具拮抗能力葉用甘藷內生細菌之鑑定與其防治青枯病之效果評估

Identification of an Antagonistic Bacterial Endophyte from Vegetable Sweet Potato and Assessment of Its Efficacy on Controlling Bacterial Wilt Disease.

陳以錚(Y. J. Chen) ； 潘蕙如(H. R. Pan) ； 林益昇(Y. S. Lin) ； 鍾文鑫(W. H. Chung)

《植物病理學會刊》 22卷1期 (2013/03) Pp. 45-56

https://doi.org/10.6649/PPB.201303_22(1).0005

摘要

田間葉用甘藷莖蔓中含有大量內生細菌，本研究嘗試自葉用甘藷中篩選具拮抗青枯病菌能力的內生細菌，並評估其防治青枯病的潛力。自大甲、大里及中埔等三處葉用甘藷栽培區共分離191株內生細菌，並於TTC培養基上經拮抗活性測試；結果顯示，SPX1、SPE001、SPE003-1、SPE003-2、SPE004及SPC029-2等6株甘藷內生細菌菌株具有顯著抑制青枯病菌能力，其中以SPX1菌株抑制效果最強（p=0.05）。進一步於PDA、NA、LB、KB、TTC和YDC等六種培養基上評估上述6菌株拮抗青枯病菌之能力，僅SPX1菌株在6種培養基中皆可顯著抑制青枯病菌的生長，抑制圈直徑介於1.5－4.0公分之間。以剪莖方式將上述6株甘藷內生細菌菌株導入甘藷後培養28天，結果顯示SPX1、SPE003-2及SPE004菌株並不影響甘藷的生長。進一步在溫室進行SPX1、SPE003-2及SPE004三菌株防治甘藷青枯病試驗，結果指出SPX1菌株較其它兩株菌株更能有效延緩青枯病發生，罹病度較對照組降低67.9%，證實SPX1具有防治青枯病的潛力。進一步以Biolog、脂肪酸圖譜及16S rRNA基因序列分析鑑定SPX1菌株後確定其為Bacillus amyloliquenfaciens。

關鍵字

葉用甘藷青枯病 ； 內生菌 ； 拮抗試驗 ； Bacillus amyloliquenfaciens

並列摘要

Field-grown vegetable sweet potato (VSP) contained plentiful bacterial endophytes in vine tissues. In this study, a total of 191 bacterial endophytes were isolated from VSP in Dali, Dajia, and Zhongpu districts and evaluated by their inhibitory effect on the growth of Ralstonia solanacearum on TTC medium and the potential on controlling bacterial wilt disease in a greenhouse. The inhibitory test indicated six bacterial endophytes could inhibit R. solanacearum growth, and SPX1 strain showed the highest inhibitory effect on the growth of R. solanacearum. Furthermore, only SPX1 showed good antagonistic activity against R. solanacearum on TTC, PDA, NA, LB, KB, and YDC media with the diameter of inhibition zones ranging from 1.5 to 4.0 cm. The six bacterial endophytic strains were introduced into VSP vines by a cut and soaking method to evaluate their effects on plant growth and the efficacy of controlling bacterial wilt disease. Our results revealed that VSP vines inoculated with SPX1, SPE003-2 and SPE004 grew as well as the water-inoculated