Please use this identifier to cite or link to this item:
標題: 輸美蝴蝶蘭園之真菌相調查暨蝴蝶蘭黃葉病菌 Fusarium solani 之研究
Fungal flora in greenhouse of Phalaenopsis and the study of Fusarium solani associated with Phalaenopsis yellowing leaf disease
作者: 陳人瑋
Chen, Jen-wei
關鍵字: Phalaenopsis;蝴蝶蘭;fungal diversity;Fusarium solani;inoculation;molecular phylogenticity;真菌相;Fusarium solani;接種試驗;分子親緣性分析
出版社: 植物病理學系所
引用: Albertini C, Gredt M, Leroux P, 1999. Mutations of the β-tubulin gene associated with different phenotypes of benzimidazole resistance in the cereal eyespot fungi Tapesia yallundae and Tapesia acuformis. Pesticide Biochemistry and Physiology 64, 17-31. Anderegg RJ, Biemann K, Buechi G, Cushman M, 1976. Malformin C, a new metabolite of Aspergillus niger. Journal of the American Chemical Society 98, 3365-70. Aoki T, O'Donnell K, Homma Y, Lattanzi RA, 2003. Sudden-death syndrome of soybean is caused by two morphologically and phylogenetically distinct species within the Fusarium solani species complex — F. virguliforme in North America and F. tucumaniae in South America. Mycologia 95, 660-84. Arya A, Arya C, 2004. Aspergillus terreus - a new fruit rot pathogen of aonla. Journal of Mycology and Plant Pathology 34, 154-5. Bartmańska A, Dmochowska-Gładysz J, 2006. Transformation of steroids by Trichoderma hamatum. Enzyme and Microbial Technology 40, 1615-21. Benyon F, Summerell BA, Burgess LW, 1996. Association of Fusarium species with root rot of Cymbidium orchids. Australasian Plant Pathology 25, 226-8. Chang, CA. 2004. Orchid virus diseases in Taiwan and their control strategies. Proceedings of 8th Asia Pacific Orchid Conference, 343-56. Chen CW, Huang JH, Hsieh TF, 2008. The disease of phalaenopsis caused by Fusarium spp. Plant Pathology Bulletin 17, 78. (abstract in chinese) Chen JW, Huang JS , Chung WC, Chung WH, 2009. Molecular characteristic and pathogenicity of Fusarium solani in Phalaenopsis. Plant Pathology Bulletin 18, 86. (abstract in chinese) Cheng ZH, Shen YJ, 2008. Isolation of Alternaria porri (Elliott) Cif. and Study on Its Culture Conditions. Actaagriculturne Boreali-occidentalis Sinica 17, 274-8. Chin KM, Chavaillaz D, Kaesbohrer M, Staub T, Felsenstein FG, 2001. Characterizing resistance risk of Erysiphe graminis f.sp. tritici to strobilurins. Crop Protection 20, 87-96. Chu CL, Chien CC, 1973. Physiological Study of Curvularia lunata and Its Pathogenicity to Rice Plant. Journal of Agricultural Research of China 22, 213-20. Darvas JM, Kotze JM, Wehner FC, 1987. Pathogenicity of fungi causing pre- and postharvest diseases of avocado fruit. Phytophylactica 19, 489-93. Erhardt FA, Stammen S, Jördening HJ, 2008. Production, characterization and (co-)immobilization of dextranase from Penicillium aculeatum. Biotechnology Letters 30, 1069-73. Farr DF, Bills GF, Chamuris GP, Rossman AY, 1989. Fungi on plants and plant products in the United States. St. Paul, MN, USA: APS Press. Felsenstein J, 1978. Cases in which parsimony and compatibility methods will be positively misleading. Systematic Zoology 27, 401-10. Fletcher JT, 1994. Fusarium stem and fruit rot of sweet peppers in the glasshouse. Plant Pathology 43, 225-7. Grünwald NJ, Coffman VA, Kraft JM, 2003. Sources of resistance to Fusarium root rot in the Pisum core collection. Plant Disease 87, 1197-200. Gugnani HC, Okeke CN, Sivanesan, N, 1990. Curvularia clavata as an aetiological agent of human skin infection. Letters in Applied Microbiology 10, 47-9. Huang JW, Sun SK, 1997. The Genus Fusarium from Taiwan. Taiwan, ROC: Shih Way Publishers. Ichikawa K, Saito H, 1998. Occurrence of dry rot on Cymbidium caused by Fusarium solani. Proceedings of Kanto-Tosan Plant Protection Society 45, 119-21. Ishii H, Fraaije BA, Sugiyama T, Noguchi K, Nishimura K, Takeda T, Amano T, Hollomon DW, 2001. Occurrence and molecular characterization of strobilurin resistance incucumber powdery mildew and downy mildew. Phytotpathology 91, 1166-71. Jackson CR. 1962. Aspergillus crown rot of peanuts in Georgia, seed treatment fungicides for control of seedborne fungi in peanut. Plant Disease Reporter 46, 888-92. Kim WG, Lee BD, Kim WS, Cho WD, 2002. Root rot of moth orchid caused by Fusarium spp. The Plant Pathology Journal 18, 225-7. Klich AM, Pitt IJ, 1988. A laboratory guide to common Aspergillus species and their teleomorphs. North Ryde, South Wales, Australlia: Commonwealth scientific and industrial organization. Kubicek PC, Gary EH, 1998. Trichoderma and Gliocladium volume 1. London UK: Taylor & Francis Ltd. Lai BZ, 2003. The report about Phalaenopsis's consignment and sale in Taiwan. Taiwan, ROC: Association for the Development of Chinese potted flower. Latiffah, Z, Nur Hayati MZ, Baharuddin S, Maziah Z, 2008. Vegetative compatibility group of Fusarium species associated with root and stem rot of orchid. Malaysian Journal of Microbiology 4, 49-52. Latiffah Z, Nur Hayati MZ, Baharuddin S, Maziah Z, 2009. Identification and pathogenicity of Fusarium species associated with root rot and stem rot of Dendrobium. Asian Journal of Plant Pathology 3, 14-21. Lee BD, Kim WG, Cho WD, Sung JM, 2002. Occurrence of dry rot on Cymbidium orchids caused by Fusarium spp. in Korea. The Plant Pathology Journal 18, 156-60. Leslie JF, Summerell BA, 2006. The Fusarium Laboratory Manual. IA, USA: Blackwell Publishing. Li CL eds, 2001. The Plant Disease and Pest Compendium Series- Compendium of Orchid Disease and Pests. Taiwan, ROC: Bureau of Animal and Plant Health Inspection and Quarantine, COA, EY. Liao LC, 2005. Practical Pesticides. Taiwan, ROC: TELI Chemical Co., Ltd. Lin QH, Lin GG, Huan YH, Zhang N, Liu FP, 2007. Identification and bionomics of leaf butt rot of Phalaenopsis. Chinese Journal of Tropical Crops 28, 93-7. Matuo T, Sakurai Y, 1965. Fusarium solani f. robiniae n. f., one of the causal fusaria of the twig blight of Robinia pseudoacacia. Annuals of Phytopathology Society of Japan 30, 31-6. Matuo T, Snyder WC, 1973. Use of morphology and mating populations in the identification of formae speciales in Fusarium solani. Phytopathology 63, 562-5. McClure TT, 1951. Fusarium foot rot of sweet-potato sprouts. Phytopathology 41, 72-7. Morita Y, Arie T, Kawarabayashi S, Suyama K, Namba S, Yamashita S, Tsuchizaki T, 1992. A new disease of Phalaenopsis and Doritaenopsis caused by Nectria haematococca. Annuals of the Phytopathological Society of Japan 58, 452-455. Nagayama K, Watanabe S, Kumakura K, Ichikawa T, Makino T, 2007. Development and commercialization of Trichoderma asperellum SKT-1 (Ecohope®), a microbial pesticide. Journal of Pesticide Science 32, 141-2. Nemec S, 1987. Fusarium solani association with branch and trunk cankers on citrus weakened by cold weather in Florida, USA. Mycopathologia 97, 143-50. Ninet B, Jan I, Bontems O, Léchenne B, Jousson O, Lew D, Schrenzel J, Panizzon RG, Monod M, 2005. Molecular identification of Fusarium species in onychomycoses. Dermatology 210, 21-5. O'Donnell K, 2000. Molecular phylogeny of the Nectria haematococca- Fusarium solani species complex. Mycologia 92, 919-38. O'Donnell K, Cigelnik E, 1997. Two divergent intragenomic rDNA ITS2 types within a monophyletic lineage of the fungus Fusarium are nonorthologous. Molecular Phylogenetics and Evolution 7, 103-16. Ofoegbu CAE, 1993. Root rot disease of cocoyam (Xanthosoma sagittifolium (L) Schott) in the south eastern state of Nigeria. Discovery and Innovation 5, 373-6. Okuda T, Yoneyama Y, Fujiwara A, Furumai T,1984. Penitricin, a new class of antibiotic produced by Penicillium aculeatum. The Journal of Antibiotics 37, 712-7. Oyeka CA, Gugnani HC, 1992. Skin infections due to Hendersonula toruloidea, Scytalidium hyalinum, Fusarium solani and dermatophytes in cement factory workers. Journal de Mycologie Medicale 2, 197-201. Pegg KG, Willingham SL, O'Brien RG, Cooke AW, Coates LM, 2002. Base rot of golden passionfruit caused by a homothallic strain of Fusarium solani. Australasian Plant Pathology 31, 305-6. Pérez L, Hernández A, Hernández L, Pérez M, 2002. Effect of trifloxystrobin and azoxystrobin on the control of black Sigatoka (Mycosphaerella fijiensis Morelet) on banana and plantain. Crop Protection 21, 17-23. Saitou N, Nei M, 1987. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution 4, 406-25. Sajise CE, 1988. Influence of cultivar inoculum density and plant age on the incidence of Fusarium root and stem rot in cowpea. Annuals of Tropical Research 10, 9-15. Sakurai Y, Matuo T, 1959. On the form name and race of Hypomyces solani (Rke. et Berth.) Snyd. et Hans. which is pathogenic to mulberry trees. Annuals of Phytopathology Society of Japan 24, 219-23. Sakurai Y, Matuo T, 1961. Taxonomy of the causal fungus of trunk-blight of Xanthoxylum piperitum and heterothallism in this fungus. Annuals of Phytopathology Society of Japan 26, 112-7. Samac DA, Leong SA, 1989. Disease development in Cucurbita maxima (squash) infected with Fusarium solani f. sp. cucurbitae. Canadian Journal of Botany 67, 3486-89. Schweigkofler W, O''Donnell K, Garbelotto M, 2004. Detection and quantification of airborne conidia of Fusarium circinatum, the causal agent of pine pitch canker, from two California sites by using a real-time PCR approach combined with a simple spore trapping method. Applied and Environmental Microbiology 70, 3512-20. Secor GA, Gudmestad NC, 1999. Managing fungal diseases of potato. Canadian Journal of Plant Pathology 21, 213-21. Silbernagel MJ, Mills LJ, 1990. Genetic and cultural control of Fusarium root rot in bush snap beans. Plant Disease 74, 61-6. Snyder WC, Hansen HN, 1941. The species in Fusarium with reference to section Martiella. American Journal of Botany 28, 738-42. Suga H, Hasegawa T, Mitsui H, 2000. Phylogenetic analysis of the phytopathogenic fungus Fusarium solani based on the rDNA-ITS region. Mycological Research 104, 1175-83. Tsai YP eds, 2002. List of plant disease in Taiwan. Taiwan, ROC: Taiwan phytopathlogy society. Tung HT, 1980. New Orchid Culture vol 2. Taiwan, ROC: San Min Book Co., Ltd. Tzean SS, Chiu SC, Chen JL, Hseu SH, Lin GH, Liou GY, Chen CC, Hsu WH, 1994. Penicillium and related teleomorphs from Taiwan. Taiwan, ROC: Food industry and development institute. Wahab S, Lal B, Jacob Z, Pandey VC, Srivastava OP, 1979. Studies on a strain of Fusarium solani (Mart.) Sacc. Isolated from a case of mycotic keratitis. Mycopathologia 68, 31-8. Wang YX, Jia FJ, Li Y, Huang JB, Breithaupt H, Hoog GS, 2007. Biological characteristics of the pathogen causing dark leaf spot on Atractylodes lancea. Plant protection 33, 107-10. White TJ, Bruns T, Lee S, Taylor JW, 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR Protocols , 315-22. Yang HC, 1998. The Flower Disease Guide. Taiwan, ROC: MAOLI Co., Ltd. Yoneda K, 2007. Phalaenopsis. Taiwan, ROC: National Chung Hsing University Agricultural Extension Center.
本研究自2007年4月27日至2008年1月9日為止,從臺中、彰化、雲林、嘉義、台南、高雄及屏東等共48家經農委會動植物防疫檢疫局檢查合格之輸美蝴蝶蘭園區分離到248株菌株。經形態學與分子生物學鑑定,可將248株菌株中的237株菌株鑑定為Alternaria、Annulohypoxylon、Ascomycetes sp.、Aspergillus、Bipolaris、Chaetomium、Colletotrichum、Curvularia、Exserohilum、Fusarium、Lasiodiplodia、Nodulisporium、Trichoderma及Penicillium等13屬真菌,而Fusarium屬有5種,Penicillium屬有7種,Aspergillus屬有3種,Trichoderma屬有4種, Alternaria屬有2種, Curvularia屬有2種,Bipolaris屬、Colletotrichum屬、Chaetomium屬、Exserohilum屬、Lasiodiplodia屬和Annulohypoxylon屬各1種。所有蒐集到菌株以菌絲塊接種後,依病斑直徑大小判斷,顯示248株菌株中具強毒性之菌株有62株,弱毒性之菌株138株,而無毒性之菌株有48株。將強毒性的菌株以105 conidia /ml孢子懸浮液接種蝴蝶蘭植株後,証實除F. solani菌株仍具毒性外,其餘菌株無法引起任何病徵。測試14種殺真菌劑對所分離菌株生長抑制效果,顯示50%撲克拉錳(Prochlorate manganece)可溼性粉劑對供試菌株之菌絲生長抑制效果較其他供試藥劑來得好,此外24.9%待克利(Difenoconazole)乳劑、23.6%百克敏(Pyraclostrobin)乳劑及5%菲克利(Hexaconazole)水懸劑等藥劑除對Fusarium屬真菌抑制效果較不明顯外,對其餘供試菌株皆有良好的抑制效果。比較施用殺真菌劑前後之蝴蝶蘭葉表真菌相變化,顯示百克敏、菲克利及撲克拉錳皆能降低蝴蝶蘭葉表真菌之種類與分離比率。本研究亦針對引起蝴蝶蘭黃葉病F. solani菌株的病原性及分子特性進行研究,結果得知引起蝴蝶蘭黃葉病之F. solani菌株無法引起虎頭蘭(Cymbidium hybrid)、四季蘭(Cymbidium ensifolium)、報歲蘭(Cymbidium sinense)、文心蘭(Oncidium sp.)、石斛蘭(Dendrobium sp.)及嘉德麗亞蘭(Cattleya sp.)產生病徵,此外將引起蝴蝶蘭黃葉病的F. solani菌株以孢子懸浮液澆灌方式接種於豌豆、洋香瓜和胡瓜的地基部與根部亦無法造成任何病徵,顯示引起蝴蝶蘭黃葉病之F. solani菌株可能具有寄主專一性。比較不同溫度對引起蝴蝶蘭黃葉病F. solani菌株病原性影響,結果指出當接種溫度超過20℃時,發病嚴重度高於16℃或18℃。於分子親緣性分析上,本研究增幅internal transcribed spacer(ITS)、intergenic spacer(IGS)及β-tubulin基因核苷酸序列,並利用近鄰結合法(Neighbor-joining method, NJ)和最大簡約法(Maximum parsimony method, MP) 進行分子系統親緣性分析,顯示引起蝴蝶蘭黃葉病之F. solani菌株可形成單一分子群(monophylogentic group),並被高的bootstrap值所支持(>84%),顯示引起蝴蝶蘭黃葉病F. solani菌株的分子親緣性與不同寄主來源的F. solani菌株有明顯差異。比較病原性測試及分子親緣性分析結果,初步認定引起蝴蝶蘭黃葉病F. solani菌株與其他來源之F. solani菌株有所不同。

A total of 248 isolates was obtained from 48 Phalaenopsis greenhouse including Taichung, Changhua, Yulin, Chiayi, Tainan, Kaohsiung and Pingtung. Based on morphological characteristics and ITS rDNA, 237 isolates were identified as Alternaria (2 species), Annulohypoxylon (1 species), Ascomycetes sp., Aspergillus (3 species), Bipolaris (1 species), Chaetomium (1 species), Colletotrichum (1 species), Curvularia (2 species), Exserohilum (1 species), Fusarium (5 species), Lasiodiplodia (1 species), Nodulisporium (1 species), Penicillium (7 species) and Trichoderma (4 species). All fungal isolates were examined for their virulence using mycelial disk. The inoculation results showed that 62 isolates were highly virulence (HV), 138 isolates were less virulence (LV), and 48 isolates were avirulence (AV). However, when 105 conidia/ml conidia suspension was used to inoculate Phalaenopsis, highly virulent isolates showed no symptom on Phalaenopsis except isolates of F. solani. To control the fungal pathogen, 14 different fungicides were examined for the efficacy of mycelial growth in Potato Dextrose Agar (PDA) medium. The results showed that the mycelial growth of fungal isolates could be inhibited when PDA was added with 10 mg/l Prochlorate manganese and Pyraclostrobin. Furthermore, 100 mg/l of Difenoconazole, Hexaconazole and Iminoctadine triacetate were effective in inhibiting mycelial growth of fungal isolates. However, other fungicides did not effectively inhibit mycelial growth of fungal isolates. Consequently, Prochlorate manganese, Pyraclostrobin and Hexaconazole were selected for further test on Phalaenopsis. The results demonstrated that 3 fungicides could reduce the fungal flora and population after spraying on Phalaenopsis lines of V3, V3-1 and Shiny-Yaun Golden Beauty. This work also studied the biological characters of F. solani associated with Phalaenopsis yellowing leaf disease. The isolates of F. solani caused Phalaenopsis yellowing leaf disease were to inoculate on five different Orchidaceae plants, including Cymbidium hybrid, Cymbidium ensifolium, Cymbidium sinense, Oncidium sp., Dendrobium sp. and Cattleya sp. The results revealed that F. solani obtained from Phalaenopsis didn't cause significant symptom on the test plants. Furthermore, the isolates of F. solani causing Phalaenopsis yellowing leaf disease did not cause any symptom on pea, cucumber and melon after inoculation. In this study, effect of temperature on disease severity of Phalaenopsis inoculated by F. solani was examined. The results indicated that the disease was more severe at 20℃ and 24℃ than those at 16℃ and 18℃. For molecular analysis, sequence of internal transcribed spacer (ITS), intergenic spacer (IGS) and β-tubulin gene were used to evaluate the phylogenic relationship by neighbor-joining method (NJ) and maximum parsimony method (MP). The results demonstrated that isolates of F. solani from Phalaenopsis formed an monophylogentic group and supported with high bootstrap values (>84%). Moreover, molecular phylogentic analysis also revealed that the isolates of F. solani causing Phalaenopsis yellowing leaf disease were molecular distinct from other F. solani from different plant hosts. According to the results of inoculation test and molecular phylogenicity, the isolates of F. solani causing Phalaenopsis yellowing leaf disease are significantly different from other F. solani isolates from other sources.
其他識別: U0005-1208200911433900
Appears in Collections:植物病理學系

Show full item record

Google ScholarTM


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.