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研究生: 劉聖文
Sheng-Wen Liu
論文名稱: 賜諾殺穩定性及東方果實蠅費洛蒙之研究
Studies on the stability of spinosad and on the pheromone of the Oriental Fruit Fly, Bactrocera dorsalis (Hendel)
指導教授: 高永旭
Yung-Hsi Kao
黃榮南
Rong-Nan Huang
口試委員:
學位類別: 碩士
Master
系所名稱: 生醫理工學院 - 生命科學系
Department of Life Science
畢業學年度: 97
語文別: 中文
論文頁數: 81
中文關鍵詞: 賜諾殺東方果實蠅費洛蒙
外文關鍵詞: spinosad, bactrocera dorsalis (Hendel), pheromone
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  • 東方果實蠅為我國最重要之果樹害蟲,防治上主要使用0.02 % 賜諾殺濃餌劑 (香兒寶) 及甲基丁香油滅雄法,然而已有許多有關害蟲對賜諾殺產生抗性之報導,此外迄今尚無一種有效果實蠅雌蟲誘引劑。故本研究主要目的為探討賜諾殺濃餌劑有效成分 (賜諾殺) 之穩定性以及果實蠅誘雌劑。HPLC 結果顯示,賜諾殺濃餌劑於溫度 20~40°C 及 pH 5~9下三週對其有效成分並無顯著影響。但在 UVC 照射下,不論是賜諾殺濃餌劑所含賜諾殺或賜諾殺標準品,皆有隨照射時間增加而產生光解的現象,此情形又以標準品更為明顯。添加化學藥品 (如碳酸銨或甲基丁香油) 會加速賜諾殺濃餌劑有效成分光解;但加入醋酸銨後,除了能增加對果實蠅誘引效果外,還有助於賜諾殺之光穩定性;進一步實驗亦發現賜諾殺濃餌劑所含賜諾殺於酸性環境中較為穩定,顯示pH 值在賜諾殺光解作用中扮演重要角色。此外,發現果實蠅雄蟲正己烷萃取物對於果實蠅雌蟲具良好誘引效果,利用固相微萃取法 (Solid-phase microextraction, SPME) 及GC-MS 鑑定分析,顯示farnesene可能為果實蠅性費洛蒙成分之一,farnesene對果實蠅雌雄蟲均有三成以上誘引力,而α-pinene最具誘雌效果,誘引力可達 35 %。以複成分配方進行誘引試驗,發現複成分 (Farnesene/Limonene/α-Pinene) 較單成分對果實蠅更具誘引性 (43.3 %) (P>0.05),推測東方果實蠅之性費洛蒙可能不只單一成分。至於雄蟲所釋放費洛蒙是否具有產卵誘引性則有待進一步確認。


    The oriental fruit fly, Bactrocera dorsalis (Hendel), is one of the major fruit pests in Taiwan. Current emphasis in area-wide management program use the spinosad-based fruit fly bait (GF-120) or methyl eugenol combined with naled to attract and kill oriental fruit fly. However, spinosad-resistance have been recorded in some pests and no effectiveness of female-based attractant have been developed. The aims of this thesis are to investigate the stability of spinosad and to identify the potential attractants for female oriental fruit fly. The HPLC results showed that the active ingredient of GF-120 was stable either at 20-40°C or at pH 5-9 for three weeks. However, both spinosad standard and GF-120 were rapidly degraded under the condition of UVC irradiation, especially in spinosad standard. The results also showed that either ammonium carbonate or methyl eugenol might enhance the degradation of GF-120, while ammonium acetate could effectively increase the attractive efficiency by decreasing the photolysis of GF-120. Furthermore, the photodegradation rate of GF-120 was slower in acidic aqueous solution than in basic condition, indicating the matrix pH may play an important role in photolysis of GF-120. In different experiments, the hexane extract of male flies exhibited attractive activity toward female flies. The components identified by SPME/GC-MS to be the candidate for the sex pheromone is farnesene. Further analysis showed that farnesene could attract both female and male flies by above 30 %, and that α-pinene attracted to female flies with 35 % attraction rate. Further experiments found the multiple components (Farnesene/Limonene/α-Pinene) were more attractive than single component (P>0.05), suggesting the multiple components of sex pheromone of oriental fruit fly. Whether the male-released attractant also showed oviposition preference for female flies need further investigation.

    目錄 ………………………………………………………………Ⅰ 圖目錄 …………………………………………………………IV 表目錄 …………………………………………………………V 縮寫與全名對照表 ……………………………………………VI 壹、前言 一、東方果實蠅 …………………………………………………1 二、果實蠅生活史 ………………………………………………1 三、現今果實蠅防治技術 (一) 使用費洛蒙誘引之滅雄技術 ………………………………3 (二) 噴灑賜諾殺濃餌劑 …………………………………6 四、研究動機 ………………………………………………………9 貮、材料與方法 一、供試果實蠅 ………………………………………………11 二、賜諾殺濃餌劑穩定性研究 …………………………………11 (一) 高效液相層析 (High performance liquid chromatography, HPLC) 分析方法 …………………………………………11 (二) 賜諾殺貯存標準液 (Standard stock solution) 之配製 …11 (三) 賜諾殺標準檢量線 (Standard calibration curve) 製作 …12 (四) 賜諾殺濃餌劑穩定性因子探討 ………………………12 三、果實蠅性費洛蒙之研究 (一) 前驅誘蟲活性檢定 ………………………………………13 (二) 費洛蒙萃取方法 ………………………………………15 (三) 化學鑑定 ………………………………………16 (四) 單成分配方對果實蠅誘引試驗 (非選擇性) …………17 (五) 複成分配方對果實蠅誘引試驗 (非選擇性) …………17 叄、結果 一、賜諾殺濃餌劑穩定性分析 …………………………………18 (一) 不同溫度及 pH 值對賜諾殺濃餌劑穩定性之影響 ……18 (二) 光照 (UVC) 對賜諾殺濃餌劑穩定性之影響 ……………18 二、賜諾殺濃餌劑對光穩定性因子探討 ………………………19 (一) 不同化學藥品對賜諾殺濃餌劑光穩定性之影響 ………19 (二) 誘引劑對賜諾殺濃餌劑光穩定性之影響 ……………19 (三) 不同 pH 值對賜諾殺濃餌劑光穩定性之影響 ………20 三、東方果實蠅揮發性成分之萃取與分析鑑定 ………………20 (一) 東方果實蠅雄蟲正己烷萃取物對果實蠅之誘引效果 …20 (二) 使用GC-MS 分析SPME 萃取之果實蠅費洛蒙成分 …21 (三) 東方果實蠅費洛蒙單成分對果實蠅之誘引效果 ……22 (四) 東方果實蠅費洛蒙複成分配方對果實蠅之誘引效果 ……22 肆、討論 一、賜諾殺濃餌劑所含賜諾殺光解作用之探討 …………………24 (一) 醋酸銨、碳酸銨或甲基丁香油對賜諾殺濃餌劑光穩定性之影 響.……………………………………………………………25 (二) 賜諾殺濃餌劑於酸性環境中 (pH 5) 對光較穩定 ………25 二、東方果實蠅費洛蒙成分分析及誘引效果之探討 ……………26 (一) 使用 SPME 方式萃取所發現之東方果實蠅已知費洛蒙成分 (二) 使用 SPME 方式萃取所發現之東方果實蠅未知費洛蒙成分 (三) 東方果實蠅費洛蒙複成分配方較單成分對果實蠅更具誘引力 圖表…………………………………………………………………30 參考文獻……………………………………………………………47 附錄…………………………………………………………………58

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