In vitro interaction of Trichoderma isolates against

Indian Journal of Fundamental and Applied Life Sciences ISSN: 2231-6345 (Online) An Online International Journal Available at http://www.cibtech.org/jls.htm 2011 Vol. 1 (3) July-September, pp. 125-128/ Agarwal et al.

Research Article

In vitro interaction of Trichoderma isolates against Aspergillus niger, Chaetomium sp. and Penicilium sp. *

Tripti Agarwal1, Abhiniti Malhotra1, Manish Biyani2 and PC Trivedi1 1 Department of Botany, University of Rajasthan, India. 2 Department of Biotechnology, BGC, Jaipur, India * Author for Correspondence

ABSTRACT A biocontrol agent that can possess many modes of action is preferable for effective control of plant pathogens. In the present study in vitro screening using the dual culture technique was undertaken to assess the potential of interaction of seven Trichoderma sp. against Aspergillus niger, Chaetomium sp. and Penicilium sp. The test pathogens were isolated from the seeds of Chickpea. The test antagonists grow faster than the pathogen and produced inhibition zones thereby limiting the growth of the pathogen. These antagonistic interactions influence the incidence and severity of the disease caused by the pathogen. The present communication describes the impact of different Trichoderma isolates on growth inhibition of seed borne pathogens under in vitro conditions. The results revealed that the bioagents used were able to inhibit the growth of the pathogen mycelia significantly. Key Words: Trichoderma, Aspergillus niger, Chaetomium and Penicilium Chickpea. INTRODUCTION Trichoderma sp. are now the most common fungal biological control agents that have been comprehensively researched and deployed throughout the world. Several fungal cell wall degrading enzymes like chitinase and glucanase seems to play an important role in the antagonistic action of Trichoderma against a wide range of fungal pathogens (Kucuk and Kivanc, 2008). Chickpea (Cicer arientium L.) which is an important legume crop of India, commonly known as ‘gram’ grown over 6.66 m ha of land (Kochhar, 2009) has been found to be attacked by 172 pathogens including 67 species of fungi (Nene et al., 1996). The disease can affect the crop at any stage of growth. Characteristic symptoms are sudden drooping of leaves and petioles, no external rotting of roots and black internal discoloration involving xylem and pith (Dubey and Singh, 2004).Considering the hazardous effects and high cost of pesticides used for the management of these fungal diseases plant pathologists are diverted to find out the alternative techniques of plant disease control which may cause little or no adverse effect on environment. (Ghaffar 1988a, b 1992) In recent times, there has been a worldwide swing to the use of eco-friendly methods for protecting the crops from pests and diseases. The first requirement of biological control is the identification and deployment of highly effective strains. The filamentous fungi, Trichoderma have attracted the

attention because of their multiprong action against various plant pathogens (Harman et al., 2004). Therefore, in the present investigation experiments on the screening of different isolates of Trichoderma were evaluated to find out the most effective fungal antagonists against Aspergillus niger, Chaetomium sp. and Penicilium notatum for their further evaluation MATERIALS AND METHODS Seed borne pathogens viz., A.niger, Penicillium sp ,Chaetomium sp were isolated from seeds of Chickpea. Various Trichoderma species were screened against test pathogens by dual culture method (Rama Bhadra Raju et al., 2000) using potato dextrose agar (PDA). A mycelial disc (5mm) obtained from the peripheral region of 5-7-day-old cultures of., A.niger,Penicillium sp ,Chaetomium sp on PDA, was placed on a fresh PDA plate (3 cm from the center) and then a 5mm- mycelial disc, obtained from the periphery of a 5-7-day-old culture of Trichoderma species was placed 3 cm away from the inoculum of the pathogen.. The radial growth of the pathogen in dual culture and control plates was measured after seven days of incubation at 28±10C and the inhibition per cent of pathogen was calculated as described by Vincent and Budge (1990). The percentage inhibition growth of tested pathogens in presence of Trichoderma strains was

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Research Article calculated over control. The growth inhibition was calculated by using the formula

100 x (r1 –r2) r1

Where, r1 = diameter of fungal colony in control r2 = diameter of fungal colony in dual inoculation The degree of antagonisms between each bioagent and test pathogen in dual culture was scored on scale of 1-5 as proposed by Bell et al. (1982), which is ; R1= complete overgrowth ; R2 = 75 % overgrowth ; R3 = 50% overgrowth; R4 = growth inhibition at line of contact; R5 = pathogen overgrowing antagonist. Microscopic examination of hyphae from the interaction zone was also carried out to find out the events of hyphal interactions meter of fungal colony in dual inoculation.

RESULTS AND DISCUSSION Our result explains that significant success in biocontrol is achieved under in vitro conditions. It was shown that Trichoderma species could restrict growth of pathogens on Potato Dextrose Agar medium in the dual culture (Table 1-3). The per cent inhibition of radial growth of tested fungi viz., A. niger (78.77%), Cheatomium sp.(41.48%) and Penicillium sp. (44.46 %) were reduced by Trichoderma harzianum (T7) with maximum inhibition of A.niger. The maximum growth inhibition of Chaetomium sp. (68.88) was by Trichoderma harzianum (T2).The growth inhibition of Penicillium sp was found to be reduced by 76.31 % Trichoderma harzianum (T8). Regarding the mechanism of biological control of the pathogen, it controls mainly being mycoparasite and an aggressive competitor with pathogens caused twisting, air bubbling and disintegration of the pathogen hyphae. Besides they may also produce antifungal phenolic compounds (Saba Banday et al., 2008). Trichoderma viride (T.V.) did not produce any signification demarcation zone as compare to others.

Table 1 Antagonistic activity of Trichoderma spp. against A. niger by dual culture method. S. No.

Test Mycoflora

Colony growth (mm) of A. niger Control (mm)

Interaction (mm)

Growth Inhibition A. niger (%)

1

Trichoderma harzianum(T1)

45

12.77

71.62

2


45

17

62.22

3

Trichoderma harzianum(T3) Trichoderma harzianum (T7)

45

15

66.66

45

11.65

74.11

5

Trichoderma harzianum (T8)

45

14.22

68.4

6

Trichoderma viride (T.V.)

45

26.66

40.75

4

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Research Article Table 2: Antagonistic activity of Trichoderma spp. against Chaetomium spp. by dual culture method S.No.

Test Mycoflora

Colony growth (mm) of Chaetomium Control (mm)

Interaction (mm)

Growth Inhibition Chaetomium (%)

1


45

18

60

2


45

14

68.88

3


45

25.33

43.71

4


45

26.33

41.48

5


45

25.66

42.97

6


45

30.33

32.6

Table 3: Antagonistic activity of Trichoderma spp. against Penicillium sp by dual culture method. S.No.

Test Mycoflora

Colony growth (mm) of Penicillium spp Control (mm)

Interaction (mm)

Growth Inhibition Penicillium spp (%)

1


45

16

64.44

2


45

20.66

54.08

3

Trichoderma harzianum(T3) Trichoderma harzianum (T7)

45

22.66

49.64

45

24.22

44.46

5


45

14

68.88

6


45

4

37.77 28

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Research Article Plant pathogenic fungi are a widespread problem and the use of chemicals is hardly successful (Anand and Jayarama, 2009).However, the high cost associated with the use of fungicides to control disease caused by soil borne fungi is a limiting factor in the profitability of crop production. In that case biological control could be the best alternative. Trichoderma is the most commonly used fungal biological control agent and have long been known as effective antagonists against plant pathogenic fungi (Chet et al., 1981; Papavizas, 1985). Some of the isolates of Trichoderma spp. included in the present study significantly inhibited several pathogens (Dubey, 2003). All the Trichoderma isolates inhibited the growth of all the three test pathogens in their own way. Thus, it is well known that all isolates collected from different samples of soil are not equally antagonistic to pathogen and searching of effective isolate to locally suit the purpose is important.

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