Archives of Phytopathology and Plant Protection

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Cultural, morphological and bio-chemical

variability of different isolates of Colletotrichum
truncatum causing anthracnose of greengram

Tanya Marak, Yashi Umbrey, Sunita Mahapatra & Srikanta Das

To cite this article: Tanya Marak, Yashi Umbrey, Sunita Mahapatra & Srikanta Das (2019)
Cultural, morphological and bio-chemical variability of different isolates of Colletotrichum�truncatum
causing anthracnose of greengram, Archives of Phytopathology and Plant Protection, 52:1-2,
141-154, DOI: 10.1080/03235408.2019.1588194

To link to this article: https://doi.org/10.1080/03235408.2019.1588194

Published online: 28 Mar 2019.

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ARCHIVES OF PHYTOPATHOLOGY AND PLANT PROTECTION
2019, VOL. 52, NOS. 1-2, 141–154
https://doi.org/10.1080/03235408.2019.1588194

Cultural, morphological and bio-chemical variability

of different isolates of Colletotrichum truncatum
causing anthracnose of greengram
Tanya Marak, Yashi Umbrey, Sunita Mahapatra and Srikanta Das
Department of Plant Pathology, Bidhan Chandra Krishi Viswavidyalaya, Nadia, India

ABSTRACT ARTICLE HISTORY

A laboratory experiment was conducted to study the vari- Received 15 September 2018
ability among the eight isolates of Colletotrichum truncatum Revised 8 February 2019
of greengram collected from different locations on the basis Accepted 18 February 2019
of cultural, morphological and biochemical characteristics by
KEYWORDS
using nine different culture media viz., Potato Dextrose Agar Antracnose; greengram;
(PDA), Potato Carrot Agar (PCA), Oat Meal Agar (OMA), Corn cultural; morphological and
Meal Agar (CMA), Carrot Agar (CA), Sabouraud’s Agar (SA), biochemical variability
Czapek’s Dox Agar (CDA), Richard’s Agar (RA) and V8 Juice
Agar. Colony colour varied in different media from white or
white with light brown centres which later changed to
black or dark to light brown with increase in the age of the
fungal cultures. Mostly, the colonies had fluffy or cottony
mycelial growth with slight variations and regular to irregu-
lar white margin. PDA, PCA, CA and RA produced maximum
mycelial growth (90 mm) at 10 DAI (days after inoculation).
Minimum growth was observed on SA (69.56 mm) and V8
juice agar media (55.42 mm) and their difference was statis-
tically significant. Morphological variability among the iso-
lates was studied by comparing their conidial length,
breadth and length and breadth of setae and their differen-
ces were statistically significant. Biochemical variability
among the isolates was based on a- and b-esterase and
peroxidise profiling. Positive activity was observed for both
a- and b-esterase. a-Esterase enzyme showed the highest
enzyme activity in terms of maximum numbers of banding
loci among the three isozymes tested. The findings of the
present study clearly revealed that cultural, morphological
and biochemical variability did exist among the different
isolates of C. truncatum.

Introduction
Greengram [Vigna radiata (L.) Wilczek] commonly known as mungbean
is an important legume of Asian origin and it is widely cultivated in
Asian, Australian and African continents (Yang et al. 2008). Greengram

CONTACT Sunita Mahapatra sunitamahapatra@yahoo.co.in Department of Plant Pathology, Bidhan

Chandra Krishi Viswavidyalaya, Mohanpur 741 252, Nadia, West Bengal, India.
ß 2019 Informa UK Limited, trading as Taylor & Francis Group
142 T. MARAK ET AL.

suffer from many diseases caused by fungi, bacteria, viruses, nematodes

and abiotic stresses. Among the fungal diseases, powdery mildew,
anthracnose, Cercospora leaf spot, web blight and dry root rot are the
most prevalent ones (Agarwal 1989). In recent years, greengram anthrac-
nose caused by Colletotrichum truncatum (Schw.) Andrus and Moore has
become one of the major disease and causes yield loss in upto 24–67%
(Deeksha and Tripathi 2002). The disease has been reported from all the
mungbean growing regions of India in mild to severe form (Anonymous
2013). In tropical and subtropical areas, it causes considerable damage by
reducing seed quality and yield every year in varying intensity. The dis-
ease causes qualitative as well as quantitative losses (Sharma et al. 1971).
The Colletotrichum genus exhibits wide genetic variability. High variabil-
ity has been observed in morphological and physiological traits, and sev-
eral of these are important for pathogen development and survival, such
as mycelial growth rate, colony diameter, sporulation capacity and per-
cent germination (Mota et al. 2016). However, the high variability of
Colletotrichum spp. under culture conditions makes this criterion not
adequate for reliable differentiation among the species (Freeman et al.
1998; Afanador-Kafuri et al. 2003). The sample survey revealed that, in
West Bengal greengram is grown with more intensive cultivation almost
in all the districts particularly more in Nadia, Murshidabad, Burdwan
and 24-Parganas. The disease severity varies in region to region. The
variation in disease severity from location to location may be attributed
to the weather factors which affected the growth and development of the
pathogen and its aggressiveness. Aggressiveness among the different iso-
lates of Colletotrichum capsici have been reported previously by Taylor
et al. (2007). Many researchers have used isoenzymes analysis to study
variation among several species of Colletotrichum (Bonde et al. 1991).
The knowledge of the genetic diversity of the populations is necessary to
understand how populations will evolve in response to different control
strategies. Therefore, present study was taken up to see the cultural, mor-
phological and biochemical variability of C. truncatum causing anthrac-
nose disease of greengram in the alluvial-gangetic plains of West Bengal.

Materials and methods

Sample collection and isolation of fungi
Fungi were isolated from symptomatic greengram leave samples having
typical dark circular sunken lesion with reddish margin from different
location of West Bengal (Table 1). The Infected along with healthy tissue
bits were surface sterilised in 0.1% mercuric chloride (HgCl2) for 30 s
and subsequently washed with sterile distilled water to remove the traces
 ARCHIVES OF PHYTOPATHOLOGY AND PLANT PROTECTION 143

Table 1. Radial growth of fungal mycelia in different media at 10 DAI.

Isolates (mm)
Sl. No. Media GGC1 GGC2 GGC3 GGC4 GGC5 GGC6 GGC7 GGC8 Mean
1 PDA 90.00 90.00 90.00 90.00 90.00 90.00 90.00 90.00 90.00
2 PCA 90.00 90.00 90.00 90.00 90.00 90.00 90.00 90.00 90.00
3 CA 90.00 90.00 90.00 90.00 90.00 90.00 90.00 90.00 90.00
4 OMA 86.00 73.67 77.00 85.17 90.00 82.17 85.00 90.00 83.63
5 CDA 87.17 68.67 80.67 79.83 86.19 81.00 85.83 87.67 82.13
6 CMA 90.00 90.00 90.00 90.00 90.00 90.00 90.00 90.00 90.00
7 RA 90.00 72.33 73.00 73.17 90.00 83.50 85.83 86.67 81.81
8 V8 62.17 57.33 56.33 47.50 52.83 48.50 63.67 55.00 55.42
9 SA 77.33 68.67 66.50 70.00 66.00 78.00 68.00 62.00 69.56
Mean 84.74 77.85 79.28 79.52 82.78 81.46 83.15 82.37
S.Em ± C.D. at 5%
Isolate 0.08 0.21
Media 0.08 0.23
Isolate x Media 0.23 0.64

of mercury and then transferred into sterilised Petri plates (1–2 leaf bits
per Petri dish) containing potato dextrose agar (PDA) for incubation at
27 ± 1
C. Single spore isolation was done for purification of the patho-
gen and pure culture was maintained in slant with frequent subculture.

Designation Place of collection/location Geographical information

GGC1 Jaguli, Nadia N 22
940 46.400
E 88
530 15.200
GGC2 Chakdaha, Nadia N23
40 42.232800
E 88
410 38.95800
GGC3 Karimpur, Nadia N 23
580 48.622800
E 88
380 16.0987200
GGC4 Jalangi, Murshidabad N 24
50 20.72400
E 88
410 45.985200
GGC5 Dhomkol, Murshidabad N 24
70 43.550400
E 88
350 21.789600
GGC6 Bagdah, North-24 Parganas N 23
120 31.348800
E 88
530 17.797200
GGC7 Raiganj, North Dinajpur N 25
610 85
E 88
120 55
GGC8 Majhian, South Dinajpur N 25
310 31
E 88
760 64

Cultural and morphological studies

Cultural variability in respect of mycelial growth pattern, colony diam-
eter, colour, zonations, type of margin and sporulation were studied by
using nine different culture media viz., PDA, Potato Carrot Agar (PCA),
Oat Meal Agar (OMA), Corn Meal Agar (CMA), Carrot Agar (CA),
Sabouraud’s Agar (SA), Czapek’s Dox Agar (CDA), Richard’s Agar (RA)
and V8 Juice Agar. 5 mm culture disc of 10 days old actively growing C.
truncatum were cut by using a sterilised metal cork borer and a single
disc was placed at the centre of the Petri plate containing different media
by using a sterilised inoculating needle. Each set of experiments were
144 T. MARAK ET AL.

replicated thrice, and the plates were incubated at 27 ± 1
C for 10 days.

The radial growth from the centre to the periphery of Petri plate was
measured for all the media used and analyzed statistically. A spore sus-
pension from the selected fungal cultures on PDA media were prepared
and observed under Phase-contrast microscope for morphological char-
acters such as length and breadth of conidia and presence or absence of
setae. Measurement of total length and breadth of conidia and setae were
done by micrometric measurement with stage and ocular scale. Fifty con-
idia from each isolate were measured with stage and ocular micrometers
(Weir et al. 2012).

Biochemical studies
Extraction of enzyme
Enzymatic extracts were obtained from the eight isolates, grown in 50 ml
of liquid medium (malt extract 30% and peptone 5%) for 10 days at
25
C without agitation. And 500 mg of freshly harvested mycelium from
actively growing culture of Colletotrichum isolates was collected on
Whatman no.1 filter paper by vacuum filtration and washed several
times with distilled water. Which was then homogenised with acetate
buffer (0.05 M, pH 4.5) in porcelain mortar and pestle. The resulting
homogenate mixture at 4
C in Heraeus Biofuge (Stratos, Biorad). The
supernatant was collected and kept in the refrigerator at 4
C and used
as enzyme source (Loureiro et al. 2011).

Electrophoresis and enzyme visualisation

Electrophoresis was done in 7.5% polyacrylamide gel (27 ml buffer B
and 3 ml buffer C with 3.25 g of acrylamide, 0.067 g bis-acrylamide, also
add 5 ml ammonium persulphate (APS) solution and 20 ml TEMED (N,
N, N0 , N0 -tetramethylethylene diamine) following method given by
Kahler and Allard (1970). After solidification samples were loaded
(50 ml sample and 10 ml of 5% dye [Bromophenol blue:Glycerol 1:1]),
and gel was run at 80 volts for 2.5–3 h. On completion of the gel run,
the gels were incubated in different substrate solutions according to the
enzyme under study i.e. a, b-esterases and peroxidase (Vallejos 1983).
For a, b-esterase, gel was stained in 100 ml buffer D containing 100 mg
Fast Blue RR salt and 0.004 g a, b-naphthyl acetate (dissolved in 1 ml of
ethyl alcohol) while for peroxidase, gel was place in 50 mg Ortho-diani-
sidine (dissolved in 1 ml acetic acid) containing 100 ml water and 1 ml
hydrogen peroxide (H2O2) (Welter and Dyck 1983). Both the gel was
incubated at 28
C in dark condition for 30 min with occasional shaking
 ARCHIVES OF PHYTOPATHOLOGY AND PLANT PROTECTION 145

Figure 1. Cultural studies of eight Collectotrichum isolates on different media (from top left
to right) PDA, PCA, SA, RA, CMA, CA, OMA, V8 juice Agar and CDA.

for development of band. After development of the bands, the gel was
washed with distilled water. The gel was transferred and photographed.
The zymograms obtained were analysed based on the procedure
described by Wendem and Weeden (1989). The band length was meas-
ured and relative mobility (Rm) value was calculated using the follow-
ing formula.
Distance of the band from origin
Rm value ¼
Distance of buffer front

Results
Cultural and morphological variability
The cultural variability which was observed with respect to morphology
of mycelium, colony colour and margin of colony on PDA, PCA, CA,
OMA, SA, RA, CMA, V8 Juice agar and CDA medium is shown in
Figure 1. It was found that the size of colony increased with increase in
incubation period. Colony colour varied from white or white with light
brown centres which later changed to black or dark or light brown with
increase in the age of the fungal cultures in every media. In RA media,
all the isolates produced cream colour colony while in other media they
produced light to dark brown colony colour. Mostly, the colonies had
fluffy or cottony mycelial growth with slight variations and regular to
irregular white margin. The results agreed with Kulkarni (2009) who
found that the growth varied from flat to fluffy with smooth or irregu-
lar margins.
146 T. MARAK ET AL.

Table 2. Variability in conidial and setae shape and size of Colletotrichum truncatum isolates
grown on PDA.
Size of conidia Size of setae
Sl. No. Isolates Length (mm) Breadth (mm) Length (mm) Breadth (mm)
1 GGC1 26.83 ± 1.73 4.24 ± 0.65 58.88 ± 2.86 5.42 ± 0.09
2 GGC2 20.94 ± 0.36 3.45 ± 0.31 57.40 ± 2.68 5.08 ± 0.11
3 GGC3 21.15 ± 0.29 3.56 ± 0.32 57.80 ± 2.79 4.95 ± 0.27
4 GGC4 22.06 ± 0.18 3.64 ± 0.32 58.03 ± 2.88 5.27 ± 0.14
5 GGC5 27.45 ± 1.30 4.50 ± 0.49 60.72 ± 2.56 5.51 ± 0.12
6 GGC6 22.78 ± 0.38 4.06 ± 0.17 60.80 ± 3.40 5.62 ± 0.23
7 GGC7 25.51 ± 0.53 4.29 ± 0.45 59.01 ± 2.90 5.67 ± 0.20
8 GGC8 28.91 ± 0.39 4.66 ± 0.53 61.03 ± 2.55 5.81 ± 0.20
S.Em ± 0.48 0.06 0.29 0.03
C.D. at 5% 1.46 0.19 2.99 0.08

Maximum mycelial growth (90 mm) at 10 DAI (days after inoculation)

was produced in PDA, PCA, CA and CMA followed by OMA
(83.63 mm), CDA (82.13 mm) and RA (81.81 mm) and their difference
were statistically significant at CD 5%. Minimum growth was observed
on SA (69.56 mm) and V8 juice agar media (55.42 mm) and their differ-
ence was statistically significant. Irrespective of the media tested, GGC1
produced maximum growth (84.74 mm) followed by GGC7 (83.15 mm),
GGC5 (82.78 mm), GGC8 (82.37 mm) and GGC6 (81.46 mm) and their
difference were statistically significant. Minimum growth was observed
in GGC4 (79.52 mm) followed by GGC3 (79.28 mm) and GGC2
(77.85 mm) and their difference were statistically significant (Table 1).

Morphological variability
Eight C. truncatum isolates showed morphological variability and their
differences were statistically significant. The maximum average conidial
length was found in GGC8 (28.91 ± 0.39 lm) statistically at par with
GGC5 (27.45 ± 1.30 lm) and minimum conidial length was noticed in
GGC2 (20.94 ± 0.36 lm). The maximum breadth of conidia was noticed
on GGC8 (4.66 ± 0.53 lm) and GGC5 (4.50 ± 0.49 lm) and minimum
breadth of conidia was noticed in GGC2 (3.45 ± 0.31 lm). The maximum
length of setae was observed in GGC8 (61.03 ± 2.55 mm) and minimum in
GGC2 (57.40 ± 2.68 mm). The maximum average breadth of setae was
observed in GGC8 (5.81 ± 0.20 mm) and minimum in GGC3
(4.95 ± 0.27 mm) (Table 2). Microscopic view of conidia, acervulus and
setae (Figure 2).
Further, a dendogram constructed based on morphological variation of
the eight isolates showed two major clusters with 25% Euclidean distance
(Figure 3). Group I comprised of four isolates GGC2, GGC3, GGC4 and
GGC6. While group II comprised of remaining four isolates GGC1,
GGC7, GGC5 and GGC8. Group I was further sub-clustered into two
 ARCHIVES OF PHYTOPATHOLOGY AND PLANT PROTECTION 147

Figure 2. Microscopic view of acervulus along with setae (left) and conidia (right) of
Collectotrichum isolates.

Figure 3. Dendogram obtained based morphological character of C. truncatum.

groups, of which first sub-cluster (group IA) had three isolates GGC2,
GGC3 and GGC4 showing their close relationship while second sub-clus-
ter (group IB) had only GGC6 which formed separate individual cluster.
Similarly, group II was further subdivided into two clusters, i.e. group
IIA, which comprised of two isolates GGC1 and GGC7 while group II B
comprises of another two isolates GGC5 and GGC8 showing their close
relationship.

Biochemical variability
Positive activity was observed for both a- and b-esterase. a-Esterase
enzyme showed the highest enzyme activity in terms of maximum num-
bers of banding loci among the three isozymes tested. Alpha esterase iso-
enzyme polymorphism showed all the isolates have different banding
148 T. MARAK ET AL.

pattern and maximum loci 5 was observed in isolate GGC2 and GGC4.
The remaining six isolates produced 4 banding patterns. Likewise, beta
esterase isoenzyme showed all the isolates have different banding pattern
and maximum loci 4 was observed on isolates GGC2 and GGC5. One
isolate produced 3 banding patterns (GGC4) and the other five isolates
produced 2 banding patterns each (GGC1, GGC3, GGC6, GGC7 and
GGC8). Among the 8 isolates, all the isolates produced loci of Rm value
0.47 except GGC6, GGC7 and GGC8. In case of peroxidase isoenzyme
also all the isolates had different banding pattern and maximum loci 3
was observed on isolates GGC1, GGC2, GGC3, GGC5, GGC6 and GGC7.
The isolates GGC4 and GGC8 produced 2 banding patterns. Among the
8 isolates, all the isolates produced loci of Rm value 0.47, 0.63 and 0.75
except GGC4, GGC5 and GGC8 (Table 3 and Figure 4).
Further dendogram was constructed for three isozymes tested respect-
ively to see the variability of test isolates based on their clustering pattern
(Figure 5).

Alpha esterase isoenzyme dendogram

A dendogram was generated by UPGMA clustering where one major
cluster was identified with 25% Euclidean distance. Group I comprised
of eight isolates GGC2, GGC3, GGC1, GGC6, GGC5, GGC8, GGC4 and
GGC7. Group I was further sub-clustered into five groups, of which first
sub-cluster (group IA) had four isolates GGC2, GGC3 and GGC1 and
isolate GGC6 was in separate individual cluster. Group IB consists of two
isolates GGC5 and GGC8 and group IC and ID consists of GGC4 and
GGC7 in individual clusters.

Beta esterase isozyme dendogram

Dendrogram identified two major clusters with 25% Euclidean distance.
One cluster (group I) comprised of seven isolates GGC6, GGC8, GGC4,
GGC7, GGC2, GGC5 and GGC1 and another cluster (group II) comprised
of one isolate GGC3. Group I was further sub-clustered into three
groups, of which first sub-cluster (group IA) had four isolates (GGC6,
GGC8, GGC4 and GGC7). Second sub-cluster (group IB) included two
isolates GGC2, and GGC5 while GGC1 formed separate individ-
ual cluster.

Peroxidase isozyme dendogram

A dendogram was generated by UPGMA clustering as presented in the
Figure 5. This dendrogram identified two major clusters with 25%
Table 3. Relative mobility (Rm) values of esterase and peroxidase in different isolates of Colletotrichum truncatum.
Isolates
GGC1 GGC2 GGC3 GGC4 GGC5 GGC6 GGC7 GGC8
No. of No. of No. of No. of No. of No. of No. of No. of
Sl. No. Isoenzymes Rm Bands Rm Bands Rm Bands Rm Bands Rm Bands Rm Bands Rm Bands Rm Bands
1 a-Esterase 0.50 4 0.50 5 0.50 4 0.50 5 0.44 4 0.49 4 0.41 4 0.43 4
0.56 0.56 0.56 0.56 0.53 0.54 0.44 0.54
0.59 0.59 0.59 0.59 0.59 0.63 0.61 0.63
0.71 0.71 0.71 0.61 0.69 0.74 0.63 0.71
0.77 0.77
2 b-Esterase 0.35 2 0.23 4 0.47 2 0.32 3 0.25 4 0.30 2 0.33 2 0.30 2
0.47 0.30 0.62 0.37 0.33 0.37 0.40 0.37
0.35 0.47 0.38
0.47 0.47
3 Peroxidase 0.58 3 0.58 3 0.58 3 0.58 2 0.52 3 0.58 3 0.58 3 0.58 2
0.63 0.63 0.63 0.75 0.65 0.63 0.63 0.75
0.75 0.75 0.75 0.75 0.75 0.75
ARCHIVES OF PHYTOPATHOLOGY AND PLANT PROTECTION
149
150 T. MARAK ET AL.

Figure 4. Polyacrylamide gel electrophoresis profile of eight Collectotrichum truncatum iso-

lates isozymes: (A) alpha esterase, (B) beta esterase and (C) peroxidase.

Euclidean distance. One cluster (group I) comprised of two isolates

GGC4 and GGC8 and another cluster (group II) comprised of six isolates
GGC6, GGC7, GGC1, GGC2, GGC3 and GGC5. Group II was further
sub-clustered into two groups, of which first sub-cluster (group IIA) had
five isolates (GGC6, GGC7, GGC1, GGC2 and GGC3) while the isolate
GGC5 were in separate individual cluster.

Discussion
In India, the greengram anthracnose was first reported from Jorhat of
Assam state in 1951 (Majid 1953). The disease has been reported from
all the mungbean growing regions of India in mild to severe form and in
tropical and subtropical areas it causes considerable damage by reducing
seed quality and yield every year in varying intensity. The disease causes
qualitative as well as quantitative losses (Sharma et al. 1971).
Anthracnose continues to be one of the major constraints in greengram
production inspite of all the management approaches viz., use of tolerant
varieties (Marak et al. 2018), application of fungicides (Ramdial et al.
2017), cultural practices (Dabbas et al. 2015) and combination of
approaches leading to integrated management of the disease (Rathaiah
and Sharma 2004; Laxman 2006). Very limited research work has been
carried out on this disease in India and in West Bengal no such works
has been conducted so far. The information on the disease reported so
far needs to be constantly improved with regards to several aspects if
any safeguard against this risk is to be developed in near future.
During the field survey, it was also observed that in most of the fields
greengram was grown as a sole crop which may lead to the increase of
the pathogen population and enhance the disease. Eight single spore-cul-
tures of Colletotrichum isolates collected from different locations showed
cultural variability in respect of morphology of mycelium, colony colour
 ARCHIVES OF PHYTOPATHOLOGY AND PLANT PROTECTION 151

Figure 5. Dendogram of eight isolates of Colletotrichum truncatum showing relationship

with three isozymes tested at 25% Euclidean distance. (i) On alpha esterase, (ii) on beta
esterase and (iii) on peroxidise isozyme.

and margin of colony on different medium. The maximum mycelial

growth at 10 DAI was observed in PDA, PCA and CA and least growth
in V8 Juice agar media. The solid media were found better for the
growth and sporulation of C. lindemuthianum reported by Kumar and
Yadav (2004); Vinod and Benagi (2009); Prema et al. (2011) and
Ashutosh et al. (2012). The present results are in similar with the results
152 T. MARAK ET AL.

obtained by Singh and Shukla (1986); Wasantha and Rawal (2008);

Vinod and Benagi (2009) and Prema et al. (2011).
Different isolates showed variability in respect to their size of conidia.
The results clearly, suggest that variability exist within the isolates.
Similar finding in case of C. capsici was reported by Sangdee et al.
(2011) and by different workers from different regions of India (Jagtap
and Sontakke 2009; Adhipathi et al. 2013).
Biochemical variability among the eight isolates of C. truncatum iso-
lates from different locations was investigated in respect to isozyme to
observe the polymorphism among the isolates and the results are dis-
cussed below. Similar work on isoenzyme reported by Serra et al. (2006)
and Kilambo et al. (2013) suggests that methods based on isozyme ana-
lysis and RAPD are effective in differentiating Colletotrichum gleospor-
oides isolates from cashew and mango trees. Furtado et al. (1999)
observed variation in the number, intensity and relative mobility of
bands formed in polyacrylamide gel, allowing the visualisation of similar-
ity groups between C. gleosporoides isolates obtained from rubber trees.
Cluster analysis of banding profiles revealed the existence of intraspecific
variability among Colletotrichum kahawae isolates (Loureiro et al. 2011).
Isoenzyme profile analysis has been useful to investigate diversity within
Colletotrichum. Despite lesser number of isolates, the isoenzyme analysis
using IEF was a useful tool to detect intraspecific variability among C.
kahawae isolates and to separate C. kahawae from C. gloeosporioides
(Loureiro et al. 2011). This study provides useful insights into the range
of variability existing across these isolates. However, to understand the
variability among the populations of C. truncatum need to work on dif-
ferent geographical and spatial scales.

Conclusion
This study clearly revealed that cultural, morphological and biochemical
variability did exist among the different isolates of C. truncatum collected
from different location of West Bengal. However, more isolates from
diverse location needs to be studied to get better understanding of vari-
ability arising among the isolates and new molecular technique can also
be used.

Disclosure statement
No potential conflict of interest was reported by the authors.
 ARCHIVES OF PHYTOPATHOLOGY AND PLANT PROTECTION 153

ORCID
Sunita Mahapatra http://orcid.org/0000-0002-8476-0490

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