Production of Biopesticide - Trichoderma viride

Aim:

To isolate, identify and Mass production of Trichoderma viride.

Introduction:

            Trichoderma viride is very promising method against soil borne plant parasitic fungi. The fungal pathogens play a major role in the development of diseases on many important field and horticultural crops; resulting in severe plant yield losses. Intensified used of fungicides has resulted in accumulation of toxic compound potentially hazardous to human and environment an also in the buildup of resistance of the pathogens. Large scale production, along with shelf life and establishment of bioagents in targeted niche, determine the success of biological control. Therefore cost effective large scale production, shelf life of formulation, establishment of bioagent in to targeted niche and consistency in disease control are the primary concern with augmentative biological control. Adaptation of technology in the biocontrol arsenal needs to be investigated. Development of acceptable easily prepared and cost effective formulations for delivery should be major goal.

Materials Required:

PDA Medium

Grains (Rice, Wheat, Pulses)

Procedure:

Isolation and Identification of T. viride

Fungal species Trichoderma viride was isolated from soil samples by using potato dextrose agar (PDA) mediumGreen conidia forming fungal bodies were selected and microscopic observation was identified to be Trichoderma viride. The culture was maintained on PDA slants.

Grain Medium

 Three Grains viz rice, wheat and pulses were used for estimating the biomass of Trichoderma viride at 26 ˚C. 20 g of each grain was washed well and boiled in distilled water for 1 hr. and then mesh properly and filter it, now makeup 1 liter with distilled water. Now these grain mediums were packed separately in individual 500 ml conical flask for Trichoderma viride. They were plugged with cotton wool and autoclaved at 15 psi for 1hr at 121˚C. After cooling, 1 gm of the fungal culture was inoculated into each flask, separately. All these action were done under laminar air flow chamber. They were incubated in BOD incubator at 26˚C for 3 weeks. To avoid clumping, after 7 days of inoculation, the flasks were shaken vigorously to separate the culture and to break the mycelial mat. After 14 days of incubation, the mycelial mat appeared in flasks. Now it was grow well for 21 days. The flasks were shaken in mechanical shaker for 10 min. The suspension was filtered through double layered muslin cloth and then taken biomass in each grain medium.

Result:

Among the grain media, pulses medium produced significantly higher of biomass production was recorded in Trichoderma viride. Abundance of minerals in the pulses medium may enhance the growth of fungi. Rice and wheat medium also supported the growth of both the tested fungi.

Ref:

Mridula et.al., 2012. Isolation, characterization & biomass production of Trichoderma viride using various agro products- A biocontrol agent. Advances in Applied Science Research, 2012, 3 (6):3950-3955.