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A Comparative Analysis of Culture Media for Optimizing the Mycelial Growth

and Sporulation of Stemphylium vesicarium Cause of White Blotch of Onion

Article in Journal of Agricultural Science and Technology A · June 2015

DOI: 10.17265/2161-6256/2015.06.006

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Journal of Agricultural Science and Technology A 5 (2015) 440-448
doi: 10.17265/2161-6256/2015.06.006
D DAVID PUBLISHING

A Comparative Analysis of Culture Media for Optimizing

the Mycelial Growth and Sporulation of Stemphylium
vesicarium Cause of White Blotch of Onion

Hosna Ara Chowdhury1, Nurul Islam2, Belal Hossain1, Moudood Ahmed1, Sayed Mohsin1 and Rafiqul Islam1
1. Department of Plant Pathology, Sher-e-Bangla Agricultural University, Dhaka 1207, Bangladesh
2. Department of Botany, University of Dhaka, Dhaka 1000, Bangladesh

Abstract: This study was conducted to identify a cheap and suitable culture medium for the mycelial growth and sporulation of
Stemphylium vesicarium and to determine the cultural and morphological variability of this pathogen. A total of 24 isolates of S.
vesicarium collected from eight different onion growing areas were characterized in terms of cultural and morphological aspects.
Front colony colors were greenish brown to dirty white, deep grey to whitish, light grey to whitish, deep greenish white, light grey
and dirty white to greenish. Reverse colony colors were brown, deep brown and light brown. Colony shapes were circular and
irregular with umbonate, raised and flat type colony elevation. Colony textures were cottony, fluffy and velvety with entire, undulate
and filiform type colony margin. Among the culture media, V-7 juice agar found to be the most suitable culture media for mycelial
growth of S. vesicarium. The sporulation of the fungus was remarkably influenced by V-7 juice mixed with potato dextrose agar
(PDA) media, this media exhibited the highest sporulation (87.76-169.0/mm2) of S. vesicariumin comparison with other media. The
minimum days (28 d to 31 d) for conidial production were observed on V-7 juice agar medium. The length of conidia varied from
14.6 µm to 30.6 µm. The maximum mean length of conidia was 29.97 µm found in isolate DSSA, while the minimum mean length
17.36 µm was found in isolate MSMM 02. The breadth of conidia ranged from 4.7 µm to 15.7 µm. The maximum mean breadth of
conidia was 12.55 µm found in the isolate DSSA, while the minimum mean breadth 9.760 µm was found in the isolate CCKH 02.
The horizontal septation varied from 1 to 3 and the longitudinal septation varied from 0 to 4.

Key words: Onion, white blotch, S. vesicarium, V-7 juice agar.

1. Introduction cepa) is one of the most important and familiar spices

crop throughout the world, so attention must be given
White blotch of onion, commonly known as
on white blotch disease. Out of 15 important
stemphylium blight which is caused by Stemphylium
vegetables and spice crops listed by FAO, onion
vesicarium, is an important disease throughout the
stands the second in terms of annual world production
world. As per literatures in Refs. [1-3], onion crop is
[6]. Among the onion producing countries, China tops
affected by 66 diseases, and among them stemphylium
the list with 205.08 million tons onion production,
blight is an important one. In Bangladesh, onion
followed by 133.72 million tons in India, 33.21
production is gradually decreasing due to this major
million tons in USA and 19.23 million tons in Iran,
disease problem. Onion seed production is also
whereas the production of onion in Bangladesh is only
severely affected by this disease, because it causes the
1,159,259 tons [7], which is very lower compared to
breaking of floral stalks [4]. About 80%-85% losses
other onion growing countries in the world. The onion
were caused by S. vesicarium affecting leaves and
production per unit area is gradually decreasing in
inflorescence stalks of onion [5]. As onion (Allium
Bangladesh due to diseases problem [8]. Stemphylium
blight has become more widespread in the onion
Corresponding author: Belal Hossain, Ph.D., associate growing region during recent years.
professor, research field: molecular biology.
 A Comparative Analysis of Culture Media for Optimizing the Mycelial Growth and Sporulation of 441
Stemphylium vesicarium Cause of White Blotch of Onion

Isolation and characterization of causal organism of cultural variability. For this target, 5 mm diameter
Stemphylium from infected onion plant or plant parts mycelial discs of 7-day-old culture were transferred to
is rather difficult. To study the Stemphylium in lab the centre of different culture media—PDA, V-7 juice
condition is a problem, particularly because of its slow agar and combined PDA + V-7 juice agar (Table 1)
growth habit on routine laboratory artificial culture and incubated at 25 ± 1 °C. Radial mycelial growth
media, like potato dextrose agar (PDA). Sometimes, it was measured after 3 d of incubation at 25 ± 1 °C.
grows as mycelium, but does not sporulate easily in Three replications were maintained for each isolate in
the culture media, and without sporulation it is a completely randomized design. The radial mycelial
difficult to identify a pathogen. Hashemi et al. [9] also growth was recorded from the 3rd days to 15th days
reported that Stemphylium sp. do not sporulate well on after inoculation. Growth per day was calculated by
ordinary synthetic media. To overcome this problem, Eq. (1):
many researchers have tried to find out the suitable Growth rate (cm/d) = (growth on a day – growth on
culture media for the mycelial growth and sporulation previous day)/2 (1)
of Stemphylium sp.. Borges et al. [10] harvested 7 × Sporulation time and conidia production were
104 conidia/mL of S. botryosum in V-8 juice agar measured and recorded by using pure culture of S.
medium from alfalfa. Salter and Leath [11] also used vesicarium grown on PDA, V-7 juice agar and
V-8 juice agar and achieved 1 × 104 conidia/mL to 5 × combined PDA + V-7 juice agar media. The conidia
104 conidia/mL. In Bangladesh, V-8 juice agar produced per unit surface area were measured using
medium is unavailable and somewhat costly. Thus, haemacytometer and digital microscope. The
conducting research on Stemphylium sp. cost effective measurement was according to Chauhan and Pandey
and easily available media is necessary. So the present [12] as Eq. (2):
study was undertaken to identify a cheap and suitable No. of conidia per mL suspension
culture medium for the mycelial growth and No. of conidia/mm2 = (2)
Total surface area of suspension
sporulation of S. vesicarium and to determine the
cultural and morphological variability of this pathogen. Finally, front and reverse colony color, shape,
elevation, margin and texture of 24 isolates of S.
2. Materials and Methods vesicarium colony were recorded on PDA for cultural
2.1 Collection, Isolation, Purification and variability.
Preservation of S. vesicarium 2.3 Morphological Variability Study of S. vesicarium
S. vesicarium was isolated by tissue planting Isolates
methods from diseased leaves of onion grown at Morphological variability of 24 isolates of S.
different locations. Purification was carried out by vesicarium in the terms of conidial shape, color, size
transferring conidia of S. vesicarium into PDA and septation were observed and recorded on PDA
medium and incubated at 25 ± 1 °C with maintaining medium. The 30-days-old, 40-days-old and
alternating dark and light period. The pure cultures of 90-days-old pure culture was used for morphological
S. Vesicarium were preserved in refrigerator at 4 °C variability.
for further use.
2.4 Experimental Design
2.2 Cultural Variability Study of S. vesicarium Isolates
The experiment was laid out in a completely
A total of 24 isolates of S. vesicarium were used for randomized design with three replications.
442 A Comparative Analysis of Culture Media for Optimizing the Mycelial Growth and Sporulation of
Stemphylium vesicarium Cause of White Blotch of Onion

Table 1 Composition of different culture media for 1 L.

Composition
Name of media
Ingredient Amount
Potato slice 200 g
Dextrose 20 g
PDA media
Agar 17 g
Distilled water 1,000 mL
Tomato 174 mL
Carrot 8 mL
Celery 4 mL
Spinach 4 mL
V-7 juice agar media Beet 4 mL
(vegetables juice agar) Lettuce 4 mL
Onion 2 mL
Agar 15 g
CaCO3 2g
Distilled water 800 mL
Components of PDA medium 200 mL
Combined PDA + V-7 juice agar media
Components of V-7 juice agar media 800 mL

2.5 Statistical Analysis and combined PDA + V-7 juice agar culture media.
The highest number (87.76-169.0/mm2) of conidia
The data were analyzed by statistical software,
was recorded on combined PDA+V-7 juice agar
MSTAT-C computer package program. The data were
media, and the lowest number (20.73-36.67/mm2) of
subjected to an analysis of variance, and least
conidia was found on PDA media, preceded by V-7
significant difference (LSD) were used to separate
juice agar (55.01 to 92.12/mm2) as clear shown in
means and compared with Dunkan’s multiple range
Fig. 2.
test (DMRT), where F values indicated significantly
3.1.3 Sporulation Time of S. vesicarium on
differences at 5% level of probability.
Different Culture Media
3. Results The effect of PDA, V-7 juice agar and combined
PDA + V-7 juice agar culture media on sporulation
3.1 Cultural Variability of S. vesicarium Isolates
time of S. vesicarium is presented in Fig. 3. The
3.1.1 Mycelial Growth of S. vesicarium on earliest (28.33-31.00 d) conidia production was
Different Culture Media observed on V-7 juice agar in respect of all isolates.
The effect of different culture media, i.e., PDA, V-7 On PDA, comparatively more days (86.33-90.00 d)
juice agar and combined PDA + V-7 juice agar, had a were required for conidia production.
significant role on per-day radial mycelial growth of S. 3.1.4 Characteristics of Colony Color, Shape,
vesicarium (Table 2 and Fig. 1). The fungus grew well Elevation, Margin and Texture on PDA
on V-7 juice agar medium and the maximum growth The isolates of S. vesicarium exhibited variations in
was recorded as 0.610-0.650 cm/d. The minimum colony characters, like color, shape, elevation, margin
radial mycelial growth (0.423-0.483 cm/d) was and texture. Front colony colors were greenish brown
recorded on PDA medium, preceded by combined to dirty white, deep grey to whitish, light grey to
PDA + V-7 juice agar medium (0.470-0.503 cm/d). whitish, deep greenish white, light grey and dirty
3.1.2 Conidium Production of S. vesicarium on white to greenish, whereas reverse colony colors were
Different Culture Media deep brown, brown and light brown. Circular and
Marked variations were observed on conidium irregular colony shapes were found in S. vesicarium,
production of S. vesicarium on PDA, V-7 juice agar respectively. Colony elevation was umbonate, raised
 A Comparative Analysis of Culture Media for Optimizing the Mycelial Growth and Sporulation of 443
Stemphylium vesicarium Cause of White Blotch of Onion

Table 2 Daily radial mycelial growths of 24 isolates of S. vesicarium on different culture media in different location.
Mean colony diameter of radial mycelia growth/day (cm)*
Location
Isolates On combined
(district/sub-district) On PDA agar On V-7 juice agar
PDA + V-7 agar
DSSA Sher-e-Bangla Nagar, Dhaka 0.460 0.616 0.473
DSTR 01 0.480 0.630 0.483
Savar, Savar
DSTR 02 0.480 0.626 0.470
MMBH BAU, Mymensingh Sadar 0.433 0.640 0.493
MTBB 01 0.476 0.633 0.490
Trishal, Trishal
MTBB 02 0.463 0.620 0.500
RBHR 01 0.466 0.650 0.486
RBHR 02 Rajshahi, Bagmara 0.453 0.626 0.480
RBHR 03 0.443 0.640 0.496
GJBS BARI, Joydebpur 0.440 0.640 0.496
GGBB 01 0.430 0.620 0.486
Gazipur Sadar
GGBB 02 0.453 0.626 0.480
CCKH 01 0.423 0.640 0.496
CCKH 02 Chandina, Comilla 0.483 0.616 0.496
CCKH 03 0.460 0.616 0.490
JJLL 01 0.466 0.636 0.486
JJLL 02 Jamalpur Sadar 0.446 0.630 0.476
JJLL 03 0.470 0.633 0.490
MSMM 01 0.460 0.620 0.503
MSMM 02 Manikganj, Shibalaya 0.470 0.633 0.486
MSMM 03 0.463 0.640 0.486
FFKU 01 0.456 0.623 0.480
FFKU 02 Faridpur, Faridpur Sadar 0.476 0.626 0.500
FFKU 03 0.453 0.610 0.483
LSD (0.05) 0.0519 0.0519 0.0519
CV 6.40% 2.72% 2.84%
*
Means of three observations for each isolate.

and flat type. Entire, undulate and filiform margins 30.6 µm. The maximum mean length (29.97 µm) was
were found in S. vesicarium colony. Colony texture recorded in DSSA isolate, and the minimum mean
was cottony, fluffy and velvety. These results are length (17.36 µm) in isolate MSMM 02. The breadth of
presented in Table 3. conidia ranged from 4.7 µm to 16.24 µm. The highest
mean breadth (12.55 µm) was recorded in isolate DSSA
3.2 Morphological Variability of S. vesicarium
and the minimum mean breadth (9.760 µm) was in
Isolates
CCKH 02 isolate. These results are presented in
3.2.1 Conidial Shape, Color and Size of S. Table 4.
vesicarium on PDA 3.2.2 Septation of S. vesicarium Conidia on PDA
Distinct variations were observed in conidial shape, Distinct variations were observed in horizontal and
color and size of 24 isolates of S. vesicarium on PDA. longitudinal conidial septation of 24 isolates of S.
Conidial shape was ovoid, oblong and ovoid to oblong vesicarium. The maximum mean number of horizontal
type, which were light brown, brown and deep brown septation was recorded in DSTR 01 (2.500) isolate
in color. The length of conidia varied from 14.6 µm to and the minimum (1.600) was in isolates of MTBB 01,
444 A Comparative Analysis of Culture Media for Optimizing the Mycelial Growth and Sporulation of
Stemphylium vesicarium Cause of White Blotch of Onion

Radial
Radialmycelial
mycelialgrowth / day (cm)
growth/day (cm)ononPDA
PDAmedium
medium Radial
Radial mycelia
mycelial Radial
Radial mycelia growth/day (cm)
mycelial
growth/day
growth/Day (cm) on
(cm) on combined
growth/Day (cm)(PDA + V-7 juice
on combined (PDA+agar)
V-7medium
juice agar) medium
onV-7
V-7juice
juice agar
agar medium
medium
Radial mycelial growth/day (cm)

0.700
0.600
0.500
0.400
0.300
0.200
0.100
0.000

Isolates

Fig. 1 Radial mycelial growth of S. vesicarium on PDA, V-7 juice agar and combined PDA + V-7 juice agar media.

Numberofofconidia/mm2
conidia/mm2 PDA medium
Number PDA medium
Numberofofconidia/mm
Number conidia/mm2
2 V‐7
V-7 juice
juice agar
agar medium
medium
Numberofofconidia/mm
Number conidia/mm2
2 Combined
combined (PDA+V‐7
(PDA juice
+ V-7 juice agar)
agar) medium
medium

180
160
140
Number of conidia/mm2

120
100
80
60
40
20
0

Isolates

Fig. 2 Conidia production of 24 isolates of S. vesicarium on PDA, V-7 juice agar and combined PDA + V-7 juice agar media.
 A Comparative Analysis of Culture Media for Optimizing the Mycelial Growth and Sporulation of 445
Stemphylium vesicarium Cause of White Blotch of Onion

Sporulation Sporulation Sporulation

time (days) in PDA time (days) in V7 time (days) in PDA+V7
100
90
Sporulation time (days)

80
70
60
50
40
30
20
10
0

Isolates

Fig. 3 Sporulation time (days) of 24 isolates of S. vesicarium on PDA, V-7 juice agar and combined PDA + V-7 juice agar
media.

Table 3 Front and reverse colony color, shape, elevation, margin and texture of 24 isolates of S. vesicarium on PDA.
Cultural variability on PDA
Isolates Reverse colony Colony Colony Colony Colony
Front colony color
color shape elevation margin texture
DSSA Greenish brown to dirty white Deep brown Irregular Raised Undulate Cottony
DSTR 01 Deep greenish white Deep brown Irregular Umbonate Entire Cottony
DSTR 02 Greenish brown to dirty white Deep brown Circular Raised Entire Cottony
MMBH Light grey Light brown Circular Raised Entire Cottony
MTBB 01 Deep grey to whitish Deep brown Irregular Raised Filiform Fluffy
MTBB 02 Deep grey to whitish Deep brown Circular Raised Entire Fluffy
RBHR 01 Dirty white to greenish Brown Circular Flat Entire Velvety
RBHR 02 Deep grey to whitish Brown Circular Raised Filiform Fluffy
RBHR 03 Greenish brown to dirty white Light brown Irregular Raised Undulate Cottony
GJBS Deep grey to whitish Brown Circular Umbonate Entire Fluffy
GGBB 01 Greenish brown to dirty white Light brown Circular Umbonate Entire Cottony
GGBB 02 Greenish brown to dirty white Light brown Circular Raised Entire Cottony
CCKH 01 Greenish brown to dirty white Light brown Circular Raised Entire Velvety
CCKH 02 Deep grey to whitish Brown Circular Umbonate Filiform Fluffy
CCKH 03 Deep grey to whitish Brown Irregular Umbonate Undulate Fluffy
JJLL 01 Deep grey to whitish Brown Circular Raised Undulate Fluffy
JJLL 02 Deep grey to whitish Brown Irregular Umbonate Filiform Fluffy
JJLL 03 Deep grey to whitish Brown Circular Raised Filiform Fluffy
MSMM 01 Deep grey to whitish Brown Irregular Raised Entire Fluffy
MSMM 02 Deep grey to whitish Brown Irregular Raised Undulate Fluffy
MSMM 03 Deep grey to whitish Deep brown Circular Raised Filiform Fluffy
FFKU 01 Deep grey to whitish Deep brown Circular Raised Filiform Fluffy
FFKU 02 Light grey to whitish Light brown Circular Raised Entire Cottony
FFKU 03 Deep grey to whitish Deep brown Irregular Umbonate Filiform Fluffy
446 A Comparative Analysis of Culture Media for Optimizing the Mycelial Growth and Sporulation of
Stemphylium vesicarium Cause of White Blotch of Onion

Table 4 Shape, color and size of conidia of 24 isolates of S. vesicarium on PDA.

Conidial size
1
Isolates Shape Color Conidial length (µm) Conidial breadth1(µm)
Range Mean Range Mean
DSSA Ovoid Deep brown 29.5-30.6 29.97 4.7-14.5 12.55
DSTR 01 Oblong Light brown 20.4-27.8 25.13 7.7-15.1 9.80
DSTR 02 Oblong Light brown 23.5-26.2 24.43 6.0-15.3 11.89
MMBH Ovoid to oblong Light brown 19.0-25.3 23.13 6.9-14.5 11.10
MTBB 01 Ovoid Deep brown 20.0-27.6 24.87 8.2-12.1 10.15
MTBB 02 Ovoid Deep brown 23.7-25.2 24.60 9.6-14.7 11.36
RBHR 01 Ovoid Brown 19.8-27.9 23.86 9.4-14.4 11.86
RBHR 02 Oblong Brown 22.4-23.1 22.67 8.1-13.8 10.81
RBHR 03 Ovoid to oblong Light brown 21.2-24.9 22.60 9.3-13.7 10.89
GJBS Ovoid to oblong Deep brown 16.9-22.7 20.74 8.1-12.1 11.12
GGBB 01 Oblong Light brown 17.6-25.7 21.27 9.0-14.6 10.98
GGBB 02 Oblong Light brown 24.9-28.1 26.80 8.2-14.7 11.73
CCKH 01 Oblong Brown 15.0-19.1 17.50 8.2-15.7 11.29
CCKH 02 Oblong Brown 20.2-27.7 25.42 6.7-16.2 9.76
CCKH 03 Oblong Light brown 20.8-28.4 25.37 7.7-14.2 10.34
JJLL 01 Ovoid Deep brown 19.9-26.4 24.02 7.9-14.9 10.54
JJLL 02 Oblong Brown 18.3-23.2 20.73 8.7-13.9 11.01
JJLL 03 Ovoid Light brown 16.3-21.4 19.47 7.3-13.3 10.95
MSMM 01 Ovoid to oblong Deep brown 15.5-22.3 18.09 7.3-12.9 9.82
MSMM 02 Ovoid to oblong Brown 14.6-20.5 17.36 7.3-14.4 10.23
MSMM 03 Oblong Brown 19.0-25.3 23.32 9.2-12.8 10.13
FFKU 01 Oblong Light brown 19.7-24.6 19.98 9.6-13.5 9.86
FFKU 02 Ovoid Brown 16.9-25.2 21.87 7.9-14.3 10.52
FFKU 03 Ovoid Deep brown 17.2-24.9 24.52 8.0-13.9 11.23
LSD (0.05) 0.563 0.395
CV 1.50% 2.23%
1
Means of 10 observations for each isolate.

RBHR 01, GGBB 01 and MSMM 01. The maximum sporulation capacity, combined PDA + V-7 juice agar
mean number of longitudinal septation (2.500) was medium exhibited the highest performance
recorded in MMBH isolate and the minimum (1.400) (87.76-169.0 conidia/mm2) in comparison to other
in GGBB 02 isolate. media used. Kumar [13] also identified a suitable
culture medium for the sporulation of S. botryosum and
4. Discussion
found that the fungus produce the highest number of
In this study, three different culture media were conidia (84.7 ± 6.0 × 104 conidia/mL) on V8 juice
used to identify which would be suitable in the terms potato dextrose agar (V8-P) medium. All the isolates
of facilitating radial mycelial growth. Results showed showed variation in respect of sporulation time on
that among the three selected culture media, V-7 juice different media. The sporulation time of S. vesicarium
agar gave the best performance, while the lowest on PDA varied from 86.33 d to 90.67 d with the
radial mycelial growth was observed on PDA medium. maximum sporulation time in isolate GGBB 02 and
Results revealed that presence of seven different MSMM 02, respectively, and the minimum in GJBS
vegetables juice in culture medium favored radial isolate. On V-7 juice agar, the sporulation time for all
mycelial growth of S. vesicarium. But in the terms of isolates varied from 28.33 d to 31.0 d with the
 A Comparative Analysis of Culture Media for Optimizing the Mycelial Growth and Sporulation of 447
Stemphylium vesicarium Cause of White Blotch of Onion

maximum sporulation time in isolate JJLL 03 and the findings agreed with the report of Ellis [18] and Koike
minimum in JJLL 01. On combined PDA + V-7 juice et al. [19], who found oblong to ovoid shaped conidia
agar, the sporulation time varied from 35.0 d to 39.67 of Stemphylium, and reported that the conidia of
d with the maximum sporulation time in isolate Stemphylium botryosum were broadly ovoid. Oblong,
CCKH 02 and the minimum in DSSA and RBHR 03 ovoid or broadly ellipsoidal shaped conidia of
isolates, respectively. The results of the present Stemphylium luffae, Stemphylium lycii and Stemphylium
investigation showed that S. vesicarium produces cucumis was also found by Pei et al. [20]. Remarkable
conidia on V-7 juice agar earlier than on PDA and variations were also observed in conidial color of 24
combined PDA + V-7 juice agar medium. isolates of S. vesicarium on PDA, where deep brown,
Colonies of S. vesicarium showed deep grey to brown and light brown color conidia were observed.
whitish, greenish brown to dirty white, light grey to The current findings were well supported by some
whitish, deep greenish white, light grey and dirty reported works [17, 19, 21], which worked on
white to greenish color from frontal view on PDA. Stemphylium solani and Stemphylium lycopersici and
The results are in agreement with Salter and Leath observed tan to light brown colored conidia. Koike et al.
[11], who found that the colony color of Stemphylium [19] and Zheng et al. [21] found brown colored conidia
solani isolated from pepper was gray in color and gray of Stemphylium botryosum and Stemphylium solani.
to light brown growth found in case of Stemphylium Marked variations were observed in conidial length
lycopersici isolated from tomato plants on PDA. and breadth of S. vesicarium on PDA. The length of
Hosen et al. [14] observed greenish brown and dirty conidia varied from 14.6 µm to 30.6 µm, and the
white color colony of Stemphylium botryosum isolated breadth of conidia ranged from 4.7 µm to 15.7 µm.
from lentil plants. Whereas, reverse colony colors The present findings agreed with the report of
were deep brown, brown and light brown. Circular Arzanlou et al. [16] and Ellis [18], who measured
and irregular shaped colonies were found in all the 25-40 μm  13-21 μm and 20-24 (-30) μm × 12-15 μm
isolates with entire, undulate and filiform margin on conidial length and breadth of S. vesicarium. Hosen et
PDA. The results are in agreement with Hosen [15], al. [14, 15], Koike et al. [19] and Simmons [22] also
who found that the colony margin of Stemphylium observed 33-35 μm  24-26 μm, 17-28 μm  13-19 μm,
botryosum isolated from lentil plants was entire and 10-25 μm  5-15 μm and 12.35-23.45 μm  10.5-15
regular. Umbonate, raised and flat type colony μm onidia of Stemphylium botryosum. In this research,
elevation were found among all the isolates with the horizontal septation of S. vesicarium conidia
cottony, fluffy and velvety texture on PDA. Arzanlou varied from 1 to 3, and the longitudinal septation
et al. [16] found flat type colony elevation of S. varied from 0 to 4. These findings collaborate with the
vesicarium grown on potato carrot agar (PCA) media. reports in other Refs. [11, 16, 18] which found that S.
The results of textural growth of S. vesicarium are in vesicarium conidia have 1-3 transverse, 1-4
agreement with Mehta [17], who found that the longitudinal or oblique septa and 1-5 transverse and
mycelium of Stemphylium solani isolated from cotton several longitudinal septa. Pei et al. [20] noted 1-3
was velvety, cottony or immersed. Similarly, Hosen et dark colored transverse septa and 2-7 longitudinal or
al. [14] also observed velvety, effuse and fluffy type oblique septa in Stemphylium solani. They also found
colony texture of Stemphylium botryosum isolated 3-5 transverse septa and 2-5 longitudinal or oblique
from lentil plants. Ovoid, oblong and ovoid to oblong septa in Stemphylium luffae [20].
shaped conidia of S. vesicarium were observed under In this research work, only three types of cultural
digital microscope in this research work. The present media were used, but for the establishment of suitable
 448 A Comparative Analysis of Culture Media for Optimizing the Mycelial Growth and Sporulation of
Stemphylium vesicarium Cause of White Blotch of Onion

cultural medium for the growth and sporulation of S. Blight of Lentil: Should You Be Worried?” Present at the
Abstract of Pulse Days 2004 on Securing Success:
vesicarium, more research work should be performed.
Shaping the Pulse Industry of the Future, Saskatchewan
Pulse Growers, January 12-13, 2004, Saskatoon,
5. Conclusions
Saskatchewan, Canada.
From the present research, it is concluded that all [10] Borges, O. L., Stanford, E. H., and Webster, R. K. 1976.
“The Host-Pathogen Interaction of Alfalfa and
isolates of S. vesicarium showed variations in the
Stemphylium botryosum.” Phytopathology 66 (6): 749-53.
terms of cultural and morphological aspects. Different [11] Salter, R., and Leath, K. 1991. “Stemphylium Leaf Spot
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