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Journal of Integrative Agriculture  2023, Vol. 22 Issue (3): 812-824    DOI: 10.1016/j.jia.2022.08.021
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Halloween genes AhCYP307A2 and AhCYP314A1 modulate last instar larva–pupa–adult transition, ovarian development and oogenesis in Agasicles hygrophila (Coleoptera: Chrysomelidae)

JIN Ji-su1*, LIU Yi-ran1*, ZHOU Zhong-shi1, WAN Fang-hao1, 2, GUO Jian-ying1#

1 State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.China

2 Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518120, P.R.China

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摘要  昆虫蜕皮激素由Halloween家族基因合成,在几个关键的发育事件中发挥重要作用,包括蜕皮和变态。然而,这些Halloween基因在莲草直胸跳甲中的功能作用仍不清楚。在本研究中,通过RT-qPCR技术检测了两个Halloween基因AhCYP307A2AhCYP314A1在不同发育阶段的表达模式。此外,利用RNA干扰(RNAi)技术探究了这两个基因的功能,并通过对莲草直胸跳甲雌性卵巢进行解剖观察其卵巢发育情况。qPCR结果显示,AhCYP307A2AhCYP314A1在末龄幼虫和雌虫中均显著高表达。此外,AhCYP307A2在卵和蛹中也有高表达,但显著低于在末龄幼虫和雌虫中的表达水平RNAi结果显示,注射dsAhCYP307A2dsAhCYP314A1显著抑制了它们的表达以及三个相关的卵黄原蛋白基因AhVgs的转录水平。敲AhCYP307A2AhCYP314A1可显著抑制幼虫蜕皮,破坏末龄幼虫--成虫的转变,延迟卵巢发育,停止产卵。以上研究结果表明,AhCYP307A2AhCYP314A1在莲草直胸跳甲末龄幼虫--成虫转换期的蜕皮发育和繁殖过程中发挥重要的调控作用。


In insects, ecdysteroids are synthesized by genes of the Halloween family and play important roles in several key developmental events, including molting and metamorphosis.  However, the roles of these genes in Agasicles hygrophila are still largely unknown.  In this study, the expression patterns of the two Halloween genes AhCYP307A2 and AhCYP314A1 were determined by quantitative PCR (qPCR) at different developmental stages.  Moreover, the functions of these two genes were explored using RNA interference (RNAi), and ovarian development was observed by dissecting the ovaries of Ahygrophila females.  The qPCR results showed that AhCYP307A2 and AhCYP314A1 were highly expressed in last instar larvae and in adult females.  In addition, AhCYP307A2 was also highly expressed in eggs and pupae but was markedly lower than in third-instar larvae and females.  The RNAi results showed that the injection of dsAhCYP307A2 or dsAhCYP314A1 markedly inhibited their expression and the transcription levels of three related AhVgs.  Knockdown of AhCYP307A2 or AhCYP314A1 significantly inhibited larval molting, impaired last instar larva–pupa–adult transition, delayed ovarian development, and stopped egg production (i.e., no eggs were laid).  These results indicate that AhCYP307A2 and AhCYP314A1 play important regulatory roles in last instar larva–pupa–adult transition and reproduction in Ahygrophila.

Keywords:  Agasicles hygrophila        ovary        oogenesis        AhCYP307A2        AhCYP314A1        RNA interference  
Received: 25 January 2022   Accepted: 09 May 2022

This work was supported by the National Natural Science Foundation of China (31572068 and 32172486).

About author:  JIN Ji-su, E-mail:; LIU Yi-ran, E-mail:; #Correspondence GUO Jian-ying, Tel: +86-10-82109572, E-mail: * These authors contributed equally to this study.

Cite this article: 

JIN Ji-su, LIU Yi-ran, ZHOU Zhong-shi, WAN Fang-hao, GUO Jian-ying. 2023. Halloween genes AhCYP307A2 and AhCYP314A1 modulate last instar larva–pupa–adult transition, ovarian development and oogenesis in Agasicles hygrophila (Coleoptera: Chrysomelidae). Journal of Integrative Agriculture, 22(3): 812-824.

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