請用此 Handle URI 來引用此文件: http://hdl.handle.net/11455/31215
標題: Bacillus mycoides 防治甘藍幼苗病害之效果評估
Evaluation for Efficacy of Bacillus mycoides on Control of Cabbage Seedling Diseases
作者: 黃靜淑
Huang, Jing-Shu
關鍵字: Bacillus mycoides
生物防治
biocontrol
cabbage
dimethyl disulfide
Pythium aphanidermatum
Rhizoctonia solani
二甲基二硫化物
甘藍
Bacillus mycoides
Pythium aphanidermatum
Rhizoctonia solani AG-4
出版社: 植物病理學系所
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摘要: 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 %。
Damping-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.
URI: http://hdl.handle.net/11455/31215
其他識別: U0005-1208200814251800
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-1208200814251800
顯示於類別:植物病理學系

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