Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/31103
標題: 利用產氣細菌防治萵苣猝倒病
Control of Lettuce Damping-off by Gas-Producing Bacteria
作者: 楊維哲
Yang, Wei-Zhe
關鍵字: Biocontrol;生物防治;Lettuce;Pythium aphanidermatum;Pythium damping-off;King's medium B;gas-producing bacteria;萵苣;萵苣猝倒病;King’s medium B;產氣細菌
出版社: 植物病理學系所
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摘要: 
萵苣猝倒病是萵苣苗期的重要病害之一,它的主要病原菌是 Pythium aphanidermatum Edson,可經由栽培介質傳播,因此如何確保栽培介質免於病原菌污染,是防治本病害的一種重要手段。本試驗主要目的在於探討產氣細菌用於防治萵苣猝倒病的可行性。從台灣中部農田與植株根圈土分離產氣菌株,比較它們產氣抑制 P. aphanidermatum PA01 菌絲生長之效果後,選取抑菌較強之 E006、E010 及 P013 菌株進行生理生化測試與 Biolog 鑑定系統測試,結果將 E006 及 E010 鑑定為 Enterobacter 屬;P013 鑑定為 Pseudomonas putida。利用培養皿盤對扣檢測法測試 King’s medium B、Nutrient agar、Yeast malt extract agar、Luria-Bertani medium 及 Potato dextrose agar 等五種培養基對 E006、E010 及 P013 等三菌株產生氣體抑制本病菌的效果,結果顯示各菌株生長在 KB 培養基上,均具有較佳的抑菌效果。進一步修正 KB 培養基配方,以測試其各組成分影響三菌株產氣之抑菌效果,結果發現配方中的 proteose peptone No.3 和 glycerol 是三產氣菌有效抑菌的關鍵成分。嘗試利用 14 種土壤添加物替代 KB 培養基中的 proteose peptone No.3,發現 2 % (w/v) 苜蓿種子粉、苜蓿乾草粉與肉骨粉能使產氣菌株產生抑制 P. aphanidermatum PA01 菌絲生長的氣體。進一步,以氣相層析質譜儀分析三菌株分別於 KB 培養基、苜蓿種子粉培養基、苜蓿乾草粉培養基或肉骨粉培養基中生長所產生主要的氣體種類,結果計有: 1-Undecene、benzoic acid, 2-hydroxy-, methyl ester、methane, thiobis-、benzyl methyl sulfide 等,其中 benzoic acid, 2-hydroxy-, methyl ester、methane, thiobis- 與 benzyl methyl sulfide 均具有抑菌的效果。在含有病原菌的介質中單獨添加 0.5 % (w/w) 苜蓿種子粉、苜蓿乾草粉及肉骨粉等添加物,或僅接種產氣細菌等處理,皆無法有效防治萵苣猝倒病;然而若將苜蓿種子粉或苜蓿乾草粉與產氣細菌 Enterobacter sp. E010 一併均勻拌入含有病原菌的介質中,然後密封燻蒸處理6 天後,再播種萵苣種子,則可達到 90 % 左右的防治率。

Damping-off caused by Pythium aphanidermatum Edson is a serious disease of lettuce seedlings grown in the culture medium in central Taiwan. The culture medium could be contaminated with the pathogen. Thus, preventing culture medium from contamination of the pathogen should be an important aflternative method for controlling the disease. Five gas-producing bacterial strains were isolated from field and rhizophere soils. Three strains E006, E010 and P013 were more effective in inhibiting mycelial growth of P. aphanidermatum PA01. Strains E006 and E010 were identified as Enterobacter spp. strain P013 was identified as Pseudomonas putida. Effective evaluations of the gas-producing bacteria on plates of King's medium B, Nutrient agar, Yeast malt extract agar, Luria-Bertani medium and Potato dextrose agar for suppression of mycelial growth of P. aphanidermatum PA01 were conducted. Three strains grown on King's medium B expressed better effectiveness in inhibiting the pathogen. Proteose peptone No.3 and glycerol were two major ingredients in King's medium B for the bacteria producing volatile compounds to inhibit the pathogen. Alfalfa seed meal (ASM), alfalfa hay meal (AHM) and bone meat meal (BMM) were respectively used to replaced proteose peptone No.3 in King's medium B. Then, the modified media were made and used to culture E006, E010 and P013 for studying their suppressive ability against the pathogen. Those released from gas-producing bacteria grown onto the King's medium B, ASM medium, AHM medium and BMM medium were analyzed by Gas Chromatography. Volatile compounds were identified as 1-Undecene, benzoic acid, 2-hydroxy-, methyl ester, methane, thiobis- and benzyl methyl sulfide. Benzoic acid, 2-hydroxy-, methyl ester, methane, thiobis- and benzyl methyl sulfide were more effective in inhibiting the pathogen. The culture media infested with the pathogen were treated with Enterobacter spp. E006, E010 and Pseudomonas putida P013 and/or organic amendments in the PE plastic bag. Enterobacter sp. E010 mixed with ASM or AHM was significantly effective in reducing 90 % disease incidence of lettuce damping-off caused by P. aphanidermatum in the greenhouse.
URI: http://hdl.handle.net/11455/31103
其他識別: U0005-2708200723000200
Appears in Collections:植物病理學系

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