Inhibition of Trichoderma Species from Growth and Zoospore Production of Phytophthora Drechsleri and Their Effects on Hydrolytic Enzymes

Document Type : Research Article


1 Faculty of Agriculture, University of Zabol, Zabol, Iran

2 Horticultural Science Research Institute, Pistachio Research Center, Agricultural Research, Education and Extension Organization (AREEO), Rafsanjan, Iran


Understanding the function of Trichoderma species in the control of Phytophthora drechsleri in pistachio orchards is very important.  In this study, the effects of liquid extra-cellular secretions and volatile compounds secreted by 27 isolates of Trichoderma harzianum, T. crassum, T. koningii, T.aureoviride, T. asperellum, T. brevicompactum, T.longibrachiatum and T. virens were investigated on Phytophthora drechsleri growth and zoospore production. Due to cell wall combination of P. drechsleri, the ability of Trichoderma isolates in the production of β-1,3 glucanase and cellulase was evaluated in media with different carbon sources. The inhibitory effects of the 16 isolates of Trichoderma from growth of P. drechsleri were examined in a dual culture test. The results showed that Trichoderma isolates had a variable effect on the growth and zoospore production of P. drechsleri. Trichoderma harzianum-136 and T. harzianum-8279 revealed the highest inhibitory effect on radial growth of P. drechsleri in 20 and 30 percent concentrations of liquid extra-cellular secretions, respectively. Both isolates also showed the highest inhibitory effect on zoospore production of P. drechsleri in 10 percent concentration of extra-cellular liquid secretions. In the volatile compounds test, T. harzianum-8279 had the highest effect on the growth of P. drechsleri. In all Trichoderma isolates, the activity of β-1,3 glucanase was higher than cellulase activity. The enzyme production was also higher in the liquid medium containing the cell wall of P. drechsleri compared to glycerol as a carbon source. The highest activity of β-1,3 glucanase and cellulase was observed in T. harzianum-8279.


Aminaee MM, Ershad J (1991) Isolation of Phytophtora drechsleri from infected pistachio trees with gummosis in Kerman. Proceedings of the 10th Iranian Plant Protection Congress. pp. 106. [In Persian].
Ashkan M, Abusaidi D, Banihashemi Z (1995) Distribution of Phytophtora species causing crown and root rot of pistachio in Rafsanjan. Proceedings of the 12th Iranian Plant Protection Congress. Pp. 218. [In Persian].
Ayobi N, Zafari D, Mirabolfathi M (2010) Effect of Trichoderma species on Zoospore production of Phytophthora sojae, disease severity and glucanase enzyme activity assay. Iranian Journal Plant Pathology. 46, 203-215. (In Persian)
Bahramsari N, Zamani M, Motallebi M (2005). β-1,3-glucanase production in Trichoderma isolates. Iranian Journal of Biology. 18, 261-271. [In Persian].
Banihashemi Z (1989) Study of pistachio gummosis in southern provinces of Iran.Proceedings of the9th Plant Protection Congress.Pp. 87. [In Persian].
Banihashemi Z (1999) Reaction of Pistacia spp. to root rot Phytophthora species. Plant Diseases. 34, 213-224.
Behboody K, Sharifi Tehrani A, Hajarood Q, Zad J (2005) Antagonistic effects of Trichoderma species on Phytophthora capsici, the causal agent of pepper root and crown rot. Iranian Journal of Plant Pathology. 41, 345-362. [In Persian].
Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein dye-banding. Annual Biochemistry.12, 951-973.
Calistru C, McLean M, Berjak P (1997) In vitro studies on the potential for biological control of Aspergillus flavus and Fusarium moniliforme by Trichoderma species. Mycopathologia. 137(2), 115-124.
Chet I (1987) Trichoderma: application, mode of action, and potential as a biocontrol agent of soilborne plant pathogenic fungi. pp. 137-160. In: Chet I (ed) Trichoderma.Wiley, New York.
Cruz J, Hidalgo-Gallego A, Lora JM, Benitez T, Pintor-Toro JA, Llobell A (1992) Isolation and characterization of three chitinases from Trichoderma harzianum . European Journal of Biochemistry. 206, 859–867.
 Cruz J, Pintor-Toro JA, Benitez T, Llobell A, Romero LC (1995) A novel endo-beta-1,3-glucanase, BGN13.1, involved in the mycoparasitism of Trichoderma harzianum . Journal of Bacteriology. 177, 6937–6945
Dennis C, Webster J (1971) Antagonistic properties of species groups of Trichoderma (Hyphal interactions). Transactions of British Mycological Society. 57, 363-369.
Dickinson ME, Selleck MA, McMahon AP, Bronner-Fraser M (1995) Dorsalization of the neural tube by the non-neural ectoderm.Development. 121(7), 2099-2106.
Dinesh R, Prateeksha M (2015) A review of interacations of Trichoderma with plants and pathogens. Research Journal Agricultural Forest Science. 3, 20-23.
Elad Y, Chet I, Boyle P, Henis Y (1983) Parasitism of Trichoderma spp. on Rhizoctonia solani and Sclerotium rolfsii-scanning electron microscopy and fluorescence microscopy. Phytopathology. 73, 85-88.
El-Katatny M, Gudelj M, Robra KH, Elnaghy M, Gübitz G (2001). Characterization of a chitinase and an endo-β-1, 3-glucanase from Trichoderma harzianum Rifai T24 involved in control of the phytopathogen Sclerotium rolfsii.Applied Microbiology and Biotechnology. 56(1-2), 137-143.
Fani SR, Mirabolfathi M, Zamanizadeh HR (2004). Etiology of pistachio gummosis in Sistan and Baloochestan province. Proceedings of the16th Iranian Plant Protection Congress. pp. 382. [In Persian].
Fani SR, Moradi Qahdarijani M, Alipour Moqadam M, Sherafati A, Mohammadi Moqaddam M, Sedaqati A, Khodaygan P (2013). Efficacy of native strains of Trichoderma harzianum in biocontrol of pistachio gummosis. Iranian Journal Plant Protection Science. 44(2), 243-252. [In Persian].
Fattahi Ardekani M, Ershad J, Mirabolfathi M (2000) Study of pistachio gummosis in Yazd province. Proceedings of the 14th Iranian Plant Protection Congress. pp. 126. [In Persian].
Ghisalberti EL, Rowland CY (1993) Antifungal metabolites from Trichoderma harzianum.Journal of Natural Products. 56(10), 1799-1804.
Haghdel M, Zafari D, Mohammadi A, Sharzee M (2009) Study of antagonistic effect of isolated fungus from pistachio nuts on toxigenic Aspergillus flavus. Proceedings of the 18th Iranian Plant Protection Congress. pp. 252. [In Persian].
Hancock JT, Jones OT (1987) The inhibition by dihenylneiodonium and its analogues of superoxide generation by macrophages. Biochemical Journal. 242, 103-107.
Haran S, Schikler H, Chet I (1996a) Molecular mechanisms of lytic enzymes involved in the biocontrol activity of Trichoderma harzianum. Microbiology. 142, 2321-2331.
Haran S, Schikler H, Oppenheim A, Chet I (1996b) Differential expression of Trichoderma harzianum chitinases during mycoparasitism. Phytopathology. 86, 981-985.
Harman GE (2006) Overview of mechanisms and uses of Trichoderma spp. Phytopathology. 96, 190–194.
Harman GE, Howell CR, Viterbo A, Chet I, Lorito M (2004) Trichoderma species-opportunistic avirulent plant symbionts. Nature Review Microbiology. 2, 43–56.
Howell CR (2003).Mechanisms employed by Trichoderma species the Biological control of plant Diseases: the history and evolution of current concepts. Plant Disease. 86, 104-10.
Jamdar Z, Mohammadi AH, Mohammadi S (2013) Study of antagonistic effects of Trichoderma species on growth of Verticillium dahliae, the causal agent of Verticillium wilt of pistachio under laboratory condition. Journal of Nuts. 4, 53-56.
Jung WJ, Jin YL, Kim KY, Park RD, Kim TH (2005) Changes in pathogenesis-related proteins in pepper plants with regard to biological
control of Phytophthora blight with Paenibacillus illinoisensis. Bio Control.50, 165-178.
Lorito M, Harman GE, Hayes CK, Broadway RM, Troncoso A, Woo SL, di Pietro A (1993) Chitynolytic enzymes produced by Trichoderma harzianum: antifungal activity of purified endochitinase and chitobiase. Phytopathology. 83, 302-307.
Lorito M, Woo SL, García Fernández I, Colucci G, Harman GE, Pintor-Toro JA, Filippone E, Muccifora S, Lawrence CB, Zoina A, Tuzun S, Scala F (1998) Genes from mycoparasitic fungi as a source for improving plant resistance to fungal pathogens. Proceedings of the National Academy of Sciences, USA. 95, 7860-7865.
Mirabolfathy M, Cooke DEL, Duncan JM, Williams NA (2001) Phytophthora pistaciae sp. Nov. and P. melonis: the principal causes of pistachio gummosis. Iranian Mycology Research. 105, 1166-1175. [In Persian].
Mirabolfathi M, Davoodi A, Yasini A [2004]. Study of pistachio gummosis in Qazvin province. Proceedings of the 16th Iranian Plant Protection Congress. pp. 375.
Mirabolfathi M, Ershad J (1986) Isolation of Phytophtora cf  megasperma from pistachio trees with gummosis. Proceedings of the of 8th Iranian Plant Protection Congress. pp. 84. [In Persian].
Mirkhani F, Alaei H, Mohammadi AH, Haghdel M, Sedaqati A (2013) Identification of dominant Trichoderma species in alkaline soils of pistachio orchards in Kerman province. Proceedings of the 1st Iranian Mycology Congress, pp. 60. [In Persian].
Mohammadi AH (2010) Interaction between Glomus mosseae and Phytophthora drechsleri on root rot of pistachio under greenhouse conditions and their mechanisms. Ph.D thesis of Plant Pathology, Faculty of Agriculture, Shiraz University. 209 p.
Mostowfizadeh-Ghalamfarsa R, Cooke DEL, Banihashemi Z (2008) Phytophthora parsiana sp. nov., a new high-temperature tolerant species. Mycological Research.112, 783-794.
Ozbay N, Newman SE (2004) Biological control with Trichoderma spp. with emphasis on
T. harzianum.Pakistan Journal of Biological Sciences.7(4), 478-484.
Papavizas GC (1985) Trichoderma and Gliocladium Biology, ecology and potential for biocontrol. Annual Reviews Phytopathology. 23, 23-45.
Ridout CJ, Coley-Smith JR, Lynch JM (1986) Enzyme activity and electrophoretic profile of extracellular protein induced in Trichoderma spp. by cell walls of Rhizoctonia solani. Microbiology.132, 2345–2352
Samuels GJ (1996) Trichoderma: a review of biology and systematics of the genus. Mycological Research. 100, 923-935.
Shen S, Chretien P, Bastien L, Slilty SN (1991) Primary structure of the glucanase gene from Oerskovia xanthineolytica expression and purification of the enzyme from Escherichia coli. Journal Biology Chemistry. 266, 1058–1063
Simon A, Sivasithamparam K (1989) Pathogen-suppression: a case study in biological suppression of Gaeumannomyces graminis var. tritici in soil.Soil Biology and Biochemistry. 21(3), 331-337.
Tapwal A, Sharma YP, Lakhanpal TN (2004) Effect of volatile componds  related by Gliocladium virens and Trichoderma spp. on  the growth of Dematophora necatrix. Journal of Mycology and Plant Pathology. 34(1), 37-40.
Tondje PR, Roberts DP, Bon MC, Widmer T, Samuels GJ, Ismaiel A, Begoude AD, Tchana T, Nyemb-Tshomb E, Ndoumbe-Nkeng M, Bateman R (2007) Isolation and identification of mycoparasitic isolates of Trichoderma asperellum with potential for suppression of black pod disease of cacao in Cameroon. Biological Control.43(2), 202-212.
Vigo C, Norman JR, Hooker JE (2000) Biocontrol of the pathogen Phytophthora parasitica by arbuscular mycorrhizal fungi is a consequence of effects on infection loci. Plant Pathology. 49, 509-514.
Yedidia I, Benhamou N, Chet I (2000) Induction of defence responses in cucumber plants (Cucumis sativus L.) by the biocontrol agent Trichoderma harzianum. Applied Environmental Microbiology. 65, 1061–1070.
Zavari F, Sahebani N, Etebarian HR (2012) Measuring of β-1,3 glucanase activity in Trichoderma virens isolates and selection of the best isolates for biological control of cucumber root rot. Journal of Stainable Agriculture and Production Science. 22(4), 150-161. [In