Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/31956
標題: 栽培介質添加蝦蟹殼粉與微生物防治西瓜幼苗蔓割病之效果評估
Evaluation for the Efficacy of Cultural Medium Amended with Shrimp and Crab Shell Powder and Microorganisms on Controlling Fusarial Wilt of Watermelon Seedlings
作者: 莊偉秀
Chuang, Wei-Shiou
關鍵字: Fusarium oxysporum f. sp. niveum
Fusarium oxysporum f. sp. niveum
香菇太空包堆肥
有機添加物
蝦蟹殼粉
Bacillus subtilis
Iturin A
spent forest mushroom compost
organic amendments
shrimp and crab shell powder
Bacillus subtilis
Iturin A
出版社: 植物病理學系所
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摘要: 西瓜蔓割病係由土媒病原菌Fusarium oxysporum f. sp. niveum (E. F. Smith) Snyder & Hansen所引起,防治頗為不易。目前台灣主要以化學藥劑或嫁接扁蒲(Lagenaria siceraria Ser.) 防治本病害。本研究為了尋找其它替代的防治方法,因此嘗試利用介質添加物與微生物,用以防治西瓜幼苗蔓割病的發生。首先以香菇太空包堆肥 (spent forest mushroom compost, SFMC) 按不同體積比例與荷蘭泥炭苔 (Bas Van Burren No. 4, BVB No.4) 混拌,結果發現兩者體積混拌比例在50:50 (SFMC50) 時,除可降低西瓜蔓割病的發生,西瓜幼苗尚可正常生長。在SFMC50栽培介質添加0.75%蝦蟹殼粉 (SC75) 或0.25%牛血粉 (SB25) 後,兩者栽培介質水萃取液皆可降低西瓜蔓割病菌分生孢子的發芽百分率;其中SC75栽培介質可顯著降低蔓割病的發生達30.48%。將不同組成的栽培介質以熱蒸氣 (100℃,30分鐘) 滅菌處理後,即喪失對於蔓割病菌分生孢子發芽的抑制作用。由SFMC50與蝦蟹殼粉中分離而得之18株細菌菌株分別於馬鈴薯牛肉煎汁瓊脂 (PNA) 平板上與蔓割病菌進行對峙培養,發現Bacillus subtilis Ju020菌株對病原菌FNH0103拮抗效果最佳,可抑制菌絲生長達51.3%。取SFMC50添加0.75%蝦蟹殼粉與Ju020 (106 cfu/mL medium) 製成SCJ20栽培介質,可顯著降低蔓割病發病率達40.3%。利用玻片洋菜薄膜培養法觀察SFMC50、SC75以及SCJ20栽培介質對於病原菌分生孢子發芽的影響,結果發現SCJ20栽培介質可使西瓜蔓割病菌FNH0103與FNP0401菌株分生孢子發芽率分別下降37.33%與36.67%,發芽管長度減少68.99±5.61 μm與104.65±1.85 μm,發芽管不正常膨大比例增加30.33%與26.00%,且分生孢子於第24小時會出現瓦解的情形,FNH0103及FNP0401菌株分生孢子瓦解率分別達45.67%與44.00%。以高壓液相層析儀 (HPLC) 分析栽培介質中可能之抑菌物質,發現有類似iturin A之波峰產生,但是不具有surfactin相似波峰。綜言之,SFMC50搭配0.75%蝦蟹殼粉與B. subtilis Ju020所製成的SCJ20栽培介質,具有防治西瓜蔓割病發生的功效。至於其防病的部分原理可歸因於SCJ20介質中存在有iturin A抑菌物質及擁有較豐富的微生物相與活性,致使西瓜蔓割病菌分生孢子發芽率下降,發芽管出現不正常膨大及瓦解等的現象,因而喪失感染植株的能力。
Fusarial wilt of watermlon is caused by soil-borne fungal Fusarium oxysporum (Schl.) f. sp. niveum (E. F. Smith) Snyder & Hansen, and it is hard to be controlled. Chemical fungicides and grafting bottle gourd (Lagenaria siceraria Ser.) are mainly used to control the disease in Taiwan. In order to search alternative methods, the purpose of this study is to use amendments and microorganism to develop a bioformulated cultural medium for controlling watermelon fusarial wilt. To compare different composition of spent forest mushroom compost (SFMC) mixed with Bas Van Burren No. 4 (BVB No.4) peat moss, it was found that the cultural medium consisted of SFMC and BVB No.4 at the ratio of 50:50 (v/v) was effective in reducing watermelon fusarial wilt, and made watermelon seedlings grown normally. The water-soluble extract of cultural medium SFMC50 amended with 0.75% shrimp and crab shell powder (SC75) or 0.25% blood meal (SB25) could reduce the conidial germination of F. oxysporum f. sp. niveum. The disease incidence of watermelon seedlings grown in cultural medium SC75 was reduced by 30.48%. Inhibition of conidial germination of the pathogen was nullified when cultural media SC75 and SB25 were steamed at 100℃ for 30 min. Eighteen bacterial isolates obtained from SFMC50 and SC75 were dually cultured with F. oxysporum f. sp. niveum on the potato nutrient agar plates. The results indicated that Bacillus subtilis Ju020 was able to inhibit the mycelial growth of F. oxysporum f. sp. niveum isolate FNH0103 up to 51.3%. The cultural medium SFMC50 amended with 0.75% (w/v) SCSP and antagonistic bacterium Ju020 (106 cfu/mL medium) (SCJ20) could reduce 40.3% of fusarial wilt on watermelon seedlings. The effect of cultural media SFMC50, SC75 and SCJ20 on the conidial germination of F. oxysporum f. sp. niveum was analyzed by slide agar-film culture method. The results showed that the conidial germination of F. oxysporum f. sp. niveum isolates FNH0103 and FNP0401 in the cultural medium SCJ20 were decreased by 37.33% and 36.67%, the length of conidial germ tube were reduced by 68.99±5.61 μm and104.65±1.85 μm, abnormal germ tubes were incited by 30.33% and 26.00%. The germ tube lysis percentage of F. oxysporum f. sp. niveum isolates FNH0103 and FNP0401 were observed by 45.67% and 44.00% 24hrs after treatment. Analyzed antifungal compounds in the cultural media by High pressure liquid chromatography (HPLC), it was found that the water-soluble extract of cultural medium SCJ20 had peaks similar to iturin A, but it didn’t have peaks similar to surfactin. The partial mechanisms for reducing fusarial wilt of watermelon seedlings with the cultural medium SCJ20 may attribute plenty of microbial activity and the antifungal activity of iturin A existed in the medium.
URI: http://hdl.handle.net/11455/31956
其他識別: U0005-0207201314093800
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-0207201314093800
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