathogenic Fungi. Plant Disease Reporter 49:159-163.
Davelos AL, Kinke LL, Samac DA (2004). Spatial Variation in Frequency and Intensity of Antibiotic Interactions Among Streptomycetes from Prairie Soil. Applied and Environmental Microbiology 70: 1051-1058.
Deshpande M (1986). Enzymatic degradation of chitin and its biological applications. Journal of Science Industry Research 45:273–281.
Elahinia SA (1999). Mycology and phytopathology. Gilan University Publisher. 3 th Edition. Iran. 
El-Mansi EMT, Bryce CFA (1999). Fermentation Microbiology and Biotechnology. Taylor and Francis UK.
Itoh Y, Takahashi K, Takizawa H, Nikaidou N, Tanaka H, Nishihashi H, Watanabe T,  Nishizawa Y (2003). Family 19 Chitinase of Streptomyces griseus HUT6037 Increases Plant Resistance to the Fungal Disease. Bioscience Biotechnology Biochemistry 67: 847-855.
Katsuichiro O, Yousuke Y, Minoru N, Noriyuki S, Isamu K, Shigeru H (2004). Molecular Cloning and Expression of the Gene Encoding Family 19 Chitinase from Streptomyces sp. J-13-3. Bioscience. Biotechnology and Biochemistry 68: 341-351.
Konishi M, ohkuma H, Matsumoto K, Saitoh K, Miyaki T, Oki T, Kawaguchi H (1991). Dynemicins New Antibiotics with the 1,5-DIYN-3-ENE and Anthraquinone Subunit I Production Iolation and Physico-Chemical Properties. The Journal of Antibiotics 44: 1300-1304.
Lee JY, Hwang BK (2002). Diversity of Antifungal Actinomycetes in Various Vegetative Soils of Korea. Canadian Journal of Microbilogy 48: 407-417.
Locci R (1989). Streptomycetes and Related Genera In: Bergey’s Manual of Systematic Bacteriology. Williams and willkins USA 4: 2451-2492.
Mansouri, S., Foroumadi, A., Ghaneie, T., Gholamhosseinian Najar, A (2001). Antibacterial Activity of the Crude Extracts and Fractionated Constituents of Myrtus communis. Pharmaceutical Biology 39: 399-401.
Ohno T, Aemand S, Hata T, Nikaidou N, Henrissat B, Mitsutomi M, Watanabe T (1996). A modular family 19 chitinase found in the prokaryotic organism Streptomyces griseus HUT 6037 J. Bacteriology 178: 5065-5070.
Rogers SO, Bendich AJ (1994). Extraction of Total Cellular DNA from Plants, Algae and Fungi. In: Gelvin, S.B. and Schilperoort, R.A. (eds), Plant Molecular Biology Manual , 2nd Edition, Kluwer Academic Press, Dordrecht, The Netherlands, D1:1-8.
Rothrock CS, Gottlieb D (1984). Importance of Antibiotic Production in Antagonism of Selected Streptomyces Species to Two Soil-Born Plant Pathogens. The Journal of Antibiotics PP: 830-834.
Saadoun IF, Gharaibeh R (2001). The Streptomyces Flora of Jordan and It’s Potential as a Source of Antibiotics Active Against Antibiotic-Resistant Gram-Negative Bacteria. World Journal of Microbiology and Biotechnology 18:465-470.
Sabaratnam S, Traquir JA (2002). Formulation of a Streptomyces Biocontrol Agent for the Suppression of Rhizoctonia Damping-off in Tomato Transplants. Biological Control 23: 245-253.
Saito A, Fujii T, Yoneyama T, Redenbach M, Ohno T, Watanable T, Miyashita K (1999). High-multiplicity of chitinase genes in Streptomyces coelicolor A3(2) Bioscience Biotechnology and Biochemistry 63: 710-718.
Sabaratnam S, Traquir JA (2002). Formulation of a Streptomyces Biocontrol Agent for the Suppression of Rhizoctonia Damping-off in Tomato Transplants. Biological Control 23: 245-253.
Sadeghi A, Hessan AR, Askari H, Aghighi S, Shahidi Bonjar GH (2006). Biological Control Potential of Two Streptomyces Isolates on Rhizoctonia solani, the Causal Agent of Damping-off of Sugar Beet. Pakistan Journal of Biological Sciences 9: 904-910.
Shahidi Bonjar GH, Karimi NA (2004). Antibacterial Activity of Some Medicinal Plants of Iran Against Pseudomonas aeruginosa and P. fluorescens. Asian Journal of plant Sciences 3: 61-64.
Shahidi Bonjar GH, Rashid Farrokhi P, Aghighi S, Shahidi Bonjar L, Aghelizadeh A (2005). Antifungal Characterization of Actinomycetes Isolated from Kerman, Iran and Their Future Prospects in Biological Control Strategies in Greenhouse and Field Conditions. Plant Pathology Journal 4: 78-84.
Shapira R, Ordentlich A, Chet I, Oppenheim AB (1989). Control of plant diseases by chitinase expressed from cloned DNA in E. coli. Phytopathology 79: 1246-1249.
Synowiecki J, Al-Khateeb N A (2003) Production, properties and some new application of chitin and its derivatives. Reviews Food Science and Nutrition 43: 145-171.
Zakir Sultan M, Ara Khatune N, Sultana Sathi Z, Shah Alam Bhuiyan MD, Golam Sadik M, Akteruzzaman Choudury M, Gafur MA, Abdur Rahman A (2002). In Vitro Antibacterial Activity of an Active Metabolite Isolated from Streptomyces Species. Biotechnology pp: 100-107.