Mass multiplication of a Biofertilizer- Trichoderma viride

## 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.

1. Mass Production of Trichoderma viride Presented By: Souvik Chaktraborty Rijhu Bediya Susmita Das

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.

3. Requirements PDB media Talc powder Autoclave Incubators Mother culture of Trichoderma

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

7. Consideration Aseptic condition Quality Control Storage condition Source: (Tjamos, K ., et al . 2012)

8. Usages Seed treatment Seedling treatment Soil application Foliar application Drench application Source: (Deepika,2023) Trichoderma: An Eco-friendly Bio-Fungal Agent For Sustainable Agriculture

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.

10. Thank You