The spotted-wing Drosophila (Drosophila suzukii) is a widespread invasive pest that causes severe economic damage to fruit crops. The early development of D. suzukii is similar to that of other Drosophilids, but the roles of individual genes must be confirmed experimentally. Cellularization genes coordinate the onset of cell division as soon as the invagination of membranes starts around the nuclei in the syncytial blastoderm. The promoters of these genes have been used in genetic pest-control systems to express transgenes that confer embryonic lethality. Such systems could be helpful in sterile insect technique applications to ensure that sterility (bi-sex embryonic lethality) or sexing (female-specific embryonic lethality) can be achieved during mass rearing. The activity of cellularization gene promoters during embryogenesis controls the timing and dose of the lethal gene product. Here, we report the isolation of the D. suzukii cellularization genes nullo, serendipity-α, bottleneck and slow-as-molasses from a laboratory strain. Conserved motifs were identified by comparing the encoded proteins with orthologs from other Drosophilids. Expression profiling confirmed that all four are zygotic genes that are strongly expressed at the early blastoderm stage. The 5′ flanking regions from these cellularization genes were isolated, incorporated into piggyBac vectors and compared in vitro for the promoter activities. The Dsnullo promoter showed the highest activity in the cell culture assays using D. melanogaster S2 cells. The similarities in the gene coding and 5′ flanking sequence as well as in the expression pattern of the four cellularization genes between D. melanogaster and D. suzukii, suggest that conserved functions may be involved in both species. The high expression level at the early blastoderm stage of the four cellularization genes were confirmed, thus their promoters can be considered in embryonic lethality systems. While the Dsnullo promoter could be a suitable candidate, all reported promoters here are subject to further in vivo analyses before constructing potential pest control systems.

中文翻译：

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四个果蝇铃木细胞化基因及其启动子的鉴定与表征

斑翅果蝇（Drosophila suzukii）是一种广泛入侵的害虫，对果农造成严重的经济损失。铃木蝇的早期发育与其他果蝇相似，但必须通过实验确定单个基因的作用。一旦细胞合胞胚层中的细胞核膜开始围绕细胞核侵入，细胞化基因就会协调细胞分裂的开始。这些基因的启动子已用于遗传虫害控制系统中，以表达赋予胚胎致死性的转基因。这样的系统在无菌昆虫技术应用中可能会有所帮助，以确保在大规模饲养过程中可以实现无菌（双性胚胎致死率）或性别（女性特异性胚胎致死率）。胚胎发生过程中细胞化基因启动子的活性控制致死基因产物的时间和剂量。在这里，我们报告了从实验室菌株中分离出的铃木D.细胞化基因nullo，偶然性α，瓶颈和糖蜜缓慢。通过将编码的蛋白质与其他果蝇的直向同源物进行比较来鉴定保守的基序。表达谱证实，所有四个都是在胚盘早期强烈表达的合子基因。分离来自这些细胞化基因的5'侧翼区域，并入piggyBac载体中，并在体外比较启动子活性。Dsnullo启动子在使用D. melanogaster S2细胞进行的细胞培养分析中显示出最高的活性。D. melanogaster和D. suzukii之间的基因编码和5'侧翼序列以及四个细胞化基因的表达模式的相似性表明，保守的功能可能与这两个物种有关。确认了四个细胞化基因在胚盘早期的高表达水平，因此可以在胚胎致死系统中考虑其启动子。尽管Dsnullo启动子可能是合适的候选者，但在构建潜在的有害生物控制系统之前，这里所有报道的启动子都需要进行进一步的体内分析。因此，它们的启动子可以在胚胎致死系统中考虑。尽管Dsnullo启动子可能是合适的候选者，但在构建潜在的有害生物控制系统之前，这里所有报道的启动子都需要进行进一步的体内分析。因此，它们的启动子可以在胚胎致死系统中考虑。尽管Dsnullo启动子可能是合适的候选者，但在构建潜在的有害生物控制系统之前，这里所有报道的启动子都需要进行进一步的体内分析。