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標題: | 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 Application of Bacillus spp. and Trichoderma spp. for growth promotion and control of anthracnose and Fusarium wilt in strawberry |
作者: | En-Sheng Tang 湯恩聖 |
指導教授: | 林乃君(Nai-Chun Lin) |
關鍵字: | 草莓炭疽病,萎凋病,芽孢桿菌,木黴菌,生物防治, strawberry anthracnose,strawberry Fusarium wilt,Bacillus sp.,Trichoderma sp.,biological control agent, |
出版年 : | 2018 |
學位: | 碩士 |
摘要: | 草莓是臺灣具高經濟價值之水果之一,主要產區位於苗栗縣大湖鄉和獅潭鄉一帶, 其育苗及本田初期歷經夏秋兩季高溫潮濕的氣候,病害問題嚴重,尤以 Colletotrichum sp. 引起之炭疽病,以及 Fusarium oxysporum f. sp. fragariae 引起之萎凋病為甚。炭疽病病勢發展快速但已有推薦藥劑,而萎凋病是近年日趨嚴重的病害,尚無推薦藥劑可使用。病害不僅增加農民在育苗期及本田初期的補植成本,產季期間由於草莓為連續採收作物,藥劑的安全採收期難以掌握,也使得農藥殘留風險提升,故開發安全防治的資材需求迫切。本研究旨在探討臺灣本土分得之二株Bacillus sp. 595 和 376 及二株 Trichoderma sp. ML1和ML56 促進草莓植株生長之潛力。此外,除上述四株微生物外,也加入 Serendipita indica 以了解其對於預防草莓炭疽病及萎凋病之效果。將 595 及 376 二菌株以 gyrB 引子對增幅之片段進行定序分析並比對美國國家生物技術資訊中心 (National Center for Biotechnology Information) 資料庫後得知 595 應可能為 Bacillus amyloliquefaciens,而 376 則最近似於 Bacillus subtilis。將 B. amyloliquefaciens 595 和 B. subtilis 376 分別以 OD600 = 0.05、0.1 和 0.2 三種濃度,而木黴菌分別以 1 × 105、1 × 106和1 × 107 conidia mL-1 三種濃度,搭配泥炭土或赤玉土兩種栽培介質,每週澆灌或隔週澆灌兩種施用頻率等四種模式,並以葉面積及植株重量作為生長評估指標,測試最適合之施用濃度與頻率。經過比較後,B. amyloliquefaciens 595 和 B. subtilis 376 之推薦施用濃度均為 OD600= 0.1, Trichoderma sp. ML1和 ML56 之推薦施用濃度分別為 1 × 105 conidia mL-1 及 1 × 107 conidia mL-1。比較不同組合處理對植株生長之影響,595+376+ML1 有助於增加植株葉面積,而595+376+ML56則是促進根系發展。;而預防草莓炭疽病之能力,以 S. indica、, Trichoderma sp. ML1 及 ML56 具有較佳之趨勢;預防草莓萎凋病之能力,以 595 及 ML56 有較佳之趨勢效果,但預防病害感染試驗結果在統計上並不顯著。綜觀上述數據根據本研究結果推薦之使用方法為育苗期接種 S. indica 並澆灌 ML56以預防作為炭疽病及萎凋病,本田初期澆灌 595+376+ML56 以促進根系發展後,再於生育期澆灌595+376+ML1 增進植株葉面積生長。 Strawberry is one of the economically important fruits in Taiwan. Due to the hot and humid weather in summer and fall, strawberry plants in the nurseries and in the early stage after transplanting, are threatened by two diseases, strawberry anthracnose caused by Colletotrichum sp. and strawberry Fusarium wilt caused by Fusarium oxysporum f. sp fragariae. The disease progression of strawberry anthracnose is very fast, and there are recommended pesticides, such as Difenconazole, Pyraclostrobin, and Thiabendazole. Fusarium wilt becomes severe during the past decade, but it just like other Fusarium disease, no recommended pesticide for it. Prevalence of anthracnose and Fusarium wilt not only increase the cost on replanting, but also increase the risk of pesticide residue, due to the continuous-harvest characteristic for strawberry production, which makes hard to keep the pre-harvest interval. Therefore, a safer control mean is an urgent demand. This study focused on investigation of the strawberry growth-promoting potential of two Bacillus species, 595 and 376, and two Trichoderma species, ML1 and ML56. In addition to these four microorganisms, Serendipita indica was also included to evaluate for their ability to prevent anthracnose and Fusarium wilt of strawberry. Based on the sequencing result of the gyrB fragment and sequencing amplified by primer gyrB and analysis using the NCBI database, 595 was is more related to Bacillus amyloliquenfaciens and 376 was identified as B. subtilis. To assess the concentration and frequency for the applications of these beneficial microorganisms regarding to their effects on leaf areas and dry weight of strawberry plants, three different concentrations for Bacillus spp., (OD600 = 0.05, 0.1, and 0.2), and three different concentrations for Trichoderma, (1 × 105, 1 × 106, and 1 × 107 conidia mL-1), in combination with two growing substrates, akadama clay granulate and peat moss, and two application frequencies, biweekly or per-weekly, were used. As the results showed, After the trial, the proper concentration is OD 600= 0.1 for B. amyloliquenfaciens 595, OD 600= 0.1 for B. subtilis 376, 1 × 105 conidia mL-1 for ML1, and 1 × 107 conidia mL-1 for ML56. For the effects on plant growth, strawberry plants treated with 595+376+ML1 have bigger leaf area while those treated with 595+376+ML56 had more root weight. S. indica, ML1 and ML56 have gave rise to more obvious trend than other treatments in the greenhouse trial. To prevent Fusarium wilt, 595 and ML56 were more effective on protection of strawberry from anthracnose whereas B. amyloliquenfaciens 595 and Trichoderma sp. ML56 were more effective on protection of strawberry from Fusarium wilt in greenhouse trials, but the result is not statistically significant. Overall speaking, a strategy includes inoculation of strawberry seedlings with S. indica and Trichoderma sp. ML56 to prevent anthracnose and Fusarium wilt in the nursery and drenching 595+376+ML56 for root development after transplanting into the field followed by drenching 595+376+ML1 for increasing leaf area is suggested for strawberry production based on the results obtained in this study. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/70429 |
DOI: | 10.6342/NTU201802078 |
全文授權: | 有償授權 |
顯示於系所單位: | 植物醫學碩士學位學程 |
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