STUDIES ON LEAF SPOT OF CHILLI (Capsicum

annuum L.) CAUSED BY Cercospora capsici Heald &

Wolf

SURESH, K. R.
PALB 1246

DEPARTMENT OF PLANT PATHOLOGY

UNIVERSITY OF AGRICULTURAL SCIENCES
BANGALORE – 560 065
2013
 STUDIES ON LEAF SPOT OF CHILLI (Capsicum
annuum L.) CAUSED BY Cercospora capsici Heald &
Wolf

SURESH, K. R.
PALB 1246

Thesis submitted to the

UNIVERSITY OF AGRICULTURAL SCIENCES,
BANGALORE
in partial fulfillment of the requirements for the degree of

MASTER OF SCIENCES (AGRICULTURE)

IN
PLANT PATHOLOGY

BANGALORE JULY, 2013

 Affectionately
Dedicated
To
My Beloved Parents,
Sister, Brothers
Brothers
and Guide
 DEPARTMENT OF PLANT PATHOLOGY
UNIVERSITY OF AGRICULTURAL SCIENCES
BANGALORE – 560 065

CERTIFICATE

This is to certify that the thesis entitled “Studies on leaf spot of

chilli (Capsicum annuum L.) caused by Cercospora capsici Heald &
Wolf ” submitted by Mr. Suresh, K. R. ID. No. PALB 1246, in partial
fulfillment of the requirements for the award of degree of MASTER OF
SCIENCES (AGRICULTURE) IN PLANT PATHOLOGY to the University of
Agricultural Sciences, GKVK, Bangalore, is a record of research work
carried out by him during the period of his study in this university under
my guidance and supervision and that no part of thesis has been
submitted for the award of any degree, diploma, associateship, fellowship
or any other similar titles.

Bangalore Dr. T. Narendrappa

Date: 3rd AUGUST, 2013 Major advisor
Professor and head
Department of Plant Pathology
College of Agriculture
UAS, GKVK, Bengaluru-65

APPROVED BY:
Chairman:
__________________________
(T. Narendrappa)

Members: 1. __________________________
(Y. M. Somasekhara)

2. __________________________
(P. Chowdappa)

3. __________________________
(K. Madhavi Reddy)
 ACKNOWLEDGEMENT

“Gratitude is the memory of the h eart”

“Gratitud

I take this wonderful opportunity to express my deep sense of gratitude for all
those who have helped me in bringing out this thesis successfully.

My diction is too poor to translate the sense of gratitude into words and heartfelt
reverence to esteemed Dr. T. Narendrappa
Narendrappa,
rendrappa Professor and Head, Department of Plant
Pathology, University of Agricultural Sciences, Bengaluru and Chairman of my advisory
committee for his sustained encouragement, valuable suggestions, and timely advice and for
providing an atmosphere of freedom of work during this investigation. Mere any words
from the vastness of literature fails to express my sense of gratitude to him, for his
continued calm endurance shown throughout my research programme and compilation of
this thesis, without which this work would not have seen the light of the day.

I wish to express my earnest and profound sense of gratitude to Dr. Y. M.

Somashekara Associate Professor, Department of Plant Pathology, GKVK, Bengaluru, Dr.
P. Chowdappa, Principal scientist IIHR Bangalore, Dr. K. Madhavi Reddy,
Reddy Principal
scientist, IIHR Bangalore, members of my advisory committee. I thank for their productive
discussion and timely suggestions; and for their valuable guidance and practical approach
that has resulted in the work presented in this thesis.

I am extremely thankful to B. M. R. Reddy

ddy, Professor and University head,
Department of Plant Pathology and all the staff members who have provided an excellent
studying atmosphere during my Masters programme.

I am thankful to the staff members of the Department of Plant Pathology, Dr. N.

Nagaraju, Dr. N. G. Rav
Ravichand
chandra, Dr. K. T. Rangaswamy, Dr. Nagaraju,
Nagaraju Dr. S. G.
Man
Mantur, and Mrs. H. A. Pra
Prameela for their valuable guidance and moral support during
my stay in this department.

But for the affection, words of encouragement, boundless love, unflagging

inspiration, interest and selfless sacrifice of my parents I would not have been what I am
today. I owe all my success to them. The special persons in my life my father Mr. Rajappa,
Rajappa,
my mother Mrs.
rs. Rathnamma,
Rathnamma my brothers Mr. Praveen,
Praveen, K.
K. R., Devaraj.
Devaraj. K.R
K.R., my sister
Sheela K. R. and Relatives without their love and blessings this study would have scarily
accomplished.

I express my sincere thanks to my seniors Basavraj and Pushpa,

Pushpa for extending help
and support during my research work.

A special thanks to my friends Niyaz, Jagadeesh, Kuldeep, Jeevan, Sanjay,

Kiran, Nirmal, Thejesh, Prasanna, Uday, Sumanth, Chandra, Ravi, Arun, Kedar, Jabbar,
Jabbar,
Pavithra, B. S, Pavithra, R. S,
S, and my seniours Manjunath, Shreesail, Dinesh Divya, Prema,
Pallavi, Shantamma and Juniors of my department for their constant encouragement and
cooperation throughout the study.

I express my special thanks to Technical staff at Department of Plant Pathology

Ramesh
Ramesh Shivakumar,
Shivakumar Puttaraju,
Puttaraju Siddaraju and Lakshmamma,
Lakshmamma, for helping me out with all
sorts of things for their support and help during my research work.

I greatly acknowledge Department of Pla

Plant Pathol
Pathology
ogy, Agriculture College, GKVK
for providing facilities to carry out my research work.

Last but not the least I thank all those who helped me directly or indirectly during
period of my stay in this UAS campus.

Bengaluru, (Suresh, K. R.)

R.)

3rd August, 2013

 STUDIES ON LEAF SPOT OF CHILLI (Capscicum annuum L.),
CAUSED BY Cercospora capsici Heald & Wolf.
SURESH, K. R.

Abstract

Cercospora leaf spot is an important disease assuming limiting factor in the

commercial production of chilli (Capsicum annuum L.) in many parts of the country,
including Karnataka. Survey was carried out in Karnataka. The highest leaf spot severity
was observed in Davanagere (14 per cent), followed by Srinivasapura (12%) and Honnali
(12%) during summer, 2013. The growth of Cercospora was highest on different nutrient
solid media viz., Carrot dextrose agar, Asthana and Hawker’s agar, Czapek’s and Oat
meal agar. The fungus was grown well at the temperature of 250C with highest dry
mycelial weight of 72.5 mg followed by 300C with dry mycelial weight of 56.9 mg. The
maximum growth (73.73mg) of Cercospora capsici was observed at pH 7.0 and the next
best at pH 6.0 (69.73mg). The eight systemic and five contact fungicides were tested in
vitro conditions at different concentrations against the pathogen. The fungicides viz.,
Propiconazole, Hexaconazole, Penconazole and Tebuconazole were best in inhibiting the
growth of the fungus up to 94.44 percent at 100, 500, 1000 and 1500 ppm concentration.
Among non-systemic fungicides, Mancozeb and Propineb were found to be effective
against the pathogen at 500, 1000, 1500 and 2000 ppm. The maximum fungal growth
inhibition was observed in Trichoderma harzianum (73%) and Trichoderma viride
(72.7%) in vitro conditions. The chilli genotypes viz., EC771550, EC771551, EC771553,
EC771544, EC771556, EC771545 and EC771548 were found resistant to Cercospora leaf
spot in laboratory condition.

Dept. of Plant Pathology Signature of the major Advisor

UAS, B. G.K.V.K. T. Narendrappa
Bangalore

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 CONTENTS

CHAPTER TITLE PAGE No.

I INTRODUCTION 1-3

II REVIEW OF LITERATURE 4-20

III MATERIAL AND METHODS 21-34

IV EXPERIMENTAL RESULTS 35-52

V DISCUSSION 53-60

VI SUMMARY 61-62

VII REFERENCES 63-74

 LIST OF TABLES

Table Page
Title
No. No.
List of fungicides used in vitro evaluation against
1. 31
Cercospora capsici.
List of bio-agents used against C. capsici and their
2. 33
sources of collection.
Survey for incidence and severity of Cercospora leaf
3. spot of chilli in Southern parts of Karnataka during 36
2013.
Colony growth of C. capsici on different solid media at
4. 39
different days of incubation.
Cultural characteristics of C. capsici on seven solid
5. 40
media.
Dry mycelial weight of C. capsici in carrot dextrose
6. 41
broth medium.
Comparison of morphological characters of C. capsici
7. 42
from chilli leaves, with the earlier report on it.
Effect of temperature on growth of C. capsici at 18 days
8. 44
Incubation.
Effect of hydrogen ion concentrations (pH) on growth of
9. 46
C. capsici at 18 days Incubation.
Per cent inhibition of mycelial growth of C. capsici by
10. 48
different Systemic fungicides.
Per cent inhibition of mycelial growth of C. capsici by
11. 49
different Contact (non- systemic) fungicides.
In vitro evaluation of bio-agents against Cercospora
12. 51
capsici.
Reaction of chilli genotypes against Cercospora leaf
13. spot under lab condition using detached leaf 52
technique.
 LIST OF FIGURES

Figure Title Between

No. Pages

1. Growth of Cercospora capsici on Carrot 39-40

dextrose broth at different incubation periods

2. Growth of C. capsici on different solid media at 41-42

different days of incubation

3. Effect of temperature on mycelial growth of C. 44-45

capsici

4. Effect of hydrogen-ion concentration on 46-47

mycelial growth of C. capsici

5. In vitro evaluation of systemic fungicides 48-49

against C. capsici

6. In vitro evaluation of non-systemic fungicides 49-50

against C. capsici

7. In vitro evaluation of bioagents against C. 51-52

capsici
 LIST OF PLATES

Plate Between
Title
Nos. Pages

1. Symptoms of Cercospora leaf spot on leaf 36-37

Microphotographs of C. capsici isolated from chilli

2. 37-38
leaf

Growth and sporulation of C. capsici on Carrot

3. 37-38
dextrose agar medium

4. Proving pathogenicity of Cercospora capsici on chilli 37-38

5. Growth of C. capsici on different solid media 39-40

6. Physiological studies of C. capsici in liquid media 44-45

In vitro evaluation of systemic fungicides against C.

7. 48-49
capsici at different concentrations (ppm)

In Vitro evaluation of non- systemic fungicides against

8. 49-50
C. capsici at Different concentrations (ppm)

9. In vitro evaluation of bioagents against C. capsici 51-52

Disease scoring scale of Cercospora leaf spot of chilli

10. 52-53
(0-9)
INTRODUCTION
 I. INTRODUCTION

Chilli (Capsicum annuum L.) is an important member of the

solanaceae family and an important spice as well as vegetable crop of
India. Chilli was known to Indians about 400 years ago, when this
crop was first introduced into India by Portuguese, towards the end of
the 15th century. Its cultivation became popular in the 17th century. It
is now grown in all parts of India covering about 9,53,800 hectares.
Chilli is valued for its diverse commercial uses.

Chilli is an important cash crop grown for its pungent fruits,

which are used as both in green and ripe form (the latter in the dried
form) to impart pungency to the food. Green chillies are rich in
Vitamin A and C, minerals and protein. Dry chillies are also rich in
Vitamin A and D. As a condiment, it has become indispensable in
every Indian home for making sauces, chutneys and pickles. Nadkarni
(1927) reported many medicinal value of chilli and used for externally
as rubefacient and also local stimulant for the tonsils in tonsillitis and
also used with many ingredients for local remedies.

India is the largest producer of chillies in the world. In India,

the maximum production level covers around 1.1 million tons
annually and also the maximum area 9.5 lakh ha dedicated to the
production of this crop (Anon., 2011). The major regions where chilli
is cultivated in India are Andhra Pradesh (1.7 lakh ha) Karnataka
(0.69 lakh ha) followed by Maharashtra, Uttar Pradesh, Punjab, Tamil
Nadu, Rajasthan, Orissa, West Bengal and Madhya Pradesh. It
occupies 49 per cent share in the Indian total production and
produces around 2.7 lakh tonnes of chillies. Karnataka follows
Andhra Pradesh, which share 14 per cent of the production in the
country.

In Karnataka, chilli is grown in larger areas in Belgaum, Haveri,

Dharwad, Chitradurga, Bellary, Raichur, Gulbarga and Bijapur
districts under rainfed and irrigation conditions. Karnataka stands
second in chilli production with 4.37 lakh tonnes which accounts for
19 per cent of total chilli production of India (Anon, 2010).

Successful production of chillies have been affected by number

of biotic constraints, among them diseases are the major ones. Chilli
is commonly affected by fungal diseases like Cercospora leaf spot,
damping off, wilt, anthracnose (dieback/fruit rot), leaf spots, powdery
mildew, bacterial diseases (soft rot and bacterial wilt) and some viral
diseases like tomato spotted wilt virus, cucumber mosaic virus,
tobacco mosaic virus, murda complex etc.

The fungus responsible for leaf spot of chilli is Cercospora

capsici it belongs to Family- Dematiaceae; Order Moniliales; Class-
Deuteromycetyes; Phylum- Ascomycota; Kingdom- Fungi. The perfect
stage was identified as Mycosphaerella capsici. The pathogen was first
isolated from bell pepper by Heald and Wolf (1911) and later the
studies on leaf spot disease of chilli were carried out by several
researchers on various aspects (Chupp, 1953; Vasudeva, 1963; Meon,
1990; Lim and Kim, 2003 and Bhat et al., 2008). Until 2000, the
cause of chilli leaf spot was related to C. capsici (Marchal and
Steyaert, 1929). However, C. capsici and C. unamunoi are considered
to be synonyms to C. capsicicola (Bhartiya et al., 2000). As such, there
is no much research information available on Cercospora leaf spot of
chilli and the fungus C. capsici. Hence, it was pertinent to confirm the
identity of fungus by using conventional and morphological
characters.

Cercospora, the leaf spot fungus, survives in or on seed as tiny

black stromata in old affected leaves in the soil. Infection occurs by
direct penetration to the leaf. Cercospora spores required high
humidity in the atmosphere for germination and penetration to the
host; however, heavy dew is sufficient for infection of the pathogen.
The disease is most severe during periods of warm temperatures and
excessive moisture (either from rain or overhead irrigation). The
fungus is spread by splashing water, wind, and leaf-to-leaf contact.
However, the pathogen does not infect fruit.

The research work on leaf spot of chilli regarding cultural,

physiological and management studies are limited. Hence, the present
investigation was undertaken for the studies on culture, physiological,
morphological with management studies by using chemical, bioagents
and host resistance with the following objectives:

1. Survey for incidence and severity of Cercospora leaf spot of chilli in

Southern parts of Karnataka

2. Isolation, identification and proving pathogenicity of the pathogen

3. Cultural, Physiological and Morphological studies on Cercospora

leaf spot

4. In vitro evaluation of fungicides and bio-agents against Cercospora

capsici

5. Evaluation of chilli genotypes against Cercospora leaf spot disease

REVIEW OF LITERATURE
 II. REVIEW OF LITERATURE

Chilli leaf spot caused by Cercospora capsici Heald and Wolf is

gaining importance in the irrigated tracts where high input cultivation
is practiced with closer plant spacing which in turn creates a micro
climate more congenial for foliar diseases such as leaf spot. As such,
there is little research information available on Cercospora leaf spot of
chilli. Hence, literatures available on Cercospora leaf spot of other
crops have been reviewed in the chapter. The literature pertains to
symptoms, survey on leaf spot disease, cultural and physiological
studies, in vitro evaluation of bio-agents and fungicides and finally
host resistance are covered in this chapter.

2.1 Survey for chilli leaf spot caused by Cercospora capsici in

southern Parts of Karnataka

Survey conducted from 1971 to 1981 in major groundnut

growing states of India to obtain information on late leaf spot and
other diseases of groundnut and also to assess their relative
importance in different regions. Late leaf spot disease was the most
important disease in all groundnut growing areas of India
Subramanyam and McDonald (1987); Kolte 1984).

Singh et al. (1994) reported, the disease severity of Cercospora

canescens causing leaf spot in greengram was observed 3.6 and 5.7
scale in 1991 and 1992 respectively in Dhiansur, Jammu and
Kashmir. Khunti et al. (2002) used 1-9 rating scale of disease severity
to calculate the disease intensity of leaf spot of greengram caused by
C. canescens which was 35.40 per cent during 1995 to 1999 in
Targhadia Gujarat.

Benagi (1995) conducted survey for late leaf spot of groundnut

in northern Karnataka and reported the high severity of late leaf spot
disease in major groundnut growing areas. The environmental factors
viz., temperature and relative humidity were played important role for
the establishment, spread and multiplication of the pathogen. Bdliya
and Kura (2007) reported that, the incidence of Cercospora leaf spot
was 61.4 and 78.9 per cent during 2002 and 2003, respectively.

Ramesh Chand et al. (2003) recorded maximum disease

intensity of (62.33 and 64.74 per cent in 1999 and 2000 respectively)
Cercospora leaf spot of greengram. Ali et al. (2011) observed the per
cent disease index (PDI) of Cercospora leaf spot of mungbean.
Recorded the lowest disease incidence 13.17, 25.45 and 34.69 per
cent (PDI) at 30, 40 and 60 DAS. The highest PDI was recorded in
control plot.

Five different variety / strain of winter mungbean viz.,

Binamoog 1, Mut 2a(5)B, MC38, MC89, MB63 were used to determine
incidence and disease severities. The incidence and severities of
Cercospora leaf spot recorded 45 days after sprouting was found to be
consistently increased with the increased maturity of the crop growth
in field and varied among the strains (Hossain et al., 2011).

Rathore (2006) reported the maximum disease incidence (35 per

cent and 21%) and low yield was observed in Cercospora leaf spot
infected greengram fields in Rajastan during 2006 and 2008 (Rathore,
2009).

Frog eye leaf spot disease on tobacco caused by Cercospora

nicotianae affected production of bidi tobacco due to severe infection
(Anon, 2004). Survey for prevalence of frog eye leaf spot disease was
conducted in seven bell pepper growing locations of Kashmir viz.,
Bugham, Gangbugh, Kanihama, Dal-area, Noorbagh, Shalimar and
Shalteng at fruit set and fruit harvest during 2003. The overall
incidence and intensity of the disease at fruit set stage ranged from
20.06 to 37.20 per cent and 7.46 to 18.33 per cent, respectively.
During fruit harvest stage, the incidence and intensity of the disease
ranged from 54.63 to 68.92 per cent and 32.46 to 44.03 per cent,
respectively. Disease incidence and intensity was recorded
significantly higher at Noorbagh and minimum at Kanihama locations
(Farooq et al., 2008).

2.2 Symptomatology

Munjal et al. (1960) described the symptoms of Cercospora

canescens on Vigna radiata as fungus produced spots on leaves,
which were first brown, later turning grey or dirty grey with narrow
reddish brown margins bearing fructification on both the surfaces.

Vasudeva (1963) in his compilation on ‘Indian Cercosporae’

described the symptoms caused by Cercospora capsici as the leaf
spots were circular to sub circular, 1- 7 mm in diameter raised on
upper surface, brown later became greyish-brown, marginated by a
very definite darker zone and outside of which is a more or less
extended yellow halo. Leaf spots of chilli caused by C. capsici are
usually amphigenous, circular 1-10 mm in size, later coalescing to
form large patches with dark brown margin. Conidiophores produced
in fascicles mainly on lower surface of infected leaves and conidia are
hyaline and indistinctly multi septate (Meon, 1990).

The typical disease symptoms of frog eye leaf spot of pepper

caused by C. capsici were observed on leaves, stem and peduncles. On
leaves, the spots appeared as necrotic, circular to sub-circular with
greyish white centre, surrounded by brown to greyish brown area and
marginated by definite darker zone. The spots enlarged up to a mean
diameter of 9.8 mm, coalesced frequently and led to defoliation with or
without yellowing. The spots become raised and resembled frog eye.
The lesions on stem, peduncle and petioles, however, were longer
rather than round (Bhat et al., 2008).

Vakili (1977) while distinguishing of C. canescens and C.

cruenta on cowpea, described symptoms of the former as,
amphigenous lesions. On leaves, generally round, orange to light
brown spindle lesions on petiole, penduncle and stem.
 Patel et al. (2001) explained the symptoms of frog eye leaf spot
of tobacco caused by Cercospora nicotianae. The spots were brown
with ash grey centres. Often the centres may turn white and dry up.
The typical spot had a white centre, surrounded by black margin
resembling the eye of frog; several spots may coalesce towards the leaf
tip and margin causing the leaf to dry up from the margin. Fungus
produced definite spots on green gram leaves which were first brown,
later turned grey to dirty grey with narrow reddish brown margin, sub
circular to irregular and 5-10 mm wide (Jamadar, 1988).

2.3 Isolation of pathogen

The fungus Cercospora capsici was first isolated from bell

pepper by Heald and Wolf (1911) and later studied by, Chupp (1953)
and Vasudeva (1963). Infection, sporulation and spore germination of
Cercospora capsici, causal agent of leaf spot on Capsicum annuum L.
was investigated by Padmavathi et al. (1993).

Nagel (1934) isolated Cercospora beticola Sacc. causing leaf spot

disease of sugarbeet by standard tissue isolation technique. Dange
and Patel (1968) isolated Cercospora beticola Sacc. from naturally
diseased spinach beet plants by standard tissue isolation method.

Siddaramaiah (1986) isolated Cercospora moricola Cooke

causing leaf spot disease of mulberry by standard tissue isolation
technique. Jamadar (1988) isolated Cercospora canesecens Ell. and
Mart. causing leaf spot disease of mungbean by following standard
tissue isolation technique. Mallappa (2007) isolated Cercospora
nicotianae Ell. and Eve. from tobacco by tissue isolation technique
and proved pathogenicity.

Pradeep (2005) isolated the fungus Cercosporidium personatum

from the infected groundnut leaves using Richard’s agar medium. He
proved the pathogenicity by inoculating the mycelium bits of the
culture on the groundnut plants.
2.3.1 Morphological studies of Cercospora

Conidia and conidiophores of Cercospora were 4-5 X 70 -25 µm

and 4.5 X 30-60 µm (Heald and Wolf, 1911). Chupp (1954) revealed
that stromata lacking, a few brown cells rarely 15-30 µm,
conidiophores geniculate, 3.5- X 20-150 µm, bases sometimes wider,
conidia hyaline, acicular, rarely obclavate, 2.5 - 4 X 30-200 µm.
Vasudeva (1963b) described the morphological characters of C. capsici
as conidiophores in clusters, tufted, amphigenous, brown, 10-15
facsciculae, septate, 30-60 X 4.5-5.5 µm. Conidia straight, clavate,
dilute brown in colour and septate.

Vasudeva (1963) in his compilation on “Indian cercosporae”

described the morphological characters of C. nicotianae as
condiophores in clusters, which were tufted, amphigenous, 75 -100 x
4-5µ in size, 2-3 times geniculate, dark brown septate and arise
through stomata. The conidia were slender, slightly curved, thin
walled, hyaline multiseptate i.e. 3-6 septe and measure 40-75µ x3-5µ
in size.

The mycelium of Cercospora capsici was irregularly branched,

septate, light brown, 3.9 to 4.8 µm in diameter. Conidiophores were
light brown, un-branched, continuous, 1 to 5 septate, straight to sub-
straight and borne on stromata in fascicles of 7 to 13. Conidia were
acicular, continuous, 1 to 13 septate, hyaline and borne solitary on
conidiophores. The conidia and conidiophore measured 3.5 - 5.2 X 25-
86 µm and 3.5 - 5.5 X 30-75 µm respectively (Bhat et al., 2008).

Gill and Singh (1962) reported that conidiophores of C. beticola

were pale brown, not branched, septate, geniculate and measured 4 x
25- 115 µ. The conidia were hyaline, septate and measured 4 x 54 –
165 µ.

Munjal et al. (1960) described the fungal morphology as the

conidiophores which were olivaceous brown to dark, septate,
geniculate, coming out from stromata in dens fasicules. They measure
3.8-5.8 X 19.3-130.9 µ. The conidia are hyaline, acicular, septate and
measure 2.9-4.8 X 73.2-231.0 µ.

Jamadar (1988) observed the conidiophores of Cercospora

canescens coming out from stomata in dense fascicles, olivaceous
brown to dark in colour, which had many septations. The conidia were
hyaline, variously curved or straight and septate.

2.4 Cultural studies of Cercospora

2.4.1 Growth characters on solid media

Mandelson (1933) and Yen (1956) grew the fungus Cercospora

nicotianae on potato dextrose agar (PDA) medium. Nagel (1934)
studied the conidial production of C. moricola on sugar beet leaf
extract agar.

Hill (1936) reported that no conidial production from single

spore culture of C. nicotianae grown on several types of media
including potato dextrose agar and plain tobacco agar. Diachun and
Valleau (1941) obtained sporulation of C. nicotianae on tobacco leaf
decoction agar medium and also reported several isolations on same
medium and most of these isolates produced conidia and
conidiophores six days after inoculation on the medium. The fungus
C. nicotianae grew well on synthetic nutrient solution with sucrose as
carbon source but sporulation was not observed (Steinberg, 1950).

Murakishi (1951) reported that C. kikuchii failed to sporulate in

culture but observed that the best mycelial growth was found on
potato dextrose agar. Kilpatric and Johnson (1956) observed good
growth and sporulation of Cercospora species on carrot leaf decoction
agar.

Stavely and Nimmo (1968) observed maximum radial growth of

C. nicotianae on agar medium containing 1.6 g DL-Leucine, 50 g
sucrose and 2-4 g yeast extract per litre and mineral nutrient and
concluded that sucrose was the best source of carbon for the growth.
Further, they tested ten different types of media and reported
maximum growth of fungus on carrot leaf decoction agar and
minimum on PDA.

Dange and Patel (1968) reported that spinach leaf spot caused
by C. beticola Sacc. grew both in complex organic and synthetic
media. Sporulation was scanty on non-synthetic media. However,
potato dextrose agar and Czepack’s (dox) agar medium were best for
the growth of the fungus. A good growth of an isolate C. arachidicola
and two isolates of C. personata were observed on potato dextrose
agar, Czapeck’s (dox) agar with yeast extract, radish dextrose agar and
carrot dextrose agar (Sulaiman and Hande, 1968).

Alasoadura and Fajola (1970) observed abundant conidia

production on tobacco decoction agar when C. nicotianae culture was
subjected to darkness and high relative humidity. Verma and
Agnihotri (1972) found maximum growth of C. cruenta and C. beticola
on Czapeck (dox) agar followed by Carrot leaf decoction agar media.
According to Kanti (1975), the isolates of C. moricola made better
growth and sporulation on potato dextrose agar. He could get
abundant sporulation on boiled potato piece left over after decantring
the extract.

Chen et al. (1979) reported that the fungus C. kikuchii can grow
most rapidly on malt extract agar or PDA and least growth occurred
on water agar. The largest numbers of conidia were formed on V-8
juice agar and carrot leaf decoction agar. No conidia developed on
water agar, malt extract agar and PDA.

Kwon and Oh (1981) obtained abundant sporulation of C.

canescens on mungbean leaf decoction-oatmeal agar. Dinesha (1984)
observed PDA supporting maximum growth of C. sorghi. Khandar et
al. (1985) reported that good radial growth of C. canescens on PDA,
Oatmeal and Anderson’s agar whereas moderate growth occurred on
standard nutrient, glucose asparagine, carrot leaf and mungbean
decoctions and Richard’s agar media.

Queiroz and Menezes (1993) obtained the sporulation of C.

nicotianae by plating a spore suspension on dishes containing
different culture media. The maximum sporulation was obtained on V-
8 juice - CaCO3 agar, followed by coconut milk agar and tomato juice -
CaCO3 agar under the alternate light regime. Sporulation was very low
on dry tobacco leaves - CaCO3 agar, dry tobacco leaves agar, PDA and
PDA-panvit media under both the light regimes tested.

Suraj (2004) observed that abundant sporulation of C.

abelmoschi causing leaf spot disease of okra on carrot leaf extract agar
and host leaf extract agar. Maximum mycelial growth was observed on
PDA, host leaf extract agar, carrot leaf extract agar and V-8 juice agar.

Prashanth (2004) observed maximum growth of C. kikuchii on

malt extract agar medium followed by PDA. Pairashi and Jahagirdar
(2007) revealed that host extract dextrose agar supported maximum
radial growth of C. nicotianae followed by Czapeck’s (dox) agar and
carrot leaf decoction agar.

Chandrashekhran and rangaswami (1960) noticed that the

pathogen Cercospora cruenta causal agent of leaf spot of cowpea grew
well on both synthetic and complex organic media. In their studies,
Czapeck’s agar was a better medium for the growth of the pathogen
than Richard’s medium.

Berger and Hanson (1963) studied the radial growth of

Cercospora zebrine on 11 solid media and could record maximum
growth on modified Richard’s agar and potato dextrose agar media. In
liquid culture, maximum growth was obtained in Cornmeal broth,
modified Richard’s broth and Oatmeal broth.

Misra and Bhattacharyya (2002) reported that the PDA was the
best for both radial growth and sporulation and carrot leaf decoction
oatmeal extract agar (COA) medium was the best for sporulation of C.
canescens among six different media used. The pathogen favoured
natural or semi-synthetic media while synthetic media was unsuitable
when grown in agar based media.

Ekpo and Esuruosa (1978) tested the growth and sporulation of

C. canescens and C. cruenta isolated from cowpea on ten different
solid media. Best mycelial growth was obtained on 1.2 per cent PDA
and V-8 juice agar for C. canescens and on V-8 juice agar and
nutrient agar for C. cruenta. Sporulation was poor or totally absent on
most of the media. C. canescens produced conidia on cowpea leaf
decotion agar and on V-8 juice agar.

Satya Prasanth (2004) observed maximum radial growth of

Cercospora kikuchii fungus on malt extract agar which was on par
with potato dextrose agar. Minimum radial growth was observed on
carrot leaf decoction agar.

Mallappa Prakash (2007) reported that host extract dextrose

agar supported maximum radial growth of C. nicotianae which was on
par with Czapeck (dox) agar.

2.4.2 Growth characters on liquid media

2.4.2.1 Growth Phase

Lily and Barnett (1951) while discussing growth pattern of fungi

outlined the following growth phases: viz., stationary phase,
accelerated phase, declining acceleration phase, maximum stationary
phase and phase of decline or autolysis. They attributed these phases
of fungus to the environmental and nutritional condition in which it
grows.

Kanti (1975) reported the maximum growth of C. moricola at 20

days after inoculation in potato dextrose broth (PDB).
Lakshminarayana (1981) obtained maximum growth of C. solani-
melongenae on 22nd day of incubation in potato dextrose broth.
Dinesha (1984) harvested maximum mycelium of C. sorghi on 16th
day of incubation in PDB.

Prashant (2004) obtained maximum growth of fungus C.

kikuchii on 22nd day after inoculation. Maximum growth of the fungus
C. nicotianae was observed on 19th day after inoculation in PDB
(Pairashi and Jahagirdar, 2007).

2.5. Physiological studies

2.5.1 Effect of temperature

Sulaiman and Hande (1968) reported that the temperature of

24-280C was found to be optimum for C. arachidicola and also for the
isolate of C. personata. Stavely and Joyce (1968) noticed that the
optimum temperature for sporulation of C. nicotianae was 180C but it
required 260C for radial growth.

Verma and Agnihotri (1972) recorded the best growth of C.

cruenta and C. beticola at a temperature of 260C. Makadia et al. (1979)
reported that 270C was the optimum temperature for maximum
growth and spore production of C. nicotianae.

Dinesha (1984) recorded maximum spore germination of C.

sorghi at 250C. Nelson and Campbell (1990) obtained the maximum
mycelial growth of C. zebrine at 240C in V-8 juice agar. Lim and Kim
(2003) reported that the optimum temperature required for mycelial
growth of C. capsici was 250C and the fungus did not grow below 50C
and over 350C. Suraj (2004) recorded the maximum mycelial growth of
C. abelmoschi at 250C.

Prashant (2004) obtained maximum mycelial growth of C.

kikuchii at temperature of 250C. Maximum mycelial growth of the
fungus C. nicotianae was at temperature 250C and growth was
minimum at temperature 50C and 400C (Pairashi, 2007).
2.5.2 Effect of Hydrogen ion concentration (pH)

Singh (1934) reported that the good growth of the two strains of
the C. indica sp. Parasitica was at optimum pH of 6.7. Chandrasekhar
and Rangaswamy (1960) reported that C. cruenta and C. beticola
growth was found best at pH 6.0. Landers (1964) obtained that best
growth of C. arachidicola at pH 4.5. Dayal and Ram (1968) recorded
the higher percentage of spore germination of C. jasminicola at pH 5.5.

Dange and Patel (1968), C. beticola grew at pH levels between

3.1 and 9.1, but the best growth occurred at pH 6.0. Sridharan and
Rangaswamy (1968) recorded the best growth of the Cercospora sp. on
Abelmoschus esculentus at pH 5.5. Stavely and Joyce (1968) found
that the optimum pH values for sporulation of C. nicotianae were 4.5
and 5.0.

Raghunathan (1969) reported two optimum pH 6.5 and 8.0 as

most favourable for the growth of C. canescens and C. dolichi,
respectively. Verma and Agnihotri (1972) reported a pH of 6.8 and pH
7.0 to be most favourable for the growth of C. cruenta and C. beticola.
Kanti (1975) recorded the growth of C. moricola in a wide range of pH
3.0 to 8.0 (optimum 6.5) and the sporulation in the pH between 5.0
and 6.5 (optimum 6.0).

Makadia et al. (1979) reported that the optimum pH for

maximum growth and sporulation of C. nicotianae was 4.5 and 6.5,
respectively. Lim and Kim (2003) observed that optimum pH for
mycelial growth of C. capsici was 4.0- 8.0. Suraj (2004) reported the
optimum pH of 5.0 for maximum mycelial growth of C. abelmoschi.

Prashant (2004) reported that the fungus C. kikuchii grew at all

pH levels but maximum mycelial growth was recorded at pH of 4.0
and optimum growth in between pH of 4.0-7.0. Pairashi (2007)
recorded maximum mycelial growth of C. nicotianae at pH of 5.0.
Growth decreased as the pH moved away from 5.0 on either side.
2.6 Management of Cercospora leaf spot of chilli

2.6.1 In vitro evaluation of fungicides

Chemical fungicides are the main sources to control most of the

plant diseases. Chemical compounds are more effective against plant
pathogens through its virtual toxicity. In recent years, farmers are
practicing an intensive use of chemical fungicides. Therefore,
fungicides continue to be our main stay for foliar disease
management.

Agarwal and Joshi (1971) reported that seed treatment with

Thiram increased the germination of purple stained soybean seed to
some extent in both laboratory and field.

Prashanth (2004) evaluated systemic and non-systemic

fungicides against C. kikuchii causing purple seed stain of soybean
under in vitro condition. Systemic fungicides Carbendazim,
Thiophanate methyl, Hexaconazole and Prochloraz (0.25, 0.05 and
0.1per cent), among non-systemic fungicides Carbendazim 12 per cent
+ Mancozeb 63 per cent (0.1, 0.2 and 0.25 per cent) Mancozeb and
Thiram (0.25 per cent) recorded 100 per cent inhibition of mycelial
growth.

Pairashi (2007) conducted in vitro evaluation of systemic and

non-systemic fungicides against C. nicotianae infecting tobacco.
Among systemic fungicides, Hexaconazole, Penconazole and Benomyl
recorded 100 per cent inhibition at all tested concentrations (0.05, 0.1
and 0.15 per cent) followed by Propiconazole and Carbendazim (0.15
per cent) and Thiophanate methyl was least inhibitor even at 0.15 per
cent. Among the non-systemic fungicides, maximum inhibition was
noticed in Carbendazim 12 per cent + Mancozeb 63 per cent (0.25 per
cent).

Adisa (1989) reported that the chemical brestan (Fentin+Maneb)

was more effective in inhibiting spore germination of C. cruenta than
Benlate (Benomyl). In another experiment involving in vitro evaluation
of chemicals on spore germination C. zinniae, Carbendazim was highly
effective in inhibiting spore germination (Reshi et al., 2001). Ruben et
al. (2007) while working on chemical control of leaf spot of husk
tomato caused by Cercospora sp. obtained 100 per cent inhibition of
conidial germination with Tebuconazole at 30 ppm.

Khalil and Jalaluddin (2004) tested the fungicidal management

strategies for Cercospora leaf spot of black gram (Cercospora cruenta)
and found that all the foliar fungicides, Carbendazim, Mancozeb,
Mancozeb + Thiophanate-methyl, Mancozeb + Metalaxyl and Sulfur
consistently reduced the incidence and severities of the disease and
increased seed yield. Among these fungicides, carbendazim was the
best in economic management of the disease.

Mukhopadhyay and Rao (1974) concluded that two sprays at

20-day intervals with the systemic fungicides, Butylcarbamoyl,
Benomyl 50 WP, Carbamate, Thiabendazole, gave better disease
control with two protective fungicidal sprays against Cercospora leaf
spot diseases.

Peterson et al. (1976) concluded that application of benomyl 2-

week before the flowering in mountain laurel (Kalmia latifolia) was
most effective against the Cercospora disease.

In laboratory tests with eight fungicides, RH-2161 and

Carbendazim completely inhibited mycelial growth of an isolate of C.
canescens from Vigna radiate, while Captafol, Captan and Ziram were
only moderately effective (Jamadar and Padaganur, 1991).

Satya prasanth (2004) evaluated four systemic and five non-

systemic fungicides under laboratory conditions for their efficacy
against C. kikuchii. Among systemic fungicides, viz., Carbendazim,
Thiophanate Methyl, Hexaconazole and Prochloraz showed cent per
cent inhibition of fungus at all the concentrations tested. In non-
systemic fungicides, cent per cent inhibition was shown by
Carbendazim (12 per cent) + Mancozeb (63 per cent) at all
concentrations tested. Complete inhibition was shown by Thiram and
Mancozeb at 0.2 per cent concentration. However the inhibition shown
by Mancozeb at 0.2 per cent was more than that shown by Thiram at
the same concentration. The least mycelial growth inhibition was
observed in case of Chlorothalonil.

Raj et al. (2011) showed the effect of fungicides on mycelial

growth of C. canescens. After 10 days incubation in Richard’s broth
medium Propiconazol and Carbendazim at their different
concentration (0.1-50mg/ml) totally inhibited the mycelial growth.
There was no growth recorded at 0.1 mg/ml concentrate in both
fungicides. Triadimefon and Copper oxy chloride inhibited mycelial
growth of C. canescens only at 50 mg/ml concentrations.

Chandra Pal Singh et al. (2011) studied the efficacy of five

fungicides viz., Thiram, Dithane-M-45, Bavistin, Blitox and Vitavax,
two antibiotics viz., Griseofulvin and Streptocycline and one
Sulphadrug viz., Sulphamethoxazole under in vitro conditions against
Cercospora traversiana. They observed that the radial growth of
pathogenic fungus was highly affected (95-100 per cent) with Bavistin
while other tested fungicides, antibiotics and Sulphadrugs moderately
inhibited the radial growth of Cercospora traversiana.

2.6.2 In vitro evaluation of bioagents

The control of plant pathogens using antagonistic

microorganisms plays an important role in plant disease management,
indicating the need for isolating and evaluating best bio control
agents. Use of these antagonist beneficial microorganisms can reduce
the polluting chemical, residual levels of fungicides in crop products,
potential environmental contamination, occurrence of fungicide
tolerance strains of pathogens and protection of beneficial organisms.
In addition, antagonistic agents are used as one of the component in
integrated disease management practices.

Siddaramaiah (1986) reported that Trichoderma harzianum

giving hyperparasitic reaction on C. moricola and Bacillus subtilis
showed high degree of inhibition to the growth of C. moricola in
bioassay studies.

Prashanth (2004) reported that Trichoderma viride showed the

maximum inhibition of C. kikuchii followed by T. koningii. Among the
bacterial bioagents Bacillus subtilis was the best inhibitor. Pairashi
(2007) reportd that T. koningii recorded maximum growth inhibition of
C. nicotianae, whereas among the bacterial bioagents, Bacillus subtilis
was best inhibitor and Pseudomonas fluroscens was least inhibitor.

The antifungal activity of the algal culture filtrate has been

attributed to the presence of bioactive compounds that is total
phenolic compounds, total saponins and alcoloidsin. The best results
were obtained by Spirulina platensis, Oscillatoria sp. and Nostoc
muscorum. The highest inhibition of mycelial growth of C. beticola was
achieved by the concentrations of 30 and 40 per cent. The fungal
spores production was completely inhibited at 300 and 400 ppm,
particularly at the concentration of 400 ppm. The same result was
obtained by Topsin M 70 (Hussein et al., 2009).

Raguchander et al. (2005) evaluated the talc formulations of two

biocontrol agents viz., Bacillus subtilis and Pseudomonas fluorescens
as seed and foliar treatments against foliar diseases of urdbean (Vigna
mungo) and found that bioagents were as effective as Carbendazim in
reducing the severity of Cercospora leaf spot and enhanced the yield.

2.7 Screening

Iqbal et al. (1992) reported that out of 87 genotypes of Vigna

radiata tested in the field under artificial inoculated conditions, only 4
genotypes were highly resistant to C. canescens, while 12 were
moderately resistant, 16 intermediate, and the remaining were
susceptible.

Cantonwin et al. (2006) evaluated the effect of integrated

management schedule against early leaf spot of groundnut caused by
Cercospora arachidicola and reported that leaf spot intensity
decreased with increased fungicide applications, use of resistant
cultivars and strip tillage.

Grover et al. (1976) found peanut variety PI 109839 was highly

resistant to early leaf spot caused by Cercospora arachidicola in
greenhouse and in five field tests. Six other peanut varieties, PI
162857, PI 350680, PI 259639, PI 259679, PI 259747 and PI 270808
were also found moderately resistant to early leaf spot.

Bashir et al. (1993) screened 20 Vigna radiata genotypes against

Cercospora canescens under field conditions and found that NCM-10
was resistant and remaining genotypes were susceptible.

Jamadar (1988) reported that out of 43 greengram varieties

screened during kharif 1987-88 under natural conditions, only CO-4
and ML-323 were found to be resistant to C. canescens.

Katna et al. (2001) evaluated forty-one genotypes of urdbean

(Vigna mungo) for their resistance to Cercospora leaf spot caused by
Cercospora canescens and C. cruenta (Mycosphaerella cruenta) under
mono and intercropping systems in a field experiment during the
kharif season and none of the genotypes were resistant or highly
resistant to the disease under both cropping systems. Under the
intercropping system, 21 genotypes exhibited moderate resistance to
the pathogen.

Raje and Roa (2002) evaluated two hundred germplasm lines

along with six commercial varieties of mungbean against Cercospora
leaf spot out of which only thirty five genotypes were found resistant.
Daisy Basandari et al. (2003) screened 250 stocks of blackgram
against Cercospora leaf spot out of which 30 genotypes showed varied
levels of resistance.

Browne and Cooke (2005) evaluated the relative resistance of 15

winter barley, three winter wheat and three winter oat cultivars and
two spring wheat cultivars using Microdochium nivale in detached leaf
assays to further understand components of partial disease resistance
(PDR) and Fusarium head blight (FHB) resistance across cereal
species. Barley cultivars showed incubation periods comparable to
latent periods longer than the most FHB resistant Irish and UK wheat
cultivars evaluated.

Brunet et al. (1973) showed effective alternative method of

screening white beans (Phaseolus vulgaris L.) for anthracnose
(Colletotrichum lindemuthianum (Sacc. & Magn.) Briosi & Cav.)
resistance using excised leaves or leaflets.

Browne et al. (2005) investigated the utility of a detached leaf

assay, inoculated using inoculum from isolates of Microdochium nivale
var. majus, to identify components of FHB resistance among 30
entries of U.S.

Twizeyimana et al. (2007) evaluated fourteen soybean

accessions and breeding lines for resistance to soybean rust caused
by the fungus Phakopsora pachyrhizi. They observed that inoculation
of detached leaves with 1 × 106 spores/ml resulted in a significantly
higher total number of pustules and spores per unit leaf area than
inoculations with lower spore concentrations.
MATERIAL AND METHODS
 III. MATERIAL AND METHODS

The experiments pertaining to the present investigation on

“Studies on leaf spot of chilli (Capscicum annuum L.), caused by
Cercospora capsici Heald and Wolf” was conducted in the Department
of Plant Pathology UAS, Gandhi Krishi Vignana Kendra, Bengaluru.
The materials used and methodology followed during the present
investigations are briefly described here.

3.1 General procedure

3.1.1 Cleaning and sterilization of materials

The materials used in this study were washed thoroughly in tap

water and rinsed with double distilled water. The plastic wares were
air-dried and the glassware’s were dried in hot air oven. The cleaned
and dried materials were wrapped in aluminium foil, covered with
paper and autoclaved at 1210C at 15 PSI for 15 minutes.

3.1.2 Sterilization of the laminar air flow chamber

Isolation and cultural studies were conducted under aseptic

conditions in the laminar air flow cabinet. The working surface of
laminar air flow was sterilized by swabbing with 70 per cent ethanol.
Any material coming from outside was also sprayed with alcohol. In
case of glassware, the mouth of the bottles etc., was flamed before and
after use. The forceps, inoculation loop etc., were sterilized by heating
in the flame and hands were sterilized with ethanol.

3.2 Survey for incidence and severity of Cercospora leaf spot of

chilli in Southern parts of Karnataka

Chilli is one of the main vegetable crops of Karnataka and it is

grown both in kharif and summer conditions under protected
irrigations. An intensive survey was conducted during 2013 to know
the incidence and severity of Cercospora leaf spot disease in chilli
growing areas. The survey was conducted in six districts viz.,
Bangalore rural, Chikkaballapur, Kolar, Ramnagar, Mandya and
Davanagere.

Survey was made in intensive chilli growing areas of above six

districts. In each district five chilli grown villages, in each village five
fields were selected. In each field 25 plants were randomly selected
and in each plant, five leaves were taken, two each from bottom and
middle and one from top were observed to determine the incidence of
Cercospora leaf spot by following 0 to 5 scale and infected leaf
samples were also collected.

Scale Description

0 = No disease symptoms

1 = A few spots towards tip covering 10 per cent leaf area

2 = Several dark brown patches covering up to 20 per cent leaf area

3 = Several patches with pale outer zone covering up to 40 per cent

leaf area

4 = Leaf blight covering up to 75 per cent leaf area or breaking of the

leaves

5 = Complete drying of the leaves or breaking of the leaves from plant

Per cent disease index (PDI) was worked out using the formula
given by Wheeler (1969)

Sum of individual rating

Per cent disease index = --------------------------------------- X 100

No. of leaves observed X Max. disease grade

3.3 Symptomotology and causal organism

An attempt was made to identify the Cercospora leaf spot

symptoms caused by Cercospora capsici in kharif season, 2012, the
chilli sown area in the G.K.V.K farm was regularly visited for the
identification and severity of Cercospora leaf spot disease.

3.4 Isolation of the pathogen

The infected leaves were cut in to small leaf bits and surface
sterilized with sodium hypochlorite 1per cent solution for two to three
minutes and further bits were washed in sterile water for three times
to clear residue toxicity. Then the infected leaf bits were transferred on
to Petri dishes (1-2 leaf bits per Petri dish) containing Carrot Dextrose
Agar medium (CDA) with the help of a sterile forceps and incubated at
250 C for 15 days. Further the culture was purified, subcultured on
CDA for further work.

3.4.1 Single spore isolation

This method was followed for obtaining pure culture of

Cercospora since, the fungus is known to be highly heterozygous.
Hyphal tip isolation was done on two per cent water agar plates.
Diluted spore suspensions were prepared in sterile distilled water.
One ml of such suspension was spread on petriplates and poured 15
ml water agar medium and allowed for few minutes for solidification.
The spores coming out from conidia was marked using marker under
microscope and were cut by using cork borer and transferred on to
the carrot dextrose agar medium plates incubated at 25±1oC to get the
pure culture. The pure culture was maintained in the laboratory
under refrigerator condition for further studies.

3.4.2 Identification and Maintenance of the culture

The study was undertaken to confirm the identity of the fungus.

Conidiophores and conidia which were observed from the fresh leaves
were recorded for the measurement of length, breath and number of
septation. The observations obtained were compared with the earlier
description of the fungus. The fungus was sub cultured on carrot
dextrose agar slants and allowed to grow at 250C for 15 days and such
slants were preserved in a refrigerator at 40C and sub cultured once in
30 days. Such culture was used throughout the study period.

3.4.3 Proving the Pathogenicity

Pathogenicity test was conducted on susceptible chilli cv.

Byadagi kaddi. Seedlings were raised in pots under glass house
condition. Thirty days old seedlings were sprayed with the suspension
containing mycelial bits of the fungus prepared in sterilized distilled
water. Such inoculated plants were covered with polythene bags and
kept in dark condition for 12 h and then transferred to a growth
chamber, which was maintained at 250C with 100 per cent relative
humidity. The pots were removed from the growth chamber after 48
hours and kept in glass house. The observations were taken for the
appearance and development of symptoms. Suitable controls were
maintained by spraying the plants only with distilled water.

3.5 Morphological studies of Cercospora capsici

Morphological characters such as mycelial width, length and

width of conidia, colony colour, and type of margin, colour of margin,
mycelial growth and sporulation were measured under 10x through a
compound microscope. The length, breadth, colour and number of
septations and the shapes of spore were also recorded.

3.6 Cultural Studies

3.6.1 Growth phase

Twenty ml of carrot dextrose broth was poured in each of the

100 ml conical flasks. These were sterilized and inoculated with five
mm mycelium disc cut from the periphery of the 15 days old culture
and incubated at 250C. A set of three flasks were harvested every 48
hours starting from second day of inoculation up to 30th day at two
days interval. Cultures were filtered through Whatman No.42 filter
paper, which were previously dried to a constant weight in hot air
oven at 600C. The mycelial mat on the filter paper was thoroughly
washed with distilled water to leach out any salts associated with the
mycelium. Subsequently, the filter paper along with the mycelial mat
were dried to a constant weight and weighed on an electronic balance.
The data was analysed statistically.

3.6.2 Growth studies on different solid media

The variation in cultural characters of Cercospora capsici was

studied on the following synthetic and non-synthetic solid media and
the best media for the fungus growth was identified.

The growth characters of the fungus were studied on seven

different solid media. All the media were sterilized at 1210C, 15
pounds pressure for 15 minutes. Twenty ml of each of the medium
was poured in to 90 mm diameter petridishes. Such plates were
inoculated with five mm disc of fungal growth and incubated at 250C.
Each treatment was replicated thrice. Colony diameter was recorded
by averaging the linear growth of the colony in in two directions for
each plate after 20th day of inoculation. The fungal colony colour,
surface elevation and sporulation were also recorded at the end of the
incubation period. The data on radial growth was analysed
statistically.

The different media used in the studies were follows:

Non-synthetic media

1. Carrot dextrose agar

2. Potato dextrose agar

3. Oat meal agar

Synthetic media

1. Asthana and Hawker's agar

2. Czapeck’s (Dox) agar

3. Richard’s agar

4. Sabourauds`s agar

The composition and preparation of the above mentioned

synthetic and nonsynthetic media were obtained from Ainsworth and
Bisby’s “Dictionary of the Fungi” by Hawksworth et al. (1983). The
compositions of media are given below.

1. Asthana and Hawker's agar

Glucose 5.00 g

Potassium nitrate 3.50 g

Potassium dihydrogen phosphate 1.70 g

Magnesium sulphate 0.75 g

Agar-agar 20.00 g

Distilled water 1000 ml (volume to make up)

Agar-agar was melted in 500 ml of distilled water. All the

ingredients were dissolved in 500 ml of distilled water. Both the
solutions were mixed thoroughly and sterilized at 1.1 kg cm-2
pressure for 20 minutes and preserved for further use.

2. Carrot dextrose agar

Peeled carrot 200 g

Dextrose 20 g

Agar-agar 20 g

Distilled water 1000 ml (volume to make up)

The 200 g of peeled carrot were cut into small pieces and boiled
in distilled water and the extract was cooled by filtering through
muslin cloth. Dextrose 20 g and agar 20 g of each were dissolved in
carrot extract and the final volume was make up to 1000 ml with
distilled water and sterilized at 1.1 kg cm-2 pressure for 20 min and
preserved for further use.

3. Czapeck’s (Dox) agar (CA)

Sucrose (C6H12O6) 30.00 g

Sodium nitrate NaNO3 20.00 g

Potassium dihydrogen phosphate (KH2PO4) 1.00 g

Magnesium sulphate (MgSO4. 7H2O) 0.50 g

Potassium chloride (KCl) 0.50 g

Ferric chloride 0.01 g

Agar–agar 20.00 g

Distilled water 1000.00 ml (volume to

make up)

All the chemical ingredients excluding agar were dissolved in

300 ml water and agar-agar was melted separately in 500 ml distilled
water. Two solutions were mixed thoroughly and the volume was
made up to 1000 ml by using distilled water and was sterilized at 1.1
kg cm-2 pressure for 20 min and preserved for further use.

4. Oat meal agar (OMA)

Oat flakes 30 g

Agar–agar 20 g

Distilled water 1000 ml (volume to make up)

Oat flakes were boiled in 500 ml distilled water for 30 minutes

and filtered through muslin cloth. Agar was melted in 500 ml distilled
water separately. Both the solutions were mixed thoroughly and the
volume was made up to 1000 ml and was sterilized at 1.1 kg cm-2
pressure for 20 min and preserved for further use.

5. Potato dextrose agar (PDA)

Peeled potato 200 g

Dextrose 20 g

Agar-agar 20 g

Distilled water 1000 ml (volume to make up)

200 g of peeled potatoes were cut into small pieces and boiled in
distilled water and the extract was cooled by filtering through muslin
cloth. Dextrose 20 g and agar 20 g of each were dissolved in potato
extract and the final volume was make up to 1000 ml with distilled
water and sterilized at 1.1 kg cm-2 pressure for 20 min and preserved
for further use.

6. Richard’s agar

Sucrose 20 g

Potassium dihydrogen phosphate (KH2PO4) 5g

Potassium nitrate (KNO3) 10 g

Magnesium sulphate (MgSO4 2H2O) 2.5 g

Ferric chloride (FeCl3. 6H2O) 1g

Agar-agar 20 g

Distilled water 1000 ml (volume to make

up)

All the ingredients except potassium dihydrogen phosphate were

dissolved in 450 ml distilled water. Agar melted in 500 ml distilled
water was mixed with the above solution. Potassium dihydrogen
phosphate was dissolved separately in 50 ml water and mixed
together at the time of pouring to plates and sterilized at 1.1 kg cm-2
pressure for 20 min and preserved for further use.

7. Sabouraud’s agar (SA)

Dextrose 40 g

Peptone 10 g

Agar-agar 20 g

Distilled water 1000 ml (volume to make up)

All the ingredients were dissolved one by one in 400 ml distilled

water and agar was dissolved separately in 500 ml distilled water and
mixed with the above solution and the volume was made up to one
litre before sterilization.

3.7 Physiological studies

3.7.1 Temperature requirement

Carrot dextrose broth was used in this experiment. Conical

flasks of 100 ml capacity containing 20 ml of each liquid medium were
inoculated and incubated at different temperature levels. The different
temperature levels tested for the growth of Cercospora capsici were 10,
15, 20, 25, 30, 35 and 400C. Each treatment was replicated thrice.
The dry mycelial weight at each temperature level was recorded after
incubating for 18 days and the data were analysed statistically.

3.7.2 hydrogen-Ion concentration (pH)

Carrot dextrose broth was used as basal medium. The pH of the

liquid medium was adjusted using 0.1 N alkali (NaOH) or 0.1 N acid
(HCL). The different hydrogen ion concentrations used were 4.0, 5.0,
6.0, 7.0, 8.0 9.0 and 10. Conical flasks of 100 ml capacity and each
containing 20 ml of the medium were inoculated at 25 0C. Each
treatment was replicated thrice. The dry mycelial weight was obtained
after incubating for 18 days as described earlier and the data were
analysed statistically.

3.8 In vitro evaluation of fungicides against Cercospora capsici

3.8.1 Poison food technique

The poison food technique (Sharvelle, 1961) was followed to

evaluate the efficacy of fungicides in inhibiting the mycelial growth of
Cercospora capsici. The systemic fungicides were tested at 100, 500,
1000 and 1500 ppm concentrations and non-systemic fungicides at
500, 1000, 1500 and 2000 ppm concentrations. The list of fungicides
used in the present studies with common names is given in Table 1.

The fungus Cercospora capsici was grown on CDA medium for

15 days prior to setting up the experiment. The CDA medium was
prepared and melted. The fungicidal suspension was added to the
melted medium to obtain the derived concentration on the basis of
active ingredient present in the chemical. Twenty ml of poisoned
medium was poured in each sterilized Petri plates. Suitable check was
maintained without addition of fungicides. The plates were then
inoculated as described earlier and incubated at 25 ±10 C. Three
replications were maintained for each treatment. The radial growth of
colony was recorded when maximum growth was observed in control
plate and per cent inhibition was calculated by using the formula
given by Vincent (1927);

C −T
I = × 100
C

Where;

I = Per cent inhibition

C = Radial growth of fungus in control

T = Radial growth of fungus in treatment

Table 1. List of fungicides used in vitro evaluation against Cercospora
capsici

Sl. Common name Chemical Name Trade Name

No.
I. Non-systemic fungicides / contact fungicides
1. Chlorothalonil Tetra chloro iso phthalo nitrate Kavach 75% WP
2. Mancozeb Co-ordination product of zinc ion
Dithane M-45
and manganese ethylene bis
75% WP
dithiocarbamate
3. Copper oxy Copper oxy chloride
Blitox 50% WP
chloride
4. Carbendazim Methyl 1-1-2 benzimidazole
12%+ carbonate +zinc ion and
SAAF 75% WP
Mancozeb 63% manganese ethylene bis dithio
carbamate.
5. captan N-trichloromethyl thio4-
Captaf 50% WP
cyclohexene-1, 2-dicarboximide
II. Systemic fungicides
1. Carbendazim Methyl -2 Benzimidazole Bavistin 50%
carbamate WP
2. Propiconazole (RS)-2-(2,4-dichlorophenyl)-1-(1H-
Tilt 25 % EC
1, 2,4-triazole-1-yl) prope-2-Ol
3. Hexaconazole (RS)-2-(2,4-dichlorophenyl)-1-(1H-
Contaf 5 % EC
1, 2,4-triazole-1-yl) hexan-2-Ol
4. Difenoconazole Cis, trans-3-chloro-4(4-methyl-2-
(1H-1, 2, 4- Traizole-1-y1, methyl)-
Score 25% EC
1, 3-dioxolan-2-y1) phenyl 4-
chlorophenyl ether
5. Tebuconazole 1-(4-chlorphenyl)-4,4-dimethyl-3-
Folicar 25.9 %
[1,2,4]triazol-1-ylmethyl-pentan-3-
EC
ol
6. Azoxystrobin Methyl (E)-2-{2-[6-(2-
Amistar 25%
cyanophenoxy) pyrimidin-4-
EC
yloxy]phenyl}-3-methoxyacrylate
7. Penconazole 1-(2,4-dichloro-b-propylphenethyl)-
Topas 10% EC
1H-1,2,4-triazole
8. Tricyclozole 5-Methyl-1,2,4-triazolo(3,4-
Baan 75 % WP
b)benzothiazole
3.9 In vitro evaluation of bio-agents against Cercospora capsici

Bio-agents obtained indigenously as well others viz.,

Trichoderma viride, T. harzianum and Bacillus subtilis were evaluated
for their efficacy under in vitro using dual culture technique against
Cercospora capsici fungus. The bio-agents and their sources are
mentioned in Table 2.

3.9.1 Dual culture test

Bio-agents were evaluated for their efficacy through dual culture

technique. The bio-agents and test fungus were inoculated side by
side on a single Petri dish containing solidified CDA medium. Three
replications were maintained for each treatment with one control by
maintaining only pathogen and bio-agent. They were incubated for 3
weeks. The colony diameter of both bio-agent and the pathogen was
measured in two directions and average was recorded. Per cent
inhibition of growth of the test pathogen was calculated by using the
formula.

3.10 Screening of chilli to Cercospora leaf spot disease.

Twenty three chilli genotypes were screened against Cercospora

chilli leaf spot pathogen in vitro by using detached leaf technique to
identify the resistance sources using 0 to 9 scale (Mayee and Datar,
1986).

Screening was done by using detached leaf technique for

detecting resistance to Cercospora capsici in chilli. Leaves from
healthy chilli plants were excised, surface sterilized for 30 sec in 0.05
per cent NaOCl3 solution, and inoculated with 0.5mm of 2 day old
Cercospora capsici culture grown on carrot dextrose agar. Inoculated
leaves were placed in petri dishes containing one sterile filter paper
and 10 ml of sterile distilled water. Petri dishes were sealed with
parafilm and incubated in a growth chamber at 27 0C with constant
fluorescent light.
Table 2. List of bio-agents used against C. capsici and their sources of
collection

Sl. No. Bio-agents Source

Bio control lab, Department of Plant

1. Bacillus subtilis Pathology, College of Agriculture, UAS,
Bengaluru

Bio control lab, Department of Plant

2. Trichoderma harzianum Pathology, College of Agriculture, UAS,
GKVK, Bengaluru

Bio control lab, Department of Plant

3. Trichoderma viride Pathology, College of Agriculture, UAS,
GKVK, Bengaluru

Indian Institute of Horticultural

4. T. harzianum
Research, Hesaraghatta, Bengaluru.

Indian Institute of Horticultural

5. T. viride
Research, Hesaraghatta, Bengaluru.
 Number of lesions per leaf and percentage of leaf area infected
were recorded daily for six days after inoculation. Genotype ranking
for lesion development and percentage of infected leaf area were
similar to know ranking of the same genotypes for field resistance.

Available chilli genotypes were screened for Cercospora leaf spot

disease. The different genotypes were screened in the laboratory
conditions to identify the resistant sources by detached leaf technique
using 0 to 9 scale (Mayee and Datar, 1986) as reported above.

Scale Description Category

0 No leaf showing any symptoms Immune

1 1% or less symptoms Resistant

3 1-10% symptoms Moderately resistant

11-20% symptoms Moderately

5
susceptible

7 21-50% symptoms Susceptible

9 51% symptoms Highly susceptible

EXPERIMENTAL RESULTS
 IV. EXPERIMENTAL RESULTS

The present investigation on leaf spot disease of chilli caused by

Cercospora capsici was conducted in Department of Plant Pathology,
College of Agriculture, University of Agricultural Sciences, Bengaluru
during 2012-13. The results of the experiments obtained thereon are
presented in this chapter.

4.1 Survey for chilli leaf spot caused by Cercospora capsici in

southern parts of Karnataka

Chilli is grown in kharif and in summer season in Southern

Karnataka, Hence, random survey was conducted during 2013 in the
following districts viz., Bengaluru Rural, Chickballapur, Kolar,
Ramnagar, Mandya and Davanagere. The disease incidence was
prevail in all the locations surveyed during cropping period and data
are presented in Table 3, Plate 1.

Cercospora capsici produced brownish spots later turn to grey

or white within a few days after infection; subsequently to dark brown
with a reddish margin. A clear to yellowish halo appear around the
reddish margin. The diseased spots dried and fall from the leaf,
leaving conspicuous holes. Severely infected leaves turn yellow and
drop from the plant. The severity of disease was calculated by using 0-
5 point scale and these scales were converted to Per cent disease
index (PDI). The PDI ranged from 5 to 20 per cent irrespective of
location and season of the year. The disease severity was highest in
Davanagere (12.66 per cent) during summer 2013, followed by Kolar
(10.81 per cent) and Chikballapur (9.00 per cent) during 2013.

The maximum disease severity was recorded in Davanagere

taluk (14 %) and minimum in Ramnagara taluk from Ramnagara
district. In Chickballapura district, the disease severity was maximum
at Gouribidanoor taluk (10 %) and minimum in Shidlaghatta taluk (8
%). In Kolar district, the disease severity was maximum at
Table 3. Survey for incidence and severity of Cercospora leaf spot of
chilli in Southern parts of Karnataka during 2013

Crop Per cent

Name of the Name of the Name of the
stage disease
district taluk village
(Days) index
Basapura 45-50 10
Gouribidanoor Mindenahalli 30-40 15
Kallinayakana halli 30-40 12
Chikballapura
Vijipura 30-40 20
Siddlaghatta I Basapura 30-40 12
Mallenahalli 30-40 10
Neltoor 45-50 10
Srinivasapura Koltoor 45-50 10
Belaganahalli 45-50 12
Kolar
Chowdaenahalli 60-70 20
Kolar Madivala 50-60 10
Narsapura 45-50 20
Aluru 30-40 10
Maddur Aluru doddi 30-40 12
Hagalahalli 45-50 10
Mandya
Ajjalli 30-40 15
Mandya Malligepura 45-50 8
Holaluru 45-50 5
Chanpattana 50-60 8
Ramnagara Ramnagara Alalli 45-50 10
Hanuman doddi 45-50 8
Sasvehalli 50-60 20
Honnali Kulagatte 45-50 15
Benakanahalli 50-60 20
Siramagondahalli 45-50 10
Davanagere Davanagere Betur 45-50 20
Bada 45-50 20
Kumarnalli 45-50 15
Changiri Nallur 45-50 10
Hatti 45-50 12
Bashetthalli 45-50 10
Bengaluru
Dodaballapur Bisuvanahalli 45-50 8
rural
sadenahalli 45-50 10
a. Leaf showing symptoms of Cercospora leaf spot

b. Infected field

Plate 1: Cercospora leaf spot disease

Shrinivaspura taluk (12.33 %) and minimum in Kolar taluk (9.30 %).
In Davanagere district, the disease severity was maximum at
Davanagere taluk (14 %) and minimum in Changiri taluk (11.6 %). In
Bangalore rural district the disease severity was (9.33 %) during 2013.

4.2 Isolation of the pathogen

The causal organism was isolated from infected chilli leaves

showing typical symptoms by following the tissue isolation technique.
The pure culture was obtained by single spore isolation method. The
pure culture was maintained in the slants and kept in the refrigerator
for further use. However, the fungi put forth only vegetative mycelium
but failed to produce spores on PDA medium.

4.2.1 Identification of the pathogen

Fungus grew very slow on Potato dextrose agar compared to

Carrot dextrose agar medium. All the morphological
characters/parameters studied are given in Table 7 and Plate 2.

The fungal pathogen produced uniform dense colonies on Carrot

dextrose agar. The colonies were whitish colour. The hypha of the
fungus was light brown in colour, septate and unbranched under
compound microscope. The fungus produced amphigenous fruiting
structures. The conidiophores were in dense fascicles, septate, shorter
with conidiophores, were medium dark in colour, multiseptate spore,
never branched, straight curved, geniculate, medium to large spores
scar at tunicate tip, Conidia were hyaline, rarely obclavate, straight to
mild curved. On the basis of these characters the pathogen was
identified as Cercospora capsici Heald & Wolf, (1911). (Plate 3)

4.2.2 Pathogenicity of the fungus

The pure culture of the fungal pathogen was used in

pathogenicity studies. The fungus was spray inoculated on leaves as
described in “Materials and Methods” to prove the Koch postulates
and symptom expression artificially. The symptoms of the disease
 a. Conidiophores

b. conidia

Plate 2: Microphotographs of conidiophore and conidia of

C. capsici
 a. Growth of C. capsici on CDA media

b. Sporulation on CDA media

Plate 3: Growth and sporulation of C. capsici on CDA media

 a. Inoculated plant

b. Healthy plant

Plate 4: Proving pathogenicity of Cercospora capsici

appeared on 10th day after inoculation. Initially, the minute reddish
brown spots appeared on the leaf which turned later as grey spots
with narrow reddish brown margin. The symptoms appeared first on
older leaves and later on younger leaves. No symptoms were observed
on any other plant part except on the leaves (plate 4). The fungus was
re-isolated from these spots and confirmed its identity as causal agent
by comparing with original culture.

4.3 Cultural Studies

4.3.1 Growth and sporulation of C. capsici on different solid

media

Cercospora capsici was grown on seven different solid media.

The radial growth of C. capsici was measured when the maximum
growth was attained in any of the media tested. Observations were
recorded as described in Table 4, Fig. 1 and Plate 5.

The maximum radial growth of C. capsici was recorded on

Carrot dextrose agar (85 mm) and Richard’s agar (83 mm) media
which were on par with each other and significantly superior over all
other media tested viz., Asthana and Hawker’s agar (79 mm),
Czapeck’s agar (77 mm), Oat meal agar (65 mm). However, the
minimum radial growth was observed in Potato dextrose agar (43 mm)
and Sabouraud’s agar (39 mm).

Of the seven solid media tested, the sporulation was observed

only in Carrot dextrose agar, Asthana and Hawker’s agar and
Richard’s agar where-in other tested media did not support any
sporulation. Abundant sporulation was obtained in Carrot dextrose
agar, moderate sporulation was observed in Asthana and Hawker’s
agar and scanty sporulation was observed in Richard’s agar. From
these observations, it was found that the fungal pathogen requires
varied nutrients for its growth and sporulation.
Table 4. Colony growth of C. capsici on different solid media at
different days of incubation

Mean colony diameter (cm)

SL.
Media
No.
5th day 10th day 15th day 20th day

Asthana and
1. 2.1 5.23 7.5 7.97
hawker’s agar

2. Czapek’s agar 1.73 4.6 7.33 7.77

3. Potato dextrose agar 1.3 3.3 4.03 4.30

4. Carrot dextrose agar 1.97 6.3 8.4 8.5

5. Richard’s agar 1.2 5.07 8.17 8.33

6. Sabouraud’s agar 1 2.17 3.50 3.90

7. Oat meal agar 1.43 3.43 5.97 6.5

S.Em ± 0.06172 0.2353 0.2814 0.2340

CD at 1% 0.25 0.99 1.1848 0.9851

CV (%) 4.97 5.48 3.59 4.00

 Mean colony diameter (cm) 5 days Mean colony diameter (cm) 10 days
Mean colony diameter (cm) 15 days Mean colony diameter (cm) 20 days
9

)7
m
(c
r6
e
t
e
m 5
a
i
d
y
n4
o
l
o
c
n3
a
e
M2

0
1 2 3 4 5 6 7

Different solid media

Fig 1. Growth of C. capsici on different solid media at different days of incubation

 1. Sabouraud’s agar
2. Oat meal agar
3. Potato dextrose agar
4. Czapek’s agar
5. Asthana and Hawker’s
6. Richard’s agar
7. Carrot dextrose agar

Plate 5: Growth of C. capsici on different solid media

Table 5. Cultural characteristics of C. capsici on different solid media
after 25 days of incubation

Sl.
Medium Growth characterists Sporulation
No.

Asthana and Good mycelial growth, white to

1. ++
hawker’s agar black colour, and raised growth.
Good mycelial growth, grey to
2. Czapek’s agar light brown colour, raised -
growth.
Slow mycelial growth, greyish
Potato dextrose
3. brown colour, slightly raised -
agar
growth with regular margin.
Good mycelial growth, black to
Carrot dextrose
4. light brown colour, raised ++++
agar
growth.
Good mycelial growth, grey
5. Richard’s agar +
colour. Raised growth.
Slow mycelial growth, greenish
6. Sabouraud’s agar -
colour with regular margin.
Good mycelial growth, grey to
7. Oat meal agar -
white colour, raised growth.

- = no sporulation, + = scanty sporulation (one to two spores per

microscopic field), ++ = moderate sporulation (up to 10 spores per
microscopic field), +++ = good sporulation (11-19 spores per
microscopic field ), ++++ = abundant sporulation (20 and above
spores per microscopic field)
Table 6. Dry mycelial weight of C. capsici in carrot dextrose broth
medium

Dry mycelial weight

Sl. No. Days after seeding
(mg)

1. 2 3.06

2. 4 5.80

3. 6 10.86

4. 8 16.33

5. 10 27.56

6. 12 38.43

7. 14 56.86
8. 16 68.20
9. 18 74.26
10. 20 72.20
11. 22 67.86

12. 24 65.00

13. 26 62.33

14. 28 61.10

15. 30 58.20

S.Em± 0.64
C.D at 1% 2.5
C.V % 2.42
 80

70
Dry mycelium weight ()mg

60

50

40

30

20

10

0
2 4 6 8 10 12 14 16 18 20 22 24 26 28 30

Incubation period (Days)

Fig 2. Growth of C. capsici on carrot dextrose broth at different incubation periods

Table 7. Comparison of morphological characters of C. capsici from
chilli leaves, with the earlier report on it

reference Morphological characters

Cercospora capsici Colonies amphigenous, consisting of

(Heald & Wolf, 1911) randomly distributed fasciculate
conidiophores. Conidiophores fasciculate,
un-branched, straight or slightly curved,
pale brown geniculate, 30 -80 µm wide,
conidial scars 1 – 2 µm wide, usually on a
small shoulder. Conidia solitary, (3-15)
septate, with a a thickened scar at the
base, 40 – 135 µm long, 3-5 µm wide.

Cercospora capsici Colonies produce shorter conidiophores,

(Under study) were medium dark colour, the conidia are
elongated , cylindrical having 3-12 septa,
narrow towards tip, multiseptate, never
branched, 3.8 -4.6 X 32.0 -86.0 µm in
width and length, geniculate, medium to
large spores, hyaline, straight to mild
curved.
4.3.2 Growth studies of Cercospora capsici on Carrot dextrose
broth at different incubation periods.

The experiment was conducted to ascertain the number of days

required for maximum growth of the fungus by monitoring the dry
mycelial weight (Table 6 and Fig. 2).

There was a significant difference between the incubation

periods. The dry mycelial weight of C. capsici gradually increased from
sixth day (10.86 mg) and reached maximum on 18th day (74.26 mg)
and was significantly superior over all other treatments. It showed
declining trend from 20th day onwards.

4.4 Morphological studies of Cercospora capsici

The fungus Cercospora capsici produces light brown colour

mycelium, mycelium is septate, and unbranched. The fungus
produces amphigenous fruting structure, the conidiophores are short,
septate, geniculate, were in dens fascicles are medium to dark brown
in colour. The Conidia of fungus were elongated, cylindrical having a
3-12 septa with hyaline colour measures 4.2-6.0 X 32.0 -86.0 µm in
width and length. Conidia were septate, narrow towards tip, obclavate
and straight to mild curved. The fungus was very poor sporulation in
culture media. The data is presented in Table 7.

4.5 Physiological studies

4.5.1 Temperature requirement

The average dry mycelial weight of Cercospora capsici after

eighteen days of inoculation at different temperatures were recorded
and presented in the Table 8, Fig. 3 and Plate 6. The fungus
Cercospora capsici was grown on carrot dextrose broth at different
temperature range viz., 10, 15, 20, 25, 30, 35 and 400C to know the
suitable temperature requirement for the maximum dry mycelial
weight.
Table 8. Effect of temperature on growth of C. capsici at 18 days
Incubation

Sl. No. Temperature Mean dry mycelial weight (mg)

1. 10 22.33

2. 15 32.00

3. 20 52.03

4. 25 72.53

5. 30 56.97

6. 35 32.33

7. 40 10.47

S.Em ± 0.67

C.D. at 1% 2.84

C.V % 2.93
 80

70
Dry mycelial weight (mg)

60

50
10
15
20
40
25
30
30
35
40
20

10

0
Temperature 0C

Fig 3. Effect of temperature on mycelial growth of C. capsici

 0
a. Effect of different temperatures ( C) on growth of C.
capsici

b. Effect of Hydrogen-ion concentration (pH) on the

growth of C. capsici

Plate 6: Physiological studies of C. capsici in liquid media

The maximum dry mycelial weight of the fungus at a temperature of
250C (72.53 mg) which was significantly superior over all other
temperatures. Lowest mean dry mycelial weight was obtained at
temperature of 400C (10.47 mg).

4.5.2 Hydrogen-Ion concentration (pH)

The effect of different pH levels on the growth of fungus

Cercospora capsici was studied. The reaction of the medium was
adjusted to required pH levels viz., 4, 5, 6, 7, 8, 9 and 10 as detailed
in ‘Material and Methods’. The data are given in Table 9, Fig. 4 and
Plate 6.

The growth of the fungus C. capsici at different pH levels

significantly differed. The maximum dry mycelial weight of the fungus
was obtained at pH 7 (73.73 mg) which was significantly more than
the growth at other pH levels followed by pH 6 (69.00 mg) and pH 8
(67.60 mg). However, the least growth of the fungus was recorded at
pH 4 (26.33 mg) and was followed by growth at pH 10 (39.30 mg).

4.6 Management of leaf spot of chilli

4.6.1 In vitro evaluation of fungicides against Cercospora capsici

In vitro evaluation of fungicides provides useful and preliminary

information regarding efficacy of fungicides against pathogen within a
short period of time and therefore, serves as a guide for field testing.
In the present investigation, eight systemic and five non systemic
fungicides were tested at four concentrations in the laboratory for
their efficacies against the pathogen.

In vitro evaluation of fungicides on inhibition of spore

germination of C. capsici was carried out as per the methodology
“Food Poison Technique” described in Material and Methods. Different
systemic fungicides viz., Carbendazim Propiconazole Hexaconazole
Difenoconazole Tebuconazole Azoxystrobin Penconazole and
Tricyclozole at various concentrations viz., 100, 500, 1000 and 1500
Table 9. Effect of hydrogen-ion concentrations (pH) on growth of C.
capsici at 18 days Incubation

Sl. No. pH of the medium Mean dry mycelial weight (mg)

1. 4 26.33

2. 5 41.37

3. 6 69.03

4. 7 73.73

5. 8 67.60

6. 9 55.27

7. 10 39.33

S.Em ± 0.64

C.D. at 1% 2.69

C.V % 2.08
Fig 4. Effect of hydrogen-ion concentration on mycelial growth of C. capsici
ppm were evaluated. The non-systemic fungicides viz., Chlorothalonil,
Mancozeb, Copper oxy chloride, Propineb and Captaf at various
concentrations viz., 500, 100, 1500 and 2000 ppm concentrations
were evaluated. The data pertaining to the results are detailed in
Tables 10 and 11.

The results revealed that, there was significant difference among

the systemic fungicides, concentration tested and their interactions.
Among the eight systemic fungicides tested Propiconazole,
Hexaconazole and Penaconazole recorded 94.44 per cent inhibition of
Cercospora capsici at all the four concentrations (100, 500, 1000 and
1500 ppm) followed by 94.44 per cent inhibition in Carbendazim,
Difenoconazole and Tebuconazole at 500, 1000, 1500 ppm and 88.88
percent at 100ppm. Tricyclozole recorded 94.44 per cent inhibition in
1000 and 1500 ppm, 88.88 and 36.66 per cent inhibition at 500 and
100 ppm, respectively. The least inhibition of mycelial growth among
the systemic fungicides was observed in Azoxystrobin (30 per cent) at
100 ppm concentration as illustrated in Table 10, Fig. 5 and Plate 7.

Among the non-systemic fungicides tested, maximum mycelial

growth inhibition (94.44 per cent) was recorded with Mancozeb and
Propineb at all the four concentrations (500, 1000, 1500 and 2000
ppm) tested, followed by 94.44 per cent inhibition at 1000, 1500 and
2000 ppm, 88.88 per cent inhibition at 500 ppm, 91.11 per cent and
87.77 per cent per cent inhibition in Captan at 1500 and 2000 ppm,
respectively. Chlorothalonil recorded 67.77 per cent inhibition at 1500
ppm concentration. The least inhibition of mycelial growth was
observed in Chlorothalonil (62.22 per cent) at 500 and 2000 ppm
concentration, as evident from the Table 11, Fig. 6 and Plate 8.

4.6.2 In vitro evaluation of biocontrol agents against C. capsici

Three biocontrol agents viz. Bacillus subtitlis, Trichoderma

harzianum and T. viride were evaluated against C. capsici and the
results are presented in Table 12, Fig 7 and Plate 9. The results
Table 10. Per cent inhibition of mycelial growth of C. capsici by
different Systemic fungicides

Per cent inhibition of mycelial growth at

Sl. different concentration (ppm)
Fungicides
No.
100 500 1000 1500
Systemic fungicides
77.78 94.44 94.44 94.44
1. Carbendazim
(53.40) (76.79) (76.79) (76.79)
94.44 94.44 94.44 94.44
2. Propiconazole
(76.79) (76.79) (76.79) (76.79)
94.44 94.44 94.44 94.44
3. Hexaconazole
(76.79) (76.79) (76.79) (76.79)
71.11 90 84.44 94.44
4. Difenoconazole
(47.04) (67.16) (60.22) (76.79)
88.88 94.44 94.44 94.44
5. Tebuconazole
(62.52) (76.79) (76.79) (76.79)
30.00 35.55 36.66 40.00
6. Azoxystrobin
(16.50) (19.94) (20.85) (22.52)
94.44 94.44 94.44 94.44
7. Penconazole
(76.79) (76.79) (76.79) (76.79)
36.66 88.88 94.44 94.44
8. Tricyclozole
(19.94) (66.18) (76.79) (76.79)
73.46 85.82 85.96 87.63
Mean
(53.72) (67.15) (67.72) (70.00)
Fungicide Conc FXC
S.Em ± 0.07 0.05 0.15
C.D at 1% 0.21 0.15 0.43
C.V % 4.36

*Values in parenthesis are arcsine transformed values

 100
Per cent inhibition of radial growth

90

80

70

60

50 100 ppm

500 ppm
40
1000 ppm
30
1500 ppm
20

10

Systemic fungicides

Fig 5. In vitro evaluation of systemic fungicides against C. capsici

 1. Carbendazim 2. Propiconazole
3. Hexaconazole 4. Difenoconazole
5. Tebuconazole 6. Azoxystrobin
7. Penaconazole 8. Tricyclozole

Plate 7: In vitro evaluation of systemic fungicides against

C.capsici at different concentrations (ppm)
Table 11. Per cent inhibition of mycelial growth of C. capsici by
different non- systemic (Contact) fungicides

Sl. Fungicides Per cent inhibition of mycelial growth at

No. different concentration (ppm)

500 1000 1500 2000

Non- systemic (Contact) fungicides

1. Chlorothalonil 62.22 63.33 67.77 62.22

(38.85) (40.17) (43.89) (38.85)

2. Copper oxy 88.88 94.44 94.44 94.44

chloride (67.10) (76.79) (76.79) (76.79)

3. Mancozeb 94.44 94.44 94.44 94.44

(76.79) (76.79) (76.79) (76.79)

4. Propineb 94.44 94.44 94.44 94.44

(76.79) (76.79) (76.79) (76.79)

5. Captan 87.77 91.11 87.77 91.11

(64.52) (69.57) (68.21) (70.08)

Mean 85.55 87.55 87.77 87.33

(64.81) (68.02) (55.27) (67.86)

Fungicide Conc. FXC

S.Em ± 0.13 0.11 0.26

C.D at 1% 0.38 0.34 0.76

C.V % 4.11

*Values in parenthesis are arcsine transformed values

 100
Per cent inhibition of radial growth

90

80

70

60 Concentration

50 500 ppm
1000 ppm
40
1500 ppm
30
2000 ppm
20

10

0
Chlorothalonil Copper oxy Mancozeb Propineb captan
chloride
Non systemic fungicides

Fig 6. In vitro evaluation of non-systemic fungicides against C. capsici

 1. Chlorothalonil
2. Copper oxy chloride
3. Mancozeb
4. Propineb
5. Captan

Plate 8: In Vitro evaluation of non-systemic fungicides against

Cercospora Capsici at different concentrations (ppm)
revealed that all the antagonists significantly reduced the growth of C.
capsici. After measuring the colony diameter of C. capsici, it was
noticed that maximum reduction in colony growth was observed in T.
harzianum (73 per cent) which was significantly superior to all other
bioagnets tested. Next best was T. viride IIHR isolate (72.77 per cent)
followed by T. viride (71.33 per cent) and T. harzianum IIHR isolate
(71.11 per cent). Least inhibition was notice in B. subtilis (23.33 per
cent).

4.7 Genotypes screening

Twenty three chilli genotypes were screened against C. capsici

during 2013 as described in “Materials and Methods” and the results
are presented in the Table 13, Plate 10.

It was observed that among the twenty three genotypes

evaluated, four genotypes viz., EC771557, EC771546, EC771538, and
EC771535 were found to be immune. Seven genotypes viz.,
EC771550, EC771551, EC771553, EC771544, EC771556, EC771545
and EC771548 were found to be resistant. Five genotypes viz.,
EC771552, EC771559, EC771560, EC771539 and EC771543 were
found to be moderately resistant. Three genotypes viz., EC771549,
EC771554 and Tiwari were found to be moderately susceptible and
EC771555, EC771547 and Gowribidanoor were found to be
susceptible and one genotype Byadagi kaddi was highly susceptible.
Table 12. In vitro evaluation of bio-agents against C. capsici

Per cent inhibition of mycelial

Biagents
growth
23.33
Bacillus subtilis
(12.33)
73.00
Trichodema harzianum
(48.33)
71.33
Trichoderma viride
(46.66)
71.11
T. harzianum (IIHR)
(46.33)
72.77
T. viride (IIHR)
(48)
S.Em ± 0.13
CD at 1% 0.52
CV % 2.94
 80
Per cent inhibition of radial growth

70

60

50

40

30

20

10

0
Bacillus subtillis Trichoderma Trichoderma Trichoderma Trichoderma
harzianum harzianum(IIHR) vride (IIHR) viride (IIHR)
Bioagents

Fig 7. In vitro evaluation of systemic fungicides against C. capsici

 1. Bacillus subtilis
2. Trichodema harzianum
3. Trichoderma viride
4. Trichodema harzianum (IIHR)
5. Trichoderma viride (IIHR)
C - Control

Plate 9: In vitro evaluation of bioagents against C. capsici

Table 13. Reaction of chilli genotypes against Cercospora leaf spot
under lab condition using detached leaf technique

Disease reaction Name of the genotypes

EC771557, EC771546, EC771538,

Immune
EC771535

EC771550, EC771551, EC771553,

Resistant EC771544, EC771556, EC771545,
EC771548

EC771552, EC771559, EC771560,

Moderately resistant
EC771539, EC771543

Moderately susceptible EC771549, EC771554, Tiwari

EC771555, EC771547,
Susceptible
Gowribidanoor

Highly susceptible Byadagi kaddi*

*Susceptible check
Plate10. Disease scoring scale of Cercospora leaf spot of chilli
(0-9 scale)
DISCUSSION
 V. DISCUSSION

5.1 Survey for the disease incidence and severity in southern

parts of Karnataka

In the present study, random survey for Cercospora leaf spot of

chilli was carried out during 2013 in major chilli growing areas of
southern Karnataka to get precise information on the severity and
incidence of the disease.

The data on survey revealed that Cercospora leaf spot severity

and incidence varied from locality to locality, because of type of variety
grown, environmental conditions, cropping pattern and build-up of
inoculum. The average disease severity varied in various locations in
different districts owing to varied agro-climatic conditions and also
different cultivars used.

The disease severity ranged from 5 to 20 per cent irrespective of

location surveyed during the year. The disease severity was highest in
Davanagere followed by Kolar. These observations are in agreement
with the reports of Singh et al. (1994), Khunti et al. (2002), Ramesh
Chand et al. (2003), Ali et al. (2011), While lower disease severity (7
per cent) recorded in Ramnagar district, due to unfavourable
environmental conditions and resistant planting materials. Further,
the disease severity varied to greater extent in different locations
indicating the probable role of environment and or existence of
physiological races. These observations are in agreement with the
earlier report of Hossain et al. (2011) on Cercospora leaf spot of chilli.

Considering the disease severity, the location viz., Davanagere,

Kolar districts are identified as hot spot areas for Cercospora leaf spot
of chilli. Intensive cultivation of chilli crop in successive years, use of
infected seed materials, non-adoption of disease management
practices, favourable weather conditions and also the cultivation of
highly susceptible varieties of chilli could be the reasons for higher
incidence and severity of disease.

5.2 Isolation, identification and pathogenicity of the pathogen

The causal organism Cercospora capsici was isolated from

infected leaf by following the standard tissue isolation method. The
pure culture obtained was sub cultured in petridishes and slants
containing Carrot dextrose agar and stored in refrigerator for use in
further studies. The culture on carrot dextrose agar produced black to
whitish colony. However, it was reported that pathogen produces toxin
under stress condition if it is grown especially on minimal media.
Various workers (Khandar et al., 1985, Chandrasekharan and
Rangaswami, 1960 and Mallappa Prakash, 2007) have isolated the
pathogen from infected leaf.

Hypha of fungus was light brown colour, septate and branched.

Growth of the fungus was slow taking more than 16 days to attain
maximum growth of 9.00 cm. the identification of fungus, was made
on the basis of important morphological characters such as; size,
shape and septations of conidiophores and conidia. The morphological
characters given by Heald and Wolf (1911) were compared with the
one under study and they were found to be identical, hence the
fungus was identified as Cercospora capsici Heald & Wolf, (1911).

The pathogenicity was studied by artificially inoculating

Cercospora capsici pathogen on chilli seedlings. In the present study,
mycelial and conidial suspension of Cercospora capsici pathogen was
sprayed on one month old chilli plants and symptoms were noticed
after 10 days of inoculation. In the beginning symptoms appeared as
minute spots later developed into circular grey to dirty grey spots with
brown margin. Infected leaves developed circular lesions varying from
pinpoint size to 0.5 cm diameter. The pathogen was reisolated from
the infected leaves and identity of causal organism was confirmed by
comparing with the original culture.
 Munjal et al. (1960) observed that, the symptoms appeared first
on older leaves and gradually progressed upward on to younger
leaves. Lakshminarayana (1981) also made similar observations and
he opined that delay in appearance on younger leaves of brinjal may
be due to the difference in amount and type of sugars present in old
and young leaves. Similar symptoms observed by many workers
(Chandrashekhran and Rangaswami, 1960; Vakili, 1977; Jamadar,
1988).

5.3 Cultural studies

Every living being requires food for its growth and reproduction
and fungi are not an exception. Fungi derive the food from the
substrate upon which they grow. To culture fungi artificially, it’s a
necessary to supplement in the medium, those essential nutrients
needed for their growth and development. So, to find out the best
sources of nutrients for the fungus, different synthetic and non-
synthetic were tested. The radial growth of the fungus was used to
determine the growth of fungus on solid media.

5.3.1 Growth and sporulation of Cercospora capsici on different

solid media

The fungus was grown on seven different media. The results

indicated that the best mycelial growth was on Carrot dextrose agar
(CDA) with the mean colony diameter of 85 mm followed by Richards
agar 83 mm, asthana and hawkers agar 79 mm and Czapeck agar 77
mm, Sabouraud’s agar (39 mm) and Potato dextrose agar (40 mm) did
not support the growth. Similar observations were made earlier by
many workers in Cercospora moricola Cooke (Kanti, 1975), C.
canescens Ell. and Mart. (Ekpo and Esuruoso, 1978) C. solani-
melongenae Chupp (Lakshminarayana, 1981), Cercospora sorghi Ell.
(Jamadar, 1988) and Cercospora canescens (Misra and Bhattacharyya,
2002).
 Of the seven solid media tested, maximum sporulation was
obtained in Carrot dextrose agar and Asthana and hawker’s agar
followed by scanty sporulation in Richard’s agar and no sporulation in
remaining four media. This confirms that Carrot dextrose agar and
Asthana and hawker’s agar are the best media for obtaining
sporulation of Cercospora species which has been generally accepted
by many workers in the past (Dange and Patel, 1968 in C. beticola;
Mew et al., 1975 in C. canescens; Khandar et al., 1985 in C. canescens
and Misra and Bhattacharyya, 2002 in C. canescens).

5.3.2 Growth phase of C. capsici on carrot dextrose broth at

different incubation periods

The growth phase of C. capsici was studied by inoculating the

fungus on carrot dextrose broth and harvesting the growth starting
from second day onwards up to 30th day. The findings of the
experiment revealed that there was increase in the dry mycelial weight
of fungus from second day onwards up to 18th day. Thereafter, there
was a decline in the dry mycelial weight up to 30th day.

The maximum dry mycelial weight of 74.26 mg was recorded on

18th day and it was considered as the optimum period for growth of
the fungus. The increase in dry mycelial weight from second day to
18th day can be attributed to presence of the nutrients in the medium
and the fungus showed gradual increase in the weight by utilising
them to the maximum extent. The decrease in the dry mycelial weight
from 20th day onwards may probably due to the mycelium and
exhaustion of nutrients in the medium.

Unlike most fungi, Cercospora species are slow growing fungi

and need a long incubation period to attain maximum growth as
evidenced in Cercospora moricola Cooke (Kanti, 1975), Cercospora
solani-melongenae Chupp (Lakshminarayana, 1981), Cercospora
sorghi Ell. and Evert. (Dinesha, 1984), Cercospora kikuchii (Satya
Prasanth, 2004) and Cercospora nicotianae (Mallappa Prakash, 2007)
which need 20, 22, 16, 22 and 19 days respectively. Similar
observations were made in case of Cercospora canescens (Jamadar,
1988, Misra and Bhattacharyya, 2002). Hence, to obtain maximum
fungal growth of C. capsici, 18 days of incubation period was found to
be optimum.

5.4 Physiological studies

5.4.1 Effect of temperature on the growth of C. capsici

Temperature plays an important role in infection and disease

development. Each fungus has its own temperature range for growth
and development. An effort was made to know the optimum
temperature for the growth. In present study, it was observed that the
maximum fungal growth was observed at 250C (72.53 mg) followed by
300C (56.97 mg). The temperature between 250C and 300C appears to
be optimum for the growth of the fungus. Least fungal growth was
observed at 400c (10.47 mg), 100c (22.33 mg) and 150c (32 mg) which
were significantly different. Similar findings were also reported by
Ekpo and Esuruoso (1978) in C. canescens, Dinesha (1984) in C.
sorghi, Khandar et al. (1985) in C. canescens, Satya Prasanth (2004)
in C. kikuchii and Mallappa Prakash (2007) in C. nicotianae. The slight
difference may be due to the change in the basal medium used by
different workers and the strain used.

In present study, the excellent fungal growth was observed at

250C followed by 300C. Hence, the temperature range of 25 to 300C
can be recommended to obtain excellent fungal growth of C. capsici.

5.4.2 Effect of Hydrogen-ion concentration (pH) on growth of C.

capsici

Any living organism requires a particular medium for the growth

and development. The pH of the medium should be optimum for
growth. A wide range of pH supported the growth of C. capsici. Good
growth was found at 7 pH. The optimum pH range was obtained
towards neutral range. The observations are in agreement with those
of Ragunathan (1969), Verma and Agnihorti (1972) and Jamadar
(1988).

In the present study, the maximum dry weight of the fungus

was recorded at 7 pH. However, the optimum growth of the fungus
was also observed at pH of 6 to 8. Hence, it can be recommended that
to obtain good growth of C. capsici, the optimum pH from 6 to 8 could
be maintained in the culture media.

5.6 Bioassay of fungicides and botanicals against C. capsici

5.6.1 In vitro evaluations of fungicides

Bioassay of fungicides provides useful for preliminary evaluation

against a pathogen with in the shortest period of time and therefore, it
will be helpful for further evaluation of fungicides in field conditions.
In the present study, eight systemic and five non-systemic fungicides
were tested against pathogen at four different concentrations.

Among the eight systemic fungicides, Propiconazole,

Hexaconazole, Penaconazole inhibited the growth of C. capsici at all
the concentrations tested; while the next best fungicides were
Tebuconazole, Difenaconazole, Tricyclozole, these fungicides inhibited
the fungal growth at 500, 1000, 1500 ppm concentration. This shows
that these fungicides tested were effective against the pathogen.
Azoxystrobin was not effective against leaf spot caused by C. capsici in
chilli.

Among the non-systemic fungicides Mancozeb, Propineb were

found to be effective against the pathogen at all the concentrations,
followed by Captan. The least inhibition of mycelial growth was
observed in copper oxy chloride.

5.6.2 In vitro evaluation of bioagents

In present day constraints in plant disease control practices

especially those on the use of pesticides, biological control is
increasingly occupying the minds of scientists all over the world as
they are eco-friendly and cost effective. In recent year, the use of
Trichoderma has gained more importance. These antagonistic
organisms act on the pathogen by different mechanisms viz.,
competition, lysis, antibiosis, siderophore production and hyper
parasitism (Vidyasekaran, 1999).

In the present study, the maximum reduction in colony growth

was observed in T. harzianum which was significantly superior to all
the bioagents tested. Next best was T. viride. There was minimum
inhibition of C. capsici by B. subtilis. Species of Trichoderma viz., T.
harzianum and T. viride showed higher inhibition of mycelial growth of
the pathogen compared to bacterial antagonists. It may be due to
production of antibiotic substance (viridin) produced by T. harzianum
as reported by Brain (1951) and also due to the penetration of the
antagonistic hyphae into hyphae of pathogen at the place of contact.
Similar results were reported by Siddaramaiah (1986) in case of leaf
spot of mulberry caused by C. moricola.

5.7 Genotypes screening

In the present investigation, 23 genotypes were screened to

identify the sources of resistance against Cercospora capsici. The
results indicated that four genotypes showed immune reaction against
the pathogen. viz., EC771557, EC771546, EC771538 and EC771535.
Seven genotypes viz., EC771550, EC771551, EC771553, EC771544,
EC771556, EC771545 and EC771548 were found resistant. Five
genotypes viz., EC771552, EC771559, EC771560, EC771539 and
EC771543 showed moderately resistant reaction. Three genotypes
viz., EC771549, EC771554 and Tiwari showed moderately susceptible
reaction. Three genotypes viz., EC771555, EC771547 and
Gowribidanoor showed Susceptible and one genotype Byadagi kaddi
showed highly susceptible reaction.
 Similar attempts were made for evaluating germplasm against
Cercospora species some of the reports were made in the past in India
and abroad by various workers (Cantonwin et al. 2006; Khandar et al.
1983; Jamadar, 1988; Grover et al. 1976; Bashir and Malik, 1993;
Katna et al. 2001; Daisy Basandari et al. 2003; Raje and Rao, 2002;
Browne and Cooke, 2005; Iqbal et al. 2004; Twizeyimana, 2007) to
locate the source of resistance against Cercospora capsici. Their
studies revealed that very few genotypes showed resistance against
Cercospora capsici.
SUMMARY
 VI. SUMMARY

Various studies with respect to Cercospora capsici causing leaf

spot of chilli viz., survey for its incidence and severity, identification
and pathogenicity along with cultural, morphological and
physiological characters of the pathogen, in vitro evaluation of
fungicides, bioagents and identification of resistance sources against
pathogen were carried out and the results obtained thereon are
summarized hereunder.

The Cercospora leaf spot disease severity on chilli was found to

be highest during the survey in Davanagere taluk (14.00 per cent),
followed by Srinivaspura and Honnali taluk (12.33 %), in summer
season 2013.

The pathogen produced small brown spots with greyish centre

on the leaves. The pathogen was isolated from infected leaf on carrot
dextrose agar by employing standard tissue isolation method and
recorded various morphological characteristics. The mycelium was
brownish, septate, fruiting structure was amphigenous, conidiophores
were brown to dark, multi septate (0-5), geniculate. Conidia were
hyaline, acircular, straight to slight curved, multiseptate and
measured 5.2-8.0 X 50.0-16.8 µm in size. The pathogen was identified
as Cercospora capsici Heald & Wolf based on these conidia and
conidiophore characters.

Out of seven solid media tested, four media viz., Carrot dextrose
agar, Asthana and Hawker’s agar, Czapek’s agar and Oat meal agar
were identified as the best media for growth of Cercospora capsici. The
present investigation optimizes 20 days of incubation for maximum
growth of the fungus on Carrot dextrose agar followed by 18 days for
liquid media.

The fungus was favoured by an optimum temperature of 250C

recording highest dry mycelial weight of 72.5 mg followed by 300C
with 56.9 mg.
 The fungus was also favoured by an optimum pH of 7.0
recording 73.73 mg dry mycelial weight followed by a pH of 6.0
recording 69.73 mg.

Eight systemic fungicides were tested in vitro conditions against

C. capsici, Propiconazole, Hexaconazole, Penconazole and
Tebuconazole were best in inhibiting (94.44%) the growth of the
fungus at all the four concentrations tested (100, 500, 1000 and 1500
ppm). The non-systemic fungicides Mancozeb and Propineb were
found to be effective at all concentrations (500, 1000, 1500 and 2000
ppm).

Among the bioagents tested in vitro conditions, Trichoderma

harzianum (GKVK) (73.00%) and Trichoderma viride (IIHR) (72.77%)
were found to inhibit the mycelial growth.

Seven genotypes out of 23 screened viz., EC771550, EC771551,

EC771553, EC771544, EC771556, EC771545 and EC771548 were
found resistant to Cercospora leaf spot and these lines could be
further evaluated under artificial inoculation conditions before using
them in breeding programme.
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