The Drosophila melanogaster β2-tubulin gene (Dm-β2t) controls the function of microtubules in the testis and sperm, and has been evaluated for use in biocontrol strategies based on the sterile insect technique, including sexing and the induction of male sterility. The spotted-wing Drosophila (Drosophila suzukii) is native to eastern Asia but has spread globally as an invasive pest of fruit crops, so biocontrol strategies are urgently required for this species. We therefore isolated the β2tubulin ortholog Ds-β2t from the USA laboratory strain of D. suzukii and confirmed the presence of functional motifs by aligning orthologs from multiple insects. The developmental expression profile of Ds-β2t was determined by RT-PCR using gene-specific primers and was similar to that of Dm-β2t. We then isolated the Ds-β2t promoter and used it to generate transgenic strains expressing a testis-specific fluorescent protein starting from the thirdinstar larvae. Efficient sexing was achieved based on fluorescence detection, and the transgenic males showed a similar survival rate to wild-type males. Fluorescence imaging and PCR were also used to confirm the insemination of wild-type females by transgenic males. We therefore confirm that D. suzukii strains expressing fluorescent markers under the control of the Ds-β2t promoter can be used for sexing and the confirmation of mating, and we discuss the wider potential of the Ds-β2t promoter in the context of genetic control strategies for D. suzukii.

在黑腹果蝇β2微管蛋白基因（DM-β2T）控制在睾丸和精子微管的功能，以及用于基于所述不育昆虫技术，包括性别鉴定和雄性不育的诱导生物防治策略使用已被评估。斑翅果蝇（Drosophila suzukii）原产于东亚，但已作为水果作物的一种侵入性害虫在全球范围内传播，因此该物种迫切需要采取生物防治策略。因此，我们从美国铃木D.实验室菌株中分离了β2微管蛋白直向同源物Ds-β2t 。并通过比对来自多个昆虫的直系同源物来证实功能性基序的存在。使用基因特异性引物通过RT-PCR确定Ds-β2t的发育表达谱，并且与Dm-β2t相似。然后，我们分离了Ds-β2t启动子，并用它来生成从第三龄幼虫开始表达睾丸特异性荧光蛋白的转基因菌株。基于荧光检测，可以实现有效的性别鉴定，转基因雄性的存活率与野生型雄性相似。荧光成像和PCR还用于确认转基因雄性对野生型雌性的授精。因此，我们确认铃木D.所述的控制下表达荧光标记物的菌株的Ds-β2T启动子可用于性别鉴定和配合的确认，并讨论了更广泛的潜在的Ds-β2T在用于遗传控制策略的上下文中的启动子D. suzukii。