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Since the accurate identification of the causal agent of plant diseases is the key information in the determination of control measures, the results of this study will be valuable for that purpose. The objectives of the study presented here was to determine what species of were associated with diseased taro corm in some subdistricts in Bogor area and the pathogen host range. This information would be valuable and may provide some understanding for strategic management of the disease. . Preliminary surveys were conducted to observed characteristic, above or below grown symptoms, and the most common cultivars planted by farmers in surveyed areas. Samples of diseased corms were surface disinfected with 70 % alcohol, and incubated in moist chamber to induce the fungal sporulation. Nine subdistricts, representing the large taro-growing areas in Bogor were sampled (Table 1). Forty individual diseased plant samples were collected from eighteen villages of nine subdistricts in Bogor area. One or both of two cultivars, and/or , were sampled depending on the planted cultivars during the study. . Individual diseased corms were surfaced disinfected by wiping with 70 % alcohol wetted cotton, sliced transversally into 5 mm thickness with sterilized knife, and then cut in small cubes (5 mm x 5 mm x 5 mm) between the rotten and healthy tissue. Five to ten pieces of the tissue were dipped in 10 % household bleach (0.525 % sodium hypochlorite) for 10 sec, rinsed in sterilized distilled water, and blotted dry on tissue paper. Pieces were placed onto potato dextrose agar (PDA) mixed with two drops of 20% lactic acid per petri dish and incubated at room temperature (± 27 °C) under near ultra violet (366 nm) with a 12 h photoperiod. After 5 days incubation, upon small pieces of colonised agar from actively growing margins where grew from the tissue were sub-cultured onto PDA. Single spore cultures Fusarium oxysporum Fusarium Bentul Hijau Fusarium MATERIALS AND METHODS Field Survey Isolation. Field Survey were established onto water agar, sub-cultured again onto PDA, and 7 to 10 days later. Identification based on conidial size, conidiogenesis, conidial form, chlamydospores formation were determined on cultures grown on carnation leaf agar (CLA) medium, and then identified according to the taxonomic system outlined by Leslie and Summerell (2006). The experiment was performed on two taro cultivars, cv. Bentul and cv. Hijau commonly planted in Bogor. The corms harvested from healthy individual plants of taro in commercial farmer field were surface disinfected with 70 % alcohol, and sliced transversally into 1.5 cm thickness with sharp sterilized knife. Each isolate tested were cultured on PDA for 7 days in room temperature under NUV with 12 h photoperiod were plugged with cork borer (Ø 5 mm) from actively growing colonies. Inoculation was performed by putting plugs of each isolate on the centre of 3 taro corm slices, in which the fungal colony faced the corm t i s s u e s . Ta r o c o r m s l i c e s i n o c u l a t e d w i t h PDA plugs only were used as control. The treated taro corm slices were placed in plastic box moist chamber by placing cotton wetted with sterile water, covered with plastic wrap, and incubated in dark room. Daily observation up to about 5 days after inoculation was done to determine the presence of rotting symptom and incubation period. Pathogenicity tests was also performed using the whole unsliced corms with the same inoculation method as mentioned previously, except for the observation period, which was conducted up to 2 weeks after inoculation. Taro slices cv. Bentul was prepared with the methods as mentioned in the previous experiment and inoculated with conidial suspension, either in singly or combination of species recovered. Conidial suspensions of and were prepared by adding sterile distilled water to 7 to 10 day's old cultures rubbing the culture surface with a sterile glass spatula, shaking thoroughly, and filtering through four sterilized layers of cheesecloth. The conidial suspensions were adjusted into 1.25 x 10 mL by means of bright line haemocytometer. The conidial suspension (0.05 mL) of fungus cultures were inoculated singly and in combination onto taro corm slices using micropipette, and placed in plastic box moist chamber, covered with plastic wrap and incubated in the dark at room temperature (± 27 °C). Five days after inoculation rotting diameter on 6 corm slices of each treatments was observed. Fusarium F. solani F. oxysporum Inoculation on Sliced Taro Corm. Inoculation on the Whole Unsliced Taro Corms. Inoculation with Single and Combination of Isolates on Sliced Taro Corm. 6 -1 Volume 5, 2011 Microbiol Indones 133 Subdistrict Number of individual plant samples Ciomas Sukaraja Bogor Timur Kemang Bogor Selatan Cijeruk Ciawi Bogor Barat Dramaga 12 4 2 3 4 4 3 4 4 Total 40 Table 1 Number and places of diseased plant samples collected Inoculation on Vegetative Propagation of Taro. Host Range Tests Inoculation on Sliced Edible Corm. Inoculation on Ornamental Inoculation on Legume Crops RESULTS Field Observation. Two cultivars of taro (cv. Hijau and Bentul) were used in the experiment. The planting medium containing soil and goat manure (4:1 v/v) was twice heat sterilized in two consecutive days, each for 30 min at 120 °C using autoclave. Conidial suspensions of was prepared using the same method as mentioned previously Ten healthy plants were selected for freedom from disease, washed the attached soils with tap water, dipped in diluted commercial clorox solution (0.525 % sodium hypochlorite) for 60 sec, and then rinsed with sterile water. Each set of test plants were dipped in conidial suspension of each isolates for 24 h, grown in pots containing about 2 L prepared medium, and fertilized with N:P:K (15:15:15) 3 weeks after planting. Control plants were inoculated with sterile water only. Development of disease symptoms was studied during 3 months. . This experiment was performed on other edible and ornamental and commonly legume crops planted by taro farmers, as follows: giant taro ( ), blue taro ( ), Chinese evergreen ( ), dumb cane ( ), angel wings ( ), nephthytis ( ), long yard bean ( ), and French bean ( ). Sliced edible corm and inoculation procedure was performed using the same methods as mentioned in previous experiment. . Some species of ornamental ( and ) were used in the experiment. Inoculation was performed on vegetative propagation using dipping method with conidial suspensions as described previously. . Long yard bean and French bean commercial seeds was first surface disinfected by dipping in the diluted commercially clorox for 60 sec, rinsed in sterile water for 2 times, and then germinated in plastic box contained heat sterilized sand. Germinated seeds were soaked in conidial suspension (1.25 x 10 mL ) for 24 h and planted in plastic pots contained heat sterilized planting medium as mentioned in previous experiment. Development of disease development was observed once a week for 2 months. Foliar symptoms include leaf curling or crinkling, leaf browning from the edge, chlorosis, shrivelling and withering of petiole, was the distinct easily detected in the fields. During the field observation, we found 2 type of above ground Fusarium species Fusarium Araceae Alocasia macrorhiza Xanthosoma violaceum Aglaonema commutatum Dieffenbachia seguine Caladium bicolor Syngonium podophyllum Vigna ungiculata Phaseolus vulgaris Araceae Araceae A. commutatum, D. seguine, C. bicolor, S. podophyllum Araceae Araceae 6 -1 symptoms, that were plant suddenly died without changing of leaf colours (Fig 1a), and preceded with leaf yellowing (Fig 1b). Advanced disease development caused wilting, stunting of individual plants which eventually die. When the diseased plant was cut transversally, an internal symptom includes vascular discoloration and corm rotting appeared (Fig 1c and 1d). In severely infected areas, many cluster-stunted plants showed the empty hollow patches (Fig 1e). Diseased plant was easily pulled out when corm have already completely rotted. The similar symptoms as observed in this study have also been described on taro in Japan (Nishimura and Kudo 1993) and on konnyaku ( ) (Shibata and Aoki 1994). , From several observations in preliminary studies with different individual diseased plant samples incubated in moist chamber, was consistently isolated. Two species of were found among 40 cultures recovered from taro corm showed wilting and/or chlorotic symptoms in above ground from 9 different sub districts in Bogor, West Java (Table 2). Two species of identified as and were recovered from 40 samples associated with taro corm rot disease in 9 different subdistricts in Bogor-West Java. was observed as the predominant species recovered from 70% of the total diseased plant samples. Both species of species were recovered from individual plant samples showing sudden death symptoms above ground, meanwhile only was detected from diseased plants with leaf yellowing symptom. The 2 species of recovered were identified as and based on their specific characters, especially microconidiophore and the formation of microconidium in false heads with long or short monophialides, respectively (Fig 2a-b and 2e-f). These 2 species, and comprised 70% and 30% of the total samples, respectively (Table 2). Colonies of identified in this study was fast growing on PDA after 2-3 days, white or cream with spares mycelium. Numerous micro- and macroconidia were formed on CLA medium. Microconidia produced on elongate monophialide conidiogenous cells (Fig 2b); ellipsoid or reniform with 0 or 1-septate (Fig 2c), 8 -16 x 4-8 µm, and formed in false heads (Fig 2a). Macroconidia wide, straight, stout with blunt apical cell, basal cell poorly developed 3-septate, 27-39 µm x 4-7 µm (Fig 2d). Chlamydospores formed abundantly, either terminally or intercalary, and rapidly within 2 weeks on CLA, 3.9-7.8 µm in diameter (Fig 2e). Amorphophallus konjach Fusarium Fusarium Fusarium F. solani F. oxysporum F. solani Fusarium F. oxysporum Fusarium F. solani F. oxysporum F. solani F. oxysporum F. solani Pathogen Isolation, Identification and Pathogenicity Test on Taro. 134 W SIDODO AND UPRAMANA Microbiol Indones d h 10 µm10 µm75 µm 50 µm a b c 20 µm 5 µm 30 µm 10 µm e g i Fusarium solani Fusarium oxysporum f j 10 µm 10 µm Fig 2 Microscopically characters of and associated with taro corm rot based on microconidium on conidiophore, microconidium and macroconidium size on CLA medium. a, Falsehead of with long microconidiophore; b, Microconidiophore; c, Microconidia; d, Macroconidia; e, Chlamydospores; f. Falsehead of with short microconidiophore; g, Microconidiophore; h, Microconidia; i, Macroconidia; j. Chlammydospores Fusarium solani F. oxysporum F. solani F. oxysporum Volume 5, 2011 Microbiol Indones 135 b c d e a Fig 1 Variation of field symptoms of taro corm rot. a, Wilting and leaf curling symptom; b, leaf chlorosis; c, corm and leaf petiole discoloration; d, corm rotting and hollowing; e, cluster symptom in the field (circled). Pure cultures of the other suspect pathogen identified as was fast growing, mycelia floccose with range in colour from pale to dark violet on PDA. Micro- and macroconidia abundantly produced on CLA medium. Microconidia abundant, formed in falsehead on short lateral monophialides (Fig 2f and 2g), ellipsoidal to cylindrical, straight or kidney shape (Fig 2h), 8-15 µm x 4-8 µm. Macroconidia straight to slightly curved, relatively slender than apical cell tapered or curved F. oxysporum F. solani, with a slight hook, 27-39 µm x 4-5 µm (Fig 2i). Chlamydospores abundantly produced on CLA medium within 2 weeks after inoculation, formed either terminally or intercalary in hyphae, with diameter in range of 3.9-7.8 µm (Fig 2j). Both species was recovered from the individual plant samples showing sudden death symptom on above ground, while isolation from the diseased plant samples with leaf yellowing only was observed. Both species Fusarium F. oxysporum Fusarium Table 3 Pathogenicity of single and mixed inoculation with and on cv. Bentul F. solani F. oxysporum Fusarium species Corm rotting diameter (cm)* F. solani F. oxysporum F. oxysporum + F. solani 4.02 ± 0.79 a** 3.80 ± 1.02 a 4.56 ± 1.07 a *5 days after inoculation, **not significantly different ( )P=0.05 a b c Fig 3 Inoculation test of and on taro (top), giant taro (middle), and blue taro (bottom) using sliced corm. a, ; b, ; c, non-inoculated F. solani F. oxysporum F.solani F. oxysporum 136 W SIDODO AND UPRAMANA Microbiol Indones Table 2 Distribution and composition of species associated with corm rot of taro in Bogor, West Java - Indonesia Fusarium Subdistrict Number of samples Number of Fusarium species recovered F. solani F. oxysporum Ciomas Sukaraja Bogor Timur Kemang Bogor Selatan Cijeruk Ciawi Bogor Barat Darmaga 12 4 2 3 4 4 3 4 4 7 3 2 3 3 3 2 2 3 5 1 0 0 1 1 1 2 1 Total isolates 28 12 Proportion (%) 70 30 were also pathogenic and caused rotting on either cv. Bentul or cv. Hijau. Mixed inoculation with both species of although tended to show larger rotting on sliced taro corm than single treatment, but was not significantly different (Table 3). In the inoculation to whole unsliced taro corm assay, corm rotting were formed after 2 weeks incubation period. Corm surface showed water soaking symptom, and when peeled rotting have spread around the inoculation site in 3 mm depth. Unfortunately, Fusarium the pathogenicity test by inoculation on vegetative propagation of taro did not show any specific above ground symptoms in 3 months after inoculation, however, corms rotting have slightly occurred and the 2 species were reisolated. Rotting on sliced giant taro ( ) and blue taro ( ) only occurred when inoculated only by (Fig 3). Three months after single inoculation with or on taro vegetative propagation, all treated plants did not show any above ground symptoms, but slight rotted on corm was formed. Above ground symptomatic also were not formed when both species inoculated onto long yard bean, bean, and some ornamental : chinese evergreen, dumb cane, angel wings, and . Although a number of species, including and are commonly known as saprophytes (Summerell 2003) from diseased roots and stem bases, our study showed that both species were capable of causing rotting symptoms on sliced taro corm. Unfortunately, inoculation assay on vegetative propagation of taro did not show any above ground symptoms after 3 months incubation period. We predicted that, it might need more than 3 months of incubation for the species to initiate the above ground symptoms. In our interview with the farmers during the field survey, the above symptoms could be detected at least 5 months after planting when they use the healthy propagations. Generally, is well known as a causal agent of wilt diseases of over 100 cultivated plant species, including tomato, potato, sugarcane, bean, cowpea, date and oil palm, as well as cooking and dessert bananas, but rotting symptom caused by this species was also reported on various plant species, such as rose (Barguil 2009), saffron (Di Primo and Cappelli 2000), and amaranths (Chen and Swart 2001) as also observed in our study. These 2 species were also revealed as causal agents on other plant, calla lily, with dwarfism, wilting, damping off, and tuber dry rotting symptoms (Ciampi 2009). Meanwhile tuber rot incited by was reported as the most important disease that affects ornamental caladium (Knauss 1975). In other root crop such as cassava, numerous and diverse species of were associated with rotted cassava roots, with the 2 largest of the AFLP groups corresponds to and (Bandyopadhyay 2006). The occurrences of the same disease of taro caused by as in our study have also been reported in Hawaii (Raabe . 1981). In Solomon Islands corm Fusarium A. macrorhiza X. violaceum F. solani F. oxysporum F. solani Fusarium Araceae Syngobium Fusarium F. solani F. oxysporum, et al. Fusarium Fusarium F. oxysporum et al. Araceae et al. Fusarium F. solani Fusarium F. oxysporum F. solani et al. F. solani et al Host Range Tests. DISCUSSION Volume 5, 2011 Microbiol Indones 137 rot of taro have also been reported with various causal agents, including , and (Jackson and Gollifer 1975), while other researchers identified more specifically, that the causal agents of corm rot on other crop ( ) was f. sp. (Nieda 1985) and f. sp. (Nishimura and Kudo 1994; Shibata and Aoki 1994). The 2 species of , and , were also recovered from storage cocoyam ( spp.) showing spoilage symptoms in Nigeria and have been confirmed as causal agents of the disease in pathogenicity tests (Ugwuanyi and Obeta 1996), while in Cameroon was one of the fungal species associated with the destructive root rot disease of cocoyam (Pacumbaba 1992). was also one of the fungal consistently reisolated from the cocoyam rotting tissue arising from inoculation among 2 others fungi, and (Maduewesi and Onyike 1981). These 2 species were also confirmed have an association with and caused rotting symptom on white yam ( ) during storage (Ezeibekwe and Ibe 2010). Other researcher determined that was reported as one of the causal agents of storage rot of taro in Guam (Wall and Cruz 1991). Although the colony between and often appear similar on PDA, but these species easily distinguished by the presence of microconidia in chains for and the presence of chlamydospores and microconidia in false heads for as indicated in our study. The consistent recovery 2 species during our preliminary observation by incubating of several diseased corms in moist chamber and pathogenicity tests in this study, we think that those 2 species are the causal agents of taro corm rot disease in Bogor. Based on host range tests, only was pathogenic to all tested edible , while was more specific to . Neither nor isolated from diseased taro caused any symptoms on the ornamental and legume crops tested. It is possible that the associated with taro corm rot might be a specific pathogen to as previously described by Nishimura and Kudo (1994) as f. sp. . Cross inoculation tests of 17 formae speciales of to taro have been conducted by other researcher (Nishimura and Kudo 1994), and they proposed for the taro isolates as Schl. f. sp. Nishimura et Kudo. Other researcher reported that was determined as causal agent of cutting rot disease of and did not show any symptom when Pythium myriotylum, P. Splendens F. oxysporum Araceae Amorphophallus F. solani radicicola F. oxysporum colocasiae Fusarium F. solani F. oxysporum Colocasia F. solani et al. F. solani Botryodiplodia theobromae Sclerotium rolfsii Fusarium Dioscorea rotundata F. proliferatum F. proliferatum F. oxysporum F. proliferatum F. oxysporum Fusarium F. solani Araceae F. oxysporum C. esculenta F. solani F. oxysporum Araceae F. oxysporum C. esculenta F. oxysporum colocasiae F. oxysporum F. oxysporum colocasiae F. solani D. maculate inoculated to other members of the family ornamental plants (Chase and El-Gholl 1982). From this report, it is also possible that the recovered from our research might be a specific pathogen to members of the family edible crops as reported by Nieda (1985) on . Further inoculation assay on living taro plants are necessary to carry out to clarify this hypothesis. We thank the Quality for Undergraduate Education (QUE) Project, Department of Plant Protection, Institut Pertanian Bogor for research grant support in conducting the studies described in this manuscript. Araceae F. solani Araceae Amorphophallus rivieri ACKNOWLEDGMENTS REFERENCES Bandyopadhyay R, Mwangi M, Aigbe SO, Leslie JF. 2006. species from the cassava root rot complex in West Africa. Phytopathology 96(6):673-676. doi:10.1094/PHYTO-96-0673. Barguil BM, Viana FMP, Anjos RM, Cardoso JE. 2009. First report of dry rot caused on rose ( spp.) in Brazil. Plant Dis 93(7):766. doi: 10.1094/PDIS-93-7-0766A. Burdani DH, Supramana, Widodo. 2001. Studi lini dasar penyakit busuk umbi pada talas ( ( L.) 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Amorphophallus konjac Fusarium oxysporum Fusarium Colocasia Lasiodiplodia theobromae Fusarium proliferatum