Compatibility of chemical pesticides and aggravation of Trichoderma spp against Pathogens  

U.N. Bhale1 , J.N. Rajkonda2
1 Research Laboratory, Department of Botany, Arts, Science and Commerce College, Naldurg Tq. Tuljapur Dist Osmanabad, India
2 Department of Botany, Yeshwantrao Chavan College, Tuljapur Dist. Osmanabad, India
Author    Correspondence author
Bioscience Methods, 2015, Vol. 6, No. 3   doi: 10.5376/bm.2015.06.0003
Received: 27 Oct., 2015    Accepted: 08 Dec., 2015    Published: 30 Dec., 2015
© 2015 BioPublisher Publishing Platform
This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:

Bhale U.N. and Rajkonda J.N., 2015, Compatibility of Fungicides and Antagonistic Activity of <i>Trichoderma</i> spp against plant Pathogens, Bioscience Methods, 6(3): 1-9

Abstract

Compatibility of fungicides viz. Mancozeb and Captan were studied with Trichoderma spp (T. viride, T. harzianum, T. koningii, T. pseudokoningii and T. virens) in vitro at different concentrations. It was indicated that lower concentrations of Mancozeb and Captan did not affect the radial growth of Trichoderma spp. However, concentration of Mancozeb above 5000 μg/ml and of Captan above 500 μg/ml were significantly reduced the radial growth of Trichoderma species. Trichoderma viride, T. harzianum, T. virens and T. koningii showed resistance to Mancozeb and Captan. Trichoderma pseudokoningii is susceptible to these fungicides. However, when increased the concentration of Mancozeb and Captan decreased the radial growth. Trichoderma species are stronger antagonistic against number of plant pathogens and their mycoparasitic activity. All Trichoderma species inhibited the mycelial growth of Alternaria alternata while maximum inhibition performed by T. viride. Trichoderma koningii was highly antagonistic over Rhizoctonia solani followed by T. viride. Radial growth of Aspergillus niger was highly inhibited by T. koningii followed by T. harzianum. Geotrichum candidum was inhibited significantly by T. viride followed by T. pseudokoningii and T. virens. Significant antagonism was exhibited by T. virens against Fusarium oxysporum f. sp. spinacae followed by T. harzianum and T. pseudokoningii. Radial growth and sporulation of Macrophomina phaseolina was antagonized significantly by T. viride followed by T. harzianum and T. pseudokoningii.

Keywords
Trichoderma spp; Compatibility Mancozeb; Captan; fungicides; Pathogenic fungi; Dual culture; Radial growth; Antagonism

Introduction
Trichoderma species have often been used in the management of crop plant diseases. Trichoderma is a genus of asexually reproducing fungi that is present in all types of soils. Some soil borne fungi are difficult to eradicate because they produce resting structures like sclerotia, chlamydospores or oospores for their survival for a longer period of time under adverse environmental conditions (Baker and Cooke, 1974). Use of fungicides for the control of soil borne diseases is costly and also produces environment and health hazards to human and adversely affects the beneficial microorganisms in soil (Dluzniewska, 2003). This was diverted the attention of plant pathologists towards alternate methods for the control of plant diseases. Trichoderma spp is a potential biocontrol agent for the management of various plant diseases like, sapota (Wagh and Bhale, 2011; Bhale, 2013),Leafy vegetables (Rajkonda et al.,2012), spinach (Bhale, 2012). Trichoderma species are known to suppress infections of soil borne pathogens like Macrophomina phaseolina, Rizoctonia solani, Fusarium species and Pythium species on various crops (Benitez et al., 2004, Adenkule et al., 2001, Ehtesham et al., 1990; Lutchmeah and Cooke, 1875; Howell, 1982). Species of Trichoderma also have growth promoting capabilities that may or may not be integral to biological control (Dubey et al., 2007; Benitez et al., 2004, Yedidia et al., 1998). The combined use of biocontrol agent and chemical pesticides has attracted much attention in order to obtain synergistic or additive effects in the control of soil borne diseases (Locke et al., 1985). Reduced amount of fungicide can stress and weaken the pathogen and render its propogules more susceptible to subsequent attack by the antagonist (Heiljord and Tronsmo, 1998). Srinivas and Ramkrishnan (2002) have reported that integration of biocontrol agents and commonly used fungicides showed positive association by reducing the seed infection compared to fungicide and the fungal antagonists’ individually. Recently, biological control combined with chemical fungicide at lower concentration is applicable. It is being a part of IPM (Integrated Plant disease Management) strategy which was applied since chemical controls have used individually cause environmental pollution.Therefore the present study, Trichoderma species viz., T. viride, T. harzianum, T. koningii, T.pseudokoningii and T. virens were tested for compatibility with fungicides like Mancozeb and Captan.
 
1 Materials And Methods
1.1 Source of Trichoderma spp
Rhizospheric soils of irrigated and non irrigated plants were collected from different parts of Marathwada region of Maharashtra, India. From the rhizosphere soil samples, Trichoderma spp were isolated by using PDA and Trichoderma selective medium (TSM) by dilution plate technique (Johnson, 1957). Theisolated specieswere identified up to species level based on colony characters, growth, structure of mycelium, conidiophores, phialides and conidia (Kubicek and Harman, 2002). All Trichoderma spp were purified by hyphal tip technique (Tuite, 1996). The isolated Trichoderma spp were maintained throughout the study by periodical transfers on PDA and TSM slants under aseptic conditions to keep the culture fresh and viable.
 
1.2 Fungicides
a) Mancozeb 75% WP (1000 – 8000 μg/ml) is a broad spectrum contact fungicide with a protective action which belongs to the dithiocarbamates (Manganese ethylene bisdithiocarbamte) family of chemicals, which also includes maneb.
 
b) Captan 50% WP (100-700 μg/ml) (N-trichlorome- thylthio-4-cyclohexene-1, 2-dicarboximide) were used for in vitro.
 
1.3 Compatibility with fungicides
Fungicides like Mancozeb and Captan was incorporated into the medium after sterilization. The fungicides in proportionate dosage were incorporated in to the molten Czapek Dox Agar (CZA) medium after sterilization and dispersed thoroughly by continuous shaking. This was poured in to 90 mm petridishes. Mycelial discs of 8 mm cut from the growing margin of 7 days old culture of Trichoderma species was inoculated at the centre of the petridish and incubated at 26 ± 2 C. The CZA plate without fungicide served as control. The diameter of the colony was measured after 7 days and compared with the control (Tronosmo, 1989).
 
1.4 Source of pathogenic fungi
The test fungi were isolated from naturally infected plants viz. leaf spot of brinjal (Solanum melongana L.) caused by Alternaria alternata, Fruit rots of sapota (Manilkar zapota L.) caused by Rhizoctonia solani, Aspergillus niger and Geotrichum candidum, leaf spot ofspinach (Spinacea oleraea L.) and fruit rot of ivy guard (Coccinia indica Wight & Arn.) caused by Macrophomina phaseolina.
 
1.5 Antagonistic Activity
Antagonistic efficacy of Trichoderma spp viz., T. viride, T. harzianum, T. koningii, T. pseudokoningii and T. virens were tested against the isolated pathogenic fungi by dual culture experiment (Morton and Stroube 1955). Trichoderma spp and test fungi were inoculated 6 cm apart. Three replicates were maintained for each treatment and incubated at 28 ± 2°C for 7 days. Monoculture plates of both served as control. Seven days after incubation (DAI), radial growth of test fungi and Trichoderma spp were measured. Colony diameter of test fungi in dual culture plate was observed and compared with control. Percentage of radial growth inhibition (%RGI) was calculated by using the formula: 100 X [C - T / C], Where C = growth in control and T = growth in treatment (Vincent, 1947).
 
The degree of antagonism between each of the Trichodema species and test pathogens in dual culture was scored on scale of R1-R5 (Bell et al., 1982).
 
2 Statistical Analyses
Statistics recitation in vitro compatibility was statistically analysed using the main factor fungicide i.e. Mancozeb and Capton and pathogenic fungi i.e. Alternaria alternate, Rhizoctonia solani, Aspergillus niger, Geotrichum candidum, Fusarium oxysporum f. sp. spinacae, Macrophomina phaseolina  and the sub-factors were the Trichoderma species. Arcsine transformation of biological control (Trichoderma species) percentage was calculated by using the following formula:
 
Y = arcsin=sin-1
Where, p is the percentage of inhibition and Y is the result of transformation
 
Statistical analysis of the experiments was performed using the Handbook of Biological Statistics (McDonald, 2008).
 
3 Results
3.1 Compatibility of Trichoderma species against fungicides
Species of Trichoderma viz., T. viride, T. harzianum, T. koningii, T. pseudokoningii and T. virens were isolated from the rhizosphere soil of different crop plants. These isolates were deposited in Research laboratory, Department of Botany, Arts Science and Commerce College Naldurg. These isolates were maintained on PDA (Potato Dextrose Agar) media slants. Species of Trichoderma were amended with fungicides and results obtained following (Table 1&2; Figure 1).
 
 

Table 1 Compatibility of Trichoderma spp with different concentrations of Mancozeb fungicide.

Note: *Significantly reduced the radial growth of mycelium.
Figures in parentheses are arcsine transformed value of radial growth of mycelium.

 

 

Table 2 Compatibility of Trichoderma spp with different concentrations of Captan fungicide.

Note: *Significantly reduced the radial growth of mycelium.

 Figures in parentheses are arcsine transformed value of radial growth of mycelium.

 

 

Figure 1 Compatibility of Trichoderma spp with Mancozeb & Captan fungicides.

Note: a) 1-T. viride(Control), 2-Mancozeb 8000 μg/ml, 3-Captan 700 μg/ml; b) 1-T. harzianum(Control), 2-Mancozeb 8000 μg/ml, 3-Captan 700 μg/ml; c) 1-T. koningii (Control), 2-Mancozeb 8000 μg/ml, 3-Captan 600 μg/ml; d) 1-T. pseudokoningii(Control), 2-Mancozeb 5000 μg/ml, 3-Captan 500 μg/ml; e) 1-T. virens(Control), 2-Mancozeb 8000 μg/ml , 3-Captan 500 μg/ml

 
a. Trichoderma viride: Trichoderma. viride showed growth on medium containing Mancozeb@1000, 2000, 3000, 4000 & 5000 μg/ml. The growth of T. viride is least on media containing Mancozeb@6000 and 7000 μg/ml, but no growth was observed when Mancozeb was used @8000 μg/ml. Trichoderma viride grow easily when Captan was used @ 100, 200, 300 and 400 μg/ml, the growth was minimum on Captan@500 and 600 μg/ml, but no growth were recorded on Captan@700 μg/ml.
 
b.  Trichoderma harzianum: Trichoderma harzianum grow at lower concentration of Mancozeb. The growth was inhibited when concentration of Mancozeb was more than 8000 μg/ml. However, increased concentration of Mancozeb decreased the radial growth. In media containing Captan@700 μg/ml, no growth was recorded. The growth of T. harzianum was observed at lower concentration from 100 to 400μg/ml.
 
c. Trichoderma koningii: Trichoderma koningii showed growth in Mancozeb@1000 to 5000 μg/ml. The growth was very low in concentration of 7000 μg/ml, no growth occurred when Mancozeb was used above the @8000 μg/ml. T. koningii showed growth at Captan @100, 200, 300 μg/ml. The growth was slow at Captan @ 500 and 600μg/ml. No growth was observed on Captan@700μg/ml.
 
d. Trichoderma pseudokoningii: Trichoderma pseudokoningii grow slow under media containing Mancozeb@1000 2000, 3000 and 4000 μg/ml, but it was inhibited completely at Mancozeb@5000μg/ml. The radial growth of T. pseudokoningii was also low at Captan@100, 200, 300 and 400μg/ml. But no growth was observed on Captan@500 μg/ml.
 
e. Trichoderma virens: Trichoderma virens grow readily under low concentration of Mancozeb(@1000 – 5000 μg/ml). The radial growth was gradually decreased at Mancozeb@5000 to 7000 μg/ml and no growth was recorded at 8000 μg/ml. Treatment of Captan @100 to 300 μg/ml, the radial growth of T. virens was recorded. However, growth was stopped on Captan @ 500 μg/ml.
 
Only, T.pseudokoningii was found susceptible at lower tested concentrations on both fungicides. The obtained results indicated that low concentration of Mancozeb and Captan does not affect the radial growth of mycelium of Trichoderma spp. However, increased concentration of fungicides were decreased the radial growth of mycelium.
 
3.2 Antagonistic activity of Trichoderma species against pathogenic fungi
Isolated plant pathogens i.e.Alternaria alternata, Rhizoctonia solani, Aspergillus niger, Geotrichum candidum, Fusarium oxysporum f. sp. spinacae and Macrophomina phaseolina were evaluated their antagonistic nature against Trichoderma species under in vitro condition. It was observed that growth of pathogenic fungi was reduced with respect to radial growth and sporulation. The mycelium of Trichoderma species when comes in contact with the test fungi it became fungistatic and the growth of test fungi were retarded (Table 3 & Figure 2). Alternaria alternata:Results indicated that Trichoderma species significantly inhibited the radial growth of Alternaria alternata incitant of leaf spot of brinjal (Solanum melangona). Maximum inhibition of A. alternata was observed with T. harzianum (90.4%) followed by T. koningii (77.7%), T. virens (74.4%) and T. viride (73.3%). However, minimum inhibition showed under the treatment of T. psudokoningii (71.1%).
 
 

Table 3 Antagonistic activity of Trichoderma species against pathogenic fungi.

Note: At-Alternaria alternata, Rs- Rhizoctonia solani, An-Aspergillus niger, Gc-Geotrichum candidum, Fo-Fusarium oxysporum f. sp. spinacae, Mp-Macrophomina phaseolina

 

 

Figure 2 Antagonistic activity of Trichoderma spp on pathogenic fungi.

Note: Row 1: Alternaria alternata (a-control, b-T. virens, c-T. koningii)
Row 2: Rhizoctonia solani (a-control, b- T. koningii, c- T. hrzianum)
Row 3: Aspergillus niger (a-control, b-T. koningii, c-T.viride)
Row 4: Geotrichum candidum (a-control, b-T. virens, c-T.koningii)
Row 5: Fusarium oxysporum f. sp.spinaciae (a-control, b-T. virens, c-T.viride)
Row 6: Macrophomina phaseolina (a-control, b-T. viride, c-T. virens)
 
a. Rhizoctonia solani: Antagonisitic nature of Trichoderma species against Rhizoctonia solani causing fruit rot of sapota (Manilkar zapota) was recorded. The radial growth of R. solani was inhibited more by T. koningii (67.7%). Antagonism of R. solani was also showed by T. viride (51.1%), T. virens (46.6%) and T. pseudokoningii (44.4%). Trichoderma harzianum showed minimum antagonism against R. solani (38.8%).
 
b. Aspergillus niger: Antagonistic activity of isolated Trichoderma species against Aspergillus niger were evaluated under in vitro condition and results were recorded. Trichoderma koningii exhibited maximum antagonism against A. niger (57.7%). However, T. harzianum also showed antagonism against A. niger (54.4%). Radial growth of A. niger was also inhibited by T. viride (44.4%), T. virens (44.4%) and T. pseudokoningii (42.2%). It was observed that the isolated Trichoderma species showed moderate inhibitory effect over A. niger.
 
c. Geotrichum candidum: Results revealed that antagonistic effect of Trichoderma species over Geotrichum candidum causal agent of fruit rot of sapota (Manilkar zapota). Maximum inhibition against G. candidum was recorded by T. viride (74.4%), followed by T. pseudokoningii (72.2%), T. virens (68.9%), T. koningii (66.7%) and T. harzianum (61.1%). The isolated Trichoderma species exhibited significant antagonism against G. candidum.
 
d. Fusarium oxysporum f. sp. spinacae: In vitro antagonistic nature of Trichoderma species against Fusarium oxysporum f. sp. spinacae causal agent of wilt of spinach (Spinacea oleracea)was tested. It was cleared that Trichoderma species inhibited the mycelial growth of F. oxysporum f. sp. spinacae. Among Trichoderma species, maximum inhibition showed by T. virens (70.0%) followed by T. harzianum (66.6%). T. pseudokoningii (62.2%) and T. viride (55.5%).
 
e. Macrophomina phaseolina: Antagonistic effect of Trichoderma species against Macrophomina phaseolina incited fruit rot of ivy guard (Coccinia indica) was recorded and the results showed that Trichoderma species were significant in reducing the radial growth of mycelium of the test fungus. The inhibitory effect of T. viride (84.4%) found maximum followed by T. harzianum (83.3%) and T. pseudokoningii (81.1%). The antagonistic effects by T. virens (57.7%) and T. koningii (55.5%) over M. phaseolina were recorded minimum but these were also significant.  
 
3.3 Bell’s Scale
According to modified Bell’s scale, T.harzianum overgrew beyond 90 percent (R1 scale).  In case of R.solani, only T.koningii overgrew beyond 60 percent (R3 scale).All Trichoderma species were failed to
progress beyond 60 percent (R3 scale) in A.niger. In G.candidum, T.pseudokoningii and T.viride overgrew at least two third of pathogen (R2 scale) but others were beyond 60 percent (R3 scale). In case of F.oxysprum f. sp.spinaceae, T. virens overgrew at least two-third pathogens (70% over growth). Except T.koningii and T. virens other Trichoderma species were overgrew at least two third of pathogen (R2 scale) in M. phaseolina (Table 4)
 
 

Table 4 Evaluation of Trichoderma spp. against pathogenic fungi by dual culture using Bell’s scale*(R).

Note: *Degree of antagonism
R1=Trichoderma completely overgrew pathogens (100% over growth);
R2=Trichoderma overgrew at least two-third pathogens (75% over growth);
R3=Trichoderma colonizes on one half of the pathogens (50% over growth);
R4=Trichoderma and the pathogens contact point after inoculation;

R5= Pathogens overgrow bioagent – Trichoderma.

 
4 Discussion
Experiments were also conducted to determine the compatibility of biocontrol agents with commercially effective chemicals against white rot of apple (Dematophora necatrix) in India (Gupta and Sharma, 2004). They concluded that Carbendazim was inhibitory to all fungal antagonists whereas Mancozeb and Phorate were least inhibitory at 200ppm. Radial growth of mycelium of Trichoderma spp was inhibited at Carbendazim @ 1ppm and Triphonate Methyl @ 10ppm (Malathi et al., 2002). Inhibitory action of Isofenphos – methyl on T. harzianum was strong and fungistatic rate was 64.37% under treatment with 200μg/ml (Ching et al., 2008). The fuingicide Iprodione and T. harzianum were not found to be compatible (Sumitra and Madhuban, 2006). The T. harzianum was least sensitive to Procymidone and Captan and most sensitive to Mancozeb, Tebuconazole and Thiram (Mclean et al., 2001). Inhibitory effect of Carbendazim and Thiophanate Methyl on T. harzianum while Captan and Thiram recorded least inhibitory effect on T. harzianum (Gowdar et al., 2006). Recently, the fungicide Bordeaux mixture 1% found highly inhibitory to Trichoderma as compared to Copper oxychloride and Mancozeb (Suseela and Joseph, 2010). Some strains of Trichoderma show compatibility with fungicides as they are tolerant of fungicides and successfully used in IPM strategy (Dutta and Chatterjee, 2004; Hetong et al., 2008). Trichoderma viride was not compatible with Dithane, Bavistin and Ridimil in any level of selected concentration (Tapwal et al., 2012)
 
Many workers in the discipline of plant pathology suggested that growth of plant pathogenic fungi were inhibited by the Trichoderma species because of some factors produced and these substances may be volatile and non volatile (Reusser, 1967).The growth inhibition of pathogenic fungi may be due to antibiotic secretion like trichodermin, trichoviridin, dermadin and sesquiterpene heptalic acid (Nakkeeran et al., 2002), nutrient impoverishment and pH alteration in the medium (Maheshwari et al., 2001). Such type of variability in antagonistic potential of Trichoderma species against plant pathogens has been also reported (Saha and Pan, 1996; Bell et al., 1982). Mathew and Gupta (1998) also reported that T. harzianum also exhibited maximum antagonistic activity causing 58% inhibition followed by T. viride (46%) and T. virens (45%). Isolates of T. viride, T. harzianum and T. virens were evaluated for their antagonism against Pythium aphanidermatium causing damping off of tomato and found that many isolates inhibited the growth of P. aphanidermatum (Kumar and Hooda, 2007).Trichoderma viride is economically important because of their mycoparasitic ability which makes them suitable for the application as biocontrol agent against soil borne plant pathogenic fungi (Manczinger et al., 2000). The effective in vitro screening test of T. viride was carried out against Rhizopus oryzae and Aspergillus flavus pathoges of post harvest cassava (Manihot esculents Crantz.), root rot and reported that T. viride was most promoting candidate for the biocontrol (Ubalua and Oti, 2007). Growth of Rhizoctonia solani, a pathogen involved in cotton seedling disease was inhibited by the strains of T. harzianum and T. longibracheatum (Arsan-Amal et al., 2005). Strains of T. koningii were used for their antagonistic nature against Rhizoctonia solani under in vitro condition and inhibited mycelial growth by producing toxic metabolites (Melo and Faull, 2000). In the dual culture experiment evaluated by Hajieghrari et al. (2008), T. virens and T. harzianum inhibited the growth of soil borne pathogenic fungi such as R. solani, M. phaseolina, Phytophthora cactorum and Fusarium graminearum forming inhibition zone without physical contact between them. In vitro antagonistic potential of T. viride against Alternaria alternata, Ulocladium botrytis, Cladosporium harbarum, Cephalosporium madurae, Penicillium chrysogonum,Fusarium oxysporum and Humicola grisa were tested and found that significant inhibition of radial growth of these fungi in dual culture experiment (Abou-Zeid et al., 2008).
 
Rajendiran et al. (2010) evaluated antagonistic effects of T. viride on post harvest pathogens of fruit and vegetables such as Aspergillus niger, A. flavus, A. fumigatus, Fusarium sp and Penicillium sp. Trichoderma viride inhibited the radial growth of A. niger (55%), A. flavus (51%), A. fumigates (52%), Fusarium sp (64%) and Penicillium sp in dual culture. The impact of isolates of T. viride, T. harzianum and T. virens on soil borne fungal pathogens such as R. solani, S. rolfsii and Sclerotinia sclerotiorum were evaluated and inhibitory effects were reported (Amin et al., 2010). Reports on antagonistic potential of T. harzianum over Fusarium oxysporum f. sp. vanilla the stem rot pathogen of vanilla was showed by Naik et al. (2010).The isolates were found fully overgrown on all corm rot pathogens of saffron (Hassan et al., 2011). Trichoderma viride was found to exhibit effective antagonistic potentiality against R. solani (Giagole et al., 2011).
 
5 Conclusion
Trichoderma species can be used together with compatible fungicides in the integrated disease management towards the control of crop plants and soil borne pathogens. It is possible to develop Trichoderma tolerant of chemical fungicides without lack of antagonistic activity. The antagonistic nature of Trichoderma species against pathogenic fungi were evaluated under in vitro condition. It was observed that growth of pathogenic fungi was reduced with respect to radial growth and sporulation. The mycelium of Trichoderma species when comes in contact with the test fungi it became fungistatic and the growth of test fungi were retarded. The antagonism was exhibited with respect to secretion of extra cellular enzymes, antibiotics and competition related food and space. Pathogenic fungi and Trichoderma species created competition and the latter found to be dominant over the pathogenic fungi. Mycoparasitic properties of Trichoderma species was found to be the main reason responsible for their antagonistic nature.
 
Acknowledgement
Authors are thankfully acknowledged to UGC, New Delhi for financial assistance of major research project.
 
References
Abou-Zeid A. M., Altahi A D., and Abd El-Fattah R. I., 2008, Fungal control of pathogenic fungi isolated from some wild plants in Taif Governorate, Saudi Arabia. Mal.J. Microbiol., 4(1):30-39.
 
Adenkule A.T.,Cardwell D.A., Florini and Ikotun T., 2001. Seed treatment with Trichoderma species for control of damping off of cowpea caused by Macrophomina phaseolina, Biocontrol Science and Technology, 11: 449-457.
http://dx.doi.org/10.1080/09583150120067481
 
Amin F., Razdan V. K., Mohoddin F. K., Bhat K. A. and Bandey S., 2010, Potential of Trichoderma species as biocontrol agents of soil borne fungal propagules, Journal of Phytology, 2(10): 38-41.
 
Arsan-Amal A., Abd-Elsalam K.A., Omar M. R., and Aly A. A., 2005, Antagonistic potential of Trichoderma spp against Rhizoctonia solani and use of M13 microsatellite-primed PCR to evaluate the antagonist genetic variation, Journal of Plant Diseases and Protection, 112(6): 550-561.
 
Baker K.F., and Cooke R.J., 1974, Biological control of plant pathogens, W.H. Freeman Press, San Fransisco.
 
Bell D.K., Wells H.D. and Markham C.R., 1982, In vitro antagonism of Trichoderma species against six fungal plant pathogens, Phytopathology, 72, 379-382.
http://dx.doi.org/10.1094/Phyto-72-379
 
Benitez T., Rincon A.M., Limon M.C., and Codon A.C., 2004, Biocontrol mechanism of Trichoderma strains,Intl. Microbiol., 7(4):249-260.
 
Bhale U.N., Ambuse M.G., Chatage V.S. and Rajkonda J.N., 2012, Bioefficacy of Trichoderma isolates against pathogenic fungi inciting spinach (Spinacea oleracea L.), JBiopest., 5(2): 222-227.
 
Bhale U.N., Wagh P.M. and Rajkonda J.N., 2013, Antagonistic confrontation of Trichoderma spp against fruit rots pathogens on Sapodilla (Manilkara zapota L.), Journal of Yeast and Fungal Research, 4(1): 5-11.
 
Ching D.M., 2008, Compatibility of mycelial growth and conidia germination of T. harzianum with several soil applied insecticides, Journal of Anhui Agricultural Sciences, pp.25 -32.
 
Dluzneiwska J.,2003, Reaction of fungi of Trichoderma genus to selected abiotic factors,Elec. J. Polish Agr. Uni. Agron, 6 (2): 239-242.
 
Dubey C.S., Suresh M. and Singh B., 2007, Evaluation of Trichoderma species against Fusarium oxysporum f.sp.ciceris for integrated management of chickpea wilt, Biological Control, 40(1):118-127.
http://dx.doi.org/10.1094/Phyto-72-379
 
Dutta S. and Chatterjee N.C., 2004,.Raising of carbendazim-tolerant mutants of Trichoderma and variations in their hydrolytic enzyme activity in relation to mycoparasitic action againts Rhizopus stolonifer,  Journal of Plant Diseases and Protection,111(6):557-565.
 
Ehteshamul H.S., Zaki M.J., and Gaffar A., 1990, Biological control of root rot disease of okra, sunflower, soybean and mungbean. Pak. J. Bot., 22: 121-124.
 
Giagole A.H., Wagh G.N., and Khadse A.C.,2011, Antifungal activity of Trichoderma species against soil borne pathogens, Asiatic J. Biotech. Res., 2(4): 461-465.
 
Gowdar S.B., Babu-Ramesh H.N., Nargund V.B., and Krishnappa M., 2006, Compatibility of fungicides with T. harzianum, Agricultural sciences Digest,26(4): 203-209.
 
Gupta V.K., and Sharma K., 2004, Integration of chemicals and biocontrol agents for managing white root rot of apple ISHS Acta Horticulturae  Eds. A. Vanacter) XXVI International Horticultural Congress: Managing soil borne pathogens: A sound Rhizosphere to improve productivity in intensive horticultural systems, Acta horticulturae, pp. 307-313.
 
Hajieghrari B., Torabi-Giglou M., Mohammadi M.R., and Davari M.,2008, Biological potential of some Iranian Trichoderma isolates in the control of soil borne plant pathogenic fungi,African Journal of Biotechnology, 7(8): 967-972.
 
Hassan M.G., Devi L.S., Ahmad S., Manojkumar V., and Williams P., 2010, The biological control of paddy disease brown spot (Bipolaris oryzae) by using Trichoderma viride in vitro condition,Journal of Biopesticides, 3(1): 93-95.
 
Hetong Y., Ryder M. and Wenghua T., 2008,Toxicity of Fungicides and Selective Medium Development for Isolatiom and Enumeration of Trichoderma spp. In: Agricultural soils. International Subcommision on Trichoderma and Hypocrea Taxonomy, China 2008.
 
Hjeljord I. and Tronsmo A., 1998, Trichoderma and Gliocladium in biological control: an overview. In: Trichoderma and Gliocladium- Enzymes, Biological control and Commercial Applications (Eds.): G.E. Herma and C.P. Kubick. Taylor & Frasncis Ltd London, Great Britain, pp.74-106.
 
Howell C.R., 1982, Effect of Gliocladium virens on Pythium ultimum, Rhizoctonia solani and damping off of cotton seedlings, Phytopathology, 74:106.
 
Johnson L.A., 1957, Effect of antibiotics on the number of bacteria and fungi isolated and fungi isolated from soil by dilution plate method, Phytopathology,47: 21-22.
 
Kubicek C.P. and Harman G.E., 2002, Trichoderma and Gliocladium (vol. I). Basic Biology,Taxonomy and Genetics. pp. 14-24. 
 
Kumar R. and Hooda I., 2007, Evaluation of antagonistic properties of Trichoderma species against Pythium aphanidermatum causing damping off of tomato, J. Mycol. Pl. Pathol., 37(2): 240-243.
 
Locke J.C., Moris J.J. and Papavizas G.C., 1985, Biological control of Fusarium wilt of greenhouse grown Chrysanthemums. Plant Dis, 69: 167-169.
http://dx.doi.org/10.1094/PD-69-167
 
Lutchmeah R.S. and Cooke R.I., 1985, Pelleting of seeds with the antagonist Pythium oligandrum for biological control of damping off,Plant Pathology,34:528 – 531.
http://dx.doi.org/10.1111/j.1365-3059.1985.tb01403.x
 
Maheshwari D.K., Dubey R.C., and Sharma V.K., 2001, Biocontrol effects of Trichoderma virens on Macrophomina phaseolina causing charcoal rot of peanut, Indian J. Microbiol., 41:251-256.
 
Malathi P., Vishwanathan R., Padmanabhan P., Mohanraj D., and Ramesh Sunder A., 2002,Compatibility of biocontrol agent with fungicides against red rot of sugarcane, Suger Tech., 4 (3-4):131-136.
http://dx.doi.org/10.1007/BF02942694
 
Manczinger L., Antal Z. and Kredics L.,2000, Ecophysiology and breeding of mycoparasitic Trichoderma strains, Acta. Microbiol. Immunol., 49: 1-14.
 
Mathew K.A., and Gupta S.K., 1998, Biological control of root rot offrenchbean caused by Rhizoctonia solani, J. Mycol. Pl. Pathol., 28: 202-205.
 
McDonald J.H., 2008, Handbook of Biological statistics. Sparky House Publishing, Baltimore,Maryland, pp.160-164.
 
Mclean K.L., Hunt J., and Stewrat A., 2001, Compatibility of Trichoderma harzianum with selected fungicides,New Zealand Plant Protection,54: 84-88.
 
Melo I.S., and Faull J.L., 2000, Parasitism of Rhizoctonia solani by strains of Trichoderma spp, Sci. Agric., 57(1): 73-77.
http://dx.doi.org/10.1590/S0103-90162000000100010
 
Naik G., Saifulla B., Nagaraja R ,and Basavraja M.K., 2010, Biological control of Fusarium oxysporum f. sp. vanillae, the causal agent of stem rot of vanilla in vitro, International Journal of Science and Nature., 1(2):259-261.
 
Nakkeeran S., Krishnamoorthy A.S., Ramamoorthy V., and Renukadevi S., 2002, Microbial inoculants in plant disease control, J. Ecobiol., 14:83-94.
 
Rajendiran R., Jagdeeshkumar D., Sureshkumar B.T. and Nisha T.,2010, In vitro assessment of antagonistic activity of Trichoderma viride against post harvest pathogens, Journal of Agricultural Technology, 6(1): 31-35.
 
Rajkonda J.N., Jadhav D.S., and Bhale U.N., 2012, Efficacy of Trichoderma species on leafy vegetable crops. Shodhankan (International Multidiciplinary referred & reviewed Research Journal, Special Issue. pp.62-67.
 
Reusser F., 1967, Biosynthesis of antibiotics. U-22, 324, a cyclic polypeptide, J. Biol. Chem., :242-243.
 
Saha D.K. and Pan S., 1996, In vitro antagonistic potential of different isolates of Gleocladium virens of West Bengal, J. Natl. Bot. Soc., 50: 13-18.
 
Srinivas P., and Ramkrishnan G., 2002, Use of native microorganism and commonly recommended fungicides in integrated management of rice seed borne pathogens, Annl Pl Protect. Sci., 10(2): 260-264.
 
Sumitra A. and Madhuban G., 2006, Bioefficacy of Iprodione against two diseases, its compatibility with T. harzianum and residues on cabbage crops, Journal of Environmental Science and Health Part B., 41( 6): 949-963.
http://dx.doi.org/10.1080/03601230600806152
 
Suseela B.R. and Joseph T., 2010, Compatibility of Trichoderma harzianum (Refai) with fungicides, insecticides and fertilizers.Indian Phytopath,63(2): 145-148.
 
Tapwal A.R. Kumar N.G. and Pandey S., 2012, Compatibility of Trichoderma viride for selected fungicides and botanicals, International Journal of Plant Pathology, 3(2):89-94.
http://dx.doi.org/10.3923/ijpp.2012.89.94
 
Tronosmo A., 1989, Effect of fungicides and insecticides on growth of Botrytis cinera, Trichoderma viride and T. harzianum, Biol.Control,1:59-62.
 
Tuite J., 1996, Plant Pathological Methods. Fungi and Bacteria, Burgess Pub. Co. Minneapolis, Minn. USA. p. 293.
 
Ubalua A.O. and Oti E., 2007, Antagonistic properties of Trichoderma viride on post harvest cassava root rot pathogens, Afr. J. Biotechnol., 6(21): 2447-2450.
 
Wagh P.M. and Bhale U.N., 2011, In vitro biocontrol activity of Trichoderma species against fungal phytopathogen Rhizoctonia solani from post-harvest sapota, Proceeding of UGC sponsored National Conference on “Role of non agricultural Institution in the important of Agricultural technology” held at Dept of Botany, Shrikrishna Mahavidyalaya gunjoti on 23-24th Jan.2011. Bionanofrontier, Special issue. pp. 36-38.
 
Yedidia I.N., Benhamou N. and Chet, I., 1999, Induction of defense responses in cucumber plants (Cucumis sativus L.) by biocontrol agent Trichoderma harzianum, Appl. Environ. Microbiol., 65: 1061 – 1070. 

 

Bioscience Methods
• Volume 6
View Options
. PDF(462KB)
. FPDF
. HTML
. Online fPDF
Associated material
. Readers' comments
Other articles by authors
pornliz suckporn sex videos bbw mom xxx big fucking arabin porn videos teen gril sex video riding hard cock woman hard vagina . U.N. Bhale
. J.N. Rajkonda
Related articles
. Trichoderma spp
. Compatibility Mancozeb
. Captan
. fungicides
. Pathogenic fungi
. Dual culture
. Radial growth
. Antagonism
Tools
. Email to a friend
. Post a comment