Mass Production Of

Verticellium lecanii

Created by: Rohit Ramkrishna Shinde

Reg No AGL/49/2012

Biopesticides
Bio pesticides are biochemical pesticides that are naturally
occurring substances that control pests by nontoxic mechanisms.
Biopesticides are certain types of pesticides derived from such
natural materials as animals, plants, bacteria, and certain
minerals.
All the living organism which are cultivated in the laboratory on
large scale and are used and exploited experimentally for the
control of harmful organism.

THE ADVANTAGES OF USING

BIOPESTICIDES
Biopesticides are usually inherently less toxic than
conventional pesticides.
Biopesticides generally affect only the target pest and
closely related organisms, in contrast to broad spectrum,
conventional pesticides that may affect organisms as
different as birds, insects, and mammals.
Biopesticides often are effective in very small
quantities and often decompose quickly, thereby
resulting in lower exposures and largely avoiding the
pollution problems caused by conventional pesticides.
When used as a component of Integrated Pest Management
(IPM) programs, biopesticides can greatly decrease the use
of conventional pesticides, while crop yields remain high.
To use biopesticides effectively, however, users need to
know a great deal about managing pests.
Cheap, renewable can be handled safely.
Difficult for insects to develop resistance to these

THE DISADVANTAGES OF USING

BIOPESTICIDES
o Slow effect
o Lack persistence and wide spectrum activity
o Rapidly degraded by UV lights so residual action
is slow.
o Seasonal availability of plants products indicates
the needs for storage.
o They are not available easily
o Poor water solubility and generally not systemic
in nature
o All products applied followed by growers have
not been scientifically verified.

Types of biopesticides
Microbial biopesticides
Plant biopesticides
Biochemical biopesticides

Mass Production of Verticellium

Lecanii

Verticillium lecanii is a unique biological

insecticide based on a selected strain of naturallyoccurring entomopathogenic fungus
Verticillium lecanii (NCIM 1312 ) that infects and
kills mealy bugs and sucking insects like thrips,
jassids , aphids, white flies and mites. Verticillium
lecanii It is formulated as Wettable Powder with
CFU count of 1 X 10 8 / g. Verticillium lecanii is
registered by Indian Pesticides Regulatory
Authority - Central Insecticides Board, Govt of
India Verticillium lecanii is approved for use in
Organic agriculture.

Commercial products

Scientific Classification
Kingdom- Fungi
Phylum- Ascomycota (anamorphic hypocreales)
Subdivision- Pezizomycotina
Class- Sordariomycetes
Order- Hypocreales
Family- Clavicipitaceae
Genus- verticilium
Spp.- lecanni

Introduction: Extensive use of chemical insecticides has leads to

disruption of ecosystem.
Biological control can be described as the limitation of
the abundance of living organism and their product by
other living organism.
Biopesticides, a contraction of Biological
pesticides include several types of pest
management intervention
Through predatory, parasitic, or chemical relationship.
Verticillium lecanni approval name of an
entomopathogenic fungus spp.
V. lecanni itself appear primarily to be a pathogen of
soft scale insect .

Procedure
Prepration of Media
Inoculation of culture
Growth of culture
Harvesting of culture
Prepration of talc formulation
Packaging
Marketing

Types of media
PDA(Potato Dextrose Agar)
PDYB(Potato Dextrose Yeast broth)
CRUSH GRAIN METHOD
IN which PDYB and Crush Grain Method is used
For mass production Of V. lecanii
Where as PDA is used to maintain the culture

PDA
Chemical compound gm / litr
Potato- 200gm
Dextrose- 20gm
Agar-15gm
Distilled water-1000ml
PH -5.7

PDYB
Chemical compound gm / litr : Potato- 200gm
Dextrose- 20gm
Yeast extract- 1.5gm
Distilled water-1000ml
PH -5.7

PDYB (Potato Dextrose Yeast Broth)

Small pieces of 200 gm potato

Boil the Potato on gas burner

20 gm of Dextrose and 1.5 gm of Yeast

Extract.

Stirring the small pieces of

Potato

Isolation and identification of v. lecanii

v. lecanii was isolated from soil sample by using PDA
Sample were inoculated over plates by multiple tube
dialution techinique
(MTDT) and the plates incubated at 26oC/4days
The fungal colonies which were picked up and purified
by stacking and incubated at26oC/7-8 days
The culture was maintained on PDA slants

PDYB( Potato Dextrose Yeast Broth)

Firstly take 200 gm of potato cut into small
piece put it into cooking pan added with
distilled water .
Put it on the gas burner allow it to boil
after few min. take whey and measured in
measuring flask level up the volume to
1000 ml with distilled water.
Add 20 gm of dextrose and 1.5 gm of
yeast extract into the media and put it on
burner with still stirring .
Remove the media from the burner and
put it in the conical flask .

 After pouring media put that media containing

culture bottle into autoclave at 121c for 15 psi
pressure.
After autoclaving it bring the media containing
culture bottles into laminar air flow for
inoculation of V. lacanii
Inoculate V. lacanii into bottle with the help of
needle.
Plug the cotton to the culture after inoculating.
Put it in growth room for further growth of
culture.

Pouring of Media in culture bottle Autoclave media at 121c for 15 psi

pressure.

Inoculation of Culture in Media

Take culture bottle in laminar air

flow for 15 min/

Crush grain method

The grain are soaked in water over night and next day
the water is decanted and crushed slightly in a
blender for a few second
200 gm of crushed grain are taken in PP bags of
(30*20cm)and sterilized at 1210 C temperature for 30
minutes.
After cooling the bags are cut open and inoculated
with 3ml of suspension (106 spors /ml)prepared from
fresh culture and the cut portion is plugged with
sterile cotton wool
The inoculated grain are spred out as thin layer in the
bags and incubeter 7 to 10 days at room temperature
or below 300 c .

CRUSH GRAIN METHOD

Polythine bag

Mat formation

Preparation of talc formulation

with sterilized talc poweder in proportion of
1:2 or 1:3 and dried under aseptic condition
for 20 hrs.
The spores load in the formulation may be 1061010 spore /gm
The formulation diluted with water to get
spore concentration of 1010 spore /liter for
foliar sprays .
Tween-80 (0.01%) should be added to the
spore suspension for uniform spread of spores
on plant surface (ramanujam , 2003).

Preparation of talc formulation

Harvesting of
mat

Crushing of mat

Mixing with talc

Biology and ecology v. lecanii

It lacks a sexual phase (perfect stage) and
produces by means of motile asexual spores called
conidia.
Germination of this conidia produces hyphae and
after subsequent growth.
It is able to grow on both living and dead
neautricious part .
V. lecanii is important entopathogenic hyphomycet
fungi occurred on coccids , aphids, thrips ,diptera ,
homoptera ,hymenoptera , Lepidoptera .

 v. lecanii infects in vertebrate host through the

external cuticle .
under good humidity condition the dead host is
coverd by the fungal spores and hyphae .
optimum temperature for spore germination and
colony growth between 15 to 250c.
at 50c spoes of v. lecanii can germinate and grow
slowly .
above 300c its germination and growth may cease .

Mode of action
This fungus produces hyphae from germinating spores
that penetrating the insect integuments .
Then fungus destroy the internal content and the insects
dies .
The fungus eventually grows out through cuticle and
sporulates outside the body .
Insect appears as white to yellowish cottony particles.
Affected insect usually appears in 7 days .
Virulence of v. lecanii has bean associated with high extra
cellular chitinane activity .

26

Conidia of V. lecanii

Verticillium lecanii
COLONY OF V. lecanii

Method of application
Mix v. lecanii @ 5 gm/liter water and spray for
control of target pest like mealy bug and
sucking insect.
The spray volume depends upon the crop
canopy .
Target pest mealy bug thripes , jassids , aphids
, white flies and mites .

Crops suitable for application

Papaya grapes , guava, custard apple ,
sapota, chilies, cotton ,sugarcane, sorghum
,cereals ,vegetables ,fruites, spices,
flowers , medicinal crop, aromatic crop,
orchids and ornamentals .
Shelf life of v. lecanii is stable for a period 12
month from the date of manufacture.

30

Result
V. lacanii on Mealybug s
V. lacanii on Aphid

Infection of V. lecaniia against Insect Pests

Visit of Dr.K.P. Vishvnatha (VC,MPKV

Rahuri

Visit of Dr.Ulmek Sir

Markrting

Thank