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螺旋粉虱Aleurodicus dispersus Russell是一种严重威胁果树、蔬菜和园林植物生产的入侵性害虫。由于该粉虱的卵和初孵若虫体型微小,与其它粉虱类害虫形态相似,且易于携带传播,难以快速准确鉴别,本研究采用特征序列扩增区域(SCAR)标记法,通过对目的片段的克隆与测序,设计螺旋粉虱特征片段扩增引物,并对该引物的种特异性和灵敏性进行检验。结果显示,利用SCAR标记法获得了螺旋粉虱特异性片段(632bp,GenBank登录号为HM240855),根据此片段的碱基序列设计1对螺旋粉虱特异性引物,其扩增片段大小为307bp;该对引物只对螺旋粉虱具有扩增能力,对近缘种、属粉虱不具有扩增效果。该引物不仅对成虫具有良好的扩增效能,对单粒卵、1~3龄若虫和拟蛹等亦具有同样的扩增能力,其最低检出阈值为1/6400头成虫。从而表明,该SCAR标记技术完全可用于螺旋粉虱的鉴定识别及其检测和监测。
Aleurodicus dispersus Russell is an invasive pest that is a serious threat to the production of fruit trees, vegetables and ornamental plants. Because the eggs and newly hatched nymphs of the whitefly are small in shape and similar in morphology to other whitefly pests and are easy to carry and spread, it is difficult to identify them rapidly and accurately. In this study, by using the characteristic sequence amplification region (SCAR) The cloning and sequencing of the primers were designed to amplify the primers of the characteristic fragment of B. tabaci. The species specificity and sensitivity of the primers were tested. The results showed that a specific fragment (632bp, GenBank accession number HM240855) was obtained by SCAR marker. Based on the nucleotide sequence of this fragment, a pair of specific primers was designed and its amplified fragment size was 307 bp. The pair of primers only has the ability to amplify the whitefly, and has no amplification effect on the related species and whitefly. The primer not only has good amplification effect on adults, but also has the same amplification ability on single egg, first to third instar nymphs and pseudo-pupae, with the minimum detection threshold of 1/6400 adult worms. Thus, the SCAR marker technology can be used to identify, detect and monitor the whitefly.