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RNA干扰介导昆士兰实蝇(双翅目:实蝇科)雄性育性基因的敲低

RNA Interference-Mediated Knockdown of Male Fertility Genes in the Queensland Fruit Fly (Diptera: Tephritidae).

作者信息

Cruz Carlos, Tayler Alison, Whyard Steve

机构信息

Department of Biological Sciences, University of Manitoba, Winnipeg, MB R3T 2N2, Canada.

出版信息

Insects. 2018 Aug 10;9(3):96. doi: 10.3390/insects9030096.

Abstract

The Queensland fruit fly, is Australia's most important horticultural pest. The Sterile Insect Technique (SIT) has been used to control this species for decades, using radiation to sterilize males before field-release. This method of sterilization can potentially reduce the insects' abilities to compete for mates. In this study, RNA interference (RNAi) techniques were examined for their potential to sterilize male without adversely affecting mating competitiveness. adults were injected or fed double-stranded RNAs (dsRNAs) targeting spermatogenesis genes (, and ); quantitative reverse-transcriptase PCR analyses confirmed that transcript levels were reduced 60⁻80% for all three genes following injections. Feeding produced a significant gene knockdown for and after three days, but interestingly, two genes ( and ) produced an excess of transcripts after 10 days of feeding. Despite these fluctuations in transcript levels, all three dsRNAs impacted the fecundity of treated males, with - and -dsRNA-treated males producing 75% fewer viable offspring than the negative controls. Mating competition assays demonstrated that dsRNA-treated males can actively compete with untreated males. These findings suggest that RNAi technology could serve as an alternative to radiation as a means of sterilizing these insects in an SIT program.

摘要

昆士兰果蝇是澳大利亚最重要的园艺害虫。几十年来,昆虫不育技术(SIT)一直被用于控制这种果蝇,即在野外释放之前利用辐射使雄性果蝇绝育。这种绝育方法可能会降低昆虫争夺配偶的能力。在本研究中,研究了RNA干扰(RNAi)技术使雄性果蝇绝育同时又不会对其交配竞争力产生不利影响的潜力。对成年果蝇注射或喂食靶向精子发生基因(、和)的双链RNA(dsRNA);定量逆转录聚合酶链反应分析证实,注射后这三个基因的转录水平均降低了60%-80%。喂食三天后,和出现了显著的基因敲低,但有趣的是,喂食10天后,两个基因(和)产生了过量的转录本。尽管转录水平存在这些波动,但所有三种dsRNA均影响了处理过的雄性果蝇的繁殖力,用-和-dsRNA处理的雄性果蝇产生的存活后代比阴性对照少75%。交配竞争试验表明,经dsRNA处理的雄性果蝇能够与未处理的雄性果蝇积极竞争。这些发现表明,在昆虫不育技术计划中,RNAi技术可以作为一种替代辐射的方法来使这些昆虫绝育。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/181b/6163526/f2ddcfb33a8a/insects-09-00096-g001.jpg

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