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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 黃榮南(Rong-Nan Huang) | |
| dc.contributor.author | Yo-Shi Liu | en |
| dc.contributor.author | 劉又溪 | zh_TW |
| dc.date.accessioned | 2021-06-15T16:11:45Z | - |
| dc.date.available | 2020-08-28 | |
| dc.date.copyright | 2015-08-28 | |
| dc.date.issued | 2015 | |
| dc.date.submitted | 2015-08-18 | |
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(2013) Insecticidal activity in plant-beneficial pseudomonads: molecular basis and ecological relevance. Ph.D. thesis No.21005, ETH Zurich, Zurich. Sadanandane, C., Reddy, C. M. R., Prabakaran, G. and Balaraman, K. (2003) Field evaluation of a formulation of Pseudomonas fluorescens against Culex quinquefasciatus larvae and pupae. Acta. Trop. 87, 341-343. Wang, L. T., Tai, C. J., Wu, Y. C., Chen, Y. B., Lee, F. L. and Wang, S. L. (2010) Pseudomonas taiwanensis sp. nov., isolated from soil. Int. J. Syst. Evol. Microbiol. 60, 2094-8. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/52318 | - |
| dc.description.abstract | 本論文分析台灣假單胞菌 (Pseudomonas taiwanensis) 對白線斑蚊 (Aedes albopictus) 幼蟲毒性；台灣假單胞菌於Luria-Bertani培養基培養36小時後，取上清液及菌體進行殺蟲活性試驗。結果顯示斑蚊及家蚊幼蟲對台灣假單胞菌上清液菌均相當敏感，而菌體本身則不具殺蟲活性，顯示台灣假單胞菌殺蟲活性物質是一種外分泌性的代謝產物。而做為對照的大腸桿菌 (Escherichia coli)，其培養上清液對白線斑蚊幼蟲也具有相當毒性，處理1小時內死亡率即可達到100%，大腸桿菌菌體本身亦不具殺蟲活性。將二齡白線斑蚊幼蟲飼養於台灣假單胞菌上清液中2小時，即可造成接近100%的死亡率；而四齡白線斑蚊幼蟲對台灣假單胞菌上清液則較不敏感，但將四齡白線斑蚊幼蟲飼養於台灣假單胞菌上清液中12小時，仍可造成45%的死亡率；飼養24小時後，幼蟲死亡率亦可達100%。雖然前人已經從台灣假單胞菌菌體內(pellet) 得到對果蠅 (Drosophila melanogaster) 與小菜蛾 (Plutella xylostella) 具有殺蟲活性的基因tccC 所產生的毒蛋白群。但是本研究顯示台灣假單胞菌對白線斑蚊幼蟲之殺蟲活性並非源自TccC毒蛋白群，因為經高溫滅菌 (121 0C) 與蛋白酶K降解後，並不會影響台灣假單胞菌培養上清液之殺白線斑蚊幼蟲活性。台灣假單胞菌培養上清液對埃及斑蚊幼蟲及熱帶家蚊也具有相同毒性。台灣假單胞菌培養上清液在經過冷凍乾燥、減壓濃縮乾燥後，其等體積還原液則會失去殺幼蟲活性。分別使用有機溶劑 (正己烷、正丁醇、乙酸乙酯) 萃取分層後，僅有正己烷分層所獲得之水層保有原先毒殺白線斑蚊幼蟲之活性。切片觀察顯示台灣假單胞菌培養上清液處理的白線斑蚊幼蟲腸道組織受損。但以液相層析 (HPLC) 分析台灣假單胞菌培養上清液，並未發現與其殺幼蟲活性相關特殊成份。因此台灣假單胞菌上清液內之可能殺蟲有效物質仍須後續探討。 | zh_TW |
| dc.description.abstract | This thesis evaluates the mosquitocidal activity of Pseudomonas taiwanensis supernatant. The supernatant derived from P. taiwanensis culture in LB (Luria-Bertani) medium for 36 hours were subjected to mosquitocidal assay against Aedes and Culex mosquitoes. The results show that all mosquito larvae are very sensitive to the supernatant (but not the pellet) of P. taiwanensis, indicating that the mosquitocidal ingredients are extracellular metabolites. The mortality of 2nd instar Ae. albopictus larvae could reach 100% after exposed to the cultural supernatant for 2 hr. The 4th instar larvae exhibited less sensitivity toward P. taiwanensis, however it still showed around 40% mortality after 12 hr exposure to the supernatant. The supernatants (but not the pellet) of Escherichia coli are also demonstrated with significant toxicity to 2nd instar Ae. albopictus larvae, reaching 100% after exposed to the cultural supernatant less than 1 hr. Though an insecticidal gene (tccC) toward Drosophila was identified and cloned from the P. taiwanensis, it is apparently not the responsible toxin for Ae. albopictus larvae, since the mosquitocidal activity is not affect by autoclave at 121 0C for 30 min. However, lyophilization and vacuum evaporation under reduce pressure will abolish the mosquitocidal activity of P. taiwanensis supernatant. The mosquitocidal activity remains in water layer derived from hexane partition after fraction the supernatant with hexane, butanol and ethyl acetate, indicating that the mosquitocidal ingredients are hydrophobic metabolites. Moreover, the anatomical sections showed that the midgut was the major tissue affected by the P. taiwanensis supernatant treatment. However, the high performance liquid chromatography (HPLC) analysis did not show any particular ingredients specifically exist in P. taiwanensis supernatant. The active ingredients of P. taiwanensis supernatant or mechanism responsible for the mosquiticidal activity remain to be solved in the future. | en |
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| dc.description.tableofcontents | 誌謝 I 中文摘要 II Abstract III 目 錄 IV 壹、前言 1 貳、前人研究 4 一、假單胞菌屬 4 二、P. fluorescens外泌性蛋白對病媒蚊之防治應用 5 三、假單胞菌分泌之生物介面活性劑與抗蟲之關係 6 參、材料與方法 8 一、試驗昆蟲飼養 8 (一)幼蟲飼養: 8 (二)蛹之收集 : 8 (三)成蟲飼養 : 8 二、試驗P. taiwanensis之培養與菌液製備 8 (一) P. taiwanensis培養 8 (二) P. taiwanesis生長特性 9 (三)實驗菌液分離與製備 9 (四) P. taiwanensis培養菌液滅菌、稀釋、3kDa分子篩過濾及Proteinase K處理 10 (五)滲透壓測量 10 (六)有機溶劑分離萃取P. taiwanensis培養液 10 (七)冷凍乾燥P. taiwanensis培養液上清液 10 三、Bioassay 試驗方法 11 四、顯微鏡解剖與觀察 11 (一)腸道解剖 11 (二)掃描式電子顯微鏡表面拍攝 11 (三)固定 11 (四)脫水 12 (五)滲膠與包埋 12 (六)切片染色與鏡檢 12 五、減壓濃縮分層毒性測試 : 12 六、液相層析分析 12 肆、結果 14 一、P. taiwanensis生長曲線及毒性的關係 14 二、P. taiwanensis對白線斑蚊幼蟲之毒性 14 三、P. taiwanensis上清液對同齡期之埃及斑蚊、白線斑蚊、熱帶家蚊幼蟲之毒性測試 15 四、P. taiwanensis對不同齡期之白線斑蚊幼蟲之毒性測試 15 五、滲透壓測量 15 六、P. taiwanensis上清液經3kDa分子篩處理後對白線斑蚊幼蟲之毒性測試 15 七、P. taiwanensis稀釋上清液對白線斑蚊幼蟲之毒性測試 16 八、P. taiwanensis上清液經熱處理 (滅菌釜) 後對白線斑蚊幼蟲之毒性測試 16 九、P. taiwanensis上清液經冷凍乾燥處理等體積回溶對白線斑蚊幼蟲之毒性測試 16 十、P. taiwanensis上清液經Proteinase K處理後對白線斑蚊幼蟲之毒性測試 16 十一、P. taiwanensis上清液以有機溶劑萃取後之毒性測試 17 十二、P. taiwanensis 上清液以減壓濃縮乾燥萃取後之毒性測試 17 十三、白線斑蚊四齡幼蟲消化道之解剖 17 十四、白線斑蚊四齡幼蟲消化道之顯微厚切片觀查 17 十五、白線斑蚊四齡幼蟲體壁之顯微構造 18 十六、液相層析分析P. taiwanensis上清液 18 伍、討論 19 一、P. taiwanensis與E. coli對斑蚊幼蟲之毒性 19 二、P. taiwanensis上清液內之有效物質特性 20 三、主要成分為生物介面活性劑之可能性探討 22 四、白線斑蚊幼蟲病理組織觀察 23 五、液相層析分析 24 陸、參考文獻 25 柒、圖 29 圖一、P. taiwanensis與大腸桿菌 (E. coli) 之生長曲線圖。 29 圖二、 P. taiwanensis 不同時間培養菌液對二齡白線斑蚊幼蟲之生物活性測試。 30 圖三、 E.coli 不同時間培養菌液對二齡白線斑蚊幼蟲之生物活性測試。 31 圖四、培養36小時之菌液對白線斑蚊二齡幼蟲之毒性。 32 圖五、P. taiwanensis 36小時培養上清液對白線斑蚊、埃及斑蚊、與熱帶家蚊二齡幼蟲之毒性。 33 圖六、P. taiwanensis 上清液對二齡與四齡白線斑蚊幼蟲之毒性比較。 34 圖七、P. taiwanensis培養液清液之滲透壓。 35 圖八、P. taiwanensis培養上清液經3kDa分子篩過濾處理後對白線斑蚊幼蟲之毒性影響。 36 圖九、P. taiwanensis培養上清液不同稀釋倍數對白線斑蚊二齡幼蟲之毒性。 37 圖十、E.coli 培養上清液不同稀釋倍數對白線斑蚊二齡幼蟲之毒性。 38 圖十一、加熱處理P. taiwanensis與E.coli之培養上清液對白線斑蚊二齡幼蟲毒性之影響。 39 圖十二、冷凍乾燥處理P. taiwanensis培養上清液對其毒殺白線斑蚊幼蟲之影響。 40 圖十三、P. taiwanensis上清液經Proteinase K處理後對白線斑蚊幼蟲之毒性影響。 41 圖十四、P. taiwanensis 上清液殺蟲活性成份分離。 42 圖十五、減壓濃縮對P. taiwanensis培養上清液毒性之影響。 43 圖十六、對照組(LB medium)處理12小時之白線斑蚊四齡幼蟲中腸塑膠切片。 44 圖十七、經P. taiwanensis上清液處理12小時之白線斑蚊四齡幼蟲中腸塑膠切片。 45 圖十八、P. taiwanensis36小時培養上清液處理之白線斑蚊四齡幼蟲消化道。 46 圖十九、掃描式電子顯微鏡觀察經P. taiwanensis上清液處理之白線斑蚊四齡幼蟲胸節表皮變化。 47 圖二十、LB medium於三種波長下之液相層析 (HPLC) 圖譜。 48 圖二十一、LB medium 經 3kDa分子篩處理後於三種波長下之液相層析 (HPLC) 圖譜。 49 圖二十二、P. taiwanensis 上清液於三種波長下之液相層析 (HPLC) 圖譜。 50 圖二十三、P. taiwanensis上清液經3kDa分子篩處理後，於三種波長下之液相層析 (HPLC) 圖譜。 51 圖二十四、E.coli上清液於三種波長下之液相層析 (HPLC) 圖譜。 52 圖二十五、LB medium培養液於HPLC全波長掃描圖譜。 53 圖二十六、P. taiwanensis上清液經正己烷分餾後之水層於HPLC全波長掃描圖譜。 53 圖二十七、P. taiwanensis上清液經正己烷分離後的水層以減壓濃縮抽乾，再等體積回溶 (原濃度) 後之HPLC全波長掃描圖譜。 54 圖二十八、P. taiwanensis上清液經正己烷分離後的水層以減壓濃縮抽乾，再回溶成兩倍濃度後，於HPLC全波長之掃描圖譜。 54 圖二十九、P. taiwanensis上清液經冷凍乾燥後之還原液於HPLC全波長之掃描圖譜。 55 | |
| dc.language.iso | zh-TW | |
| dc.subject | 生物殺蟲劑 | zh_TW |
| dc.subject | 白線斑蚊 | zh_TW |
| dc.subject | 台灣假單胞菌 | zh_TW |
| dc.subject | 大腸桿菌 | zh_TW |
| dc.subject | 菌液上清代謝物 | zh_TW |
| dc.subject | Pseudomonas taiwanensis | en |
| dc.subject | Aedes albopictus | en |
| dc.subject | bio-pesticide | en |
| dc.subject | extracellular metabolites | en |
| dc.subject | Escherichia coli | en |
| dc.title | 台灣假單胞菌與大腸桿菌培養液對蚊子幼蟲毒性研究 | zh_TW |
| dc.title | The toxicity of Pseudomonas taiwanensis and Escherichia coli culture supernatant against mosquito larvae | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 103-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 許如君(Ju-Chun Hsu),蔡坤憲(Kun-Hsien Tsai),劉?睿(Je-Ruei Liu) | |
| dc.subject.keyword | 白線斑蚊,台灣假單胞菌,大腸桿菌,菌液上清代謝物,生物殺蟲劑, | zh_TW |
| dc.subject.keyword | Aedes albopictus,Pseudomonas taiwanensis,Escherichia coli,extracellular metabolites,bio-pesticide, | en |
| dc.relation.page | 55 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2015-08-18 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 昆蟲學研究所 | zh_TW |
| 顯示於系所單位： | 昆蟲學系 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-104-1.pdf 未授權公開取用 | 2.12 MB | Adobe PDF |
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