## Talc-based Mass Multiplication of Trichoderma viridea
**Trichoderma viridea**, a beneficial fungus known for its biocontrol properties, can be mass-produced using talc as a carrier material. This method is relatively simple and inexpensive, making it suitable for small-scale production and field applications.
Here's a detailed breakdown of the process:
**Materials:**
* **Mother culture of Trichoderma viridea:** This can be obtained from research institutes, biocontrol laboratories, or online suppliers.
* **Talc powder:** Agricultural grade talc with a high surface area is preferred.
* **Rice/Wheat/Sorghum/Maize:** Grains serve as the substrate for Trichoderma growth.
* **Plastic bags:** 8" x 12" size is commonly used.
* **Cotton:** For plugging the bag opening.
* **Rubber bands:** For securing the bags.
* **Pressure cooker:** For sterilizing the grains.
* **Heating system:** Gas or electric stove.
* **Fresh water:** For preparing the media and washing the grains.
* **Spoon:** For transferring the mother culture.
* **Candle:** For providing a sterile environment during inoculation.
**Procedure:**
**1. Grain preparation:**
* Wash the chosen grains twice with clean water to remove dust.
* Weigh 200g of grains and add them to a plastic bag.
* Add 200ml of fresh water to the bag and mix well.
* Place a plastic pipe (or bamboo) in the center of the bag with the opening facing upwards.
* Plug the pipe opening tightly with cotton and secure it with a rubber band.
* Cover the cotton plug with paper and secure it again with a rubber band.
**2. Sterilization:**
* Place a thick paper inside a pressure cooker to prevent direct contact with the heat source.
* Add water to the cooker just below the level of the paper.
* Place the prepared bag inside the cooker and cover it.
* Heat the cooker until it releases steam three times (3 whistles).
* Turn off the heat and let the pressure release naturally.
* Remove the bag once it cools down completely.
**3. Inoculation:**
* Wash your hands and the spoon with Dettol or another disinfectant.
* Light a candle to provide a sterile environment.
* Open the paper cover from the bag and remove the cotton plug using the opposite end of the spoon.
* Transfer a small amount of mother culture (talc-based) into the bag using the spoon.
* Replace the cotton plug immediately and secure it with the rubber band.
* Keep the bag at room temperature (around 25°C) for 10-12 days.
**4. Monitoring and harvest:**
* During this period, observe the bag for fungal growth.
* The entire grain-based medium should turn green due to Trichoderma sporulation.
* Once the sporulation is complete, the product is ready for use.
**5. Storage:**
* The talc-based Trichoderma viridea can be stored in a cool, dry place (preferably a refrigerator) for up to 6 months.
* Avoid direct sunlight to maintain its viability.
2. About Trichoderma
• Trichoderma spp. are free-living fangi, mycoparasitic of several
soilborne plant pathogens.
• Reduce root rot, collar rot, damping off etc.
• Release metabolites that improves the growth of seedling.
• Sweet or coconut odour.
4. Requirements of PDB media
• 200gm of Potato
• Dextrose 20 g
• Distilled Water 1 lit.
5. Preparation PDB media
Cut Potatoes --- Boil until soft---
Stain water--- Mix dextrose---
Sterilize at 15 psi for 45 min----
6. Mass multiplication of Trichoderma V.
• Sterilize both PDB and Talc
• Inoculate Trichoderma spores at a desired level.
• Transfer the inoculated culture to sterile flasks
• Keep it in BOD at 25-30 degrees centigrade for 7-10 days.
• Collect the bio-mass
• Mix with talc powder at 1:2
• Dry until 8% moisture
• Pack and seal
9. Successful Application
• (Pascale et al.,2017) Two stains of Trichoderma – Showed suppression
of powdery mildew by secondary metabolites.
• (Amin et al.,2010) shows T. viride and T. harzianum shows 65.71 and
60.51 percent inhibition over control. Where as mixture of it able to
control 71.41% of fungi.
