臺灣博碩士論文加值系統

2006年於台北縣樹林鎮、永和市首先發現所種植之葉用甘藷發生植株葉片萎凋、部分莖組織褪色的病徵，嚴重者莖基部黑色軟腐、全株產生缺水枯死、根部腐敗，最後造成全株死亡。詢問當地農友得知，該病害的歷史可追溯到十年之前。經柯霍氏法則確認該病原為細菌。經形態觀察、革蘭氏染色測定、Biolog鑑定及NGM選擇性培養基鑑定，確認該病原為Erwinia chrysanthemi。該病原主要危害葉用甘藷的莖部與根部，造成莖基部黑腐與根部褐化軟腐，進而引起植株萎凋枯死，此與國外甘藷細菌性根莖腐病病徵極為相似且病原菌相同，因此稱之為葉用甘藷根莖腐病(root and stem rot)。由E. chrysanthemi引起之葉用甘藷細菌性根莖腐病為台灣之首次記錄。在品種抗性測定方面發現供試四種葉用甘藷品種中以永和種1號最為抗病，桃園2號、台農71號及永和種2號皆具感染性。以濾紙圓盤法測試市售9種藥劑在3種不同稀釋倍數下對根莖腐病菌菌株生長之抑制效果，顯示四環黴素對葉用甘藷根莖腐病之菌生長之抑制效果最佳，而嘉賜黴素對E. chrysanthemi無任何防治效果，其餘7種防治藥劑包括銅快得寧、氫氧化銅、鏈四環黴素、多保鏈黴素、鏈黴素、維利黴素、嘉賜黴素等實驗藥劑均能抑制E. chrysanthemi根莖腐病菌之生長。

Severe root and stem rot of sweet potato was first found in Yong-He and Shu-Lin, Taipei county, in Taiwan in 2006. The disease symptoms include wilting shoot, pale leaf, black stem, root rot and death. The occurrence of this disease can be traced back to at least ten years ago as described by the farmers. The causal agent of root and stem rot of sweet potato is a pathogenic bacterium according to Koch’s postulates. The pathogen was identified as Erwinia chrysanthemi based on the morphology, Gram’s stain test, Bilolog identification, and NGM differential medium test. The pathogen mainly attacked the root and stem and caused black rot of stem base, brown rot of root, and death. The symptoms were similar to those of the bacterial stem and root rot reported in United States, although in United States it majorly attacked the roots. This is the first report of this disease in Taiwan. Four varieties including “Taoyuan#2”, “Tainong#71”, “YongHe# 1” and “YongHe#2” were screened for the disease resistance “YongHe#1” had the highest resistance, while “Taoyuan#2”, “Tainong#71” and “YongHe#2” were sensitive to E. chrysanthemi. Nine bactericides at three concentrations were tested for their control effects on E.chrysanthemi by paper disc diffusion method. Among them, tetracycline has been proven to be the most effective, while oxine-copper＋copper hydroxide, cupric hydroxide, streptomycin sulfate + tetracycline, kasugamycin +copper oxychloride, thiophanate-methyl + streptomycin, streptomycin and validanmycin could inhibit the growth of E. chrysanthemi. However, kasugamycin alone was no effective to control this bacterial disease.

口試委員會審定書……………………………………………………… ...i
誌謝………………………………………………………………………...ii
中文摘要………………………………………………………………….i ii
英文摘要…………………………………………………………………..iv
第一章 緒論 (Introductuon)…………………………………………......1
第二章 前人研究 (Literature Review) ………………………………....3
一、甘藷介紹………………………………………………………….........3
二、甘藷的病害…………………………………………………………….3
(一) 國外…………………………………………………………..……3
(二) 國內………………………………………………………………..4
三、細菌性軟腐病…………………………………………………………..5
四、Erwinia chrysanthemi的特性…………………………………………….6
(一) E. chrysanthemi的分類地位………………………………………......6
(二) E. chrysanthemi的分布範圍………………………………………......6
(三) E. chrysanthemi的寄主範圍、生理特性與形態………………………...6
(四) E. chrysanthemi在甘藷上所造成的病徵……………………………….7
五、甘藷細菌性根莖腐病相關之研究…………………………………..……8
(一)發病生態…………………………………………………………….8
(二)防治方法…………………………………………………………….8
第三章 材料與方法(materials and Methods)……………………………9
一、供試植物……………………………………………………………….9
(一) 供試植物之介紹…………………………………………………….9
1. 葉用甘藷桃園2號…………………………………………………..9
2. 葉用甘藷台農71號………………………………………………....9
3. 葉用甘藷永和種1號……………………………………………......9
4. 葉用甘藷永和種2號……………………………………………....10
(二) 供試植物之栽培與管理……………………………….……..……..10
二、葉用甘藷根莖腐病之觀察與記錄……………………………….……...10
(一) 田間發病之觀察……………………………………………...……10
(二) 溫室發病之觀察……………………………………………...……11
(三) 葉用甘藷根莖腐病病徵之觀察……………………………………..11
(四) 顯微鏡下葉用甘藷根莖腐病病徵之觀察………………………….....11
(五) 病害傳染方式之初步觀察與測試…………………………………...11
三、可疑病原菌株之分離…………………………………………………..12
四、可疑病原菌之鑑定………………………………………………...…..12
(一) 真菌型態鑑定…………………………………………………......12
(二) 細菌之革蘭氏染色(Gram’s stain)…………………………………….12
(三) 細菌疑似病原之BIOLOG系統鑑定…………………………………13
(四) NGM鑑別性培養基鑑定……………………………………………13
五、分離株之病原性測定.………………………………………………….14
(一) 注射接種………………………………………………………….14
(二) 剪根接種………………………………………………………….14
(三) 剪莖接種1………………………………………………………...14
(四) 剪莖接種2………………………………………………………...15
(五) 嫁接接種………………………………………………………….15
(六) 穿刺接種………………………………………………………….15
六、葉用甘藷不同品種之抗病性測試………………………………………..16
七、防治藥劑之感受性測試………………………………………………..16
八、無菌種苗之製備………………………………………………………17
第四章 結果(Result)……………………………………………………25
一、葉用甘藷根莖腐病之觀察與記錄………………………………………25
(一) 田間主要病蟲害之觀察……………………………………………25
(二) 台大溫室中葉用甘藷重要病蟲害之觀察……………………………25
(三) 葉用甘藷根莖腐病病徵之觀察…………………………………......25
(四) 利用顯微鏡檢葉用甘藷根莖腐病之結果……………………………26

(五) 病害傳染方式初步觀察及測試結果.………………………………..27
二、可疑病原菌株之分離結果……………………………………………...27
三、可疑病原菌之鑑定…………………………………………………….27
(一) 真菌型態鑑定…………………………………………………......27
(二) 細菌菌種鑑定…………………………………………………......28
四、病原性測定…………………………………………………………...28
五、葉用甘藷不同品種之抗病性測試………………….….………………..29
六、藥劑感受性測試………………………………………………………30
七、無菌種苗之製備………………………………………………………30
第五章 討論(Discussion)………………………………...……………..63
一、葉用甘藷根莖腐病之觀察……………………………………………...63
二、可疑病原菌株之分離結果……………………………………………..65
三、病原菌之鑑定……………………………………………...…………65
四、分離株之病原性測定…………………………………………………..65
五、葉用甘藷不同品種之抗病性測試………………………………………66
六、藥劑感受性測試………………………………………………...…….66
七、無菌種苗之製備………………………………………………………67
參考文獻(References)………………………………………………...…..68

1.辛仲文、姜金龍、林維和。1996。 葉菜甘藷。少量多樣化雜糧作物栽培手冊。台灣省政府農林廳編印。
2.徐世典、張東柱、張清安、蔡進來、蔡東篡。2002。台灣植物病害名彙第四版。中華民國植物病理學會。台中。386頁。
3.許秀惠、宋秉峰、吳峻瑋、施淑晴、林俊義。2004。向日葵細菌性軸腐病之特性、品種抗性及藥劑篩選。植物保護學會會刊。46:367-378。
4.黃秀華。1994。國蘭病害之簡介。台中區農推專訊 138期。
5.賴永昌。2005。甘藷。台灣農家要覽 (一)。行政院農業委員會。57-68頁。
6.謝照申、王明果。1995。主要土壤分布與特性。台灣農業統計要覽農業篇(一)。豐年社。台北。305頁。
7.蘇宗振。2001。國產精緻雜糧專櫃系列介紹。雜糧與畜產。行政院農業委員會農糧署。台北。
8.行政院農業委員會及所屬各機關人事室。2002。臺灣農業年報。臺灣省政府農林廳編印。424頁。
9.Adaskaveg, J. E., and Hine, R. B. 1985. Copper tolerance and zinc sensitivity of Mexican strains of Xanthiminas campestris pv. vesicatoria, causal agent of bacterial spot of pepper Plant. Dis. 69:993-996.
10.Agrios, G. N. 2005. Plant Pathology. 5th ed., Academic Press, Elsevier Burlington, MA, 922 pp.
11.Barnett, H. I., and Hunter, B. B. 1973. Illustrated Genera of Imperfect Fungi. APS Press. St. Paul, Minn. 241pp.
12.Bortels, H., and Sauthoff, W. 1965. A bacteriosis of dieffenbachia schott in germany. Phytopathologische Zeitschrift 54: 285-297.
13.Bradbury, J.F. 1986. Erwinia. In: Guide to plant pathogenic bacteria, pp. 61-79. CAB International, Wallingford, UK.
14.Burkholder, W. H., McFadden, L. H., and Dimock, A.W. 1953. A bacterial blight of chrysanthemums. Phytopathology 43, 522-525.
15.Clark, C. A. 1980. End rot, surface rot, and stem lesions caused on sweet potato selections to Fusarium root and stem canker caused by Fusarium solani. Plant Dis. 70:869-871.
16.Clark, C. A., and Moyer, J. W. 1988. Compendium of Sweet Potato Diseases. American Phytopathological Society, St. Paul, MN. 74 pp.
17.Clark, C. A., Hoy, M. W., Bond, J. P., Chen, C., Goh, Y.-K., Liang, X., Liu, X., and Lotrakul, P. 1989. Report of Erwinia cartovora subsp. cartovora causing bacterial root rot of sweet potato (Ipomoea batatas) in Louisiana. Plant Dis. 82:129.
18.CMI (1967) Distribution Maps of Plant Diseases No. 425 (edition 1). CAB International, Wallingford, UK.
19.CMI (1987) Distribution Maps of Plant Diseases No. 576 (edition 1). CAB International, Wallingford, UK.
20.Collmer, A., and Keen, N. T. 1986. The role of pectic enzymes in plant pathogenesis. Annual Review of Phytopathology. 24:383–409.
21.Deboer, S. H., and Kelman, A. 1978. Influence of oxygen concentration and storage factors on susceptibility of potato tubers to bacterial soft rot (Erwinia chrysanthemi). Potato Res. 21:65-80.
22.Dickey, R. S., and Victoria, J. I. 1980. Taxonomy and emended description of strains of Erwinia isolated from Musa paradisiaca Linnaeus. International Journal of Systematic Bacteriology. 30:129-134.
23.Dickey, R.S. 1981. Erwinia chrysanthemi: reaction of eight plants to strains from several hosts and to strains of other Erwinia species. Phytopathology. 71:23-29.
24.Doi, Y., Teranaka, M., Yora, K., and Asuyama, H. 1967. Mycoplasma or PLT group-like microorganisms found in the phloem elements of plants infected with mulberry dwarf, potato withes’-broom, aster yellows, or Paulownia witches’-broom. Ann. Phytopathol. Soc. Jpn. 33:259-266.
25.Duarte, V., and Clark, C. A. 1992. Presence on sweet potato through the growing season of Erwinia chrysanthemi, cause of stem and root rot. Plant Dis. 76:67-71.
26.Fernandez-Borrero, O., and Lopez-Duque, S. 1970. Soft rot of the pseudostem of banana (Musa paradisiaca) caused by Erwinia paradisiaca n. sp. Cenicafe. 1:3-44.
27.Halsted, B. D. 1890. Some Fungous Diseases of the Sweet Potato. N. J. Agric. Exp. Stn., Bull. 76. 32 pp.
28.Harrison, H. F., Peterson, J. K., and Snook, M. E. 2006. Sweet potato storage root phenolics inhibit in-vitro growth of Erwinia chrysanthemi. Allelopathy Journal 17 (1): 81-87
29.Harter, L. L. 1913. The foot-rot of the sweet potato. J. Agric. Res. 1:251-273.
30.Harter, L. L., and Weimer, J. L. 1919. The surface rot of sweet potatoes. Phytopathology 9:465-469.
31.Harter, L. L., Weimer, L. J., and Adams, J. M. R. 1918. Sweet potato storage rots. J. Agric. Res. 15:337-368.
32.Huang, T. C., and Lee, H. L. 1988. Identification and control of soft-rotting Erwinia from Phalaenopsis. Plant Prot. Bull. 30 : 416.
33.Jackson, C. R., and G. F. Weber. 1959. Morphology and taxonomy of Alternaria cucumerina. Mycologia 51: 401- 408.
34.Kays, S. J. 1985. The Physiology of Yield in the Sweet Potato. Pages 80-132 in: Sweet Potao Products: A Natural Resource for the Tropics. J. C. Bouwkamp, ed. CRC Press, Boca Raton, FL.
35.Lauritzen, J. I. 1926. A strain of Yellow Jersey sweet potato resistant to surface rot (Fusarium oxysporum W. and C.). J. Agric. Res. 33:1091-1094.
36.Lee, Y.-A., and Cheng, P.-Y. 2006. A differential medium for the isolation and rapid identification of a plant soft rot pathogen, Erwinia chrysanthemi. Journal of Microbiological Methods 64:200-206.
37.Lelliott, R. A. 1956. Slow wilt of carnation caused by a species of Erwinia. Plant Pathology 5:9-23.
38.Liao, C.-H., McCallus, D. E., and Wells, J. M. 1993. Calcium-dependent pectate lyase production in the soft-rotting bacterium Pseudomonas fluorescens. Phytopathology 83:813-818.
39.Martin, W.J., and Dukes, P. D. 1977. Bacteria stem and root rot of sweet potato. Plant Dis. Rep. 61:158-161
40.Moyer, J. W., and Cali, B. B. 1985. Properties of sweet potato feathery mottle virus RNA and capsid protein. J. Gen. Virol. 66:1185-1189.
41.Neergaar, P. 1945. Danish species of Alternria and Stemphyllium. Taxonomy, parasitism and economic significance. Einar Munksgaard, Copenhagen.
42.Nielsen, L. W.1969. Relationship of storage temperatures, fungicides, and varietal resistance to the infection of sweet potato roots by the Fusarium wilt fungus. Phytopathology 59:508-510.
43.Palacio-Bielsa, A., Cambra, M. A. and López, M. M. 2006. First report of bacterial soft rot on onion caused by Dickeya sp. (ex Pectobacterium chrysanthemi) in Spain. New disease reports.
44.Palava, T. K., et al. 1993. Induction of plant defense response by exoenzymes of Erwinia cartovora subsp. cartovora. Mol. Plant-Microb. Interact. 6:190-196.
45.Perobelon, M. C. M., and Kelman. A. 1980. Ecology of the soft rot Erwinia. Ann. Rev. Phyotopathol. 18 : 361-387.
46.Pérombelon, M. C. M. 1992. Potato blackleg: epidemiology, host–pathogen interaction and control. Netherlands Journal of Plant Pathology 98:135–46.
47.Pérombelon, M. C. M. 2002. Potato diseases caused by soft rot Erwinias: an overview of pathogenesis. Plant Pathology 51:1–12.
48.Pérombelon, M. C. M., and Kelman, A. 1980. Ecology of the soft rot Erwinias. Annu. Rev. Phytopathol. 18:361-387.
49.Pérombelon, M. C.M., Lumb, V. M., and Zutra, D. 1987. Pathogenicity of soft rot Erwinias to potato plants in Scotland and Israel. Journal of Applied Bacteriology 63: 73–84.
50.Person, L. H., and Martin, W.J. 1940. Soil rot of sweet potatoes in Louisiana. Phytopathology 30:913-926.
51.Samson R., Legendre J. B., and Christen R. 2005. Transfer of Pectobacterium chrysanthemi (Burkholder et al. 1953) Brenner et al. 1973 and Brenneria paradisiaca to the genus Dickeya gen. nov as Dickeya chrysanthemi comb. nov and Dickeya paradisiaca comb. nov and delineation of four novel species, Dickeya dadantii sp nov., Dickeya dianthicola sp nov., Dickeya dieffenbachiae sp nov and Dickeya zeae sp nov. International Journal of Systematic and Evolutionary Microbiology 55:1415-1427.
52.Schaad, N. W., and Brenner, D. 1997. A bacterial wilt and root rot of sweet potato Caused by Erwinia chrysanthemi. Phytopathology. 67:302-308.
53.Sivaprakasam, K., Krishnamohan, G., and Kandasway, T. K. 1997. A new leaf spot disease of sweet potato. Sci. Cult. 43:325-326.
54.Smith, C., and Bartz, J. A. 1990. Variation in the pathgenicity and aggressiveness of strains of Erwinia cartovora subsp. cartovora isolated from different hosts. Plant Dis. 74:505-509.
55.Toussoun, T. A., and P. E. Nelson. 1968. A Pictorial Guide to the Identification of Fusarium species. Pennsylvania State University Press.
56.Van Bruggen, A. H. C. 1984. Sweet potato stem blight caused by Alternaria spp.:A new disease in Ethiopia. Neth. J. Plant Pathol. 90:155-164.