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中文摘要：
【目的】针对粉虱类害虫种类多、体型微小、形态相似、难以准确快速识别的问题,以新入侵我国大陆的甘蓝粉虱Aleyrodes proletella（L.）为靶标,以田间常见的其他10种/隐种粉虱为参照,采用基于线粒体DNA细胞色素C氧化酶亚基I（mitochondrial DNA cytochrome c oxidase subunit I,mt DNA COI）基因的种特异性（species-specific COI,SS-COI）PCR方法,研究其快速分子检测技术。【方法】利用mt DNA COI基因通用型引物LCO-1490/HCO-2198获得甘蓝粉虱及其他常见粉虱的COI序列,根据测序结果设计种特异性SS-COI引物1对（APZYJF/APZYJR）,其扩增片段大小为384 bp,同时对该对引物的种特异性及灵敏性进行检测。【结果】种特异性检验结果显示,该对引物仅对甘蓝粉虱的mt DNA COI基因具有扩增效果,对我国常见的其他种类的粉虱包括温室粉虱Trialeurodes vaporariorum（Westwood）、柑橘粉虱Dialeurodes citri（Ashmead）、螺旋粉虱Aleurodicus disperses（Russell）、双钩巢粉虱Paraleyrodes pseudonaranjae Martin、非洲伯粉虱Bemisia afer（Priesner et Hosny）以及烟粉虱B.tabaci（Gennadius）5个隐种（MED,Asia I,Asia II 1,Asia II 3和China 1）等不具有交叉反应扩增能力。灵敏性检验结果显示,该对引物不仅对不同性别的成虫具有良好的扩增效能,对2-4龄若虫甚至单粒卵或单头初孵若虫亦具有同样的扩增能力,其最低检测阈值为14.00±0.37 pg/μL（相当于1/25 600头雌性成虫）。【结论】该技术体系完全可用于甘蓝粉虱的快速准确识别及检测监测,对有效阻截其进一步扩张蔓延意义重大。
英文摘要：
【Aim】Morphological identification of Aleyrodes proletella（ L.） is limited by small size,high degree of similarity to related species and polymorphism. This study aims to develop a technique based on mitochondrial DNA cytochrome c oxidase subunit I（ mt DNA COI） gene sequence for the rapid identification of A. proletella,an invasive species newly found in mainland China. 【Methods 】 The fragments of mt DNA COI gene of A. proletella and ten other whitefly species or cryptic species were amplified and sequenced using COI gene universal primers LCO-1490 / HCO-2198. One pair of speciesspecific COI（ SS-COI） primers,APZYJF and APZYJR,was designed. The target fragment amplified by the SS-COI primers（ APZYJF / APZYJR） is 384 bp in length. The species specificity and sensitivity ofthe primer pair were tested. 【Results】The species specificity of the primer pair was validated using ten other whitefly species and cryptic species,i. e.,Trialeurodes vaporariorum（ Westwood）,Dialeurodes citri（ Ashmead）, Aleurodicus disperses（ Russell）, Paraleyrodes pseudonaranjae Martin, Bemisia afer（ Priesner et Hosny）,and five cryptic species of B. tabaci（ Gennadius）（ MED,Asia I,Asia II 1,Asia II 3 and China 1）. All A. proletella specimens were successfully detected,and no cross reaction with other whitefly species or cryptic species was observed. The method was tested using individual male or female adult,individual 2nd instar,3rd instar or red-eyed nymph,and even one grain of egg or single newly emerged 1st instar nymph,and proved to be applicable for all of these life stages. Sensitivity test results demonstrated that successful amplification could be obtained with the concentration of template DNA as low as 14. 00 ± 0. 37 pg / μL,equal to 1 /25 600 of a whole female adult of A. proletella.【Conclusion】The SS-COI method developed here provides a quick,simple and reliable molecular technique for the identification and monitoring of A. proletella,which would be useful in intercepting and b
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