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National Chung Hsing University Institutional Repository - NCHUIR > 學術研究中心 > 生物科技發展中心 > 依資料類型分類 > 期刊論文 >  Display female-specific doublesex RNA interference in early generations of transformed oriental fruit fly, Bactrocera dorsalis (Hendel)

Please use this identifier to cite or link to this item: http://nchuir.lib.nchu.edu.tw/handle/309270000/126091

標題: Display female-specific doublesex RNA interference in early generations of transformed oriental fruit fly, Bactrocera dorsalis (Hendel)
作者: Chen, Shiu-Ling;Lu, Kuang-Hui;Dai, Shu-Mei;Li, Chia-Hsin;Shieh, Chwen-Jen;Chang, Cheng
Contributors: 國立中興大學生物科技發展中心
National Chung Hsing University,Biotechnology Center
Miao-zhen Luo
關鍵字: doublesex gene;Bactrocera dorsalis;RNA interference;short hairpin;RNA;reverse transcription real-time PCR;sex-determining gene;drosophila-melanogaster;dipteran insects;mammalian-cells;musca-domestica;polymerase-iii;evolution;sirna;tephritidae;expression
日期: 2011-4
Issue Date: 2012-10-19 15:15:23 (UTC+8)
摘要: BACKGROUND: The oriental fruit fly, Bactrocera dorsalis (Hendel), is one of the most destructive pests in many Asian countries. An effective strategy to reduce fly density in the field is urgently required. Recently, the doublesex of B. dorsalis (Bddsx(f)) has been cloned, and RNA interference (RNAi) indicates that it can reduce the offspring in vitro. In this study, a piggyBac-based construct that generates short hairpin RNA (shRNA) against the female-specific region of Bddsx was introduced into the pest to test the RNAi effects on reproductive functions in vivo. RESULTS: After embryonic injection and backcross, 21 transgenic lines with germline transformation were identified. Genomic DNA analysis showed that the exogenous transgene including short hairpin Bddsx(f) and a DsRed marker had integrated into the genomes of 11 transformed lines. Northern blot analysis indicated the presence of Bddsx(f) short interfering RNA (siRNA) under the control of a U6 promoter in transformed flies. As expected, the specific effects of RNAi led to the delay of egg maturation, and the offspring was significantly reduced. Reverse transcription real-time PCR further demonstrated that in vivo interference not only specifically inhibited the Bddsxf transcript but also repressed expression of the downstream yolk protein gene (Bdyp1). CONCLUSION: The results clearly indicate that RNAi is heritable through the expression of specific siRNA in early generations of transformed oriental fruit fly. These results can broaden the understanding of sex-related developmental mechanisms in the fly, and also offer a possible molecular approach for pest control in the future. (C) 2011 Society of Chemical Industry
Relation: Pest Management Science, Volume 67, Issue 4, Page(s) 466-473.
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