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Fungus BioEkologi Antraknosa (Colletotrichum spp.) Origin Crops Papaya (Carica papaya L.

) 1

(Bioecology of anthracnose pathogen (Colletotrichum spp.) Origin of Plant Papaya (Carica papaya
L.) 2

Eryna Elfasari Rangkuti3, Widodo4, Suryo Wiyono4

ABSTRACT
Papaya has many beneficial effects on human utility that is produce food substance.
Colletotrichum spp. is one of limiting factor of papaya production that cause anthracnose. This
pathogens can infect different organs of the papaya plant, so it is Necessary to detect the presence
of the fungus in the field and to identify morphological characteristics as well as the ability to
infect in various organs of the plant. The aims of studying bioechology of anthracnose
(Colletotrichum spp.) Crops origin papaya (Carica papaya L.) in West Java is will be conducted
using methods of isolation, identification, pathogenicity test, cross pathogenicity in various
different organs of the plant papaya and the ability of transmitted in seed. This experiment uses a
completely randomized design (CRD) then conducted a further test of Duncan's Multiple Range
Test (DMRT). The expected benefits from this research are to provide scientific information in
particular Offender agricultural cultivation horticulture and ecology of the species origin papaya
fruit anthracnose fungi will be used in controlling anthracnose.

Keywords: Colletotrichum, morphology, papaya, pathogenicity, seed treatment

PRELIMINARY

Background
Papaya (Carica papaya L.) is one of the fruits are highly favored by the people, including
Indonesia, it is seen from the increasing per capita consumption of papayas from year to year.
Consumption of papaya in 2002 was nearly 2:24 kg per capita with a population in 2002 amounted
to 212 million. If the estimated 2025 total population of 315 million inhabitants, the consumption
of papaya Indonesia reached 705 600 tonnes. Indonesia ranks fifth largest papaya producer world
with a production of 621 524 tonnes (FAO 2007). The amount of production in Indonesia because
papaya papaya has some privileges compared with other types of fruit, because it is easily
cultivated, fast to produce and can be grown throughout the year. One of the limiting factors in the
increased production of both quality and quantity of papaya fruit is anthracnose pathogen that
attacks when post-harvest. Pathogens that infect the fleshy fruit is caused by the fungus
Colletotrichum spp. during the pre-harvest and remain latent infections develop into symptoms of
the disease in the post-harvest.
According to Bailey and Jeger (1992), anthracnose caused by Colletotrichum species are important
pathogens that affect the economies of tropical and sub-tropical areas that can infect the host in
such a wide range of horticultural crops and food. Within 5 years in Mexico, the spread of
anthracnose in the field or in storage can reduce yields by 50% although it has been repeated
fungicide applications. Kader (2000) also states that anthracnose caused by Colletotrichum
gloeosporioides is a major cause of yield loss on papaya fruit after harvest in California. The
disease is caused by a species of C. capsici and C. gloeosporioides in Mexico, USA, Trinidad and
Tobago (Tapia-Tussell et al., 2008; Tarnowski and Ploetz 2010; Rampersad, 2011), while in
Malaysia (Sepiah et al. 1991) report that pathogens are important causes anthracnose on papaya is
C. capsici. In Indonesia, the papaya fruit anthracnose pathogen is C. gloeosporioides (Penz) Sacc
that the perfect stage known as Glomerella cingulata (solution would et al. 1991). The symptoms
appear when the fruit is in the transport, marketing or storage while in the field is still in a state of
latent pathogens. Symptoms caused by Colletotrichum spp. initially formed small brownish-black
spots on the fruit, yellowish edges and widened in humid conditions. By the time the symptoms
last more symptomatic section will form the body of the fruit in concentric circles forming pink
spore masses (Semangun 2007), the symptoms will appear at 5 days after infection. Conidia were
spherical to elongated spherical, not septate and sometimes contain 1-2 dots of different sizes.
Fungus conidia size varies by an average of 12-16 x 4-6 μm (Singh 1991).
Relatively fast development of anthracnose disease was suspected by the air humidity is high
enough 78.5% -82.95%, rainfall of 2937.1 mm (Romlawati 1999) and the optimum temperature
for growth of C. gloeosporioides is 28 ºC (AVRDC 1988). The distribution is relatively fast
because on the outside of the spores of this fungus has an adhesive so that it can easily be attached
by sprinkling water on the target (Suryaningsih et al. 1996) so that the spores will germinate
quickly when you've found the right host.
Generally the species Colletotrichum spp. can be identified from a range of growth and
morphological characteristics such as morphology of conidia, presence or absence of seta,
fungicide sensitivity, colony color and growth rate (Adaskaveg and Hartin 1997). For that to
identify the causes anthracnose fungus to determine the similarity between plant species and the
effect of temperature on the growth of pathogens. It can be used as supporting information to
determine the appropriate method in controlling anthracnose.
Formulation of the problem
Anthracnose fungus infects papaya fruit when post-harvest but can also attack during the process
of planting in the field. Further research to explore the causes anthracnose and identification of
species from various parts of the papaya plant.

Research purposes
This study aims to identify the anthracnose pathogen in various parts of the plant papaya (Carica
papaya L.), and compare the species Colletotrichum spp. in a variety of areas that include
pathogenicity test, the effect of temperature on the growth, and test seed-borne pathogens.

hypothesis
The species of fungi Colletotrichum spp. which infects the fruit, leaves, petioles and stems are the
same. This is associated with the expression of symptoms and morphological similarity fungi on
each part of the plant in various areas as well as the differences in growth rate of pathogens on
variations in temperature, and the ability to carry over seed.

Benefits of research
This research is expected to provide scientific information in particular offender agricultural
cultivation of horticultural ecology on the type and origin of papaya fruit anthracnose fungi for the
preparation of appropriate strategies in the control of anthracnose.

MATERIALS AND METHODS


Time and Location
This study will be conducted in September 2015 until June 2016 in Plant Mycology Laboratory,
Department of Plant Protection, Faculty of Agriculture, Bogor Agricultural University. Sampling
was carried out at the Center for Tropical Horticulture (PKHT) Sand Riding, Bogor, West Java
and several gardens in Bogor papaya, papaya orchard in Kebumen, Central Java.

Tools and materials


The tools used in this study is a petri dish, the petri dish disposable, test tubes, object glass, cover
glass, erlenmeyer, measuring cup, beaker glass, cork borer, autoclave, haemositometer,
microscopes, Bunsen, laminar air flow, mikrotip, incubators, shaker, refrigerator, water bath, ose
bent, spatula, tube ependof.
Materials used are part of papaya plants such as stems, leaves, leaf stems and fruit are symptomatic
anthracnose, fruit and papaya seeds genotype Calina, isolates of Colletotrichum spp., NaOCl,
Potato Dextrose Agar (PDA), media sterile soil, 70% alcohol, sterile distilled water, spirit,
laktofenol, plastic, plastic tray, cling wrap, aluminum foil.

Method
Isolation of Colletotrichum spp.
The fungus was isolated from the stem, branches, fruit and papaya on each section showing typical
symptoms of anthracnose. Isolation of fungi carried out in the laboratory using the method of
planting media network on potato dextrose agar (PDA) and incubation at 28 ° C. Pure culture
conidia harvested after 7 days. Harvesting is done by adding sterile water to the cup so submerged
(± 18 mL). Conidia were harvested using a spatula to manufacture a single spore cultures. Single
conidia obtained culture propagated back on PDA and incubated at room dark by irradiating 12
hours and 12 hours of light under near ultraviolet rays for seven days.

Identification Causes Antraknosa on Papaya


Identification of pathogens is done using the guidelines Barnet and Hunter (1972) is through
morphological observation conidia include color, shape, and width of conidia, seta and aservulus.
Morphological observation done on pure cultures as well as signs of disease on the infected papaya
fruit.

Effect of Temperature on Growth Response Colletotrichum spp.


Isolates of Colletotrichum spp. used in this test was the result of previously identified isolates.
Isolates were used previously grown on PDA for two weeks at room temperature. Of each culture
and then made a small piece measuring 0.4 cm in diameter. Pieces cultures would then be
transferred into the new PDA media on Petri dishes of 9 cm diameter. Pieces cultured fungus were
placed in a petri dish that is at the center of the petri dish. Petri dish cultures of the fungus are
stored at various temperatures, namely 16 oC, 24 oC, 32 oC. The design used was completely
randomized design (CRD). Variables measured is the average rate of growth of fungi. Observation
diameter growth of fungus carried out since the first day of inoculation at a certain temperature for
five days in a row, do repeat 2 times (Smith and Black 1990).
Pathogens pathogenicity test Antraknosa on Papaya
Isolate pure cultures of origin papaya works in the collection are used in the pathogenicity test.
The method used is the method of injecting the papaya fruit (Swart 1999). Papaya fruit that is used
in the pathogenicity test is a genotype that is susceptible to anthracnose fruit papaya varieties
namely Calina. Papaya fruit which will be used in testing beforehand by sterilization treatment.
The surface of the fruit soaked in 2% NaOCl for 30 seconds and then rinsed with sterile water.
Inoculation point on the surface of the papaya fruit is made as much as two points per fruit. The
density of spore suspension are used for inoculation was 106 spores / mL, the number of spores is
calculated using haemositometer. Furthermore, fruit inoculated and placed in plastic tubs covered
with transparent plastic. At the four corners of the tub, put wet cotton to retain moisture, then a
plastic tub incubated at room temperature. The design is used to refer to (Kanchana-Umdokan et
al. 2004), where each isolate was tested in 3 pieces with 2 repetitions at the point of inoculation.
The test results pathogenicity of Colletotrichum spp. the papaya can be seen in Figure 1.
The variables measured were:
1. Emerging whether symptoms: observations done every day after inoculation with a view typical
symptoms of anthracnose.
2. The average incubation period: the average period of days after inoculation until the appearance
of symptoms.
3. The average diameter of symptoms: symptoms diameter was measured at seven days after
inoculation.

Figure 1. Symptoms shown in papaya anthracnose pathogenicity test (a-c) Colletotrichum


gloeosporioides, (d-f) Colletotrichum capsici, (b) the mass of conidia, (e) aservuli with seta
(Calzada et al. 2012)

Cross Phatogenicity Against Fruit, Twig, Leaf and Stem Papaya


Inoculation method is carried out by inoculation method refers to the method of attachment culture
(Serra et al. 2006). Inoculation is done by using isolated fungi were identified from various parts
of the papaya plant, then testing cross pathogenicity isolates derived from the fruit of the stem,
leaf, petiole against papaya varieties Calina susceptible to anthracnose and performed well on each
piece of papaya plants others from different regions. Pieces cultured fungus measuring 0.4 cm in
diameter attached to the surface of plant tissues. Observations were made every day until the
symptoms of anthracnose.

Capability Study Carried Seed Pathogens Colletotrichum spp.


Seed infected by anthracnose in the field are separated, terraced sterilized with NaOCl, 70%
alcohol, and sterile water. A total of 5 seeds incubated in a Petri dish containing filter paper,
incubated at room temperature with 12-hour lighting of dark and light. Symptoms appear after 7
days after incubation of seeds pathogens will be observed microscopically. To test the transmission
of the seed in the field, the seed planted in the media sterile soil and sand in the ratio 1: 1. Seed
derived from papaya afflicted by anthracnose and seeds that have been coating with conidia of
Colletotrichum spp are used as treatments while papaya seeds are derived from healthy papaya is
used as a positive control. Each treatment and control using 100 papaya seeds. ± incubated for 14
days in a greenhouse. Part of papaya plants growing in a sterile cut, after which part of the plant
back on a moistened filter paper soaked in sterile water, is observed every day until the symptoms
of anthracnose (Yesuf and Sangchote 2005)

Data analysis
The experiments were performed using a completely randomized design (CRD) and descriptive
data analysis, the results of experiments analyzed using ANOVA with SAS 9.1. The treatment that
significantly tested further by Duncan's multiple range test at 5% significance level.

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