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Effectiveness of cultural parameters on the growth and sporulation of Colletotrichum gloeosporioides causing anthracnose disease of mango (Mangifera indica L.)

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Colletotrichum gloeosporioides causing anthracnose which is a serious post harvest disease in mango accounting for 15-20% loss. The variation in nutritional and physiological characteristics among five isolates of C. gloeosporioides collected from different agro-climatic regions of India was investigated. All the isolates showed differential response in requirements of media, temperature and media pH for growth and sporulation. Malt Extract Agar (MEA) medium was best suited for growth in terms of radial mycelial diameter for all the isolates. Among the studied isolates, Cg 72 (from Maharashtra) showed more virulence and maximum sporulation (137.5×103 mL-1) at 28°C and media pH 6. Maximum growth and virulence at 28°C was observed with Cg 62 isolate. Media of pH 6 was found to be most suitable for the growth of respective isolates (s), but Cg 62 which was collected from Bihar found most virulent in this experiment.
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OnLine Journal of Biological Sciences, 2012, 12 (4), 123-133
ISSN: 1608-4217
© 2012 A. Pandey et al., This open access article is distributed under a Creative Commons Attribution
(CC-BY) 3.0 license
doi:10.3844/ojbssp.2012.123.133 Published Online 12 (4) 2012 (http://www.thescipub.com/ojbs.toc)
Corresponding Author: Ashutosh Pandey, Molecular Plant Pathology Laboratory, Central Institute for Subtropical Horticulture,
Lucknow- 227 107, Uttar Pradesh, India Tel: +91 933 616 2218, Fax: +91 5278 265201
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Effectiveness of Cultural Parameters on the Growth and Sporulation
of Colletotrichum gloeosporioides Causing Anthracnose Disease of
Mango (Mangifera indica L.)
1Ashutosh Pandey, 1Lallan Prasad Yadava, 1Muthukumar Manoharan,
2Ugam Kumari Chauhan and 1Brajesh Kumar Pandey
1Central Institute for Subtropical Horticulture,
Molecular Plant Pathology Laboratory, Lucknow-227 107, Uttar Pradesh, India
2Center for Biotechnology, School of Environmental Biology,
A.P.S. University, Rewa- 486 003, Madhya Pradesh, India
ABSTRACT
Colletotrichum gloeosporioides causing anthracnose which is a serious post harvest disease in mango
accounting for 15-20% loss. The variation in nutritional and physiological characteristics among five
isolates of C. gloeosporioides collected from different agro-climatic regions of India was investigated.
All the isolates showed differential response in requirements of media, temperature and media pH for
growth and sporulation. Malt Extract Agar (MEA) medium was best suited for growth in terms of
radial mycelial diameter for all the isolates. Among the studied isolates, Cg 72 (from Maharashtra)
showed more virulence and maximum sporulation (137.5×103 mL1) at 28°C and media pH 6.
Maximum growth and virulence at 28°C was observed with Cg 62 isolate. Media of pH 6 was found to
be most suitable for the growth of respective isolates (s), but Cg 62 which was collected from Bihar
found most virulent in this experiment.
Keywords: Colletotrichum gloeosporioides, Mangifera Indica L., Radial Growth, Sporulation, Culture
Media, Temperature, Media pH
1. INTRODUCTION
Mango (Mangifera indica L.) is one of the most
popular seasonal fruit found mainly in the tropical and
subtropical countries (Shad et al., 2002). It is widely
grown in different countries of the world and is attacked
by a number of diseases of which anthracnose is one of the
most common especially in India and loss due to this
disease is substantial (Slade et al., 1987). Anthracnose
caused by C. gloeosporioides is reported on a wide variety
of crop, including almond, avocado, apple, arabica coffee,
guava, mango, dragon fruit, cassava, sorghum and
strawberry (Amusa et al., 2005; Masyahit et al., 2009;
Owolade et al., 2009; Erpelding, 2010).
More recent reports on C. gloeosporioides revealed
that ambient temperature, pH, free water or relative
humidity above 95% were required for conidial
germination and aspersorium formation (Shih et al.,
2000; Yakoby et al., 2000). The infection was favoured
at temperatures ranging from 20 to 30°C. Temperature
and moisture requirements for infection are used to build
forecasting systems for mango anthracnose a vital
component for the disease management (Prakash and
Srivastava, 1987). The fungus produces good aerial
mycelium in Richard’s and Brown’s agar and profusely
sporulates on oat meal and corn meal agar along
with abundant development of acervuli in rings
and few setae (Prakash and Srivastava, 1987).
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The optimum pH was between 5.8 and 6.5 and
temperature was optimum at 25°C for growth but it
ceased beyond 35°C. The fungus grew vigorously on
starch and peptone. Glutamic acid and alanine supported
best growth and sporulation (Prakash and Srivastava,
1987). Keeping in view of the above facts the present
investigation was performed to assess the effect of
media, temperature and media pH on the growth and
sporulation of isolates of C. gloeosporioides collected
from Andhra Pradesh, Madhya Pradesh, Bihar,
Maharashtra and Uttar Pradesh.
2. MATERIALS AND METHODS
2.1. Collection and Isolation of Pathogens
The isolates of C. gloeosporioides were isolated from
the samples collected from Chittoor (Andhra Pradesh),
Rewa (Madhya Pradesh), Lucknow (Uttar Pradesh),
Muzaffarpur (Bihar) and Dapoli (Maharashtra). A small
section of anthracnose infected leaf was surface
sterilized with 0.1% HgCl2 and washed thoroughly with
sterile distilled water. It was then inoculated on Potato
Dextrose Agar (PDA) medium (Potato 200 g, Glucose 20
g, Agar 15 g, distilled water 1000 mL) and incubated at
28±2°C for 6 days.
A total of 5 C. gloeosporioides isolates (Cg 1, Cg 19,
Cg 30, Cg 62 and Cg 72) were used in this study. These
isolates were isolated, purified and maintained on Potato
Dextrose Agar (PDA) slants under controlled
temperature. Pathogenicity of these isolates was also
confirmed suggested by Bhuvanaeswari and Rao (2001).
All the collected isolates of C. gloeosporioides were
submitted to National Agriculture Important Microbial
Culture Collection (NAIMCC), Mau, Uttar Pradesh,
India and accessions number were allotted (Table 1).
2.2. Preparation of Different Media and
Inoculation
The fungal pathogen was inoculated on various types
of media to identify the best suited media for its growth
and sporulation. In this experiment, fourteen media
(Himedia, Mumbai), viz., Potato Dextrose Agar (PDA),
Corn Meal Agar (CMA), Cooke rose Bengal agar base
(CRBA), Czapek Dox Agar (CDA), Pseudomonas Agar
(PA), Limabean Agar (LA), Conn’s Agar (CA), Yeast
Dextrose Agar (YDA), Corn Meal Peptone Yeast Agar
(CMPYA), Modified Czapex Dox Agar (MCDA), V-8
Juice Agar (VJA), Potato carrot agar (PCA), Malt extract
agar base Mycological peptone (MEA) and Oat Meal
Agar (OMA) were used 39, 17, 36.54, 49, 37.3, 23, 38,
35, 64, 45.36, 44.3, 24, 50 and 72.5 gram per liter
respectively. All these were autoclaved at 121°C under
15 psi for 20 min. Five mm diameter identical fresh
culture discs of different isolates (Cg 1, Cg 19, Cg 30,
Cg 62 and Cg 72) were grown for 7 days old culture at
27±2°C dissolve in 1 ml distilled water and count.
2.3. Preparation of Media at Different level of
pH and Inoculation
To assess the optimum media pH for the growth of C.
gloeosporioides the fresh culture on Malt Extract Agar
(MEA) medium of different pH level (5.5, 6.0, 6.5, 7.0,
7.5 and 8.0) was used in experiment. The media pH of
the medium was adjusted with 0.1N NaOH or 0.1N HCl
(Naik et al., 1988). The medium was buffered with
disodium hydrogen phosphate citric acid buffer
according to the protocol (Vogel, 1951). For inoculation
same method was adopted as mentioned earlier. The
diameter of colony was observed at 3rd, 5th and 7th days
after inoculation while the level of sporulation was
recorded at 7th Day After Inoculation (DAI) by
modifying the methods (Tastwal and Enagi, 2009).
2.4. Incubation at Different Temperature
Regime
To study the growth and sporulation of C.
gloeosporioides at different temperature regime fresh
culture was prepared in the solid media viz. Malt Extract
Agar (MEA). For inoculation same method was
adopted as mentioned earlier and culture of different
isolates were placed at different temperature regimes
to study the best suited temperature level viz. (12, 16,
20, 24, 28 and 32°C). Data were recorded with the
method (Kumara and Rawal, 2008).
Table 1. Morphological characteristic of different isolates from different geographical location of Colletotrichum gloeosporioides penz
Culture Culture Geographical Symptom of Mango Percent Sporu Mean spore size Colour of
Code accession no. location anthracnose substrate infection lation (Length × breadth) (µm) mycelial mat
Cg 1 NAIMCC-F-02694 Chittoor (Andhra Pradesh) Partial Leaf 50 ** 13.49×4.75 White black
Cg 19 NAIMCC-F-02707 Rewa (Madhya Pradesh) Typical Leaf 100 *** 14.20×5.25 Yellow white
Cg 30 NAIMCC-F-02719 Lucknow (Utter Pradesh) Typical Fruit 70 Nil 16.15×7.28 Grayish white
Cg 62 NAIMCC-F-02730 Muzaffarpur (Bihar) Typical Leaf 100 *** 15.78×4.38 White
Cg 72 NAIMCC-F-02735 Dapoli (Maharashtra) Partial Leaf 75 *** 16.87×4.57 Cottony white
(*; = Less sporulation, **; = Moderate sporulation, ***; = Profuse sporulation)
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2.5. Statistical Analysis
All treatments were designed in three factors factorial
Completely Randomized Design (CRD) with five
replications. Experimental data was statistically analyzed
using O. P. Sheoran software version 1.0 (CCS HAU,
Hisar). The Dendrogram was generated by Un-weighted
Pair-Group Arithmetic Mean (UPGMA) using FREE
TREE software version 0.9.1.50.
3. RESULTS
The effect of various factors such as media, media
pH, temperature regimes and their combinations with
days after inoculation on the growth (Table 2) and
sporulation of C. gloeosporioides were studied for 5
isolates representing different agro climatic zones.
3.1. Effect of Media on Growth and Sporulation
of C. Gloeosporioides Isolates
Malt Extract Agar (MEA) showed higher mean value
(54.98 mm) of mycelia growth as compared to other
evaluated media followed by Modified Czapex Dox agar
medium MCDA (53.01 mm). Cooke rose Bengal agar
base (CRBA) media exhibited the least mycelial
diameter (18.29 mm). Pseudomonas Agar (PA) and
Limabean Agar (LA) medium also recorded lower
growth next to CRBA medium but both of them were
found to be similar in response (Fig. 1A).
Maximum (46.12 mm) growth was noticed with Cg
72 isolate in over all media while the least growth was
recorded in Cg 1 (36.34 mm) (Fig. 1B). Cg 1 showed
comparatively slow growth in all the media evaluated for
the radial growth. As far as the effects of interaction
between media and isolates are concerned, maximum
(60.88 mm) growth was recorded with Cg 72 isolate in
the MEA media followed by MCDA (60.22 mm).
A significant interaction effect of media and
incubation period revealed highest growth in MEA
medium followed by MCDA both at 5 and 7th day after
inoculation CDA and OMA medium were also at par
with MEA medium at 7th days after inoculation. The
period of incubation clearly indicated that radial growth
of the mycelia increased with increase in incubation
period (days after inoculation) (Fig. 1C). The data of
interaction between isolates and incubation period
clearly showed that mycelial growth in all the isolates
increased with advancement of incubation period. CRBA
medium did not follow significantly for growth
enhancement throughout the incubation period. With
respect to the isolate effect, maximum radial growth was
recorded at 7th day in Cg 72 (71.27 mm) while minimum
was found in Cg 1 isolate at 3rd day after inoculation. It
is also evident that mycelial growth of C.
gloeosporioides was significantly influenced by
interaction of media × isolates × incubation period.
Higher radial growth (87.67 mm) was observed in Cg 19
isolate in Oat Meal Agar (OMA) medium at 7th day of
incubation followed by the same isolates in Malt extract
agar (86.33 mm), Cg 72 (85.33 mm) and Cg 19 (85.17
mm) in CDA, Cg 72 in MEA (84.0 mm) and MCDA
(83.67 mm) at 7th day of incubation.
In the experiment of sporulation in different isolates
was also significantly varied by media (Table 3).
Highest spore count (137.5×103 mL1 of culture
suspension) was recorded in Cg 72 isolate on MEA
medium followed by Cg 19 on CDA medium (102.5×103
mL1 of culture suspension).
3.2. Effect of Media pH on Growth and
Sporulation of C. Gloeosporioides Isolates
The media pH is another important factor like
temperature and media that influences growth and
sporulation significantly. The effect of different media
pH on the growth was observed considering test isolates
at different incubation period.
It is evident from the data, that mycelial growth in
different isolates significantly differed by pH range of
the media. The media pH of 6.0 exhibited higher
mycelial growth (54.43 mm) over other level of pH
studied (Fig. 2A). Among the isolates, Cg 62 resulted
in highest growth (43.86 mm) at this pH followed by
Cg 1 (Fig. 2B).
The effect of media pH × incubation period
interaction with respect to radial mycelial growth was
significant. The radial growth pattern was single sigmoid
with respect to media pH with peak inferring highest
radial growth at 7th day of incubation in media pH 6. Cg
62 and Cg 1 isolate showed the highest radial growth
(67.60 mm) at the 7th day of incubation, while minimum
was found with Cg 19 isolate at 3rd day of incubation. It
is interesting to note that radial growth of all the isolates
was at par with each other on 5th day of incubation.
However, radial growth of all the isolates increased with
the advancement of incubation period (Fig. 2C).
Three factors interactions among media pH, isolates
and incubation period revealed that mycelial growth of
all the test isolates increased with incubation period in
corresponding order. Higher radial growth (83.20 mm)
was recorded with Cg 1 isolate followed by Cg 62 (82.40
mm) and Cg 30 (81.40 mm) grown in the media 6.0 pH
at 7th day of incubation.
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Table 2. ANOVA for different isolates of C. gloeosporioides under different media, media pH and temperature regimes
Source Degree Sum of Mean of sum F- Critical
of variance of freedom squires of squires calculated P value Deference (C.D.)
Media 13 53381.45 4106.27 393.35 0.00001 1.339
Isolates 4 8248.73 2062.18 197.54 0.00001 0.800
Day after inoculation 2 231091.70 115545.80 11068.40 0.00001 0.620
Media × isolates 52 6161.30 118.49 11.35 0.00001 2.994
Media × Day after inoculation 26 30117.31 1158.36 110.96 0.00001 2.319
Isolates × Day after inoculation 8 2282.16 285.27 27.33 0.00001 1.386
Media × isolates × Day after inoculation 104 5671.09 54.53 5.22 0.00001 5.186
Media pH 5 26177.96 5235.59 2008.54 0.00001 0.519
Isolates 4 1685.27 421.32 161.63 0.00001 0.473
Days after inoculation 2 178894.50 89447.26 34314.81 0.00001 0.367
Media pH × isolates 20 740.12 37.01 14.20 0.00001 1.159
Media pH × Days after inoculation 10 5125.64 512.56 196.64 0.00001 0.898
Isolates × Days after inoculation 8 338.65 42.33 16.24 0.00001 0.820
Media pH × isolates × Days after inoculation 40 1641.72 41.04 15.75 0.00001 2.008
Temperature regime 5 33325.10 6665.02 4697.35 0.00001 0.383
Isolates 4 659.96 164.99 116.28 0.00001 0.349
Days after inoculation 2 176546.00 88272.98 62212.75 0.00001 0.271
Temperature regime × isolates 20 102.86 5.14 3.62 0.00001 0.855
Temperature regime × Days after inoculation 10 6349.33 634.93 447.49 0.00001 0.663
Isolates × Days after inoculation 8 32.38 4.05 2.85 0.00437 0.605
Temperature regime × 40 118.47 2.96 2.09 0.00023 1.482
isolates × days after inoculation
Table 3. Sporulation of different Colletotrichum gloeosporioides isolates under different media pH
pH ranges Cg 1 Cg 19 Cg 30 Cg 62 Cg 72 Mean
5.5 74.0000 151.00 200.3300 80.00 135.0000 128.06
6 51.0000 65.00 101.0000 63.00 94.0000 74.80
6.5 12.0000 26.00 40.0000 34.00 21.0000 26.60
7 11.0000 11.00 9.0000 15.00 21.0000 13.40
7.5 7.0000 7.00 10.0000 11.00 15.0000 10.00
8 7.0000 4.33 9.0000 3.66 1.6600 5.13
Mean 27.0000 44.05 61.5500 34.44 47.9400
Factors C.D. SE(d) SE(m)
pH ranges 0.6973 0.3485 0.2465
Isolates 0.6366 0.3182 0.2250
pH ranges x isolates 1.5590 0.7794 0.5511
Table 4. Sporulation of different Colletotrichum gloeosporioides isolates under different temperature regimes
Temperature regimes (°C) Cg 1 Cg 19 Cg 30 Cg 62 Cg 72 Mean
12 2.00 4.0000 3.33 2.0000 7.3300 3.73
16 6.33 8.0000 9.00 3.0000 12.0000 7.66
20 20.00 16.0000 15.00 15.0000 12.0000 15.60
24 51.00 43.0000 34.66 30.0000 42.0000 40.13
28 103.00 99.0000 111.00 126.0000 114.0000 110.60
32 76.33 59.0000 66.66 60.0000 62.6600 64.93
Mean 43.11 38.1600 39.94 39.3300 41.6600
Factors C.D. SE(d) SE(m)
Temperature regimes 0.7466 0.3732 0.2639
Isolates 0.6816 0.3407 0.2409
Temperature regimes x isolates 1.6690 0.8344 0.5900
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(A)
(B)
(C)
Fig. 1. (A) Radial growth of Colletotrichum gloeosporioides on the different media (B) Radial growth of different Colletotrichum
gloeosporioides isolates under different media (c)Radial growth of Colletotrichum gloeosporioides at different incubation
period under different media
(A)
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(B)
(C)
Fig. 2. (A) Radial growth of Colletotrichum gloeosporioides on the different media pH (B) Radial growth of different
Colletotrichum gloeosporioides isolates under different media pH (C) Radial growth of Colletotrichum gloeosporioides at
different incubation period under different media pH
Table 5. Sporulation of different Colletotrichum gloeosporioides isolates under different media
Media Cg 1 Cg 19 Cg 30 Cg 62 Cg 72 Mean
PDA 6.0000 5.00 1.3300 7.0000 8.0000 5.46
OMA 3.3300 37.00 81.0000 56.0000 73.3300 50.13
CDA 31.6600 103.00 14.0000 33.6600 22.0000 40.86
CRBA 2.0000 67.00 56.0000 2.3300 41.0000 33.66
CMA 23.0000 5.00 20.0000 21.0000 43.0000 22.40
MEA 20.3300 12.00 3.0000 18.0000 137.6600 38.20
PCA 67.3300 30.33 27.0000 56.0000 90.3300 54.20
V-8 4.000 6.00 12.0000 18.0000 3.0000 8.60
MCDA 3.0000 19.00 4.0000 2.3300 90.3300 23.73
CMPYA 34.0000 51.33 3.0000 12.0000 10.0000 22.06
PA 4.0000 2.33 3.6600 4.0000 5.3300 3.86
LA 3.0000 3.00 2.3300 3.0000 5.0000 3.26
YDA 3.0000 8.00 5.3300 8.0000 2.0000 5.26
CA 21.0000 13.33 2.3300 3.3300 3.0000 8.60
Mean 16.1100 25.88 16.7800 17.4700 38.1400
Factors C.D. SE(d) SE(m)
Media 0.7401 0.3723 0.2632
Isolates 0.6413 0.3116 0.2217
Media x isolates 1.6426 0.8208 0.5814
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(A)
(B)
(C)
Fig. 3. (A) Radial growth of Colletotrichum gloeosporioides on different temperature regimes (B) Radial growth of different
Colletotrichum gloeosporioides isolates under different temperature regimes (C) Radial growth of Colletotrichum
gloeosporioides at different incubation period under different temperature regimes
Fig. 4. Dendrogram generated on the basis of different media, media pH and temperature regimes data by Un-weighted Pair-Group
Arithmetic Mean (UPGMA) method from the cultural variability of five Colletotrichum gloeosporioides isolates
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Sporulation was highest at media pH 5.5 in all the
isolates followed by media pH 6. However, as media pH
increased the sporulation was increased consistently in
all the test isolates (Table 4).
3.3. Effect of Temperature Regimes on Growth
and Sporulation of C. Gloeosporioides Isolates
It was observed that growth of mycelia in different
isolates significantly varied with temperature regimes
and incubation period. Higher (53.17 mm) radial growth
of mycelia was observed at 28°C while lesser growth
(28.03 mm) was at 12°C (Fig. 3A). Among the isolate
Cg 62 showed highest growth (43.36 mm) than other
isolates, whereas least growth (40.07 mm) was observed
in Cg 19 (Fig. 3B). A significant effect of temperature x
isolates interaction on the radial growth of mycelia was
noticed where in maximum growth was recorded with
Cg 62 isolate followed by isolate Cg 72 at 28°C.
The radial growth of mycelia increased with
advancement of incubation period of the culture (Fig. 3C).
The interaction between temperature and incubation period
of culture also showed significant effect on mycelial
growth. Statistically significant average growth (82.24 mm)
was observed at 28°C after 7th days of incubation.
It is evident from the data that interaction between
isolates and incubation period exhibited significant
difference in mycelial growth of all the isolates.
Maximum radial growth was recorded at 7th days of
incubation with Cg 62 isolate (69.80 mm) while
minimum was with Cg 19 isolate (18.57 mm) at 3rd day
of incubation. Radial growth of mycelia was
significantly influenced by interaction of temperature x
isolates x incubation period. Higher radial growth was
observed with Cg 62 isolate (84.60 mm) followed by Cg
72 isolate (84.20 mm) at 7th day of incubation in 28°C.
Temperature showed marked effect on the
sporulation in the culture of test isolates. The best
sporulation was recorded (125×103 mL1 of culture
suspension) in the culture of Cg 62 isolate at 28°C.
Although, it was observed that sporulation in all the
culture of test isolates increased in temperature regime
upto 28°C, but rate of sporulation decreased when
temperature regime of the culture increased from 28° to
32°C during experimentation (Table 5).
The dendrogram profile showed that the isolates were
grouped into two major cluster, sharing 72% maximum
similarity. The entire two cluster consisted test isolates
collected from different state showed higher degree of
cultural variability. Avery close association was found
between isolates Cg 1 and Cg 30 collected from Chittoor
(Andhra Pradesh) and Lucknow (Uttar Pradesh)
respectively in the first cluster. Second cluster contains
Cg 19, Cg 62 and Cg 72 collecting from Rewa (Madhya
Pradesh), Muzaffarpur (Bihar) and Dapoli (Maharashtra)
respectively showing more than 56% similarity with
each other (Fig. 4).
4. DISCUSSION
Colletotrichum gloeosporioides, a filamentous
fungus, causing anthracnose disease in fruit crops, is
reported to exhibit different requirements of nutrients
and optimum conditions either for growth or sporulation
(Shin et al., 2000). There is, therefore, a need to study
these parameters for mango anthracnose pathogen in
order to establish the survivability of C. gloeosporioides
in soil (Green, 1994). The present study has focused on
resolving these issues pertaining to optimum conditions
for growth and sporulation of C. gloeosporioides.
Growth of mycelium and sporulation are influenced
by the medium, pH and temperature (Kumara and Rawal,
2008). These factors independently and or in
combination have positive and negative effects in most
of the fungi have been reported by several workers.
Media containing carbohydrates, lipids, proteins and
elements are basic requirements and needed by the
microorganisms as these nutrients provide energy for
biosynthesis and cell maintenance (Hilton, 1999).
Production of biomass in fungi and growth-associated
products requires nutrient-balanced media (Hilton,
1999). Some dimorphic fungi require optimal nutrition to
produce high biomass, but for sporulation require
nutritionally poor media which trigger differentiation of
conidia from vegetative growth (Vega et al., 2003).
The effects of medium composition, concentration
and temperature on SCC and microcycle conidiation by
C. gloeosporioides were studied on solid media. Among
the evaluated media, MEA was found to be most suitable
over other media which showed higher growth of the
respective isolates, corroborating with the results
(Sudhakar, 2000; Rani and Murthy, 2004). In the other
study, it have been noticed that sporulation of C.
gloeosporioides isolated from capsicum was maximum
in oat medium at 25°C (Mello et al., 2004). Earlier,
reports showed that PDA and CWA was ideal medium
for growth of C. gloeosporioides, as the coconut watery
endosperm contains considerable amounts of lipids
(1.26%), proteins (2.1%), carbohydrates and minerals
suitable for maximum growth (Santoso et al., 1996). In the
line of above reports also suggested CWA suitable medium
for the growth of C. gloeosporioides (Marikar, 2009).
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In a study on growth of C. gloeosporioides under in
vitro conditions, maximum growth was obtained after
10th day of incubation on potato dextrose broth, with
optimum temperature in the range of 25-35°C (Hegde et al.,
1990). Temperature affects almost every function of the
fungi including sporulation (Lilly and Barnett, 1951). A
temperature range of 15-35°C was also suggested to be
most suitable for maximum sporulation (Sattar and Malik,
1939). Meanwhile, 15-20°C favoured the conidia
formation by C. lindemuthianum in culture (Mathur et al.,
1950). Further they reported that the sporulation was less
at 25°C and ceased at 30°C. Ideally, 20 and 25°C was
reported as the most favorable temperature for colony
growth and sporulation in many fungi. Recently,
maximum growth of C. gloeosporioides isolates (from
Dapoli, Hessarghatta and Tumkur) at 28°C was identified
while, 30°C supported growth of Hassan and Raichur
isolates (Sangeetha and Rawal, 2009). Earlier, recorded
maximum growth of different isolates of C.
gloeosporioides at a temperature range of 25-30°C in the
mango but sporulation was at an optimum range of 25-28°C
(Sangeetha, 2003). In a study C. gloeosporioides produced
maximum radial mycelial growth at 25°C after 6 days
(Prabakar et al., 2003). But in the present study, maximum
growth was achieved on 7th day after inoculation.
In the present study, maximum sporulation was
observed at 28°C in all five C. gloeosporioides isolates
which was in conformity with the reports (Banik et al.,
1998). The sporulation of fungi of the genus
Colletotrichum is favored by temperatures in the range of
20-24°C, while temperatures above 30°C may have an
inhibitory effect, in total agreement with the results
obtained in the present study (Slade et al., 1987). The
isolates of Colletotrichum respond differently in their
growth and sporulation when exposed to different
temperature conditions (Jayasinghe and Fernando, 1998).
The pH of the medium determines the rate and
amount of growth including many other life processes of
fungi (Lilly and Barnett, 1951). A medium with a
specific pH which favours the growth but be unfavorable
for sporulation or other processes. A medium having pH
values between 5 and 6 at the time of inoculation was
suitable for sporulation in most fungi which are also in
accordance with the present study (Lilly and Barnett,
1951). According to them, fungi generally tolerate more
acid than alkali. Similar observations were also recorded
by some other workers with various species of
Colletotrichum (Naik et al., 1988). Similarly, the optimal
growth pH of 6, are in agreement with temperature and
pH optima reported for this species (Wastie, 1972). In
papaya fruit crop C. gloeosporioides grew well in a
medium of pH 5 (Kumara and Rawal, 2008).
5. CONCLUSION
Isolates of C. gloeosporioides have shown
differential response for the parameters viz. media, media
pH and temperature regimes in respect of growth and
sporulation. The optimum temperatures for maximum
growth of C. gloeosporioides were 28°C followed by
32°C with 6.0 media pH. Thus, the C. gloeosporioides
pathogen can grow maximum under the temperature
ranging 28 to 32°C with media pH of 5.5 to 6.0. Thus it
may be concluded that the temperature and media pH are
the critical factors for the growth of pathogen, which
might be the main reason for the expression of mango
anthracnose symptoms under field conditions in the
Northern parts of India.
6. ACKNOWLEDGEMENT
Researchers are thankful to Director of Central
Institute for Subtropical Horticulture (CISH), Lucknow
for providing necessary research facilities and research
grant in the form of ICAR-SRF.
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Kopyor coconuts (matured coconuts with broken meat particles in the watery endosperm due to abnormal formation of the kernel during the development of the fruits) were analysed for their proximate composition, minerals, vitamins, dietary fibres, sugars, organic acids, fatty acid composition and amino acid profile. The chief constituent of kopyor water was sucrose (about 92% of the total sugar); in contrast, the young or normal-mature nut water contained glucose and fructose as the main sugars. α-Tocopherol was detectable in the kopyor water. Total amino acid content of kopyor water was higher than that of the young or normal-mature water. Like the young or normal-mature nut water, kopyor water seemed to be a good source of dietary minerals, with potassium as the predominant one.The relatively high contents of sucrose, glucose, fructose, citric and malic acids might contribute to the deliciousness of kopyor meat. The lipid content of kopyor meat was lower than that of the normal-mature meat, but fatty acid profiles were similar.
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Problem statement: Sorghum anthracnose ( Colletotrichum sublineolum ) is a highly variable pathogen and new sources of host plant resistance are required for the development of resistant sorghum varieties. Germplasm collections are an important source of host plant resistance and screening germplasm will be essential to identify new sources of resistance. Approach: The sorghum ( Sorghum bicolor ) collection from the Mopti region of Mali was inoculated with Colletotrichum sublineolum and evaluated for foliar anthracnose disease response in Isabela, Puerto Rico during the 2004 and 2005 growing seasons using a partially balanced lattice design with three replications. Results: A resistant response was observed for 45 of the 97 accessions in the collection and mean infection severity for the 52 susceptible accessions was 27.6%. An association was observed between resistance and the administrative district where the germplasm was collected. More than 50% of the accessions from the Bandiagara and Bankass districts showed a resistant response. The lowest frequency of resistant germplasm was observed for the Mopti district with 25% of the accessions showing a resistant response. The susceptible accessions from the Mopti district, however, showed the lowest mean infection severity. Approximately 44% of the accessions from the Douentza district showed a resistant response with the susceptible accessions showing the highest mean infection severity. These results suggest an association between annual rainfall and anthracnose resistance, with sorghum accessions from drier regions showing greater susceptibility. Anthracnose resistance also showed an association with sorghum race classification and race guinea accessions were more frequently resistant as compared to accessions classified as race durra or durra-bicolor. Conclusion: The results indicated that anthracnose resistant sorghum germplasm is frequent in the Mopti region of Mali and that ecogeographic origin and sorghum characterization information can be used to aid in germplasm selection or germplasm acquisition to identify anthracnose resistant sources.
Article
Quick decline disease causes collapse of healthy mango plants within a few days. The menace has been observed in the entire mango growing areas of Pakistan. The leaf samples of healthy and diseased mango plants of two major varieties viz. chaunsa and langra were analyzed to asses the variations in the contents of sugars, free amino acids and metals in relation to quick decline. In general, the contents of total sugars, reducing sugars, non-reducing sugars, free amino acids and minerals were found to be decreased in the diseased leaves as compared to that of healthy leaves of both the mango varieties. Quantitative analysis of macro and micro-nutrients was carried out by atomic absorption and flame emission spectrometry to find the status of these elements in healthy and diseased leaves. Mg contents were found to be decreased in the diseased leaves of both the mango varieties whereas Cu and Zn contents were found to be increased. No consistent trend was observed in case of other metals.
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An alternative culture medium to grow Colletotrichum gloeosporioides on coconut watery endosperm agar and broth was developed. The medium is based upon the watery endosperm from Cocos nucifera, which contains nutrients promoting mycelial growth and sporulation. Sucrose (40%) further enhanced fungal growth in the broth. Dilution of watery endosperm reduced mycelial growth rate by 50%. The coconut watery endosperm is cheaper and more easily available than commercial potato dextrose agar medium in tropical regions.