- 學位論文
利用 Bacillus spp. 防治草莓炭疽病及其可能機制探討
Study of Bacillus spp. on the control of strawberry anthracnose and possible mechanisms involved
摘要
草莓炭疽病菌會造成草莓冠部腐爛，為嚴重影響草莓生產的真菌性病害。應用有益微生物來防治此病為可行之策略。本研究以 Bacillus spp. 來防治草莓炭疽病並探討可能參與機制。首先，以平板對峙試驗分析 Bacillus spp. PMB03、PMB04 及 PMB05 對草莓炭疽病菌菌絲生長情形的結果顯示此三菌株可抑制草莓炭疽病菌之生長。進而探討上述菌株之培養過濾液對草莓炭疽病菌孢子發芽之影響，顯示 PMB03 及 PMB05 之濾液雖不抑制孢子發芽，但可抑制發芽管長度。且 PMB03 可顯著進一步造成孢子發生細胞凋亡後菌絲膨大死亡之現象。PMB04 濾液處理後，孢子則完全喪失發芽能力。由上述結果說明三個菌株均可能具防治草莓炭疽病之潛力。隨後，以草莓果實進行接種之結果顯示，PMB03、PMB04 及 PMB05 之菌液均可顯著抑制草莓炭疽病菌的發生，其中 PMB04 菌液處理之防治效果最顯著。此外，為了解 PMB05 不具有造成草莓炭疽病菌死亡之情形，在草莓果實上卻具有顯著防治之效果，便進一步探討此些 Bacillus spp. 菌株是否具有強化草莓細胞免疫反應之效果。免疫反應分別以激活化氧及癒傷葡聚醣之累積這兩項指標訊號進行分析，結果顯示確實僅 PMB05 菌液可增加草莓細胞辨識炭疽病菌後所誘發激活化氧及癒傷葡聚醣之累積。最後，為確認上述不同機制之拮抗微生物何者於防治草莓炭疽病上扮演重要之角色。進一步以草莓植株進行接種二週後之結果說明僅 PMB04 及 PMB05 可降低病害之發生，其中 PMB05在市售草莓苗上接種草莓炭疽病菌四週之防治率更高達 63.4%。本研究推測 PMB04 造成孢子無法發芽及 PMB05 強化植物免疫分別扮演於採收後草莓果實及草莓苗防治草莓炭疽病之主要關鍵。未來將可以應用混合不同機制之 Bacillus spp. 菌株來防治草莓炭菌病的發生。
並列摘要
Anthracnose is a devastating fungal disease on strawberry production caused by Colletotrichum gloeoesporioides. Developing antagonistic microorganism to control this disease is a valuable strategy to reduce pesticide application. In this research, we attend to screen antagonistic Bacillus spp. strains on control strawberry anthracnose and possible mechanisms involved were also discussed. Firstly, in vitro inhibitory assay revealed that the hyphal growth of C. gloeosporioide SC01was inhibited by Bacillus spp. PMB03, PMB04 and PMB05. We further found that the cultural filtrate from PMB04 was lethal to the conidia of SC01. Growth of germ tube was suppressed by the filtrates from PMB03 and PMB05, and PMB03 filtrate further induced cell death via apoptosis. These results suggested that PMB03, PMB04, and PMB05 would be applied to control anthracnose on strawberry. Biocontrol efficacy on strawberry fruits exhibited the symptoms was significant suppressed by these three strains, especially the treatments of PMB04 and PMB05. To gain more insight of these antagonistic on plant immune system, the intensification of PAMP-triggered immunity was assayed through observing rapid ROS generation and callose deposition. Result showed the immunity was only intensified by the cells of PMB05 upon the inoculation of SC01. Moreover, the disease severity on strawberry seedlings was strongly reduced by PMB04 and PMB05. Especially, control efficacy of PMB05 on strawberry anthracnose reached 63.4% over one month. In addition, plant growth of strawberry seedlings were improved by PMB04 and PMB05. We concluded that lethal on conidia and intensification on plant immune responses were two important mechanisms to control strawberry anthracnose caused by Colletotrichum gloeoesporioides.
參考文獻
延伸閱讀
- 鐘珮哲、吳竑毅、曹嘉惠（2020）。偵測潛伏感染期草莓炭疽病病原菌技術之研發。苗栗區農業改良場研究彙報，(9)，46-57。https://www.airitilibrary.com/Article/Detail?DocID=P20130930009-202006-202011200016-202011200016-46-57
- 鐘珮哲、吳岱融（2016）。草莓炭疽病現勘及座談會。苗栗區農情月刊，(194)，4-4。https://www.airitilibrary.com/Article/Detail?DocID=P20130930007-201601-201606160016-201606160016-4-4
- 彭淑貞（2006）。草莓炭疽病之發生及防治要領。苗栗區農業專訊，(33)，10-11。https://doi.org/10.29551/ZHWHGX.200603.0005
- 湯恩聖（2018）。應用芽孢桿菌及木黴菌對促進草莓生長及防治炭疽病與萎凋病之研究〔碩士論文，國立臺灣大學〕。華藝線上圖書館。https://doi.org/10.6342/NTU201802078
- 莊雅竹（2021）。Evaluation of the Effects of Endophytic Bacteria on Strawberry Heat Tolerance〔碩士論文，國立臺灣大學〕。華藝線上圖書館。https://doi.org/10.6342/NTU202100882