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題 名 | 篩選抑制番茄萎凋病菌厚膜孢子發芽之拮抗細菌=Screening Antogonistic Microorganisms That Inhibit Chlamydospore Germination of Fusarium Oxysporum f. sp. Lycopersici |
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作 者 | 陳媛鈺; 林宗俊; 鍾文鑫; 王智立; | 書刊名 | 植物病理學會刊 |
卷 期 | 24:3/4 2015.12[民104.12] |
頁 次 | 頁251-266 |
分類號 | 435.272 |
關鍵詞 | 番茄萎凋病; 厚膜孢子; 栽培介質萃取液; 短芽孢桿菌; 枯草桿菌; Fusarium wilt of tomato; Chlamydospore; Potting soil broth; Brevibacillus; Bacillus subtilis; |
語 文 | 中文(Chinese) |
中文摘要 | 番茄萎凋病是由病原真菌Fusarium oxysporum f. sp. lycopersici 所造成的土壤傳播性病害,此病原菌可產生大分生孢子、小分生孢子及厚膜孢子。一般認為由於厚膜孢子可於土壤中長期殘存,導致根除此病害較困難。過去多數生物防治試驗都針對病原菌的菌絲或分生孢子進行抑制作用的評估,再對製備於土壤環境中的厚膜孢子進行評估,此篩選方式耗時且未能直接了解對厚膜孢子的影響。本研究利用栽培介質之萃取液,作為誘導番茄萎凋病菌產生厚膜孢子的基質,並以純培養產生之厚膜孢子,直接與土壤微生物混合後進行拮抗能力篩選。經測試11 種市售的園藝栽培介質,以BVB 和KEKKILÄ 栽培介質之萃取液,分別能生成最大量的厚膜孢子,濃度可達5×10^5 chlamydospores/ ml以上。此外,添加0.03%(w/v)的硫酸銨至BVB 介質萃取液中,可明顯提升厚膜孢子的數量,並降低分生孢子的產生。將厚膜孢子與分離自土壤的微生物之上清培養液或沉澱菌體共同培養,其中16 個拮抗微生物菌株可以抑制厚膜孢子的發芽率至40%以下,並以菌株PL T-1 N-5.5 的效果為最佳,厚膜孢子於其上清培養液或菌體處理下幾乎無法發芽,亦能顯著減少厚膜孢子形成菌落,但其培養濾液處理則對厚膜孢子沒有抑制效果。選擇抑制效果較佳的菌株PLT-1 N-5.5、34-5N-6 及36-4N-1 經16S rDNA、脂肪酸甲酯及Biolog 鑑定,結果顯示Brevibacillus sp.(PL T-1 N-5.5)及Bacillus subtilis(34-5N-6 及36-4N-1)的菌株可應用於抑制番茄萎凋病菌厚膜孢子的發芽。上述結果顯示,我們已建立一個可直接篩選對厚膜孢子具有抑制作用的拮抗微生物的方法。 |
英文摘要 | Fusarium oxysporum f. sp. lycopersici causes the soilborne disease of tomato. The pathogen produces macroconidia, microconidia and chlamydospores. Chlamydospores could survive in soil for a long time, which makes the elimination of the disease difficult. Most biocontrol assays, however, used mycelia or conidia to screen antagonistic microorganisms at the first step, and then challenge with chlamydospores in the complicated soil environment. This process is time consuming and not straightforward. In this study, potting soil broths were applied to induce the production of chlamydospores of F. oxysporum f. sp. lycopersici. Among 11 tested commercial potting soils, soil broths made from the BVB and KEKKILÄ could produce chlamydospores more than the concentration of 5×10^5 chlamydospores/ml. In addition, the BVB potting soil broth amended with 0.03% (w/v) of (NH_4)_2SO_4 significantly increased the amount of chlamydospores and reduced conidium production. When chlamydospores were cultured in the supernatants or pellets of soil microorganisms, 16 antagonistic microorganisms could reduce chlamydospore germination to less than 40%. Strain PL T-1 N-5.5 was the most effective strain that almost completely inhibited the germination of chlamydospores in the supernatant or pellet treatment and greatly decreased colonies formed from chlamydospores after the treatments. In contrast, its filtrate lost the inhibitory effect on chlamydospores. The three most effective strains, PL T-1 N-5.5, 34-5N-6 and 36-4N-1, were identified by 16S rDNA, FAMA and Biolog analysis. The results suggested that strains of Brevibacillus sp. and Bacillus subtilis may be good candidates for inhibiting chlamydospore germination of F. oxysporum f. sp. lycopersici. In this study, we established a method to easily screen microorganisms that showed directly inhibitory effects on chlamydospores. |
本系統中英文摘要資訊取自各篇刊載內容。