Bio Control Production Unit

Biological Control of Plant Pests, Disease and Weeds

While pursuing research on control of pests, diseases and weeds of economically important crops steady efforts have been made to develop cost-effective, eco-friendly, commercially viable mass production technologies of various biocontrol agents and improved formulations for use under the Integrated pest management (IPM). Antagonistic Fungal and Bacterial Biopesticides - Trichoderma viride and Pseudomonas fluorescens are being produced and supplied for crop pests and Diseases Management.
This laboratory produces & provides different biological products such as bio pesticides on no profit & no loss basis. Trichoderma spp. act as a biofungicide against many plant pathogenic fungi and have been found to induce systemic resistance in plants. The field applications of Trichoderma spp. require mass multiplication which can be done using solid as well as liquid state fermentation. Several strains of Trichoderma have been developed as biocontrol agents against fungal diseases of plants. The various mechanisms include antibiosis, parasitism, inducing host-plant resistance, and competition. Most biocontrol agents are from the species T. harzianumT. viride and T. hamatum. The biocontrol agent generally grows in its natural habitat on the root surface, and so affects root disease in particular, but can also be effective against foliar diseases.
P. fluorescens strains present biocontrol properties, protecting the roots of some plant species against parasitic fungi such as Fusarium or Pythium, as well as some phytophagous nematodesThe bacteria might induce systemic resistance in the host plant, so it can better resist attack by a true pathogen. The bacteria might outcompete other (pathogenic) soil microbes, e.g., bysiderophores, giving a competitive advantage at scavenging for iron. The bacteria might produce compounds antagonistic to other soil microbes, such as phenazine-type antibiotics or hydrogen cyanide

Mass multiplication of Trichoderma viride
Preparation of mother culture
Molasses yeast medium is prepared as detailed below.
Molasses : 30 g 
Yeast : 5 g 
Distiller water:1000 ml
The medium is prepared and dispensed into conical flasks and sterilized at 15 lb pressure for 15 minutes in an autoclave. After the medium is cooled it is in inoculated with 10 days old fungal disc of T. viride and then incubated for 10 days for fungal growth. This serves as mother culture.

Mass multiplication:
            Molasses yeast medium is prepared in fermentor and sterilized as described earlier. Then after the medium is cooled, the mother culture is added to the fermentor @ 1.5 lit / 50 lit of the medium and incubated at room temperature for 10 days.Then the incubated broth containing the fungal culture is used for commercial formulation preparation using talc powder.

Mass production of Pseudomonas fluorescens
Preparation of mother culture
Mother culture is prepared by using the king’s B medium
Peptone : 20.0 g
K2HPO4 : 1.5 g
Mg SO4 : 1.5 g
Glycerol : 10 ml
Distilled water : 1000 ml
The above broth is dispersed into conical flasks and autoclaved at 15 lb pressure for 15 minutes and cooled and inoculated with a loop of  P.fluorescens and incubated for 2 days.

Mass multiplication:
            The kings B medium is prepared and poured into the fermentor and sterilized at 15 lb pressure for 15 minutes. After the broth has cooled below the mother culture of P.fluorescens is added to the king’s B medium in the fermentor at the rate of 3 lit for 40 lit of the broth. Then it is incubated in the fermentor for 2 days with frequent mixing of the broth by operating the stirrer. Then the broth containing the bacterial growth is collected in plastic buckets and used for mixing with talc powder for commercial formulation.

PREPARATION OF TALC BASED PRODUCTS, AIR DRYING OF FORMULATION AND ESTIMATION OF MOISTURE CONTENT

Tichoderma viride:

The fungal biomass collected from fermentor is mixed with talc powder at 1:2 ratio. The mixture is air dried in shade and mixed with carboxy methyl cellulose (CMC) @ 5 g / kg the product. It is packed in polythene bags and should be used within 4 months.

      Quality control parameters:

  1. Fresh product should contain hot less than 28 x 106 cfu / g
  2. After 4 months of storage at room temperature, the population should be 20 x 106 cfu / g.
  3. Maximum storage period in talc is 4 months.
  4. The talc size should be 500 microns
  5. The product should be packed in polythene bags
  6. Moisture content of the final product should not be more than 20%

Pseudomones fluorescens

The broth containing the bacterial growth is collected from fermentor and added @ 400 ml / kg of talc powder. Then CMC is added @ 5 g /kg mixed well air dried to 20% moisture level and packed in polythene bags.

     Quality control parameters:

  1. Fresh produce should contain 2.5 x 108 cfu/g
  2. After 3 months of storage at room temperature the population should be 8-9 x 107 cfu/g
  3. Storage period is  3-4 months
  4. Minimum population load should be 1.0 x108 cfu /g
  5. Moisture content should not exceed 20% in the final product
  6. Population per ml of the broth should be 2 x 108 cfu /g