Mass Cultivation of Trichoderma viride

By,
Dhanishta.N (Reg No:192018010)
Department of Zoology
An Interinstitutional Project
Submitted under DBT Star College Scheme,
Department Of Biotechnology,
New Delhi.
AIM OF THIS PROJECT:

🞭 To isolate the Trichoderma species

🞭 To provide sustainability to farmers
🞭 To create awareness among young
researcher to do project on microbes
WHAT IS BIOFUNGICIDE?
🞭 Biofungicide are formulations of living
organisms that are used to control the
activity of plant pathogenic fungi and
bacteria.
🞭 The concept of fungicides is based upon
observations of natural processes where
beneficial microorganisms, usually
isolated from soil, hinder the activity of
plant pathogens.
WHY TRICHODERMA IS SELECTED FOR AGRICULTURE?

🞭 Trichoderma are fast-growing fungi that aggressively

colonize on various substrates, compete for ecological
niche, and eliminate pathogens (Bailey and Lumsden
1998).
🞭 Trichoderma, when applied as soil treatment or
coated on seeds, create a zone of protection around
plants and provide resistance against pathogens.
🞭 Thus it farmers are ready to choose it for
agriculture.
MEDIUM PREPARATION:
Czapek’s medium:
For 1 Liter:
Sugar/Glucose-30 gm
Dipotassium- 1gm
Magnesium Sulphate-0.5 gm
Potassium Chloride-0.5 gm
Ferrous Sulphate-0.01 gm
Agar(Solid medium)-15 gm.
All the ingredients were added to the distilled water upto 1000 ml
in 2000 ml conical flask
Add agar, shake and autoclaved 121*C for 15 mins. By using a
measuring cylinder, the medium was transferred into bottles for
mass multiplication in low cost production.
ISOLATION AND IDENTIFICATION OF TRICHODERMA
VIRIDE

●Soil samples are collected, air dried, and ground into powder. Stock solution of sample is
prepared by dissolving 10 g of powdered soil sample into 90 mL of distilled water. Next,
serial dilution of samples were prepared as 10−1, 10−2…10−5. One milliliter of each of
the prepared dilution is spread evenly on a suitable medium on a petri dish at 28 ± 1°C
for 7 days
●This allowed colony formation with mycelial growth.
●The number of colonies obtained in the plates were counted.
●Smooth surface, cottony white mycelial mat
●with aerial hyphae was formed on the colonies . Based on these isolates were identified
as Trichoderma viride.
●Using streak method, the colonies are inoculated into new Czapek’s mediumin petri plates
and incubated for 7 days to isolate the pure culture of Trichoderma viride .
●Thisprocedure was repeated until pure culture with high numbers of Trichoderma viride
wasobtained. The strains which showed clear zones are inoculated into the nutrient broth
for mass multiplication.
BY POUR METHOD BY STREAK METHOD
DISC DIFFUSION TEST:
Collect vermicompost and Trichoderma Viride as samples and serial dilution method take
place.Pour the Czapek’s medium into the petri dish and dip the diluted samples and place it
on the petri dish. We can see some zone of growth inhibition in Vermicompost because the
vermicompost has higher amounts of PGPR but in Trichoderma Viride no growth of
inhibition can be due to the low amount of phosphate solubilizing bacteria present here. So
Trichoderma Viride acts as a bio- fungicide.
MASS CULTIVATION OF TRICHODERMA VIRIDE

Take 100 g of Groundnut oil cake powder in a polyethylene bag and place a
PVC Pipe . One end should be placed towards the cover and the other end
should be covered with a cotton plug. Sterilize it for 15 mins and pour
liquid medium into the sterilized bag and cover it with a cotton plug.
Observe the growth of fungi in polyethylene bags.
Cost of the Tray- 20 rupee
Length - 17.5 Cm
Breadth- 14.1 Cm
Height-5 Cm
Width-14 Cm
Area of the Tray- 563 .6 sq. cm2

Volume of the Tray-672 cubic cm

Weight of the ground nut cake- 231.4 gm
ADVANTAGES….

● Culture can grow in fixed substrate

● Equal grow rate of fungal culture can
be obtained
● Can be harvest for many times
Conclusion:

● Trichoderma spp. is one of the frequently isolated fungal genera from soil
and plant roots that have been extensively studied for their vast
metabolites with various applications (agricultural, industrial, health, etc.).
● In the field of agriculture, Trichoderma are suitable antimicrobial agents
against pathogenic bacteria, fungi, and yeast.
● Similarly, they play a vital role in improving the vegetative growth of
plants and nutrient content of soil through decomposition and
biodegradation. It is a safe, cost-effective, and environmentally benign
technology to attain sustainable agriculture.