DataSheet_1_Transcriptome Analysis of Ostrinia furnacalis Female Pheromone Gland: Esters Biosynthesis and Requirement for Mating Success.xlsx

Female moths use sex pheromones to attract males, and corresponding regulatory mechanism underlying sex pheromone biosynthesis is species-dependent. However, the detailed mechanism involved in sex pheromone biosynthesis in Ostrinia furnacalis has not yet been fully addressed. In the present study, transcriptome sequencing of O. furnacalis pheromone glands screened a serials of candidate genes involved in sex pheromone biosynthesis. Our analysis showed that sex pheromone release in O. furnacalis females arrives its peak at the 2nd scotophase, consistent with its mating behavior. Pheromone biosynthesis-activating neuropeptide (PBAN) was confirmed to regulate sex pheromone biosynthesis, and Ca2+ is the secondary messenger of PBAN signaling in O. furnacalis. The functional analysis of candidate genes demonstrated that the decreased mRNA levels or activities of calcineurin (CaN) and acetyl-CoA carboxylase (ACC) led to significant decrease in sex pheromone production and female capability to attract males, as demonstrated by RNAi-mediated knockdown and pharmacological inhibitor assay. Most importantly, the activities of CaN and ACC depend on the activation of PBAN/PBANR/Ca2+. Furthermore, fatty-acyl reductase 14 was involved in PBAN-mediated sex pheromone biosynthesis. Altogether, our results demonstrated that PBAN regulates sex pheromone biosynthesis through PBANR/Ca2+/CaN/ACC pathway to promote sex pheromone biosynthesis in O. furnacalis and provided a reference for non-model organism to study neuropeptide signal transduction.

雌蛾通过分泌性信息素吸引雄蛾，而性信息素生物合成的调控机制具有物种特异性。然而，亚洲玉米螟（Ostrinia furnacalis）性信息素生物合成的详细分子机制尚未完全阐明。本研究通过对亚洲玉米螟信息素腺进行转录组测序，筛选出一系列参与性信息素生物合成的候选基因。分析结果显示，亚洲玉米螟雌蛾的性信息素释放量在第二个暗期达到峰值，这与其交配行为相契合。性信息素生物合成激活神经肽（Pheromone biosynthesis-activating neuropeptide, PBAN）被证实可调控亚洲玉米螟的性信息素生物合成，且Ca²+是该虫PBAN信号通路的第二信使。对候选基因的功能分析表明，通过RNA干扰（RNAi）介导的敲低实验与药物抑制剂实验验证，钙调神经磷酸酶（Calcineurin, CaN）与乙酰辅酶A羧化酶（Acetyl-CoA carboxylase, ACC）的mRNA表达水平或酶活性降低后，亚洲玉米螟的性信息素合成量以及雌蛾吸引雄蛾的能力均显著下降。尤为关键的是，CaN与ACC的活性依赖于PBAN/PBANR/Ca²+信号通路的激活。此外，脂肪酰还原酶14也参与了PBAN介导的性信息素生物合成过程。综上，本研究结果证实，在亚洲玉米螟中，PBAN可通过PBANR/Ca²+/CaN/ACC通路调控性信息素生物合成，进而促进性信息素的产生；本研究同时为非模式生物的神经肽信号转导研究提供了参考依据。