草莓灰黴病生物防治之應用研究

Abstract

本研究探討分離自草莓植株之類芽孢桿菌菌株TP3的生物防治活性及施用方法。菌株TP3與灰黴病菌共培養於馬鈴薯葡萄糖瓊脂培養基上，可形成明顯的抑制圈，菌株TP3具有溶磷的能力，施用於草莓幼苗根圈不影響植株的生長。進行溫室及田間試驗，發現施用菌株TP3營養細胞或內孢子懸浮液於草莓植株地上部表面，對草莓灰黴病具有良好的防治效果，保護率達67.7 ~ 79.3%；此外，澆灌菌株TP3營養細胞或內孢子懸浮液於草莓植株根圈，亦可抑制草莓葉灰黴病的發病程度。含有幾丁聚醣之馬鈴薯葡萄糖培養基並不影響草莓灰黴病菌菌絲的生長，但若將濃度5×107 CFU/ml之菌株TP3營養細胞懸浮液與幾丁聚醣共同施用於草莓植株表面，可抑制灰黴病的發病程度，且在田間試驗施用菌株TP3 (2×107 CFU/ml)混合幾丁聚醣可促進草莓果實的產量。除了草莓灰黴病之外，菌株TP3營養細胞懸浮液施用於玫瑰花及百合葉上，亦可抑制灰黴病的病徵發展。菌株TP3液態培養之上清液對灰黴病菌孢子發芽具有顯著的抑制效果，進一步利用DEAE FF離子交換管柱進行層析純化及LC-MS/MS分析，自馬鈴薯葡萄糖培養液中獲得部分純化之菌株TP3外泌抗菌物質，推測其為新穎蛋白質。

In this research, Paenibacillus sp. TP3 screened from strawberry plants showed antagonistic activity against Btrytis cinerea in dual culture assay. Paenibacillus sp. TP3 showed phosphate-solubilizing activity and did not have negative effect on the growth of strawberry plantlet by soil drench. Pot assays and field trials by foliar spray with vegetative cell suspension of Paenibacillus sp. TP3 showed 67.7 ~ 79.3% inhibition of gray mold disease in strawberry. Furthermore, soil drench of TP3 vegetative cell or endospore suspension also suppressed gray mold disease in strawberry. Although hyphal growth of B. cinerea was not inhibited on chitosan-containing PDA medium, treatment with Paenibacillus sp. TP3 suspension of 5×107 CFU/ml combining with chitosan on strawberry plantlets suppressed gray mold development. Moreover, treatment with Paenibacillus sp. TP3 of 2×107 CFU/ml in combination with chitosan improved the fruit production of strawberry in field trials. Besides, treatment with vegetative cell suspension on rose flower and lily leaves suppressed gray mold symptoms. Since bacteria-free culture supernatant showed significant suppression on spore germination, the anti-Botrytis acting compounds were further purified by DEAE ion exchange chromatography, and performed LC-MS/MS analysis. According to the result, the anti-Botrytis substances produced by Paenibacillus sp. TP3 were presumed to be novel proteins.