WHY THIS ASSIGNMENT?

In ‘Vigyan Ashram’s ‘ microbiology lab, Tricoderma viridi is multiplied, made powder of it, packed and sold to the Farmers & also used by the agriculture department students within the Vigyan Ashram. Tricoderma is an very effective bio control agent for the crops. Hence it is in demand among Farmers, Gardeners and all people involved in agricultural Section.

  1. In order to produce Tricoderma Powder, first we have to inoculate Tricoderma on the sterile substrate, allow it to multiply on it. For inoculation we require pure Tricoderma viridi strain.
  2. When we plated the Tricoderma powder which was made in Ashram on PDA; it was observed that along with Greenish growth of Tricoderma viridae, some unwanted growth (unknown) was observed. So the need was raised to isolate pure form of Tricoderma viridae from the powder.

OBJECTIVES:

  1. To isolate Tricoderma viridi from the given powder sample by Spreading method.
  2. To learn and improve the plating skills.

REQUIREMENTS:

  1. PDA media
  2. Petri plates
  3. Spreader
  4. Test tubes
  5. Conical flask
  6. Parafilm
  7. Autoclave
  8. Incubator
  9. LAF

PROCEDURE:

  1. Prepared PDA media .
  2. Autoclaved media,petri plates,distilled water,test tubes,spreader,microtips at 121C temp and 15 lbp pressure for 20 min.
  3. For Maintaining asceptic condition : At the same time, clean LAF(laminar air flow) using aceton /80% ethanol and ON UV light for 15-20 min.after 20 min OFF UV light and and ON blower and day light.
  4. Then all remaining procedure carried out under asceptic contion
  5. After sterilization, media poured into petri plates (approximately 30 ml/plate)
  6. Serial dilution: for serial dilution ,weigh 1 gm of trichoderma powder.Take 10 sterile test tube and filled it with 9 ml distilled water each.Dissolve 1 gm trichoderma powder in it and subsequently transfer 1 ml aliquot from test tube 1 to last.Then finally we have concentration of trichoderma would be 10-1,10-2,…..10-9
Image 1: Serial dilution of Trichoderma powder.
  1. After solidification of media, add 0.2 ml of aliquot from test tube (with concentration 10-4 ) on petri plate.And spread equally on whole petri plate.
Image 2 : Innoculation of Trichoderma powder on media.
  1. Follow same protocol for all the test tubes with different concentration and innoculate on the different petri plate.
  2. Packed petri plates with parafilm.And kept it in incubator at 37 C temp for 3/4 days.