First report of Colletotrichumspp. causing diseases on Capsicum spp. in Sabah, Borneo, Malaysia
Ha Kwan Yun 1, Abdul Hamid Ahmad 2, Sepiah
Muid 3 & Jaya Seelan Sathiya Seelan 4
1,2,4 Institute for Tropical Biology and
Conservation, Locked bag 2073, Universiti Malaysia Sabah, 88999, Kota Kinabalu,
Sabah, Malaysia
3 Department of Plant Science and Environmental Ecology, Faculty
of Resource Science and Technology, Universiti Malaysia Sarawak, 94300 Kota
Samarahan, Sarawak, Malaysia
Email:1 hky@yahoo.com ; 4 avinash80us@yahoo.com (Corresponding author)
Date of publication (online): 26
August 2009
Date of publication (print): 26
August 2009
ISSN 0974-7907 (online) | 0974-7893
(print)
Editor: V.B. Hosagoudar
Manuscript details:
Ms # o2273
Received 26 May 2009
Final received 08 July 2009
Finally accepted 21 August 2009
Citation: Yun, H.K., A.H.
Ahmad, S. Muid & J.S.S. Seelan (2009). First report of Colletotrichumspp. causing diseases on Capsicum spp. in Sabah, Borneo, Malaysia. Journal
of Threatened Taxa 1(8): 419-424.
Copyright: © Ha Kwan Yun,
Abdul Hamid Ahmad, Sepiah Muid & Jaya Seelan Sathiya Seelan 2009. Creative
Commons Attribution 3.0 Unported License. JoTT allows unrestricted use of this
article in any medium for non-profit purposes, reproduction and distribution by
providing adequate credit to the authors and the source of publication.
Author Details: Author Details: Ha Kwan Yun completed her BSc (Hons) in Conservation Biology
at the Institute for Tropical Biology and Conservation, Universiti Malaysia
Sabah, Malaysia. Her thesis was on
Morphological and Physiological Characterization of Colletotrichum spp.
Isolated from Capsicum spp. (Chillies) in Sabah.
Abdul
Hamid Ahmad is
the Director of the Institute for Tropical Biology and Conservation, Universiti
Malaysia Sabah.
Sepiah
Muidis a senior lecturer from Universiti Malaysia Sarawak. She has been working on taxonomy and
systematics of Borneon fungi and mushrooms for the past 20 years.
Jaya
Seelan Sathiya Seelanis presently working as a researcher/tutor at the Institute for Tropical
Biology and Conservation. He has been
involved in mushroom and fungi related research for the past seven years and
graduated in the field of fungal biotechnology from Universiti Malaysia
Sarawak, Malaysia.
Author Contribution: The
first author designed and conducted this study as a partial fullfilment of her
BSc (Hons) degree, while the fourth author supervised her research work and
gave technical advice. The second and third authors commented on the manuscript
and provided extensive information during research.
Acknowledgement: The authors are
grateful to Associate Professor Dr. Michael Wong, Mr. Liew Thor Seng, Mr. Lau
Foo Chwang, Madam Lam Nyee Fan, Mr. Azrie Ahmad, Mr. Cornelius, Madam Fatimah
Jumat and Miss Doreen Juhan for their kind help during this research.
Abstract: Blackish
or orange liquid-like spots were found on (n=100) fruits of chillies (Capsicum)
sold in five local markets in Kota Kinabalu, Sabah, Malaysia. Colletotrichum gloeosporioides and C. capsici were identified as the causal
agents of an anthracnose disease. This
is the first report of Colletotrichum spp. as the causal agent of
anthracnose infected chillies in Sabah.
Keywords:Anthracnose, Colletotrichum, chillies, morphology
For Figure & Images –
click here
Introduction
The genus Capsicum comprises several
species of chillies belonging to the family of Solanaceae. There are five important domesticated chilli
species i.e. C. annuum, C. chinense, C. baccatum, C.
frutescens and C. pubescens. This genus is native to the Americas
and is now cultivated worldwide (Sanogo 2003). In Sabah, chillies are planted intensively at Kundasang, Keningau and
many other areas. Chillies are important ingredients in local food and are used
regularly. Anthracnose is a common
disease attacking chillies in Sabah but the causal agents, the species of the
genus Colletotrichum have have not been identified so far. According to Poonpolgul & Kumphai (2007),
Thailand had encountered severe losses up to 80% due to a great fall in chilli
production affected by Colletotrichum spp. whereas losses which were
greater than 30% in plant production occurred in the United Stated due to
anthracnose (Howard et al. 1992; Wilson et al. 1992).
More seriously, five species of Colletotrichum,namely Colletotrichum coccodes, Colletotrichum crassipes, Colletotrichum
dematium, Colletotrichum gloeosporioides and Colletotrichum
graminicola have been reported to cause infection in humans also. These infections are keratitis following
traumatic implantation, subcutaneous and systemic infections among immunosuppressed
patients (Liesegang & Forster 1980; Liao et al. 1983; Shukla 1983;
Matsuzaki et al. 1988; Ritterband et al. 1997; Guarro et al. 1998; De Hoog et
al. 2000; Castro et al. 2001; Yamamoto et al. 2001; Fernandez et al. 2002; Cano
et al. 2004). The five species of the
genus Colletotrichum known to cause similar symptoms on chillies are C.
gloeosporioides, C. acutatum, C. capsici, C. cocodesand C. dematium (Hong & Hwang 1998; Gopinath et al. 2006). This study is aimed to report the causal
agents of anthracnose disease in chillies for the first time in Sabah, Borneo.
Material and Methods
Sampling: Samples were
collected from five local markets in Kota Kinabalu, Sabah, Malaysia, i.e.
Central Market, Menggatal Market, Donggongon Market, Lido Market and Wong Kwok
Market.
Fungi isolation: The samples were
brought to laboratory in a sealed plastic bag. Lesions were excised, surface
sterilized with 70% ethanol for 30s rinsed a few times with distilled water and
dried on filter papers. They were cut into
smaller pieces of approximately 5x5 mm size, cultured in petri dishes
containing potato dextrose agar (PDA) and incubated at room temperature, in the
dark, for 48 to 72 hr.
Morphological identification: An inoculum loop
was used to scratch the mycelium of the colonies and mounted on the slides.
Compound microscope and scanning electron microscope were used to observe the
colony colour and form, appressorium features, shape, size and colour of spores
from different isolates. Identification was made by comparing the
characteristics of fungi following Charlie et al. (2001) and Deacon (1998).
Results
One-hundred infected chillies with
symptoms of anthracnose caused by Colletotrichum spp. were collected
from the five local markets of Kundasang, Ranau areas, Papar and rural areas
(Image 1). The lesions gradually turned
darker, contained fluid as they matured. Lesions occured at different parts of
the fruits as well as panicle in different sizes.
Out of 100 chilli samples collected, 50
isolates of Colletotrichum from the lesions, were two species i.e. Colletotrichum
gloeosporioides and Colletotrichum capsici from five local markets
in Sabah. Overall, the Central Market
recorded the highest percentage of Colletotrichum spp. This was followed by the Wong Kwok Market and
Donggongon Market which documented 24% and 20% of the total number of Colletotrichumisolates, whereas the least number of Colletotrichum isolates were
from the Menggatal Market (Fig. 1). Totally 49 isolates of Colletotrichum gloeosporioides and one
isolate of Colletotrichum capsici were successfully obtained. Colletotrichum
gloeosporioides was mainly isolated from the Central Market followed by
Wong Kwok Market. In contrast, this species was rarely found from the
Menggatal Market (Table 1). Thus, the
samples indicate infection mainly by Colletotrichum gloeosporioides and Colletotrichum
capsici.
Observations of morphological
characteristics of the isolates showed similarities with published descriptions
of Colletotrichum spp. (Cano et al. 2004). However in this study, Colletotrichum
gloeosporioides strains indicated differences in growth patterns (Image 2).
The colour of conidia obtained varied
between olive and white while the reverse colour for olive colonies varied from
dark olive-grey to brown. For
white-coloured colonies, the reverse colour was whitish with a little olive
(Image 2B). The olive coloured colony
produced abundant sporulation while the white possessed sparse mycelium. Setae developed a dense, cylindrical layer
around the conidiamata of the olive-coloured colonies, which gradually turned
into dark olive-grey, as maturation progressed. White colonies retained their colour as they aged. Colonies of Colletotrichum spp. grew
at a moderate rate, 14 days to cover the whole surface of the petri dishes.
After a long-term storage, masses of conidia increased and aged into darker
grayish-olive. The conidia of Colletotrichum gloeosporioides were straight with obtuse apex, hyaline, cylindrical to
clavate and sometimes fusiform like Colletotrichum acutatum (Image
3). Clavate and lobed appresoria were
mainly present in matured colonies. The
spore measurement of Colletotrichum gloeosporioides varied
between 3.93-12.14 µm long and 1.43-2.14 µm wide.
Colletotrichum capsici
The isolates
showed grayish-olive colonies while the reverse color was darker olive (Image
4). Sporulation of this species was
sparse and the acervuli were scattered. This species formed smooth, circular
margin in the colony. The gray-whitish
mycelium of Colletotrichum capsici gradually developed from the isolates
from the second day of culture. Colour
of the colonies of Colletotrichum capsici is similar to Colletotrichum
gloeosporioides. However, both of
these Colletotrichum species showed distinct difference in conidia and
appresoria shapes. The conidia of Colletotrichum capsici were falcate in
shape while the appresoria were round to ovate in shape. The spore measurement of this species varied
between 13.21-16.21 µm long and 1.79-3.28 µm wide. The conidiophores of all the colonies
branched below the surface of media as they developed.
Discussion
In this study, the commonly available Capsicum
annuum, C. baccatum and C. frutescens collected from the
local markets in Kota Kinabalu, Sabah were studied. Occurrence of Colletotrichum spp.
induced chilli anthracnose at the Central Market and Wong Kwok Market was
higher when compared to the other three sites, most probably due to high
humidity and wetness. This is consistent
with the assumption of Arauz (2000) that excess moisture enables spores to be
released from acervuli. On the other hand, the Menggatal Market and Lido Market
smaller in size, and the stalls at Lido Market grouped into compartments
according to types of goods sold decreased the percentage of humidity reduced the occurrence of chilli
anthracnose.
Arauz (2000) stated that 20-30 0C
range is the optimal condition for Colletotrichum gloeosporioides to
form appresoria and germinate). In this
study, Colletotrichum gloeosporioides was found to be the most dominant
pathogen (98%), significantly more than Colletotrichum capsici , in
agreement with the study of Ellett (1989) and Manandhar & Hartman (1995).
This study reveals agreement with Black
& Wang (1993-2002) that Colletotrichum gloeosporiodes and Colletotrichum
capsici were mostly found in ripe chillies collected from Kota Kinabalu,
Sabah. Prusky (1996) postulated that
fruits are attacked by the pathogen early in their development. The fungi remain as germinated appressorium during quiescent
period ‘develop brown-black spots on the pericarp and soft rot in the mesocarp
when fruit ripens for harvest. Infection of chillies by Colletotrichumspp. is mainly due to ample nutrients and limited antifungal compounds formed
during fruit ripening (Prusky 1996).
Conclusion
The two species of Colletotrichum spp.,
i.e. Colletotrichum gleoesporioides and Colletotrichum capsiciwere isolated from the chilli lesions collected from local markets in Kota
Kinabalu, Sabah. Although Colletotrichum
acutatum is reported as one of the common pathogens on chilli lesions, it
was not found in this study. The conidia
of Colletotrichum gloeosporioides were straight with obtuse apex,
hyaline, cylindrical to clavate, and sometimes fusiform like Colletotrichum
acutatum. Appresoria were mainly
present in mature colonies. They were
clavate and lobed. The spore measurement for this species ranged within
3.93-12.14 µm long and 1.43-2.14 µm wide. Isolates from Colletotrichum capsici, had similar olive color of Colletotrichum
gloeosporioides,. However, they possessed falcate conidia distinct from Colletotrichum
gloeosporioides. In addition, Colletotrichum capsici formed round to
ovate appresoria. The spore measurement
of this species was 13.21-16.21 µm length and 1.79-3.28 µm width. Colletotrichum
gloeosporioides and Colletotrichum capsici grew better on Potato
Dextrose Agar (PDA) than in Malt Extract
Agar (MEA) and Yeast Extract Agar (YEA). Growth rates of Colletotrichum spp. were higher under light
exposure than in dark. The samples are
stored as stock culture in the Institute for Tropical Biology and Conservation
(ITBC), Universiti Malaysia Sabah.
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