Compatibility of Trichoderma viride for Selected Fungicides and Botanicals
Trichoderma viride can thrive in diverse environmental conditions as aggressive colonizers of soil and the roots of plants and act as natural bioagent to protect plants from infection by soil-borne fungal pathogens. Laboratory experiments were conducted to test the possibility of combining fungicides and botanicals with Trichoderma viride to work out their compatibility to devise a suitable integrated management of soil borne plant diseases. Five fungicides viz., dithane M-45, ridomil, captaf, blue copper, bavistin and five botanicals viz., Parthenium hysterophorus, Urtica dioeca, Cannabis sativa, Polystichum squarrosum and Adiantum venustum were evaluated at different concentration. Among fungicides only captaf and blue copper had recorded compatiblility to some extent with T. viride. While the water extracts of the tested botanicals were quite compatible with Trichoderma except for C. sativa, which have some inhibitory effect on the growth of pathogens. Present investigation suggests that compatible fungicides and botanicals can be used with Trichoderma in an IDM package to control soil borne plant pathogens.
Received: January 30, 2012;
Accepted: March 24, 2012;
Published: June 05, 2012
Soil-borne diseases are consequence from the reduction of biodiversity of soil
antagonistic organisms. Fungicide applications to soil, kills important beneficial
fungi and also weakens the natural antagonistic activity (Lenteren
and Woets, 1988). Inspite of well known side effects of chemicals on environment,
they are continuously used to control soil borne plant pathogens. To reduce
the use of pesticides, biological control method has been considered as more
natural and environmentally acceptable approach (Bagwan,
2010). Several species of Trichoderma are well documented mycoparasites
and have been used successfully against certain pathogenic fungi. Trichoderma
strains are the key antagonists for the eco-friendly management of plant
diseases. Significant growth inhibition by Trichoderma has been reported
for Armillaria mellea (Tapwal et al., 2004),
Dematophora necatrix (Tapwal et al., 2005),
Phytophthora cinnamomi (Singh et al., 2010),
Fusarium oxysporum and Rhizoctonia solani (Dar
et al., 2011), Sclerotium rolfsii (Jegathambigai
et al., 2010) and Fusarium oxysporum f.sp. psidii (Jegathambigai
et al., 2009; Srivastava et al., 2011).
Many other workers (Salehpour et al., 2005; Abdollahzadeh
et al., 2006; Mir et al., 2011; Osman
et al., 2011) utilised Trichoderma species as a potential
biological control agent. In an IDM package, incorporation of natural products
provides a viable solution to the environmental problems caused by synthetic
pesticides. Identification of these compounds and their further testing may
be an effective approach to minimise the use of hazardous chemicals (Duke,
To develop an effective disease management programme, the compatibility of
potential bioagents with fungicides and botanicals is essential. Combination
of chemicals and compatible bioagents in an IDM strategy protects the seeds
and seedlings from soil-borne and seed-borne inoculum (Dubey
and Patil, 2001). Integration of compatible bioagent with pesticides, may
enhance the effectiveness of disease control and provide better management of
soil borne diseases (Papavizas and Lewis, 1981). The combination
of biological control agents with fungicides would provide similar disease suppression
as achieved with higher fungicide use (Monte, 2001).
Combining antagonists with synthetic and non synthetic chemicals eliminates
the chance of resistance development and reduces the fungicide application.
In view of this, laboratory experiments were conducted to test the possibility
of combining Trichoderma viride with fungicides and botanicals. The long
term goal is to develop an effective IDM package for managing soil borne plant
diseases as well as to prevent the resistance development in pathogens to chemicals.
Integrating chemical resistant Trichoderma strains has an importance
in the framework of integrated disease management. Disease prevention can be
increased by using such tolerant strains that keeps pathogens under sufficient
pressure so that they cannot thrive.
MATERIALS AND METHODS
Pure culture of T. viride was collected from the Department of Botany, Shoolini Institute of Life Sciences and Business Management, Solan, Himachal Pradesh. Compatibility tests were conducted under in vitro condition to find out safer fungicides and botanicals against Trichoderma. Five fungicides viz., Dithane M-45, Ridomil, Captaf, Blue Copper, Bavistin and were evaluated against Trichoderma by food poisoning technique. Fungicides were added to molten PDA just before pouring from the common stock solution to get final concentrations of 50, 100, 200, 300 ppm, respectively. Parthenium hysterophorus, Urtica dioeca, Cannabis sativa, Polystichum squarrosum and Adiantum venustum were collected from the undisturbed habitats of Solan district, Himachal Pradesh (India). Fresh leaves of healthy plant species were washed thoroughly with tap water and air dried. One hundred grams of plant tissue was ground using pestle and mortar by adding equal amount (100 mL) of sterilized distilled water (1:1 w/v). The pulverized mass was squeezed through cheese cloth and the extracts were centrifuged at 10000 rpm for 5-10 min. The supernatant was filtered through millipore filters (45 μm) using vacuum pump assembly under aseptic conditions to avoid contamination. A requisite amount of the filtrate was mixed in PDA just before pouring to get desired concentrations of 5, 10, 15 and 20% and gently shaken for thorough mixing of the extract.
The PDA plates amended with fungicides and plant extracts were inoculated aseptically
with Trichoderma by transferring five mm diameter agar disc from fresh
cultures. Three replications were maintained for each treatment. Unamended PDA
served as the control. Inoculated petri plates were incubated at 25±1°C.
The radial growth of T. viride was measured in all treatments after three
days and compared with control. The percent growth inhibition of pathogen was
estimated by using the formula following Vincent (1947)
and converted into percent compatibility:
||Percent growth inhibition
|| Colony diameter in control
|| Colony diameter in treatment
The data was recorded in triplicates and subjected to statistical analysis and conclusions were drawn on the basis of analysis of variance. The calculated value of F was compared with the tabulated values at 5% level of significance for an appropriate degree of freedom.
RESULTS AND DISCUSSIONS
Laboratory experiments were conducted to observe the compatibility of T.
viride with fungicides and botanicals. The results revealed that at the
selected concentrations of fungicides, only blue copper and Captaf were compatible
to some extent (Table 1). The compatibility index of Blue
copper with T. viride at different concentrations ranged in between 34.9-97.9%,
followed by Captaf (16.7-25.0%). The percent compatibility decreased with an
increase in the concentration of fungicide. Trichoderma viride was not
compatible with Dithane, Bavistin and Ridomil in any level of selected concentration.
The statistical analysis revealed that only blue copper has recorded significant
differences in comparison to control and Captaf (SEM± = 3.10, CD (p =
0.05) = 9.31). Bagwan (2010) reported that thiram (0.2%),
copper oxychloride (0.2%) and mancozeb (0.2%) are compatible with Trichoderma
harzianum and Trichoderma viride. Trichoderma was most sensitive
to captan, tebuconazole, vitavax, propiconazole and chlorothalonil. In a similar
study, T. harzianum was found highly sensitive to mancozeb, tebuconazole
and thiram, less sensitive to benomyl, triadimenol and dichlofluanid are relatively
insensitive to procymidone and captan (Mclean et al.,
2001). In the Present study, Trichoderma was found insensitive to
blue copper and captaf and highly sensitive to dithane, bavistin and ridomil.
Botanicals are an important component of IPM. The aqueous extracts of tested
plant species were quite compatible with T. viride (Table
2). The results revealed that extracts of Parthenium, Adiantum and
Urtica recorded absolute compatibility at tested concentrations. The results
revealed that extracts of Parthenium, Adiantum and Urtica recorded
absolute compatibility at tested concentrations. This is followed by Polystichum
recorded 100% compatibility at 5% concentration of aqueous extract and 90-95%
compatibility at 10-20 concentration. The minimum compatibility was observed
by Cannabis in the range of 40-77.5% at different concentration of phytoextract.
The percent compatibility decreased with increase in the concentration of phytoextract.
The statistical analysis revealed that the only Cannabis had recorded
significant differences (Sem± = 0.96, CD (p = 0.05) = 2.89). Leaf extract
of Parthenium, Urtica and Adiantum, were found effective against
A. solani, A. zinnia, R. solani, F. oxysporum and C. lunata (Tapwal
et al., 2011).
|| In vitro compatibility of selected fungicides with
|SEM±: 3.10, CD (p = 0.05): 9.31
|| In vitro compatibility of selected botanicals with
|SEM±: 0.96, CD (p = 0.05): 2.89
Leaf Extracts of Parthenium, Adiantum and Urtica also showed
absolute compatibility with Trichoderma in the present study. Similarly,
Vanitha (2010) reported that wintergreen oil, lemongrass
oil and their combination under in vitro conditions did not inhibit the
growth of Trichoderma.
Antagonistic activity of biocontrol agents might be effective if it is integrated
with other control practice and may result in acceptable levels of disease control
with reduced level of chemicals use (Latorre et al.,
1997). The present investigations provide evidence for the compatibility
of Trichoderma with synthetic and natural chemicals. Curl
et al. (1976) were of opinion that combined application of PCNB with
T. harzianum effectively controlled Rhizoctonia solani in cotton
seedlings than T. harzianum alone in greenhouse studies. Similar report
of integration of biological agent and chemicals was reported by Henis
et al. (1978).
Besides having great antagonistic potential, Trichoderma has the capability
of degradading xenobiotic compounds and can survive in environments with remnants
of fungicide molecules (Chaparro et al., 2011).
Present finding indicates that seed treatment or soil application of Trichoderma would be compatible with blue copper fungicide and plant extracts viz., Parthenium, Adiantum and Urtica for the integrated management of soil borne diseases. T. viride can be combined with seed treatment fungicides like blue copper and captaf at lower concentrations. Our future studies are directed to determine the compatibility of Trichoderma and chemicals in managing soil borne diseases of various crops under greenhouse and field conditions. Long term goal is to develop an integrated disease management strategy by combing Trichoderma and chemicals so as to prevent pathogen from gaining resistance as well as in building up of Trichoderma population levels in the soil that will be effective on a long term basis.
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