1. Department of Plant Pathology, University of Agriculture, Faisalabad
EXPLOITATION OF THE ANTAGONISTIC POTENTIAL OF TRICHODERMA ISOLATES AND AGRICULTURAL COMPOSTS AGAINST FUSARIUM WILT OF TOMATO
Anam Moosa
Department of Plant Pathology, University of Agriculture, Faisalabad
ABSTRACT
MATERIALS AND METHODS a
Table 1. Inhibitory effect of Trichoderma isolates by Dual Culture Technique.
Table 3: Inhibitory Effect of Non-Volatile Metabolites.
Table 2: Inhibitory Effect of Volatile metabolites.
1. In vitro evaluation of Trichoderma isolates and Compost water extract against F. oxysporum f. sp. Lycopersici
1.1. Effect of Trichoderma isolates on F. oxysporum f.sp lycopersici
Dual Culture Technique:
Antagonistic potential of Trichoderma isolates against the pathogen was evaluated by Dual culture technique given by (Morton and Stroube, 1955). I = (C-T)/C × 100
1.1.2.Volatile Metabolites
The effect of volatile metabolites produced by Trichoderma isolates was assessed against Fusarium oxysporum f. sp lycopersici by inverted plate technique (Dennis and Webster, 1971b). I = (C-T)/C × 100
Non-Volatile Metabolites
The effect of non-volatile metabolites produced by Trichoderma isolates on linear growth of pathogen was studied following the method devised by (Dennis and Webster, 1971a) I = (C-T)/C × 100
1.2. Effect of Compost Water Extracts on F. oxysporum f. sp. lycopersici
Pouring method
Inhibitory effect of four different compost water extacts (CWEs) against Fusarium oxysporum f. sp lycopersici was evaluated according to the procedure given by (EL-Masry, et al. 2002).
I = (C-T)/C × 100
2. In vivo evaluation of compost in combination with Trichoderma isolates against F. oxysporum f. sp lycopersici.
On the basis of in vitro experiments best compost 100% Natural Dried Cow compost was selected and applied in combination with both antagonistic isolates (T.viride and T.harzianum) for its evalution in field trial. Experiment comprised of following treatments,
T1 Soil Infested with FOL only.  T2 Healthy control (No inoculum added).T3 T. viride only. T4 T. harzianum only. T5 Compost only.
T6 Soil infested with pathogen amended with natural cow compost and T. viridi. T7 Soil infested with pathogen amended with compost and T. harzianum. T8 Soil infested with pathogen amended with T. viridi only. T9 Soil infested with pathogen amended with T. harzianum only. T10 Soil infested with pathogen amended with compost only. T11 Soil infested with pathogen amended with T. viridi, T. harzianum and compost. T12Soil amended with T. viridi, T. harzianum and compost without pathogen.
Fusarium oxysporum f.sp lycopersici induces wilt disease in tomato which frequently appears worldwide causing huge amount of losses both in quality and quantity of tomato production. In vivo and in vitro studies were conducted to evaluate the effect of four natural cheaply available composts (100% Natural Dried Cow compost, 100% Natural Dried Horse compost, 100% Natural Dried Chicken compost, 100% Natural Dried Sheep compost) and two antagonistic isolates of Trichoderma i.e., Trichoderma viride and Trichoderma harzianum against Fusarium oxysporum f.sp lycopersici on a local susceptible variety of tomato. In vitro studies revealed that antagonistic Trichoderma viride and 100% Natural Cow compost showed maximum inhibition of the pathogen. Both isolates of Trichoderma and 100% Natural cow manure were evaluated in field experiment individually and in combination. Combination treatment comprising of both Trichoderma isolates and compost performed best with 0% disease incidence and disease severity 1.
No.
Treatments
Colony Diameter (cm)
Inhibition (%) 1
Control
4.10 a 0% 2
T. harzianum
2.90 b
29.27% 3
T. viride
2.67 c
34.96%
No.
Treatment
Colony Diameter (cm)
Inhibition (%) 1
Control
3.41 a
0.00 2
T. harzianum
2.21 b
35.19 3
T. viride
1.88 c
44.86
No.
Treatment
Colony Diameter (cm)
Inhibition (%) 1
Control
4.14 a
0.00 2
T. harzianum
2.08 b
49.76 3
T. viride
1.52 c
63.28
Table 4: In vivo efficacy of Trichoderma Isolates and selected
Compost against FOL.
Table 5. Effect of Trichoderma isolates and compost alone or in combination on plant growth parameters
Treatment
21ST DAY
45TH DAY
Disease Severity
Disease Incidence (%)
Healthy Control
0.0 1
FOL only
50.0
83.3
4.3
T.V only
T.H only
Compost Only
FOL+COMPOST
33.3
66.7
3.5
FOL+T.V
16.7
2.0
FOL+T.H
2.3
T.V+COMPOST+FOL
1.3
T.H+COMPOST+FOL
1.7
COMPOST+T.H+T.V
COMPOST+TV+T.H+FOL
Treatments
TPL
TPFW
TPDW
Healthy Control
18.33 h
8.90 h
2.53 g
FOL only
10.73 i
5.50 i
2.13 h
T.V only
27.67 cd
14.50 bc
3.30 c
T.H only
26.67 d
13.33 cd
3.13 d
Compost only
24.67 e
12.27 de
3.03 de
FOL+COMPOST
20.17 g
9.67 gh
2.63 g
FOL+T.V
11.37 ef
2.97 e
FOL+T.H
22.87 f
10.67 fg
2.80 f
T.V+COMPOST+FOL
28.87 c
12.91 de
3.33 c
T.H+COMPOST+FOL
26.90 d
12.37 de
T.V+T.H+COMPOST
35.67a
17.67 a
4.00 a
COMPOST+T.V+T.H+FOL
31.67 b
15.23 b
3.77 b
INTRODUTION
Fusarium oxysporum f.sp lycopersici causes wilt disease in tomato which is one of the most destructive diseases of tomato worldwide (Walker, 1971). Currently, the most effective mode of managing this disease is by application of fungicides. However, chemical management is undesirable due to its deleterious effects on other living organisms, soil microflora and environment (Sundaramoorthy and Balabaskar, 2013). Therefore, public concern has been shifted towards alternative management strategies (Sutton, 1996). Biological control have appeared as an aspiring management strategy for future use (Spadaro and Gullino, 2005). Trichoderma species are the most widely used antagonistic agents perhaps due to their wide host range, development and growth (Whipps and Lumsden, 2001). Several species of Trichoderma have been reported to have in vivo and in vitro antagonistic action against many soil-borne plant pathogenic fungi (Akrami et al. 2009; Morsy et al. 2009). Antagonistic action of Trichoderma species is due to the cell wall degrading enzymes (Chitinase and Glucanase) secreted by them which play a key role in suppressing the attack of many fungal pathogens (Kucuk and Kivanc, 2008). They secrete volatile and non-volatile compounds possessing antibiotic properties (Papavizas, 1985; Dubey et al., 2007; Ajith and Lakshmidevi, 2010). Soil and other potting material used as compost have a natural ability to reduce disease incidence because they have natural microflora which interacts with pathogen (Alabouvette, 1999). The natural suppressive ability of soils and compost media is due to the interactions between microflora naturally inhabiting them and abiotic properties of medium (Alabouvette, 1986; Workneh and van Bruggen, 1994). The use of biological control agents in combination with compost media provides a viable control strategy for many diseases. The efficacy of composts improves after amendment with specific antagonistic microbes (Hoitink et al. 1997; Dukare et al. 2011). Trichoderma species alone or in combination with organic amendments are known to suppress tomato wilt (Cotxarrera et al., 2002; Noble and Coventry, 2005).
The present study plan was designed to evaluate the potential of different natural composts in combination with two isolates of Trichoderma in vivo and in vitro to suppress the incidence of Fusarium wilt of tomato.
DISCUSSION
The present work plan was designed to test the efficacy of Trichoderma isolates i.e., T.viride and T.harzianum in combination with cheaply available natural composts. Trends for in vitro evaluation of Trichoderma isolates have been well defined in literature previously (Jee and Kim, 1987). The present results contradicted many previous findings (Devi et al. 2012; Javaid et al. 2014; Yasmin et al. 2014) where dual culture screening revealed promising results. Trichoderma inhibited the growth of pathogen due to their fast growing potential, mycoparasitism and competition with the pathogen for space and nutrition (Devi et al. 2012). The results for evaluation of the inhibitory potential of volatile and non-volatile metabolites produced by Trichoderma isolates are in conformity with the findings of (Padmodaya and Reddy, 1996). Several reports have documented the antibiosis due to non-volatile compounds produced in culture filtrate of T.viride (Ooka et al., 1966). They produce a number of antibiotic such as Trichodermin, Harzianum A and Trichodermol (Kucuk and Kivanc, 2004; Nakkeeran et al., 2002).
In vitro inhibitory potential of CWE’s revealed maximum inhibition by un-autoclaved 100% NDCM while autoclaved CWE’s had significantly lost their inhibitory potential. The results of previous studies have revealed that CWE’s have been successfully evaluated both in vitro and in vivo for their promising inhibitory effect against different pathogens (McQuilken et al., 1994).
The results of in vivo experiment are in line with the studies of (Abada et al. 2014; Pugliese et al. 2011). Trichoderma combat with the pathogen via different mechanisms which include competition for space and nutrients, induced resistance, mycoparasitism and production of enzymes (Kapulnik and Chet, 2000). Plant growth parameters were significantly enhanced compared to control in Trichoderma and compost amended treatments. Similar results were obtained by (Sundaramoorthy and Balabaskar, 2013). Trichoderma isolates and compost are associated with the promotion of plant growth parameters.
RESULTS
Fig. 1 Inhibitory effect of compost water effect against F. oxysporum f. sp. Lycopersici
CONCLUSION
REFERENCES
The results of present research work indicate that the addition of specific biological control agents to compost can lead to a substrate with a broader suppressive effect.
The level of suppressive properties of compost can be increased upon antagonist enrichment.
Morton, D.J. and W.H. Stroube. (1955). Antagonistic and stimulating effects of soil microorganism of Sclerotium. Phytopathol. 45:
Dennis, C., and Webster, J. (1971a). Antagonistic properties of specific group of Trichoderma. I. Production of non-volatile antibiotics. Trans Br Mycol Soc, 57:
EL-Masry, H. M. Khalil, I. A. Hassouna, S. M. and Ibrahim, H. A. H. (2002). In situ and in vitro suppressive effect of agricultural composts and their water extracts on some phytopathogenic fungi. World Journal of Microbiology & Biotechnology, 18: 551–558.
Special Thanks to my supervisor Prof. S. T. Sahi, Dr. S. Aleem and Dr. Imran-ul-Haq for their guidance, encouragement and \support