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標題: 台灣重要作物害蟲對有機磷劑抗藥性發生之基因體學及其功能分析-以基因轉殖與RNA默化技術探討抗有機磷殺蟲劑,福木松與乃力松,之東方果實蠅體內抗性基因之功能
Functional Analysis of Genes Responsible for the Resistance to Orgnophosphorus Pesticides, Formothion and Naled, in the Oriental Fruit Fly, Bactrocera Dorsalis, Using Techniques of Rna Interference and Insect Transgenesis
作者: 路光暉
關鍵字: 植物保護類;基礎研究;organophosphorus insecticides;insecticide resistance;high throughput DNA sequencing;transgenesis;RNA interference;Bactrocera dorsalis
施用化學藥劑仍是目前防治經濟害蟲之主要方法,但害蟲抗藥性之產生往往導致施藥量增加、藥劑殘留和環境污染等問題。以往相關研究常是針對單一農藥類型產生抗藥性之特定基因(如乙烯膽鹼脂酶對有機磷類藥劑之感受性與其酵素活性)進行探討,再廣泛以此標定基因之表現,測試該基因/酵素活性對其他農業用藥之感受性。單一項面之研究方式使得我們對抗藥性的發生和害蟲生理機制的改變等相關資訊單純化並忽略其他抗藥性機制存在之可能性。為跳脫既定之研究模式,引入新穎分析模式,本計畫擬以巨觀角度著手,配合迅速與精準之高通量定序技術,全面性解序抗性與感性害蟲品系之轉錄組序列,建立抗藥性基因資料庫;再依此資訊分別進行:1)特定抗性基因之篩選並以基因轉殖或RNA靜默技術進行功能上的研究;2)生物資訊模擬酵素與藥劑之作用機制分析,預期依此等結果作為現有藥劑之施用原則並開發新式殺蟲藥劑之依據。本計畫擬以對福木松和乃力松產生抗藥性之東方果實蠅(Bactrocera dorsalis (Hendel))品系與感性品系為對象,利用抗藥性機制之發生為研究切入點,於三年期間,分年進行下列研究:(一)將整個轉錄組(transcriptome)以新一代高通量定序法解序,以建立抗藥性相關基因之資料庫;(二)以晶片篩選與減除反應(subtraction)所得之基因表現資訊,篩選特定抗性基因,進而由基因表現與蛋白質活性兩方面探討其與感性品系之差異;(三)以基因轉殖配合RNA靜默或其他相關技術進行特定基因功能之探討。

Chemical insecticides are still a major method used to control economicpests; however, the occurrence of pest resistance has often led to more seriousproblems such as pesticide residues, and environmental pollution. Traditionalresearches on pesticide resistant mechanisms in insects are often focused on asingle target gene, for example acetylcholine esterase (AchE), and extensivelyinvestigated the involvement of the gene in resistance to the pesticide. In thisproject, we intend to apply a new concept which is to comprehensively screenresistance-related genes by analysis of the whole gene population containing inthe transcriptomes of resistant and susceptible insects and to establish thedatabases of the resistant-related genes. Base on these information, and then wewill further intensively screen for specific resistance genes and reveal theirpossible roles in the pesticide resistance using techniques such as RNAinterference (RNAi) and insect transgenesis; moreover, we will also usebioinformatics and protein simulation systems to analyze the enzyme functionsof targeted resistant proteins. We expect that such results should be able toprovide certain biological information for instructing effective pesticideapplication and even for developing new insecticides. In this three-year period,we plan to select the Formothion- and Naled-resistant oriental fruit flyBactrocera dorsalis as the model insects to carry out an in depth study whatgenes are involved in their resistance and what the molecular mechanismsshould be; and the project will include: (1) to sequence the whole expressedgenes of the transcriptomes in the resistant and susceptible flies and use thesedata to establish and analyze the database for further use, (2) to detect andanalyze the function of several candidate genes which are obtained fromscreening with microarray chips and DNA subtraction, and (3) to reveal thepossible roles of the examined genes on the pesticide resistance using RNAinterference (RNAi) and insect transgenic technique.
其他識別: NSC100-2313-B005-011-MY3
Appears in Collections:昆蟲學系

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