Anastrepha fraterculus sp. 1 is considered a quarantine pest in several American countries. Since chemical control applied in an integrated pest management program is the only strategy utilized against this pest, the development of pesticide-free methods, such as the Sterile Insect Technique, is being considered. The search for genes involved in sex-determination and differentiation, and in metabolic pathways associated with communication and mating behaviour, contributes with key information to the development of genetic control strategies. The aims of this work were to perform a comprehensive analysis of A. fraterculus sp. 1 transcriptome and to obtain an initial evaluation of genes associated with main metabolic pathways by the expression analysis of specific transcripts identified in embryos and adults. Sexually mature adults of both sexes and 72 h embryos were considered for transcriptome analysis. The de novo transcriptome assembly was fairly complete (62.9% complete BUSCO orthologs detected) with a total of 86,925 transcripts assembled and 28,756 GO annotated sequences. Paired-comparisons between libraries showed 319 transcripts differently expressed between embryos and females, 1242 between embryos and males, and 464 between sexes. Using this information and genes searches based on published studies from other tephritid species, we evaluated a set of transcripts involved in development, courtship and metabolic pathways. The qPCR analysis evidenced that the early genes serendipity alpha and transformer-2 displayed similar expression levels in the analyzed stages, while heat shock protein 27 is over-expressed in embryos and females in comparison to males. The expression of genes associated with courtship (takeout-like, odorant-binding protein 50a1) differed between males and females, independently of their reproductive status (virgin vs mated individuals). Genes associated with metabolic pathways (maltase 2-like, androgen-induced gene 1) showed differential expression between embryos and adults. Furthermore, 14,262 microsatellite motifs were identified, with 11,208 transcripts containing at least one simple sequence repeat, including 48% of di/trinucleotide motifs. Our results significantly expand the available gene space of A. fraterculus sp. 1, contributing with a fairly complete transcript database of embryos and adults. The expression analysis of the selected candidate genes, along with a set of microsatellite markers, provides a valuable resource for further genetic characterization of A. fraterculus sp. 1 and supports the development of specific genetic control strategies.

中文翻译：

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Anastrepha fraterculus sp 的转录组分析。1 雄性、雌性和胚胎：深入了解发育、求偶和繁殖

按实蝇 sp. 1 在美国的几个国家被视为检疫性害虫。由于综合害虫管理计划中应用的化学防治是对抗这种害虫的唯一策略，因此正在考虑开发无农药方法，例如昆虫不育技术。寻找参与性别决定和分化以及与交流和交配行为相关的代谢途径的基因，为遗传控制策略的发展提供了关键信息。这项工作的目的是对 A. fraterculus sp. 进行全面分析。1 转录组，并通过对胚胎和成人中鉴定的特定转录本的表达分析来获得与主要代谢途径相关的基因的初步评估。考虑对两性的性成熟成虫和 72 小时胚胎进行转录组分析。从头转录组组装相当完整（检测到 62.9% 完整的 BUSCO 直向同源物），总共组装了 86,925 个转录本和 28,756 个 GO 注释序列。文库之间的配对比较显示，胚胎和女性之间有 319 个转录本差异表达，胚胎和男性之间有 1242 个转录本差异表达，性别之间有 464 个转录本差异表达。利用这些信息和基于其他实蝇物种已发表研究的基因搜索，我们评估了一组涉及发育、求偶和代谢途径的转录本。qPCR 分析证明，早期基因 Serendipity α 和 Transformer-2 在分析阶段表现出相似的表达水平，而与雄性相比，热休克蛋白 27 在胚胎和雌性中过度表达。与求偶相关的基因（类似外卖的气味结合蛋白 50a1）的表达在雄性和雌性之间存在差异，与它们的生殖状态（处女与交配个体）无关。与代谢途径相关的基因（麦芽糖酶 2 样、雄激素诱导基因 1）在胚胎和成人之间表现出差异表达。此外，还鉴定了 14,262 个微卫星基序，其中 11,208 个转录物包含至少一个简单的序列重复，其中包括 48% 的二/三核苷酸基序。我们的结果显着扩展了 A. fraterculus sp 的可用基因空间。1，贡献了相当完整的胚胎和成人转录数据库。所选候选基因的表达分析以及一组微卫星标记为进一步鉴定 A. fraterculus sp 的遗传特征提供了宝贵的资源。1并支持特定遗传控制策略的制定。