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標題: 番茄萎凋病之生物防治菌的鑑定與防病潛力評估
Identification for biocontrol agents of tomato Fusarium wilt andevaluation for their potential biocontrol ability.
作者: 丁姵分
Ding, Pen-Fen
關鍵字: tomato;番茄;Fusarium oxysporum f. sp. lycopersici;antagonism;番茄鎌孢菌萎凋病;拮抗作用
出版社: 植物病理學系所
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Fusarium oxysporum f. sp. lycopersici引起的番茄萎凋病,是台灣夏季番茄生產的主要限制因子之一。從農作物植株體內及根圈土壤分離到140株菌株,分別導入栽培介質中培育番茄幼苗,結果總共有17% 的菌株可促進植株生長;15% 的菌株可以有效防治番茄萎凋病。選出CHT2401、CHT2402與 CHR001三菌株進行生理生化測試、培養形態觀察、Biolog 快速鑑定系統測試及gyrB 基因序列、ITS序列分別與GenBank資料庫比對,結果鑑定三菌株為Bacillus mycoides Flügge。三菌株適合生長溫度為28-36 ℃,在pH 4-8均生長良好。研究不同碳氮素源對菌株生長的影響,發現半纖維糖、蔗糖及硫酸銨可促進B. mycoides CHT2401菌株的生長;澱粉、葡萄糖、核醣、丙胺酸、穀氨酸、穀氨醯酸可促進B. mycoides CHT2402菌株的生長;而澱粉、葡萄糖、核醣、麥芽糖、精氨酸、穀氨酸、穀氨醯酸、賴胺酸及鳥胺酸則可促進B. mycoides CHR001菌株生長。在不同基質的培養基上測試三菌株對Fusarium oxysporum f. sp. lycopersici的拮抗作用,結果發現適合細菌生長的培養基NA(nutrient agar)及SA〔soy agar:5%(w/v)soy powder,2%(w/v)agar〕可使三菌株表現有較佳的拮抗作用。至於三菌株在PDA(potato dextrose agar)、CMA(corn meal agar)及MEA(malt extract agar)等平板上對番茄萎凋病菌的拮抗效果不佳。以不同濃度黃豆粉與玉米粉培養CHT2402菌株用以防治番茄萎凋病,發現5%玉米粉與5%黃豆粉培養的菌液,具有最佳的防病效果。近ㄧ步將番茄幼苗分別浸根處理於NB、PDB、5%(w/v)soybean meal(SM)及5%(w/v)corn meal(CM)培養的菌液後,比較它們防治番茄委凋病的效果,結果發現PDB培養的菌液有加速病害發生的現象;NB培養之菌液的處理可降低罹病度36.3 - 72.67%;5% SM培養的菌液可降低罹病度54.5- 63.67%,綜合結果顯示營養液會影響本生物防治菌的防病功效。

Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici has become one of the most severe diseases of tomato in Taiwan during summer. One hundred and forty bacterial isolates were isolated from tomato crop and its rhizosphere soil. Their abilities of controlling disease and promoting the growth of tomato seedling were evaluated. Three isolates CHT2401, CHT2402 and CHR001 with the best efficacy on control of tomato Fusarium wilt were obtained. They were identified as Bacillus mycoides by morphological and physiological characteristics, and Biolog system, and comparing the gene sequence of gyrB and ITS region with isolates available in the GenBank data bank. Temperatures ranged from 28- 36 ℃ were optimal for the growth of the three isolates. There was no difference in the growth of isolates among pH4~pH8. Cellobiose, sucrose and (NH4)2SO4 were effective in enhancing the growth of Bacillus mycoides isolate CHT2401. Starch, glucose, ribose, alinine, glutamic acid and glutamine were able to increase the growth of Bacillus mycoides isolate CHT2402. Starch, glucose, ribose, maltose, arginine, glutamic acid, glutamine, lysine and ornithine could marked by stimulate the growth of Bacillus mycoides isolate CHR001. The antagonism of B. mycoides isolates CHT2401, CHT2402 and CHR001 against Fusarium oxysporum f. sp. lycopersici was affected by different regimes. The antagonistic ability of the bacteria cultured in NA(nutrient agar)and SA〔5%(w/v)soy powder, 2%(w/v)agar〕 were stronger than ones in other media such as PDA:potato dextrose agar, CMA:corn meal agar and MEA:malt extract agar. The concentration of soybean meal and corn meal did also affect efficacy of the bacteria on controlling tomato Fusarium wilt. B. mycoides CHT2402 cultured with 5%(w/v)soybean meal or 5%(w/v)corn meal could reduce 67% disease severity. The tomato seedlings were respectively dipped into culture solutions of B. mycoides CHT2401, CHT2402 and CHR001 grown in NB, PDB, 5%(w/v)soybean meal, 5%(w/v)corn meal and bacterial cell suspension to control tomato Fusarium wilt. The seedlings that were dipped in PDB culture solutions of three bacterial isolates could be enhanced the disease severity. However, the seedlings that were dipped in NB and 5%(w/v)soybean meal culture solutions of three bacterial isolates were most effective in reducing disease severity of 36-73% and 55-64% respectively as compared to nontreatment as a control. The results indicated that B. mycoides isolates CHT2401, CHT2402 and CHR001 were potential biocontrol agents in controlling tomato Fusarium wilt.
其他識別: U0005-2508200615142300
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