Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/30959
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dc.contributor.advisor林益昇zh_TW
dc.contributor.advisorYi-Sheng Linen_US
dc.contributor.author陳以錚zh_TW
dc.contributor.authorChen, Yi-Jengen_US
dc.date2005zh_TW
dc.date.accessioned2014-06-06T07:40:38Z-
dc.date.available2014-06-06T07:40:38Z-
dc.identifier.urihttp://hdl.handle.net/11455/30959-
dc.description.abstract摘 要 本論文的目的,在調查與鑑定台灣羅勒發生的兩個新病害與描述一個未確定病害。2003年夏,台中大里地區羅勒於田間發生一萎凋型不明病害,罹病植株初期呈現矮化及落葉,接著嫩芽及莖頂出現壞疽。壞疽沿莖頂向下發展至全株,根部及莖部也會出現腐敗,罹病莖部表面偶有白色菌絲及孢子褥纏據;部分罹病植物呈半側萎凋;幼苗期受感染的植物會在短期內死亡,成株則可持續較長的時間。罹病株根莖維管束會有不連續褐化,尤其著生葉柄之莖部維管束褐化最為嚴重,呈黑褐色。從罹病組織可以分離得到一種Fusarium oxysporum Schlechtend,其菌落在PDA上呈白色並有橙紅色孢子褥以及藍或褐色菌核。孢子區分為三型。大孢子無色,鐮刀型,3到5隔膜,基部有足細胞,大小19.8-49.5×3.9-5.4μm;小孢子無色長橢圓形,單胞,大小5.9-16.3×2.4-4.9μm;厚膜孢子圓形或近圓形,壁厚,顏色較深,直徑7.4-12.3μm。病原性測試以剪根接種及混土接種接種於羅勒 (白骨羅勒) 幼苗上,可使其在溫室產生與田間相同病徵。將接種後之罹病株再分離,可分離得到與供試菌相同之病原菌,以此完成克霍氏法則 (Koch’s postulates )。供試菌株僅在唇形花科之羅勒 (白骨、紅骨) 造成感染,對同科及其他科的供試作物則無;又將採自羅勒不確定分化型之菌株與番茄、豌豆等六種不同分化型的F. oxysporum病原菌接種於羅勒,結果僅來自羅勒的病原菌可在羅勒上造成病徵,以此鑑定本病害是由Fusarium oxysporum f. sp. basilici (Dzidzariya) Armst. & Armst所引起,而正式稱該病害為羅勒萎凋病;病原菌於PDA上之菌絲生長溫度為8-36 ℃,但以24-28 ℃為最適;發病溫度在15 -35 ℃之間,且30-35 ℃時發病最嚴重;當接種源濃度在102 propagules/g soil或 102 spores/ml以上始發病;病原菌可經由孢子懸浮液於葉面接種而感染羅勒植株。另外,本研究也鑑定一種新的羅勒葉部病害,該病害會在初期於羅勒葉部邊緣形成黑色病斑,與健部有明顯界線,病斑隨後擴大,造成葉片畸形,嚴重時沿葉柄危害莖部;菌落在PDA上成白色,逐漸變為橄欖綠,而深黑色。分生孢子褐色,長橢圓形,鏈生於分生孢子柄頂,具2-5橫格及0-4縱格,大小為 21-56 × 7.2-16 μm,喙長2-6 μm ,經鑑定為Alternaria alternata (Fries : Fries) von Keissler,並將該病害定名為羅勒葉枯病。除此之外,一種不明原因的病害會在田間造成羅勒幼苗不分枝,莖部褐化水浸狀壞疽並且有根腐現象,在褐化部份出現白色菌絲纏聚或是著生黑色菌核,受感染植株會在兩週內死亡,其病徵類似羅勒菌核病。病原菌在28℃以下可以在PDA上產生菌核,菌絲生長溫度則以20℃為最適,發病溫度在24℃以下,其中在20℃為最嚴重。因本病原菌未在培養下觀察到其任何形式的孢子或菌核發芽行為,故先將其稱為羅勒疑似菌核病。zh_TW
dc.description.abstractAbstract This study was aimed at determining and describing the Fusarium wilt and other diseases of basil in Taiwan. During 2003-2004, an undiagnosed wilt-type disease of basil was observed in central Taiwan. The first symptoms appeared as defoliation and stunting of the diseased plants, and then the buds and stems showed necrosis. Necrosis developed basipetally from the apex to the entire plant. Sometimes infected plants remained lateral wilt and stems and roots were rot. Vascular bundles showed discoloration. Infected seedlings died rapidly within a few days, and old plants might survive longer. Sometimes white mycelia and spore masses covered rotted stems and buds. A kind of fungus was consistently isolated from diseased tissue. The fungus produced 3 kinds of asexual spores. Microconidia produced abundantly from monophialides on potato dextrose agar (PDA); single-celled, hyaline, elliptic, or ovoid; 5.9-16.3×2.4-4.9μm. Macroconidia were the type “ Fusarium” spores. They were hyaline; 1-4 septates; 19.8-49.5×3.9-5.4μm. Chlamydospores were spherical to ovoid, smooth-surfaced, intercalary or terminal; 7.4-12.3×7.4-12.3μm. The fungus was determined as Fusarium oxysporum Schlechtend:Fr. It was pathogenic to basil 'White stem' in greenhouse inoculation experiments and produced symptoms similar to those observed in the fields. Host range tests in greenhouse indicated that this fungus could not attack tested Labiatae or other family plants except some cultivars of basil. Moreover, six different forma specialis of F. oxysporum derived from other host plants showed no pathogenicity to basil. Therefore, the pathogen was indentifield as Fusarium oxysporum f. sp. basilici (Dzidzariya) Armst. & Armst. and this disease was be named Fusarium wilt and crown rot of basil. Tested basil ‘White stem' also appeared symptoms by using suspension inoculation spraying method. The optimal temperature for the mycelial growth was at 28℃. Inoculation tests revealed that the optimal infection temperature for this disease were between 25-30℃. Another undiagnosed disease with leaf blight symptoms also occurred in basil fields during 2003-2004. The symptoms appeared black spot first on leaf margin, then leaf deformed. Finally, the symptoms extended from leaf to stem and plants showed leaf and stem blight. Conidia of the pathogen were brown, ellipsoidal, 21-56 × 7.2-16μm in size, short-beaked (2-6μm), with 2-5 transverse septa and 0-4 longitudinal septa, and catenulated at the apex of the conidiophores. The pathogen was identified as Alternaria alternata (Fries: Fries) von Keissler. Both of the optimal temperature for the mycelial growth and conidia germination were at 24℃. A stem rot of basil was also observed in fields. The infected plants appeared stem and root rot and they didn't branch. Sometimes white mycelia covered stem and sclerotia were produced on diseased leaf or stem tissue of basil. The symptoms seemed to be the same with Sclerotinia rot of basil. However, the sclerotia never apothecium germinated on artifical medium in laboratory. The optimal temperature for the mycelial growth was at 20℃. Sclerotia could be produced on PDA at or below 28℃, and the pathogen could infected the basil ‘White stem' at 16-24℃.en_US
dc.description.tableofcontents目 錄 中文摘要........................................................................................................................I 英文摘要......................................................................................................................III 誌謝...............................................................................................................................V 目錄............................................................................................................................VII 附表目錄.......................................................................................................................X 附圖目錄....................................................................................................................XII 前言................................................................................................................................1 前人研究........................................................................................................................3 材料與方法....................................................................................................................7 一、田間病害調查與病徵觀察................................................................................7 二、病原菌的分離與鑑定.......................................................................................7 三、供試植物之來源與培育...................................................................................7 四、羅勒萎凋病 (Fusarium wilt)............................................................................9 1)接種方法...............................................................................................................9 剪根接種..............................................................................................................9 混土接種..............................................................................................................9 粉衣接種..............................................................................................................10 噴霧接種法..........................................................................................................10 2)寄主範圍測試.......................................................................................................10 3)不同來源之F. oxysporum 分化型菌株對羅勒的病原性...................................11 4)溫度對羅勒萎凋病的影響...................................................................................11 溫度對分生孢子發芽的影響...............................................................................11 菌絲最適生長溫度...............................................................................................11 溫度與病害發生的關係.......................................................................................12 5)接種源濃度與病害發生的關係...........................................................................12 6)羅勒萎凋病菌培養濾液對羅勒生長的影響.......................................................12 7)羅勒品種對萎凋病之抗病檢定...........................................................................13 五、葉枯病 (Leaf blight).........................................................................................13 1)病原性測定...........................................................................................................13 2)溫度病原菌生長的影響.......................................................................................14 溫度對菌絲生長的影響......................................................................................14 溫度對分生孢子發芽的影響..............................................................................14 六、疑似菌核病 (Stem rot)....................................................................................14 1)病原性測試...........................................................................................................14 菌絲塊接種...........................................................................................................14 混土接種...............................................................................................................15 2)溫度對病原菌生長及病勢發展的影響...............................................................15 溫度對菌絲生長的影響.......................................................................................15 溫度對菌核形成的影響.......................................................................................16 溫度對菌核發芽的影響.......................................................................................16 a.不同溫度產生的菌核的發芽能力....................................................................16 b.不同溫度對菌核發芽的影響............................................................................16 溫度對病勢發展的影響.......................................................................................16 結果..............................................................................................................................18 一、田間病害調查與病原菌的分離.......................................................................18 1.田間病害種類調查及病徵描述...........................................................................18 1)萎凋病 (Fusarium wilt)........................................................................................18 2)葉枯病 (Leaf blight)............................................................................................18 3)疑似菌核病 (Stem rot)........................................................................................19 4)疑似疫病 (Phytophthora root rot).......................................................................19 5)灰黴病 (Gray mold).............................................................................................19 2.病原菌的分離與鑑定...........................................................................................20 1)萎凋病...................................................................................................................20 2)葉枯病...................................................................................................................20 3)疑似菌核病...........................................................................................................20 二、羅勒萎凋病的鑑定與發病特性.......................................................................21 1. 羅勒萎凋病的鑑定.............................................................................................21 病原性測定..............................................................................................................21 寄主範圍測定..........................................................................................................21 不同來源之F. oxysporum 之分化型菌株對羅勒的病原性..................................22 2. 溫度對病原菌生長及病勢發展的影響.............................................................22 3. 接種源濃度與羅勒萎凋病罹病度的關係.........................................................22 4. 羅勒萎凋病的葉面接種.....................................................................................23 5. 羅勒萎凋病菌的濾液對羅勒生長的影響.........................................................23 6. 羅勒品種對萎凋病的抗病檢定.........................................................................23 三、葉枯病...............................................................................................................24 1. 病原性測定.........................................................................................................24 2. 溫度對Alternaria alternata AB01菌絲生長及分生孢子發芽的影響.............24 四、疑似菌核病.......................................................................................................24 1. 病原性測定.........................................................................................................25 2. 溫度對菌絲生長的影響.....................................................................................25 3. 溫度對菌核產生的影響.....................................................................................25 4. 發病溫度.............................................................................................................25 討論..............................................................................................................................26 參考文獻......................................................................................................................31 附表目錄 表一、台灣地區羅勒病害之田間調查 (2003-2004) Table 1. Field survey of diseases of basil in Taiwan during 2003-2004......................39 表二、 分離自羅勒之Fusarium oxysporum 分離株之病原性測定 Table 2. Pathogenicity tests of Fusarium oxysporum isolated from diseased basil.....40 表三、Fusarium oxysporum FOB07 的寄主範圍測試 Table 3. Host range of F. oxysporum FOB07 isolated from diseased basil..................42 表四、源自不同寄主作物的不同F. oxysporum分化型對羅勒及原寄主的病原性測定 Table 4. Pathogenicity tests of seven formae specials of Fusarium oxysporm to their original hosts and basil.................................................................................................43 表五、接種源濃度對羅勒萎凋病罹病度的影響 Table 5. Effect of inoculum density on the disease severity of basil inoculated with Fusarium oxysporum f. sp. basilici FOB07.................................................................44 表六、接種源濃度對葉面接種羅勒萎凋病的影響 Table 6. Effect of inoculum density on disease severity of Fusarium wilt of basil by using suspension inoculation spraying method............................................................45 表七、羅勒萎凋病菌之培養濾液對羅勒生長的影響 Table 7. Effect of autoclaved cultural filtrate of F. oxysporum FOB07 on the growth and disease severity of Fusarium wilt of basil.............................................................46 表八、羅勒品種對羅勒萎凋病之抗病檢定 Table 8. Reaction of different cultivars of Ocimum basilicum, O. ctridorum and O. sanctum to Fusarium oxysporum f. sp. basilici FOB07..............................................47 表九、分離自羅勒之Alternaria alternata 對羅勒的病原性測定 Table 9. Pathogenicity test of different isolates of Alternaria alternata isolated from basil to basil.................................................................................................................48 表十、分離自羅勒疑似菌核病罹病組織之真菌分離株之病原性測定 Table 10. Patogenicity tests of fungal isolates obtained from Stem rot of basil..........49 表十一、土壤菌核濃度對羅勒疑似菌核病的影響 Table 11. Effect of sclerotium density of fungal isolate SSB04 on disease severity of Stem rot........................................................................................................................50 表十二、溫度對羅勒疑似菌核病病原菌 SSB04菌核產生的影響 Table 12. Effect of temperature on the formation of sclerotia of fungal isolate SSB04 on potato dextrose agar.................................................................................................51 附圖目錄 圖一、羅勒萎凋病的病徵:(A)植株於田間半側萎凋、落葉;(B)稍枯及莖枯;(C)維管束褐化,且落葉處特別嚴重;(D)根腐。 Fig. 1. The symptoms of Fusarium wilt of basil: (A) Lateral wilt of the infected plants and defoliation ;(B) die back of the stem ;(C) vascular discoloration , and (D)root rot. ......................................................................................................................................52 圖二、羅勒葉枯病病徵:(A)(B)、葉片自葉緣產生黑色病斑並逐漸變大,與健部有明顯界線;(C)罹病葉片扭曲變形、褐化;(D)嚴重時,病斑沿葉柄感染莖部。 Fig. 2. The symptoms of Leaf blight of basil caused by Alternaria alternata: (A,B) Black spot first appeared on leaf margin, (C) browing and distortion of the infected leaf; and (D) leaf and stem blight. ...............................................................................53 圖三、羅勒疑似菌核病病徵:(A)罹病田產生缺株;(B)病原菌於罹病植株患部產生黑色菌核;(C)罹病植株不分枝,嚴重時在褐化部份出現白色菌絲纏聚並且死亡 Fig. 3. The symptoms of Stem rot of basil : (A) Poor stands of basil at a field, (B) sclerotium produced on diseased leaf of basil and (C) diseased plants with stem, root rot, no branch and sometimes white mycelia covered stem surface. ...................54 圖四、羅勒疑似疫病病徵:(A)羅勒在田間有缺株情形;(B)罹病株地上部呈現衰弱病徵;(C)根部褐化、壞疽。 Fig 4. The symptoms of Phytophthora root rot of basil:(A) The disease caused damage at a field; (B) poor growth of the infected plants; (C) root rot. .....................55 圖五、羅勒萎凋病菌形態特徵:(A)在PDA上之菌落形態,(B)大分生孢子,(C)小分生孢子與 (D)厚膜孢子。 Fig.5. Morphological characteristics of Fusarium oxysporum from basil : (A) Colonies on PDA , (B) macroconidia, (C) microconidia and (D) chlamydospore. (Bar = 10 μm)...................................................................................................................56 圖六、羅勒葉枯病病原菌形態特徵:(A)在PDA上之菌落形態;(B)分生孢子以鏈生方式著生在罹病組織周圍;(C)(D)分生孢子鏈生具2-5橫格及0-4縱格。 Fig.6 Morphological characteristics of Alternaria alternata : (A) Colonies on PDA, (B) conidia formed in chain and (C)(D) morphology of conidia. (Bar = 50 μm)...57 圖七、羅勒疑似菌核病病原菌形態:(A)病原菌菌落於PDA上成白色,並產生黑色菌核;(B)菌核具黑色外皮及白色的髓 (橫切面)。 Fig. 7. Morphological characteristics of the pathogen of Stem rot of basil : (A) The colony on PDA with white mycelia, and black sclerotia, and (B) the sclerotium has black rind and white medulla (cross section). .............................................................58 圖八、羅勒幼苗接種萎凋病菌,引起幼苗猝倒的病徵:A. 羅勒幼苗根系稀疏,下胚軸由上至下縊縮褐化,以至於死亡;B. 幼苗猝倒;C. 在潮濕環境下,下胚軸表面有白色菌絲及孢子縟覆蓋 Fig. 8. The symptoms of Fusarium wilt of basil on basil seedlings : (A) The infected seedlings showed poor root systems and hypocotyl rot, (B) seedling damping-off, and (C) sometimes white mycelia and spore masses covered hypocotyls. .......................59 圖九、溫度對羅勒萎凋病菌菌絲生長的影響。 Fig. 9. Effect of temperature on mycelial growth of Fusarium oxysporum f. sp. basilici FOB07 after inoculation on potato dextrose agar. ..........................................60 圖十、溫度對Fusarium oxysporum f.sp. basillici FOB07分生孢子發芽的影響。 Fig. 10. Effect of temperature on the conidial germination of Fusarium oxysporum f.sp. basillici FOB07. ..................................................................................................61 圖十一、溫度對羅勒萎凋病罹病度的影響。 Fig. 11. Effect of temperature on the disease severity of Fusarium wilt of basil inoculated with isolate FOB07. ...................................................................................62 圖十二、以葉面接種羅勒萎凋病之病徵:(A) 葉腋褐化;(B) 維管束自葉腋褐化處向上褐化;(C) 受感染枝條發生落葉萎凋病徵。 Fig. 12. The inoculation symptoms of Fusarium wilt of basil by using suspension spraying method : (A) The base of leaf petiole showed discoloration, (B) discoloration of vascular bundles from the infected leaf, and (C) the infected branch remained defoliation. ...................................................................................................................63 圖十三、溫度對羅勒葉枯病病菌Alternaria alternata AB01 菌絲生長的影響。 Fig.13. Effect of temperature on the mycelial growth of Alternaria alternata AB01, the causal agent of Leaf blight of basil after inoculation on potato dextrose agar. .....64 圖十四、溫度對羅勒葉枯病病菌Alternaria alternata AB01分生孢子發芽率的影響。 Fig.14 Effect of temperature on conidial germination of Alternaria alternata AB01, the causal agent of Leaf blight of basil. .....................................................................65 圖十五、溫度對羅勒疑似菌核病病原菌SSB04菌絲生長的影響。 Fig. 15 Effect of temperature on the mycelial growth of the pathogen of Stem rot of basil after inoculation on potato dextrose agar. .........................................................66 圖十六、溫度對羅勒接種疑似菌核病後罹病率的影響。 Fig. 16 Effect of temperature on disease severity of Stem rot of basil inoculated with isolate SSB04. .............................................................................................................67zh_TW
dc.language.isoen_USzh_TW
dc.publisher植物病理學系zh_TW
dc.title台灣羅勒萎凋病及其他病害的發生zh_TW
dc.titleOccurrence of Fusarium Wilt and Other Diseases of Basil in Taiwanen_US
dc.typeThesis and Dissertationzh_TW
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.languageiso639-1en_US-
item.grantfulltextnone-
item.cerifentitytypePublications-
item.openairetypeThesis and Dissertation-
item.fulltextno fulltext-
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