Subscribe Now Subscribe Today
Research Article

Antagonistic Potential of Native Isolates of Trichoderma viride on Corm Rot Pathogen Complex of Saffron (Crocus sativus) in Kashmir

G. Hassan Mir, L.S. Devi, S. Ahmad, V. Manoj Kumar and P. Williams

Investigation was undertaken to screen the potential native isolate of Trichoderma viride for bio suppression of corm rot pathogen complex, as Trichoderma viride are the most successful and widely used biocontrol agents. Taking the advantage and constraints of Trichoderma viride into consideration, efforts were made to encourage the native isolate against corm rot pathogens. Nine isolates of Trichoderma viride namely TK1, TK3, TK4, TK6, TK8, TK9, TK10, TK11 and TK15 were isolated from soils of different orchard plantations of Kashmir valley on modified Trichoderma Specific Medium (TSM). The isolates were studied for their cultural, morphometric characters and antagonistic potential against six newly recorded major fungal pathogens of saffron viz. sterile Basidiomycetes fungus, Rhizoctonia solani, Phytophthora sp., Fusarium oxysporum f. sp. gladioli, F. oxysporum and F. solani individually on Potato Dextrose Agar, the culture morphology of all the isolates was found to be similar. The isolate TK1, TK3, TK4, TK8, TK9, TK11 and TK15, were found fully overgrown on all corm rot Pathogens of saffron, where as the isolates TK13 failed to inhibit the Phytophthora sp. Efforts are onto evaluate the performance of promising isolate in field by soil and seed application methods.

Related Articles in ASCI
Similar Articles in this Journal
Search in Google Scholar
View Citation
Report Citation

  How to cite this article:

G. Hassan Mir, L.S. Devi, S. Ahmad, V. Manoj Kumar and P. Williams, 2011. Antagonistic Potential of Native Isolates of Trichoderma viride on Corm Rot Pathogen Complex of Saffron (Crocus sativus) in Kashmir. Plant Pathology Journal, 10: 73-78.

DOI: 10.3923/ppj.2011.73.78

Received: February 25, 2011; Accepted: June 22, 2011; Published: July 20, 2011


During last fifteen years, saffron crop has been affected by severe rotting caused by sterile Basidiomycetes fungus, Rhizoctonia solani, Fusarium f. sp. gladioli, Fusarium oxysporum and Fusarium solani (Mir and Devi, 2004) and reduction in yield has been reported. In 1980 the yield per hectare was 5.66 kg ha-1 (Mir, 1992) and now its present productivity is 1.53 kg ha-1 (Anonymous, 2009) which is the lowest in the world.

In recent years, attempts were also made to use a consortium of biocontrol agents to get persistent control of plant pathogens (Chaube and Sharma, 2002). Biological control, therefore, holds promise as a strategy for disease management and it is environment friendly too. Antagonistic fungi - 38 - especially Trichoderma spp. has been widely used against a number of phytopathogens (Rini and Sulochana, 2006) and parasitized hyphae of other fungi in vitro and brought about several morphological changes during destruction (Anitha and Murugesan, 2001). Screening of potential Trichoderma strains was done by Bandopadhyay et al. (2003) against major root pathogens and it was found that more or less all the strains checked the growth of the pathogen and stimulate plant defensive mechanisms (Hanson and Howell, 2004; Harman et al., 2004; Yadav et al., 2011).

Trichoderma harzianum is one efficient biocontrol that is commercially produced to prevent development of several soil pathogenic fungi (Jegathambigai et al., 2009). Biocontrol is an important approach for plant disease management under changing food habits and commercialization of agriculture (Manczinger et al., 2002).

Therefore, keeping in view medicinal importance and to remove the pesticidal residue of such valuable medicinal crop, the present study was undertaken for screening of several local antagonistic isolates of T. viride, obtained from different orchards of Kashmir valley, under in vitro conditions against few pathogens sterile Basidiomycetes fungus, Rhizoctonia solani, Fusarium oxysporum f. sp., gladioli, Fusarium gladioli, Fusarium solani, Phytophthora sp. causing corm rot syndrome of saffron.


Collection of pathogen: Four pathogenic isolates namely sterile Basidiomycetes fungus, Rhizoctonia solani, Fusarium oxysporum f. sp. gladioli, Fusarium gladioli, Fusarium solani and Phytophthora sp. were isolated, from infected corms from Kashmir valley from saffron growing area. The pathogens were maintained on PDA medium at 4°C.

Isolation of Trichoderma spp. (TK1, TK3, TK4, TK6, TK8, TK9, TK10, TK11 and TK15) was done from randomly collected soils from different vegetable fields and orchards of Kashmir valley by dilution plate technique using Trichoderma specific medium TSM (Elad et al., 1981) modified by Saha and Pan (1997).

Antagonistic potential of Trichoderma viride isolates on saffron pathogens: The antagonistic properties of fifteen isolates of Trichoderma viride were tested on PDA by dual culture plate technique. Paired cultures were observed for a total of 12 days before being discarded. All the ratings were done after contact between pathogen and the antagonist using a Bells scale (Bell et al., 1982) which is slightly modified (Class 1-7) as follows.

S1=The pathogen and the antagonism locked at the point of contact
S2=The antagonism starts overgrowth on pathogen.
S3=The pathogens starts overgrowth on mycoparasite
S4=The antagonist overgrew at least 15% of pathogen
S5=The antagonist overgrew of least 30% of pathogen
S6=The antagonist overgrew at least 60% of pathogen
S7=The antagonist completely overgrew the pathogen (100% overgrowth)


Identity of isolates of Trichoderma spp.: In general, colony morphology of all the isolates was more or less similar showing sparse to thin colony mycelial mass with whitish border in some cases. Sporulation started after 48h of incubation at 28±1°C for all the isolates (Table 2).

Micrometric measurements of Trichoderma viride (Table 1) showed that phialospore length ranged between 2.98-5.52 μm and, breadth ranged from 2.71-4.6 μm and phialides length 9.22-12.56 and breadth 1.3-2.5. These characteristics, particularly the trifid phialophore with short phialides clearly resembled the identical characters of Trichoderma viride (Rifai, 1969).

Antagonistic potential of Trichoderma viride isolates against corm rot pathogens of saffron.

Phytophthora sp.: The results showed that isolate TK1, TK8, TK4 and TK11 were antagonistic to Phytopthora by totally overgrowing the pathogen within seven, nine and eleven day respectively. Isolate TK10, TK15 and TK3, TK6 and TK9 were antagonistic to Phytopthora overgrowing 75, 45 and 60%, respectively.

F. oxysporum f. sp. gladioli: The results showed that isolate TK1, TK4, TK6, TK8, TK9 and TK11 were antagonistic to F. oxysporum f. sp., gladioli by totally overgrowing the pathogen within 8 to 12 days. Isolates TK3 and TK10, were overgrowing the pathogen 90 and 45%, respectively.

Sterile Basidiomycetes fungus: The results against Basidiomycetes fungus showed that five isolates TK1, TK8, TK15 and TK9 and TK6, totally overgrowing within nine, eight and 12 day, respectively. The remaining isolate TK11, TK4, TK3 and TK10, overgrew 90, 75, 45 and 15%, respectively.

Table 1: Micrometric measurement of phialospores, phialides and chlamydospores of isolate

Table 2: Colony characters of Trichoderma viride isolates

Table 3: Hyperparasitic potential of T. viride wild isolates on fungal pathogens of saffron
D: Days before contact, R: Rating, **: An average of five individual observation. *: The numerical value represents the days required for attaining S1 to S7 stage of modified Bell’s scale

Rhizoctonia solani: The results showed that all isolates were antagonistic to Rhizoctonia solani by totally overgrowing the pathogen with six to nine day except isolate TK15 it overgrew only 75% even after day.

F. oxysporum: The results showed that isolate TK1, TK3, TK4, TK8 and TK15 were antagonistic to F. oxysporum by totally overgrowing the pathogen within 6 to 9 days. Isolates TK9, TK10 and TK11 did not progress beyond 30% ever after day. The remaining isolate TK6 totally fails to overgrow the host pathogen even upto 12 days of inoculation inspite of attaining the point of contact of the third day.

F. solani: The result shows that five isolates TK1, TK3, TK4, TK8 and TK15 were highly antagonistic to Fusarium solani, totally overgrowing the pathogen within 6 to 11 days. Isolates TK9 and TK11 were overgrew the pathogen 30% whereas TK10 15% and TK6 failed to overgrew the host pathogen even after 12 days of inoculation, in spite of attaining the point of contact on the 4th day of inoculation.

The overview of the results (Table 3) showed that the isolates TK1, TK3, TK4, TK8, TK9, TK11 and TK15, were found fully overgrown on all corm rot Pathogens of saffron, where as the isolates TK13 failed to inhibit the Phytophthora sp. To identify then, isolates of Trichoderma spp. have been listed in the tables that reached class-I (S7) stage within 6-11 days of inoculation. However, based on this information the antagonistic Trichoderma viride did not allow an early selection of isolates, as the variability in the antagonistic characteristic within the isolate and isolate-pathogen interaction was very high.

The above observations established the fact that Trichoderma isolates existing in their natural conditions in natural ecosystem do differ with respect to their growth and antagonistic potential. Similarly Li et al. (2001) studied eighteen isolates of Trichoderma spp. of these isolates, TR13 showed greatest antagonists effects against Rhizoctonia solani. Several research papers that have appeared in the literature do reveal the fact that various species and isolates of fungal antagonist Trichoderma suppress mycelial growth, reduce root rots, increase plant growth and induce resistance in various crops with which Sclerotium rolfsii (Tian et al., 2001; Das and Dutta, 2002; Palomar et al., 2002), Rhizoctonia solani (Li et al., 2001; Burgess and Hepworth, 1996; Zapata et al., 2001; Ziedan and Mahmoud, 2002; Gaikwad and Nimbalkar, 2003; Yossen et al., 2003; Fravel and Lewis, 2004; Hajlaoui et al., 2001; Singh et al., 2003; Huang and Erickson, 2004; Salehpour et al., 2005) are associated. It is clear that the success of bioagents introduces in soil does not guarantee the control the target pathogen(s) because plants, physicochemical and biological factors of soil affect establishment, proliferation and antagonistic activities of the introduced bioagents. It is necessary that, identified antagonist efficiency against foot, root rot and damping off should be investigatited and examined in vivo conditions also, the results of such survey would be reported by the authers in near future (Shaigan et al., 2008).

It is in this context that to ensure success of introduced bioagents, they should be isolated for the local areas where they exit. Since, they have already faced various processes of evaluation, their application would be feasible and result oriented. We reviewed the literature to find out that have others worked on these aspects. Literature analysis revealed that comparative studies have been done with various species (Kucuk and Kivanen, 2003; Chang et al., 2006) studied Trichoderma isolates from different soil sampled and grouped them according to their antagonistic potential and chitin utilization.


The overgrowth by the antagonist under in vitro conditions may be good criteria of selecting an isolate shows good performance under in vitro conditions. The trend of the results also indicated that there was not only variability amongst the isolates of Trichoderma viride with differential degree of a ntagonism towards a single pathogen but also towards different pathogens.

The results of the study are the pointer to the fact that the antagonists should be isolated from different systems and locations to create a huge genetic pool and tested for their antagonistic potential against variety of the targeted plant pathogens and recommended specifically for different locations and systems. The present study clearly indicates the high potential of biocontrol agent, Trichoderma viride isolates for different plant pathogens. Efforts are onto evaluate the performance of promising isolate in field by soil and seed application methods.


I am greatly thankful to Department of Science and Technology, New Delhi for providing fully financial assistance under Young Scientist Scheme to carry out the work. I am grateful to DADA for his technical help during the research equally I wish to thank my advisor Prof. L.S. Devi and my dear teachers Prof. P. Gupta, Prof. P. Williams, Prof. Shafat Ahmad, Dr. V. Manoj Kumar, Prof. G. Abraham and Prof. Ram Lal and also appreciate Mithai Lal jee, Maqsood sahib, Amrit Lal jee, Subash jee, Gyan Chand jee, Rajkaran jee and Kanaya lal jee, AAI-DU, Allahabad, U.P. India, for their support through out the research period.

Anitha, R. and K. Murugesan, 2001. Mechanism of action of Gliocladium virens on Alternaria helianthi. Indian Phytopathol., 54: 449-452.
Direct Link  |  

Anonymous, 2009. Directorate of Agriculture. Lalmandi, Srinagar, Kashmir.

Bandopadhyay, S., N.D. Sharma and S. Dutta, 2003. Screening of potential Trichoderma strains against major root pathogens. Ann. Plant Protec. Sci., 11: 163-163.

Bell, D.K., H.D. Wells and C.R. Markham, 1982. In vitro antagonism of Trichoderma species against six fungal plant pathogens. Phytopathology, 72: 379-382.
CrossRef  |  Direct Link  |  

Burgess, D.R. and G. Hepworth, 1996. Biocontrol of sclerotinia stem rot (Sclerotinia minor) in sunflower by seed treatment with Gliocladium virens. Plant Pathol., 45: 583-592.

Chang, K.F., S.F. Hwang, H. Wang, G. Turnbull and R. Howard, 2006. Etiology and biological control of sclerotinia blight of coneflower using Trichoderma species. Plant Pathol. J., 5: 15-19.
CrossRef  |  Direct Link  |  

Chaube, H.S. and J. Sharma, 2002. Integration and interaction of solarization and fungal and bacterial bioagents on disease incidence and plant growth response of some horticultural crops. Plant Dis. Res., 17: 201-201.

Das, B.C. and P. Dutta, 2002. Management of collar rot of tomato by Trichoderma spp. and chemicals. Indian Phytopathol., 55: 235-237.
Direct Link  |  

Elad, Y., I. Chet and Y. Henis, 1981. A selective medium for improving quantitative isolation of Trichoderma spp. from soil. Phytoparasitica, 9: 59-67.
CrossRef  |  Direct Link  |  

Fravel, D.R. and J.A. Lewis, 2004. Effect of label and sub label rates of metamsodium in combination with Trichodema amatum, T. harzianum, T. virens, T. viride on survival and growth of Rhizoctonia solani. Phytoparasitica, 32: 111-118.
CrossRef  |  

Gaikwad, A.P. and C.A. Nimbalkar, 2003. Mangement of collar and root rot (Rhizoctonia solani) of bell pepper with bioagent (Trichoderma spp.) and fungicides. J. Maharastra Agric. Univ., 28: 270-273.
Direct Link  |  

Hajlaoui, M.R., D. Diop and M. Cherif, 2001. Contribution to biological control of Sclerotinia blight cause by Sclerotinia sclerotiorum (Lib.) de Bary. Al Awamia, 104: 85-101.

Hanson, L.E. and C.R. Howell, 2004. Elicitors of plant defence responses from biocontrol strains of Trichoderma virens. Phytopathology, 94: 171-176.

Harman, G.E., C.R. Howell, A. Viterbo, I. Chet and M. Lorito, 2004. Trichoderma species-opportunistic, avirulent plant symbionts. Nat. Rev. Microbiol., 2: 43-56.
CrossRef  |  PubMed  |  Direct Link  |  

Huang, H.C. and R.S. Erickson, 2004. Effect of soil treatment of fungal agents on control of apothecia of Sclerotium sclerotiorum in canola and safflower fields. Plant Pathol. Bull., 13: 1-6.
Direct Link  |  

Jegathambigai, V., R.S.W. Wijeratnam and R.L.C. Wijesundera, 2009. Control of Fusarium oxysporum wilts disease of Crossandra infundibuliformis var. Danica by Trichoderma viride and Trichoderma harzianum. Asian J. Plant Pathol., 3: 50-60.
CrossRef  |  Direct Link  |  

Kucuk, C. and M. Kivanc, 2003. Isolation of Trichoderma spp. and determination of their antifungal, biochemical and physiological features. Turk. J. Biol., 27: 247-253.
Direct Link  |  

Li, M.Y., G.J. Wang, T.F. Li and K. Liu, 2001. Selection for Trichoderma isolates applicable in biocontrol of major fungal diseases of tobacco. J. Southwest Agric. Univ., 23: 10-12.

Manczinger, L., Z. Antal and L. Kredics, 2002. Ecophysiology and breeding of mycoparasitic Trichoderma strains. Acta Microbiologica Immunologica Hungarica, 49: 1-14.
CrossRef  |  PubMed  |  Direct Link  |  

Mir, G.H. and L.S. Devi, 2004. Saffron corm rot and their management. Proceedings of the National Symposium on Detection and Management of Plant Diseases using Conventional and Modern Tools and IPS Zonal Meeting (MEZ), Dec. 31, Lucknow, pp: 20-20.

Mir, G.M., 1992. Saffron Agronomy in Kashmir a Study in Habitat, Economy and Society. Gulshan Publishers, Srinagar, Kashmir, India.

Palomar, M.K., Y.C. Mangaoang, V.G. Palermo, G.E. Escuadra and M.B. Posas, 2002. Biocontrol of root crop diseases through microbial antagonism. Proceedings of the 4th Asia-Pacific Biotechnology Congress and 30th Annual Convention of the PSM, May 16-18, 2001, PSMI, College, Laguna, Philippines, pp: 56-62.

Rifai, M.A., 1969. A revision of the genus Trichoderma. Mycological Pap., 116: 1-56.
Direct Link  |  

Rini, C.R. and K.K. Sulochana, 2006. Management of seedling rot of chilli (Capsicum annuum L.) using Trichoderma spp. and fluorescent pseudomonads (Pseudomonas fluorescens). J. Trop. Agric., 44: 79-82.
Direct Link  |  

Saha, D.K. and S. Pan, 1997. Quantitative evaluation of some specific media of Trichoderma and Gliocladium spp. J. Mycolopathol. Res., 35: 7-13.

Salehpour, M., H.R. Etebarian, A. Roustaei, G. Khodakaramian and H. Aminian, 2005. Biological control of common root rot of wheat (Bipolaris sorokiniana) by trichoderma isolates. Plant Pathol. J., 4: 85-90.
CrossRef  |  Direct Link  |  

Shaigan, S., A. Seraji and S.A.M. Moghaddam, 2008. Identification and investigation on antagonistic effect of Trichoderma spp. on tea seedlings white foot and root rot (Sclerotium rolfsii Sacc.) in vitro condition. Pak. J. Biol. Sci., 11: 2346-2350.
CrossRef  |  PubMed  |  Direct Link  |  

Singh, R., U. Narain and R. Palat, 2003. Evaluation of bioagents against Sclerotinia stem rot of ajowan. Annal. Plant Prot. Sci., 11: 386-386.
Direct Link  |  

Tian, L.S., W.H. Wang, W.L. Shi, S.S. Li, Y.M. Shi, G.W. Zhang and L.P. Zhang, 2001. Studies on mechanisms of antagonism of Trichoderma viride to Fusarium oxysporum f. sp. lycopersici and its effect of biocontrol. Plant Protect., 27: 47-48.

Yadav, J., J.P. Verma and K.N. Tiwari, 2011. Plant growth promoting activities of fungi and their effect on chickpea plant growth. Asian J. Biol. Sci., 4: 291-299.
CrossRef  |  Direct Link  |  

Yossen, N.A., G.S. Vargas, M. Diaz-P-del and C. Olmos, 2003. Compost and Trichoderma harzianum as suppresors of Rhizoctonia solani and promoters of let tuce growth. Manejo Integrado Plagasy Agroecol., 68: 19-25.

Zapata, R.L., H.E. Palmucci, V. Blanco-Murray and M.V. Lopez, 2001. Biological trials to control damping-off in eggplant (Solanum melongena) with fluorescent Pseudomonas and Trichoderma harzianum. Rev. Fac. Agron. Univ. Buenos Aires, 21: 207-211.

Ziedan, E.H. and S.Y.M. Mahmoud, 2002. Calcium and sulfur soil treatment to improve biological control with Trichoderma harzianum for root rot disease control of bean. Assiut. J. Agric. Sci., 33: 149-160.

©  2020 Science Alert. All Rights Reserved