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Sunita Devi
S-2010-40-04
Department of Microbiology
College of Basic Sciences
CSK Himachal Pradesh Krishi Vishvavidyalaya
Palampur-176062
 K
DUC
K
 mong the greatest hazards in the crop production, diseases
caused by phytopathogens are one of the major threats to
sustainable food production.
 For combating phytopathogens, many successful measures
including chemical control have been developed

hese chemicals have resulted in a variety of harmful and
undesirable effects on man, wildlife and ecosystem.
 Biological control seems to be a potent means of reducing the
damage caused by phytopathogens which can be achieved by
making use of various types of microbes including bacteria,
fungi and viruses.
 Why use biological control?
 Biological control agents are
Expensive
Labor intensive
Host specific
 Chemical pesticides are:
cost-effective
easy to apply
Broad spectrum
 Contd.
WKLL:
 Chemical pesticides
Kmplicated in ecological, environmental, and human health problems
equire yearly treatments
Broad spectrum

oxic to both beneficial and pathogenic species
BU
:
 Biological control agents
on-toxic to human
ot a water contaminant concern
nce colonized may last for years
Host specific
 nly effect one or few species
 rganic farming is becoming popular due to its eco-friendly
nature and quality of the produce.
he application of
buttermilk is one of the organic inputs being used in the
process of organic farming to control some of the diseases.

he microbial diversity of this organic input has been studied
in the dept. of Microbiology, where various types of microbes
possessing some of the biocontrol properties have been
isolated.
 ne of such microbes which is gram positive, spore former
and exhibiting fairly a good amount of biocontrol activity
against some of the fungal pathogens has also been isolated.

his particular indigenous microbe needs further systematic
evaluation of its biocontrol potential with regards to the type
of metabolites involved in biocontrol activity.
 BJEC
KVES:

o examine the antagonistic activity of
indigenous bacterial isolate against different
phytopathogenic fungi and nematode

o optimize different parameters to augment
production of biocontrol agent(s)

o purify and characterize the biocontrol
compounds produced by the above
indigenous isolate
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he bacterial isolate will be tested for its antagonistic activity against
five fungal pathogens, Y
., 
 
 ,    


  
      and    


by dual culture technique (Ganesan and Gnanamanickam, 1987).

Each fungal pathogen will be grown on a PD plate till it covers the
whole surface of the agar. With the help of a sterile cork borer, a disc
of fungal growth will be taken and placed at the center of a fresh PD
plate.
24 hour old culture of bacterial isolate will be then streaked on one
side of the fungal disc.

he plates will be kept for incubation at 30°C for 2-7 days.

Knhibition of growth of fungal pathogen will be then recorded with

respect to control plate, inoculated with the fungal pathogen only.
 gainst nematode pathogen

he culture filtrate and the whole cell culture of the

bacterial isolate will be examined for their ability to
immobilize or kill root knot nematode, °  
 , under laboratory condition (Jayakumar  .
2002).
 100 ml of 24 hour old bacterial suspension

25 ml will be emaining broth culture

centrifuged at 8000 rpm will be used as whole
for 10 minutes cell culture for bioassay.

Supernatant will be
filtered

Filterate will be used for

bioassay against root knot
nematode
 Bioassay

he egg masses of root knot nematode, °     will be made to hatch in to juveniles .Population
of juveniles present in water suspension will be counted under stereomicroscope and used for bioassay test

he plates
containing Kn another set of plates

hree ml of the nematode suspension containing
nematode containing nematode
about 50 juveniles will be taken in a petri plate to
suspension and juveniles, 10 ml of
which 10 ml of culture filtrate will be added and
plain broth will whole cell culture will be
kept for 72 hours for incubation.
serve as control added.

t 24 hours, 48 hours and 72 hours of incubation, observations for immobilized and killed
nematodes will be recorded in each plate.

he revival test will also be conducted by transferring the immobilized nematodes into
fresh water to confirm mortality in both the sets of plates

he following parameters will be optimize to augment
production of biocontrol agent(s) using esponse
surface methodology:
 Carbon sources
 itrogen sources
 PH
 eration

he partial identification of biocontrol agent(s) will be
done using suitable solvent systems by
LC and HPLC
based detection methods.
MLDKLCMSGC-MS or M based study will be
done to elucidate the structure of biocontrol agent(s)
.
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