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Journal of Integrative Agriculture  2024, Vol. 23 Issue (8): 2735-2751    DOI: 10.1016/j.jia.2023.09.001
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The Ca2+/CaN/ACC and cAMP/PKA/HK signal pathways are required for PBAN-mediated sex pheromone biosynthesis in Conogethes punctiferalis

Yao Zhang, Zelong She, Ruolan He, Shuangyan Yao, Xiang Li, Xiaoguang Liu, Xinming Yin, Jizhen Wei, Mengfang Du, Shiheng An#

State Key Laboratory of Wheat and Maize Crop Science/Henan International Laboratory for Green Pest Control/College of Plant Protection, Henan Agricultural University, Zhengzhou 450046, China

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摘要  

雌蛾释放的性信息素在其择偶中扮演着重要角色。尽管PBAN调节性信息素生物合成过程具有物种依赖性,但其细节调控的分子机制已在许多蛾类物种中阐明。桃蛀螟一种重要的农业害虫,给农业生产造成了严重的经济损失。然而,该害虫成虫性信息素生物合成的分子机制尚不清楚。本研究深入研究了PBAN调控桃蛀螟雌蛾性信息素生物合成的分子机制。通过对桃蛀螟性信息素腺体进行转录组测序,获得了参与性信息素生物合成的候选基因。结果表明,与性信息素释放节律一致,雌蛾从第一个暗周期开始交配,在第二个暗周期至第五个暗周期达到交配高峰。利用RNAi、药理学抑制剂和行为测定分析发现,PBAN是利用Ca2+cAMP作为第二信使来调控性信息素的生物合成。随后的研究发现,PBAN/Ca2+信号能够激活钙调神经磷酸酶和乙酰辅酶a羧化酶的活性;利用RNAi敲降CaNACC转录水平的表达显著后,性信息素的合成量也显著下降,导致雌蛾吸引雄蛾的能力显著降低。此外,RNAi、药理学抑制剂和行为测定分析发现,己糖激酶响应PBAN/cAMP/PKA信号通路参与调控性信息素的生物合成。随后研究发现Far2和去饱和酶1参与了PBAN调控的性信息素生物合成。总之,本研究结果表明,PBAN通过PBANR/Ca2+/CaN/ACCPBANR/cAMP/PKA/HK信号途径调控桃蛀螟性信息素的生物合成,该研究丰富了对蛾性信息素生物合成细节的理解。



Abstract  
Conogethes punctiferalis is a crop and fruit pest that has caused serious economic losses to agricultural production.  This pest relies heavily on its sex pheromone to ensure sexual encounters and subsequent mating success.  However, the molecular mechanism underlying sex pheromone biosynthesis in this species remains elusive.  The present study investigated the detailed mechanism underlying PBAN-regulated sex pheromone biosynthesis in Cpunctiferalis by transcriptome sequencing of the Cpunctiferalis pheromone glands (PGs) and subsequent functional identification of the target genes.  The results showed that female mating started from the first scotophase, and peaked at the second to fifth scotophases in accordance with the release of sex pheromones.  PBAN regulated sex pheromone biosynthesis by employing Ca2+ and cAMP as secondary messengers, as demonstrated by RNA interference (RNAi), pharmacological inhibitors, and behavioral assays.  Further investigation revealed that calcineurin (CaN) and acetyl-CoA carboxylase (ACC) were activated by PBAN/Ca2+ signaling, and the RNAi-mediated knockdown of CaN and ACC transcripts significantly reduced sex pheromone production, ultimately leading to a significantly reduced ability of females to attract males.  Importantly, hexokinase (HK) was found to regulate sex pheromone biosynthesis in response to the PBAN/cAMP/PKA signaling pathway, as demonstrated by RNAi, enzyme activity, and pharmacological inhibitor assays.  Furthermore, Far2 and Desaturase1 were found to participate in PBAN-regulated sex pheromone biosynthesis.  Altogether, our findings revealed that PBAN regulates sex pheromone biosynthesis through the PBANR/Ca2+/CaN/ACC and PBANR/cAMP/PKA/HK pathways in Cpunctiferalis, which enriches our comprehension of the details of sex pheromone biosynthesis in moths.


Keywords:  Conogethes punctiferalis        sex pheromone        cAMP/PKA        calcineurin        acetyl-CoA carboxylase        hexokinase  
Received: 05 June 2023   Accepted: 28 July 2023
Fund: 
This study was supported by the National Natural Science Foundation of China (31970472, 32272547), the National Science Fund of Henan Province for Distinguished Young Scholars, China (202300410191), the Basic Research Project of the Key Scientific Research Projects of Universities in Henan Province, China (21zx013), the Henan Agricultural Research System, China (HARS-22-09-G3), the Henan Special Support for High-Level Talents Central Plains Science and Technology Innovation Leading Talents, China (224200510018) and the earmarked fund for China Agricultural Research System (CARS-27).
About author:  Yao Zhang, E-mail: zhangyao0327@163.com; #Correspondence Shiheng An, E-mail: anshiheng@aliyun.com

Cite this article: 

Yao Zhang, Zelong She, Ruolan He, Shuangyan Yao, Xiang Li, Xiaoguang Liu, Xinming Yin, Jizhen Wei, Mengfang Du, Shiheng An. 2024. The Ca2+/CaN/ACC and cAMP/PKA/HK signal pathways are required for PBAN-mediated sex pheromone biosynthesis in Conogethes punctiferalis. Journal of Integrative Agriculture, 23(8): 2735-2751.

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