Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/31215
DC FieldValueLanguage
dc.contributor鍾文全zh_TW
dc.contributor張碧芳zh_TW
dc.contributor.advisor黃振文zh_TW
dc.contributor.advisorJenn-Wen Huangen_US
dc.contributor.author黃靜淑zh_TW
dc.contributor.authorHuang, Jing-Shuen_US
dc.contributor.other中興大學zh_TW
dc.date2009zh_TW
dc.date.accessioned2014-06-06T07:41:20Z-
dc.date.available2014-06-06T07:41:20Z-
dc.identifierU0005-1208200814251800zh_TW
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dc.identifier.urihttp://hdl.handle.net/11455/31215-
dc.description.abstract立枯絲核菌 (Rhizoctonia solani kühn AG-4) 與腐霉菌 (Pythium aphanidermatum Edson) 皆可以引起甘藍幼苗猝倒與死亡的現象,是甘藍苗床期的重要病原菌。過去防治這類土傳性病害大多使用土壤燻蒸的方式來作為病害防治的策略,但隨著許多化學藥劑的弊端浮現,尋找替代的防治策略成為刻不容緩的課題。本研究主要目的在於探討Bacillus mycoides CHT2401與CHT2402菌株之抑菌能力,進而評估它們防治甘藍幼苗病害的效果。首先利用培養皿對扣法測試King’s B培養基、Luria-Bertani medium (LB)、Nutrient agar (NA)、Potato dextrose agar (PDA)、Soy powder milk agar (SPMA) 及Tryptic soy agar (TSA) 等六種培養基培養CHT2401與CHT2402後,其產生之氣體對R. solani AG-4與P. aphanidermatum菌絲生長的影響,結果發現CHT2401與CHT2402兩菌株培養在TSA與SPMA上所產生之氣體對此二種病原菌皆具有較優異的抑制效果。進一步以氣相層析質譜儀與氨氣檢知器分析兩菌株培養於TSA與SPMA上,皆可產生二甲基二硫化物 (dimethyl disulfide) 與氨氣,其中二甲基二硫化物與氨氣皆可顯著抑制R. solani AG-4與P. aphanidermatum菌絲的生長。利用掃描式與穿透式電子顯微鏡觀察B. mycoides CHT2402培養於TSA上釋放出之氣體與二甲基二硫化物標準品燻蒸處理過的R. solani AG-4與P. aphanidermatum菌絲,結果發現兩者處理過之菌絲的外觀皆出現皺縮的現象。此外,細胞內的胞器數量皆有顯著減少的情況。溫室試驗中,利用黃豆粉培養B. mycoides CHT2401與CHT2402所獲得的細菌培養液,以5 % (v/v) 混拌至栽培介質中,分別種植甘藍、油菜、豇豆、萵苣及番茄,結果顯示兩菌株皆可促進植物生長,其中又以B. mycoides CHT2402菌株對於甘藍鮮重的促進效果最佳,較對照組提高約2倍。進一步將兩菌株的培養液以5 % (v/v) 混拌至含有R. solaniAG-4 與P. aphanidermatum的栽培介質中,分別密封放置一天後播種甘藍,結果顯示兩菌株之細菌培養液皆可以促進甘藍植株生長外,尚可降低P. aphanidermatum引起的甘藍幼苗猝倒病達 28 % 左右;然而對於防治R. solani AG-4危害植株的罹病率卻無顯著的差異。此外,若以同樣處理,將CHT2401與CHT2402兩菌株的培養液混拌至含有P. aphanidermatum的栽培介質中,密封燻蒸處理六天後播種甘藍,其防治率分別可達到 44-45 %。zh_TW
dc.description.abstractDamping-off caused by Rhizoctonia solani kühn AG-4 or Pythium aphanidermatum Edson is a serious disease of cabbage seedlings, especially in culture medium in Taiwan. Thus, preventing culture medium from contamination of these pathogens should be an important method for controlling the disease. Effective evaluations of the volatile compounds produced by Bacillus mycoides isolates CHT2401 and CHT2402 grown on plates of King's medium B, Luria-Bertani medium, nutrient agar, potato dextrose agar, soy powder milk agar, and tryptic soy agar for suppression of mycelial growth of R. solani RST-04 and P. aphanidermatum Pa01 were conducted. The two bacterial isolates grown on soy powder milk agar and tryptic soy agar expressed better effectiveness in inhibiting the pathogens. The volatile compounds produced by the two bacterial isolates were identified using ammonia detector tube and Gas Chromatography - Mass Spectrometry (GC-MS). The major volatile compounds produced by both isolates were identified as ammonia and dimethyl disulfide that were much more effective in inhibiting mycelial growth of R. solani RST-04 and P. aphanidermatum Pa01. The mycelial morphology of R. solani RST-04 and P. aphanidermatum Pa01 treated with dimethyl disulfide and the volatile compounds produced by B. mycoides CHT2402 respectively were observed under scanning electron microscope and transmission electron microscope. The hyphal structures of R. solani RST-04 and P. aphanidermatum Pa01 after treatments were malformed and the numbers of the organelles in their hyphal cells significantly decreased. These results demonstrated that dimethyl disulfide, ammonia, and the volatile compounds produced by B. mycoides CHT2402 could be harmful to the mycelia of the pathogens. In the greenhouse, the culture media were individually treated with the culture solutions of B. mycoides CHT2401and CHT2402 grown in soy powder milk. The culture solutions of the Bacillus isolates could increase the plant seedlings biomass of asparagus bean, cabbage, edible rape, lettuce, and tomato compared to the untreated control. Furthermore, the culture solutions of the Bacillus isolates were separately tested for controlling damping-off of cabbage seedlings caused by R. solani RST-04 or P aphanidermatum Pa01. The culture solutions of the two bacterial strains could respectively reduce 28% and 27% disease incidence of cabbage seedlings caused by P. aphanidermatum Pa01, but not significantly reduce the disease incidence of seedlings damping-off caused by R. solani RST-04. The results suggested that B. mycoides isolates CHT2401 and CHT2402 were potential biocontrol agents on controlling damping-off of cabbage seedlings caused by P. aphanidermatum Pa01.en_US
dc.description.tableofcontents目次 中文摘要...................................................i 英文摘要.................................................iii 目次.......................................................v 表次索引................................................viii 圖次索引..................................................ix 壹、前言...................................................1 貳、材料與方法.............................................5 一、供試植株...............................................5 二、供試病原菌菌株來源.....................................5 三、病原菌接種源製備.......................................6 (一)立枯絲核菌接種源製備.................................6 (二)腐霉菌接種源製備.....................................6 四、拮抗細菌來源與菌液製備.................................6 (一)拮抗菌來源...........................................6 (二)拮抗菌菌液製備.......................................7 五、不同培養基對Bacillus mycoides產氣抑菌之效果影響........7 六、Bacillus mycoides產氣處理過之病原菌菌絲活力測試........8 七、不同培養基培養Bacillus mycoides所產生之氣體對腐霉菌產生游走子的影響...............................................9 八、Bacillus mycoides產生氣體之種類分析....................9 (一)氣相層析質譜儀分析B. mycoides所生氣體種類............9 (二)B. mycoides菌株產生氨氣濃度之分析...................10 九、氣體標準品對病原菌菌絲生長的影響......................10 (一)二甲基二硫化物對病原菌菌絲生長影響..................10 (二)氨氣對病原菌菌絲生長影響............................11 十、Bacillus mycoides 產生的氣體、二甲基二硫化物與氨氣標準品對於病原菌菌絲外觀與內部構造的影響........................12 (一)利用Cryo-transfer系統觀察菌絲外觀之變化.............12 (二)利用超薄切片法觀察菌絲內部構造......................13 十一、Bacillus mycoides對不同植物生長的影響...............13 十二、不同濃度的Bacillus mycoides培養液對甘藍植株生長的影響........................................................14 十三、甘藍苗期病害之防治試驗..............................14 (一)甘藍立枯病之防治試驗................................14 (二)甘藍猝倒病之防治試驗................................15 1. Bacillus mycoides防治病害之效果評估....................15 2.栽培介質處理Bacillus mycoides不同天數之防病效果評估.....15 叁、結果..................................................16 一、不同培養基對Bacillus mycoides產氣抑菌之效果影響.......16 二、Bacillus mycoides產氣處理過之病原菌菌絲活力測試.......16 三、不同培養基培養Bacillus mycoides所產生之氣體對腐霉菌產生游走子的影響..............................................17 四、Bacillus mycoides產生氣體之種類分析...................17 (一)氣相層析質譜儀分析B. mycoides產生氣體的種類.........17 (二)Bacillus mycoides產生氨氣濃度之分析.................17 五、氣體標準品對病原菌生長的影響..........................18 六、Bacillus mycoides 產生的氣體、二甲基二硫化物與氨氣標準品對於病原菌菌絲外觀與內部構造的影響........................18 (一)利用Cryo-transfer系統觀察菌絲外觀之變化.............18 (二)利用超薄切片法觀察菌絲內部構造......................19 七、黃豆粉培養Bacillus mycoides對不同植物生長的影響.......19 八、不同濃度的Bacillus mycoides培養液對甘藍植株生長的影響........................................................19 九、甘藍苗期病害之防治試驗................................20 (一)甘藍立枯病之防治試驗................................20 (二)甘藍猝倒病之防治試驗................................20 1. Bacillus mycoides防治病害之效果評估....................20 2. 栽培介質處理Bacillus mycoides不同天數之防病效果評估....21 肆、討論..................................................22 伍、引用文獻..............................................26 表次索引 表一、不同培養基培養Bacillus mycoides CHT2401與CHT2402菌株所產生之氣體對腐霉菌 (Pythium aphanidermatum Pa01) 產生游走子的影響....................................................31 表二、不同培養基培養Bacillus mycoides CHT2401與CHT2402菌株所產生的氨氣濃度............................................32 表三、在溫室中黃豆粉培養Bacillus mycoides CHT2401與CHT2402菌株對不同 蔬菜植株生長的影響 .............................33 圖次索引 圖一、不同培養基培養Bacillus mycoides CHT2401與CHT2402菌株所產生之氣體對立枯絲核菌 (Rhizoctonia solani RST-04) 與腐霉菌 (Pythium aphanidermatum Pa01) 菌絲生長的影響..............34 圖二、不同培養基培養Bacillus mycoides CHT2401與CHT2402菌株所產生之氣體對立枯絲核菌 (Rhizoctonia solani RST-04) 與腐霉菌 (Pythium aphanidermatum Pa01) 菌落的影響..................36 圖三、利用不同培養基培養Bacillus mycoides CHT2401與CHT2402菌株所產生的氣體燻蒸立枯絲核菌 (Rhizoctonia solani RST-04) 與腐霉菌 (Pythium aphanidermatum Pa01) 菌絲之後,其再重新生長的情形....................................................37 圖四、胰蛋白大豆瓊脂培養基培養Bacillus mycoides CHT2401與CHT2402所產生之氣體的氣相層析質譜圖.......................38 圖五、黃豆粉瓊脂培養基培養Bacillus mycoides CHT2401與CHT2402所產生之氣體的氣相層析質譜圖..............................40 圖六、二甲基二硫化物 (dimethyl disulfide) 的質譜圖........42 圖七、不同培養基培養Bacillus mycoides CHT2401與CHT2402菌株所產生氨氣的情形............................................44 圖八、二甲基二硫化物 (dimethyl disulfide) 標準品對立枯絲核菌 (Rhizoctonia solani RST-04) 與腐霉菌 (Pythium aphanidermatum Pa01) 菌絲生長的抑制效果...................45 圖九、氨氣標準品 (28% ammonia water) 對立枯絲核菌 (Rhizoctonia solani RST-04) 與腐霉菌 (Pythium aphanidermatum Pa01) 菌絲生長的抑制效果...................46 圖十、胰蛋白大豆瓊脂培養基培養Bacillus mycoides CHT2402所產生之氣體、二甲基二硫化物與氨氣標準品對立枯絲核菌 (Rhizoctonia solani RST-04) 菌絲外觀形態的影響............47 圖十一、胰蛋白大豆瓊脂培養基培養Bacillus mycoides CHT2402所產生之氣體、二甲基二硫化物與氨氣標準品對腐霉菌 (Pythium aphanidermatum Pa01) 菌絲外觀形態的影響...................48 圖十二、不同培養基培養Bacillus mycoides CHT2402所產生之氣體與二甲基二硫化物對立枯絲核菌 (Rhizoctonia solani RST-04) 菌絲內部形態構造的影響......................................49 圖十三、不同培養基培養Bacillus mycoides CHT2402所產生之氣體與二甲基二硫化物對腐霉菌 (Pythium aphanidermatum Pa01) 菌絲內部形態構造的影響........................................50 圖十四、在溫室中黃豆粉培養Bacillus mycoides CHT2401與CHT2402 之菌液對不同蔬菜幼苗生長的影響....................51 圖十五、不同濃度黃豆粉培養Bacillus mycoides CHT2401與CHT2402之菌液對甘藍植株鮮重的影響................................53 圖十六、在溫室中黃豆粉培養Bacillus mycoides CHT2401與CHT2402 之菌液對甘藍幼苗立枯病的防治的效果................54 圖十七、在溫室中黃豆粉培養Bacillus mycoides CHT2401與CHT2402 之菌液對接種及未接種Pythium aphanidermatum Pa01的甘藍植株地上部鮮重與根重的影響............................. 55 圖十八、在溫室中黃豆粉培養Bacillus mycoides CHT2401與CHT2402 之菌液對甘藍幼苗猝倒病的防治效果..................56 圖十九、在溫室中黃豆粉培養Bacillus mycoides CHT2401與CHT2402 之菌液對防治甘藍幼苗猝倒病的效果..................57 圖二十、處理不同天數黃豆粉培養Bacillus mycoides CHT2401與CHT2402 的菌液對甘藍幼苗猝倒病的防治效果..................58zh_TW
dc.language.isoen_USzh_TW
dc.publisher植物病理學系所zh_TW
dc.relation.urihttp://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-1208200814251800en_US
dc.subjectBacillus mycoidesen_US
dc.subject生物防治zh_TW
dc.subjectbiocontrolen_US
dc.subjectcabbageen_US
dc.subjectdimethyl disulfideen_US
dc.subjectPythium aphanidermatumen_US
dc.subjectRhizoctonia solanien_US
dc.subject二甲基二硫化物zh_TW
dc.subject甘藍zh_TW
dc.subjectBacillus mycoideszh_TW
dc.subjectPythium aphanidermatumzh_TW
dc.subjectRhizoctonia solani AG-4zh_TW
dc.titleBacillus mycoides 防治甘藍幼苗病害之效果評估zh_TW
dc.titleEvaluation for Efficacy of Bacillus mycoides on Control of Cabbage Seedling Diseasesen_US
dc.typeThesis and Dissertationzh_TW
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.fulltextno fulltext-
item.grantfulltextnone-
item.cerifentitytypePublications-
item.languageiso639-1en_US-
item.openairetypeThesis and Dissertation-
Appears in Collections:植物病理學系
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