Zhao W, Li L, Zhang Y, Liu X, Wei J, Xie Y, Du M, An S
State Key Laboratory of Wheat and Maize Crop Science, College of Plant Protection, Henan Agricultural University, Zhengzhou, China.
High & New Technology Research Centre, Henan Academy of Sciences, Zhengzhou, China.
Insect Mol Biol. 2018 Jun;27(3):373-382. doi: 10.1111/imb.12379. Epub 2018 Feb 21.
Lepidoptera sex pheromone biosynthesis is regulated by pheromone biosynthesis activating neuropeptide (PBAN). PBAN regulates not only female sex pheromone biosynthesis but also male sex pheromone biosynthesis. Previous research has confirmed that PBAN regulates sex pheromone biosynthesis using Ca as a secondary messenger in all examined species to date. However, the downstream signal of Ca has remained elusive. In the present study, calcineurin A (CNA), a downstream signal of Ca , was discovered in Helicoverpa armigera male hairpencil and named HaCNA. Sequence analysis demonstrated that the open reading frame of HaCNA contains 1488 nucleotides encoding 495 amino acid residues. A homology search revealed that HaCNA shares a high amino acid identity with the CNA of other insects. Developmental and spatial expression analyses revealed that the mRNA levels of HaCNA peaked at 24 h after emergence and that HaCNA expression was ubiquitous in all examined tissues. Activity analysis revealed that PBAN activates HaCNA, and a Ca inhibitor, Lacl , attenuated the effect of PBAN by decreasing HaCNA activity. Pharmacological inhibitor and RNA interference-mediated knockdown assays revealed that both activity inhibition and decreased mRNA levels of HaCNA led to a significant decrease in the production of the male sex pheromone components [octadecanol and (Z)-11 hexadecanol)] and in the efficacy of female mating acceptance. Our results demonstrate that HaCNA acts as downstream signal of PBAN/Ca and plays an important role in PBAN-induced male sex pheromone biosynthesis and female mating acceptance.
鳞翅目昆虫的性信息素生物合成受性信息素生物合成激活神经肽（PBAN）调控。PBAN不仅调控雌性性信息素的生物合成，也调控雄性性信息素的生物合成。此前的研究已证实，在迄今为止所有被检测的物种中，PBAN利用Ca作为第二信使来调控性信息素的生物合成。然而，Ca的下游信号仍不明确。在本研究中，在棉铃虫雄性发香器中发现了Ca的下游信号钙调神经磷酸酶A（CNA），并将其命名为HaCNA。序列分析表明，HaCNA的开放阅读框包含1488个核苷酸，编码495个氨基酸残基。同源性搜索显示，HaCNA与其他昆虫的CNA具有高度的氨基酸同一性。发育和空间表达分析表明，HaCNA的mRNA水平在羽化后24小时达到峰值，且HaCNA在所有被检测组织中均有普遍表达。活性分析表明，PBAN可激活HaCNA，而Ca抑制剂Lacl可通过降低HaCNA活性来减弱PBAN的作用。药理学抑制剂和RNA干扰介导的敲低试验表明，HaCNA活性的抑制和mRNA水平的降低均导致雄性性信息素成分[十八烷醇和（Z）-11-十六碳烯醇]的产量以及雌性交配接受能力显著下降。我们的结果表明，HaCNA作为PBAN/Ca的下游信号，在PBAN诱导的雄性性信息素生物合成和雌性交配接受过程中发挥重要作用。
