IJCBS RESEARCH PAPER VOL.

1 [ISSUE 1] April, 2014 ISSN:- 23492724

Virtu and Foi
1
www.ijcbs.org
Antagonistic Activity of Trichoderma Spp, (A Bio-
Control Agent) Against Isolated and Identified Plant
Pathogens
1*
Meenakshi Kushwaha (Pursuing PhD)
Biological Sciences Department
National Botanical Research Institute
Lucknow, India
Email: meena.abc7@gmail.com
2
Ajay Kumar Verma (M.Tech.)
Civil Engineering Department
Madan Mohan Malveeya Engineering College
Gorakhpur, India
Email: ajaycrazy100@gmail.com

ABSTRACT: There is a great problem of farmers to protect their food crops from pathogenic microbes.
Some fungi play a major role to have an effect on the crops yield and survival. On visiting farmers field in
bakshi ka talab lucknow, some diseased crop plant and their fruits have been found. After screening the
diseased area of sampled crop plants causal fungi get isolated and identified morphologically in the
laboratory. The isolated and purified pathogenic fungi are Chollatotrichum from red rot of sugarcane,
Alternaria from brown-black discoloration of cauliflower, early blight of tomato and black round spot of
apple, Paracercospora from leaf spot disease of banana and Fusarium from wilt root of pegion pea and
black margine of curled potato leaf. Some species of Trichoderma like T. hargianum, T. viride, T. hamantum
have a potential to inhibit the growth of pathogenic fungi. Trichoderma had shown significant antagonistic
activity against isolated pathogenic fungi Chollatotrichum, Alternaria, Paracercospora and Fusarium
species. From this result we have found the trichoderma as a potential biocontrol agent against
pathogenic fungi.

Keywords: Trichoderma; antagonistic activity; fungal diseases of plants; Biological control; horticulture
crop diseases; bio-control agent.

1. INTRODUCTION
India with diverse soil and climate comprising
several agro-ecological regions provides ample
opportunity to grow a variety of horticulture
crops. Cultivation of these crops is labor
intensive and as such they generate lot of
employment opportunities for the rural
population. Fruits and vegetables are also rich
source of vitamins, minerals, proteins, and
carbohydrates etc. which are essential in human
nutrition. Hence, these are referred to as
protective foods and assumed great importance
as nutritional security of the people. India is the
second largest vegetable producer in the world,
next only to China with an annual production of
81 million tons from 5.1 million hectares of land.
However, the major factors responsible for low
production of fruits, cucurbitaceous,
brassicaceous and solanaceous vegetables are
the diseases caused by Pathogenic fungi. The
disease development is so fast that whole crop is
lost in a few days. Therefore, the problem
deserves immediate and effective measures of
control (Neeraj, et.al.2010). Fruit plant diseases
often are the worst natural hazards in
horticulture. New diseases and new biotypes of
existing disease-producing organisms appear
from time to time in a more virulent form. With

the liberalization of import and export of large
quantities of horticultural planting materials,
the risk of spread of diseases into new areas has
also increased. The most startling aspect of fruit
plant diseases is that their control cost us a huge
sum every year with serious environmental
consequenes.
Therefore, integrated disease management
practices need to be adopted to reduce the
losses Fruits and vegetables are highly
perishable products, especially during the
postharvest phase, when considerable. Losses
due to microbiological diseases, disorders,
transpiration and senescence can occur. A
number of microorganisms, which effectively
control postharvest pathogens, have been
identified for post-harvest control (Wilson and
Wisniewski, 1989). Biological control of plant
pathogens by microorganisms has been
considered a more natural and environmentally
acceptable alternative to the existing chemical
treatment methods (Baker and Paulitz, 1996).
Trichoderma spp. is now the most common
fungal biological control agents that have been
comprehensively researched and deployed
throughout the world. Several fungal cell wall
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degrading enzymes, amongst them chitinase and
glucanase, which seem to play an important role
in the antagonistic action of Trichoderma against
a wide range of fungal plant pathogens (Kucuk
and Kivanc, 2008). T. harzianum shows
antagonistic effects against, Alternaria spp.
Colletotrichum spp.,Paracercospora spp.,
Fusarium spp. and, causative fungi of stem end
rot, anthracnose and brown spot of rambutan
respectively and also retained the overall quality
and color of the fruits (Siva Kumar et al.,2000).
The present study aimed to find out the
efficiency of Trichoderma spp. against some
post-harvest pathogens.
MATERIALS AND METHODS
2.1 Survey and collection of variable
diseased crop samples
We have collected the following infected
horticulture crop samples during the survey of
farmers field at Bakshi Ka Talab, Lucknow and
after collection we observed their symptom. In
marigold leaf red & dry leaves, black margins on
potato leaves, in pigeon pea root wilt root with
black xylem, cauliflower with brown-black
discoloration, in tomato fruit velvety green mat
which is also known as early Blight of tomato, in
banana leaves brown spots with yellow margin,
black streaks & gray spot with black margin on
different leaves of banana, leaf red thick streaks
on sugarcane, which is also called red rot of
sugarcane, apple fruit (post-harvest, from
market) several black round spot with brown rot
on side of tip fruit we have observed with naked
eyes. These infected plant samples kept in sterile
zipper lock plastic bags and carried to the
laboratory where we have stored those samples
at 4C.
2.2 Isolation and pure culture of pathogens
Fungal pathogen was separated from infected
area of plant sample and pure cultured on PDA
plates in the laminar air flow. For preparation of
300 ml Synthetic PDA medium 39 gm. of PDA
powder (Himedia laboratories Pvt. Ltd.) is used
for making 1 ltr. of PDA medium as
recommended. After autoclave at 121 C for 15
20 minutes, cool the media up to tolerable
temperature then pouring was done into sterile
petri dishes under the flaming area in the
laminar air flow. The PDA plates were
successfully prepared. Taking small part of the
infected part of each sample separately, surface
washing of inoculums with sterile water was
done. Now in the laminar air flow, the
inoculation loop is taken and kept on the flame
for a while so as to kill all the microbes present
over its surface, it is then dipped in ethanol and
again placed over the flame, but for a short
while. It is held for a few seconds to cool it down.
Then sample was taken and the loop was
scrapped on the infected part of the samples so
as to pick a good amount of inoculums,
inoculation done properly with the help of loop.
After that the loop is taken out and the PDA plate
is covered. The whole process of inoculation was
repeated with each sample. The inoculated PDA
plates were incubated in BOD at 26 C for 72 hrs.
with one PDA plate without inoculum taking as
control. Pure colonies are isolated from
inoculated petri plates separately in aseptic
condition. Pure cultures are made with the help
of sterilized inoculation loop on PDA plate. The
isolated pure cultures on separated PDA plates
are then kept on BOD incubator for incubation at
26 C for 72 hrs. Observation was taken daily to
record the growth of microbes.
2.3 Microscopic identification
The pure colonies were picked and identified
under high quality microscope.

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2.4 Management of isolated disease causing
fungus
Antagonistic activity
We have performed the antagonistic activity of
Trichoderma against Colletotrichum, Alternaria,
Paracercospora & Fusarium.
Switch on the UV of laminar air flow. After
switching off, switch on the air flow. Take 24
sterile petriplates and pour the PDA medium in
each petriplate and keep it for solidification.
Keep 4 pour plates as control (pure media) for
checking the medium and inoculate 8 petriplates
with Trichoderma and Alternaria, Trichoderma
and Colletotrichum, Trichoderma and
Paracercospora, Trichoderma and Fusarium
respectively which worked as control 2. In other
12 petriplates inoculate Trichoderma and just
opposite of this inoculation, inoculate Alternaria
Colletotrichum, Paracercospora & Fusarium,
Replicate it three times. We Kept all the
petriplates in the BOD incubator at 27
o
C for 96
hours and taken records after every 24 hours.
Table No. 1

Table No. 2

Table No. 3


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Table No. 4


3. RESULT
The samples that we were collected from
farmers field at BKT have following diseases
which are wilt root (black xylem) of pigeon pea
root, brown spots with yellow margin of banana
leaf, bacterial watery marks of cauliflower, red
leaves of marigold leaf, red rot of sugarcane of
sugarcane leaf, black margins of potato leaf,
black round spot of apple fruit, early blight of
tomato.
Table No. 5: Isolated fungi from each infected
plant samples

Trichoderma show strong antagonistic activity
against Colletotrichum causing red rot of
sugarcane, Alternaria spp. causing Alternaria
leaf spot disease, Paracercospora spp. causing
leaf spot disease of banana, Fusarium spp.
causing wilt disease in pigeonpea.

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CONCLUSION
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 bio
control agents are from the species T. harzianum
, T. viride and T. hamatun. The biocontrol agent
generally grows in its natural habitat on the root
surface and so affects root disease in particular
but can also be affected against foliar diseases.
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