JIA-2022-0121 Halloween基因AhCYP307A2和AhCYP314A1调控莲草直胸跳甲(鞘翅目:叶甲科)末龄幼虫-蛹-成虫转变，卵巢发育和卵子发生

doi:10.1016/j.jia.2022.08.021

Journal of Integrative Agriculture ›› 2023, Vol. 22 ›› Issue (3): 812-824.DOI: 10.1016/j.jia.2022.08.021

JIA-2022-0121 Halloween基因AhCYP307A2和AhCYP314A1调控莲草直胸跳甲(鞘翅目:叶甲科)末龄幼虫-蛹-成虫转变，卵巢发育和卵子发生

收稿日期:2022-01-25 接受日期:2022-05-09 出版日期:2023-03-20 发布日期:2022-05-09

Halloween genes AhCYP307A2 and AhCYP314A1 modulate last instar larva–pupa–adult transition, ovarian development and oogenesis in Agasicles hygrophila (Coleoptera: Chrysomelidae)

JIN Ji-su^1*, LIU Yi-ran^1*, ZHOU Zhong-shi¹, WAN Fang-hao^{1, 2}, GUO Jian-ying^1#

¹State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.China
²Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518120, P.R.China

Received:2022-01-25 Accepted:2022-05-09 Online:2023-03-20 Published:2022-05-09
About author:JIN Ji-su, E-mail: jinjisu9110@163.com; LIU Yi-ran, E-mail: liuyiran0907@163.com; #Correspondence GUO Jian-ying, Tel: +86-10-82109572, E-mail: guojianying@caas.cn * These authors contributed equally to this study.
Supported by:
This work was supported by the National Natural Science Foundation of China (31572068 and 32172486).

摘要/Abstract

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AhCYP307A2和AhCYP314A1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，AhCYP307A2和AhCYP314A1在末龄幼虫和雌虫中均显著高表达。此外，AhCYP307A2在卵期和蛹中也有高表达，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dsAhCYP307A2和dsAhCYP314A1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。敲除AhCYP307A2和AhCYP314A1可显著抑制幼虫蜕皮，破坏末龄幼虫-蛹-成虫的转变，延迟卵巢发育，停止产卵。以上研究结果表明，AhCYP307A2和AhCYP314A1在莲草直胸跳甲末龄幼虫-蛹-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。

Abstract:

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 A. hygrophila 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 A. hygrophila.

Key words: Agasicles hygrophila , ovary , oogenesis , AhCYP307A2 , AhCYP314A1 , RNA interference

. JIA-2022-0121 Halloween基因AhCYP307A2和AhCYP314A1调控莲草直胸跳甲(鞘翅目:叶甲科)末龄幼虫-蛹-成虫转变，卵巢发育和卵子发生[J]. Journal of Integrative Agriculture, 2023, 22(3): 812-824.

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

参考文献

Buckingham G R. 1996. Biological control of alligator weed, Alternanthera philoxeroides, the world’s first aquatic weed success story. Castanea, 61, 232–243.
Butenandt A, Karlson P. 1954. Über die isolierung eines metamorphose hormons der insekten in kristallisierter form. Zeitschrift für Naturforschung, 9, 389–391. (in German)
Chanchay P, Vongsangnak W, Thancharoen A, Sriboonlert A. 2019. Reconstruction of insect hormone pathways in an aquatic firefly, Sclerotia aquatilis (Coleoptera: Lampyridae), using RNA-seq. PeerJ, 7, e7428.
Chávez V M, Marqués G, Delbecque J P, Kobayashi K, Hollingsworth M, Burr J, Natzle J E, O’Connor M B. 2000. The Drosophila disembodied gene controls late embryonic morphogenesis and codes for a cytochrome P450 enzyme that regulates embryonic ecdysone levels. Development, 127, 4115–4126.
Chen L, Cai D C, Chen Q, Tang C, Feng G, Peng Z Q, Jin Q A, Wen H B. 2009. The reproductive system and reproductive biology of the alligator weed flea beetle, Agasicles hygrophila (Coleoptera: Chrysomelidae). Acta Entomologica Sinica, 52, 1255–1260. (in Chinese)
Cheng D J, Li Z Q, Meng M, Peng J, Qian W L, Kang L X, Xia Q Y. 2014. Characterization of cytochrome P450 genes involving in ecdysteroidogenesis in silkworm (Bombyx mori). Scientia Agricultura Sinica, 47, 594–604. (in Chinese)
Christesen D, Yang Y T, Somers J, Robin C, Sztal T, Batterham P, Perry T. 2017. Transcriptome analysis of Drosophila melanogaster third instar larval ring glands points to novel functions and uncovers a cytochrome P450 required for development. Genes, Genomes, Genetics, 7, 467–479.
Christiaens O, Iga M, Velarde R A, Rougé P, Smagghe G. 2010. Halloween genes and nuclear receptors in ecdysteroid biosynthesis and signalling in the pea aphid. Insect Molecular Biology, 19, 187–200.
Cong L, Jiang X Z, Yang W J, Xu K K, Dou W, Ran C, Wang J J. 2015. Identification of ecdysone synthesis pathway genes and analysis on the impact of food deprivation on larvae development of Bactrocera dorsalis Hendel. Scientia Agricultura Sinica, 48, 4469–4482. (in Chinese)
Fu L J, Wu Z Q, Fu Z J. 1999. Studies on the ovary development and emergence rhythm of Agasicles hygrophila. Wuyi Science Journal, 15, 74–78. (in Chinese)
Ghanim M, Kontsedalov S, Czosnek H. 2007. Tissue-specific gene silencing by RNA interference in the whitefly Bemisia tabaci (Gennadius). Insect Biochemistry and Molecular Biology, 37, 732–738.
Gilbert L I. 2004. Halloween genes encode P450 enzymes that mediate steroid hormone biosynthesis in Drosophila melanogaster. Molecular and Cellular Endocrinology, 215, 1–10.
Gilbert L I, Warren J T. 2005. A molecular genetic approach to the biosynthesis of the insect steroid molting hormone. Vitamins and Hormones, 73, 31–57.
Guo J Y, Fu J W, Xian X Q, Ma M, Wan F H. 2012. Performance of Agasicles hygrophila (Coleoptera: Chrysomelidae), a biological control agent of invasive alligator weed, at low non-freezing temperatures. Biological Invasions, 14, 1597–1608.
Hentze J L, Moeller M E, Jørgensen A F, Bengtsson M S, Bordoy A M, Warren J T, Gilbert L I, Ole A, Rewitz K F, Reddy P S. 2013. Accessory gland as a site for prothoracicotropic hormone controlled ecdysone synthesis in adult male insects. PLoS ONE, 8, e55131.
Iga M, Smagghe G. 2010. Identification and expression profile of Halloween genes involved in ecdysteroid biosynthesis in Spodoptera littoralis. Peptides, 31, 456–467.
Jin J S, Zhao M T, Zhang H, Liu Y R, Wan F H, Zhou Z S, Guo J Y. 2021. Impact of summer temperatures on Agasicles hygrophila, a key biocontrol agent of the invasive weed Alternanthera philoxeroides in Hunan Province, China. Entomologia generalias, 41, 59–70.
Khalid M Z, Ahmad S, Ngegba P M, Zhong G H. 2021. Role of endocrine system in the regulation of female insect reproduction. Biology, 10, 614.
Kubo I, Klocke J A, Asano S. 1983. Effects of ingested phytoecdysteroids on the growth and development of two Lepidopterous larvae. Journal of Insect Physiology, 29, 307–316.
Lee C Y, Cooksey B A K, Baehrecke E H. 2002. Steroid regulation of midgut cell death during Drosophila development. Developmental Biology, 250, 101–111.
Lenaerts C, Van Wielendaele P, Peeters P, Vanden Broeck J, Marchal E. 2016. Ecdysteroid signalling components in metamorphosis and development of the desert locust, Schistocerca gregaria. Insect Biochemistry and Molecular Biology, 75, 10–23.
Li G, Niu J Z, Zotti M, Sun Q Z, Zhu L, Zhang J, Liao C, Dou W, Wei D, Wang J. 2017. Characterization and expression patterns of key ecdysteroid biosynthesis and signaling genes in a spider mite (Panonychus citri). Insect Biochemistry and Molecular Biology, 87, 136–146.
Li Y N, Guo J Y, Zhu X, Huang S, Wan F H. 2011. Effects of release density on the population dynamics of the biocontrol agent, Agasicles hygrophila (Coleoptera: Chrysomelidae). Journal of Biosafety, 20, 275–280. (in Chinese)
Liu Y R, Zhang H, Jin J S, Zhou Z S, Guo J Y. 2020. Identification and expression analysis of the Halloween gene family in Agasicles hygrophila. Scientia Agricultura Sinica, 53, 2009–2019. (in Chinese)
Livak K J, Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2–∆∆CT method. Methods, 25, 402–408.
Ma R Y, Wang R, Ding J Q. 2003. Classical biological control of exotic weeds. Acta Ecologica Sinica, 12, 2677–2688. (in Chinese)
Maeda S, Nakashima A, Yamada R, Hara N, Fujimoto Y, Ito Y, Sonobe H. 2008. Molecular cloning of ecdysone 20-hydroxylase and expression pattern of the enzyme during embryonic development of silkworm Bombyx mori. Comparative Biochemistry and Physiology (B: Biochemistry and Molecular Biology), 149, 507–516.
Meng X K, Zhang N, Yang X M, Miao L J, Jiang H, Ji C H, Xu B B, Qian K, Wang J J. 2020. Sublethal effects of chlorantraniliprole on molting hormone levels and mRNA expressions of three Halloween genes in the rice stem borer, Chilo suppressalis. Chemosphere, 238, 124676.
Niwa R, Matsuda T, Yoshiyama T, Namiki T, Mita K, Fujimoto Y, Kataoka H. 2004. CYP306A1, a cytochrome P450 enzyme, is essential for ecdysteroid biosynthesis in the prothoracic glands of Bombyx and Drosophila. Journal of Biological Chemistry, 279, 35942–35949.
Niwa R, Niwa Y S. 2014. Enzymes for ecdysteroid biosynthesis: Their biological functions in insects and beyond. Bioscience, Biotechnology, and Biochemistry, 78, 1283–1292.
Ono H, Rewitz K F, Shinoda T, Itoyama K, Petryk A, Rybczynski R, Jarcho M, Warren J T, Marques G, Shimell M J, Gilbert L I, O’Connor M B. 2006. Spook and Spookier code for stage-specific components of the ecdysone biosynthetic pathway in Diptera. Developmental Biology, 298, 555–570.
Ou Q X, King-Jones K. 2013. What goes up must come down: Transcription factors have their say in making ecdysone pulses. Current Topics in Developmental Biology, 103, 35–71.
Pan X Y, Geng Y, Zhang W J, Li B, Chen J K. 2006. The influence of abiotic stress and phenotypic plasticity on the distribution of invasive Alternanthera philoxeroides along a riparian zone. Acta Oecologica, 30, 333–341.
Parthasarathy R, Sun Z Y, Bai H, Palli S R. 2010. Juvenile hormone regulation of vitellogenin synthesis in the red flour beetle, Tribolium castaneum. Insect Biochemistry and Molecular Biology, 40, 405–414.
Petryk A, Warren J T, Marqués G, Jarcho M P, Gilbert L I, Kahler J, Parvy J P, Li Y, Dauphin-Villemant C, O’Connor M B. 2003. Shade is the Drosophila P450 enzyme that mediates the hydroxylation of ecdysone to the steroid insect molting hormone 20-hydroxyecdysone. Proceedings of the National Academy of Sciences of the United States of America, 100, 13773–13778.
Raabe M. 1987. Insect reproduction: regulation of successive steps. Advances in Insect Physiology, 19, 29–154.
Raikhel A S, Brown M R, Belles X. 2005. Hormonal control of reproductive processes. Comprehensive Molecular Insect Science, 3, 433–491.
Rewitz K F, Gilbert L I. 2008. Daphnia Halloween genes that encode cytochrome P450s mediating the synthesis of the arthropod molting hormone: evolutionary implications. BMC Evolutionary Biology, 8, 60.
Rewitz K F, O’Connor M B, Gilbert L I. 2007. Molecular evolution of the insect Halloween family of cytochrome P450s: Phylogeny, gene organization and functional conservation. Insect Biochemistry and Molecular Biology, 37, 741–753.
Rewitz K F, Rybczynski R, Warren J T, Gilbert L I. 2006a. Developmental expression of Manduca shade, the P450 mediating the final step in molting hormone synthesis. Molecular and Cellular Endocrinology, 247, 166–174.
Rewitz K F, Rybczynski R, Warren J T, Gilbert L I. 2006b. Identification, characterization and developmental expression of Halloween genes encoding P450 enzymes mediating ecdysone biosynthesis in the tobacco hornworm, Manduca sexta. Insect Biochemistry and Molecular Biology, 36, 188–199.
Rharrabe K, Bouayad N, Sayah F. 2009. Effects of ingested 20-hydroxyecdysone on development and midgut epithelial cells of Plodia interpunctella (Lepidoptera, Pyralidae). Pesticide Biochemistry and Physiology, 93, 112–119.
Riddiford L M, Hiruma K, Zhou X F, Nelson C A. 2003. Insights into the molecular basis of the hormonal control of molting and metamorphosis from Manduca sexta and Drosophila melanogaster. Insect Biochemistry and Molecular Biology, 33, 1327–1338.
Roy S, Saha T T, Zou Z, Raikhel A S. 2018. Regulatory pathways controlling female insect reproduction. Annual Review of Entomology, 63, 489–511.
SEPAC, CAS (State Environmental Protection Administration of China, Chinese Academy of Sciences). 2003. The Announcement About the First List if Invasive Species in China. Chinese Academy of Sciences, Beijing. (in Chinese)
Shi J F, Mu L L, Chen X, Guo W C, Li G Q. 2016. RNA interference of chitin synthase genes inhibits chitin biosynthesis and affects larval performance in Leptinotarsa decemlineata (Say). International Journal of Biological Sciences, 12, 1319–1331.
Sieglaff D H, Duncan K A, Brown M R. 2005. Expression of genes encoding proteins involved in ecdysteroidogenesis in the female mosquito, Aedes aegypti. Insect Biochemistry and Molecular Biology, 35, 471–490.
Wang R. 1989. Biological control of weeds in China: A status report. In: Proceedings of the Seventh International Symposium on Biological Control of Weeds. Commonwealth Scientific and Industrial Research Organisation, Melbourne. pp. 689–693.
Warren J T, Petryk A, Marques G, Jarcho M, Parvy J P, Dauphin-Villemant C, O’Connor M B, Gilbert L I. 2002. Molecular and biochemical characterization of two P450 enzymes in the ecdysteroidogenic pathway of Drosophila melanogaster. Proceedings of the National Academy of Sciences of the United States of America, 99, 11043–11048.
Warren J T, Petryk A G, Marqués G, Parvy J P, Shinoda T, Itoyama K, Kobayashi J, Jarcho M, Li Y, O’Connor M B. 2004. Phantom encodes the 25-hydroxylase of Drosophila melanogaster and Bombyx mori: A P450 enzyme critical in ecdysone biosynthesis. Insect Biochemistry and Molecular Biology, 34, 991–1010.
Wu X F, Xu J L. 1992. Ecdysone’s influence on biosynthesis and accumulation of yolk proteins and on ovary development of the Bombyx mori. Acta Sericologica Sinica, 18, 20–24. (in Chinese)
Yamanaka N, Rewitz K F, O’Connor M B. 2013. Ecdysone control of developmental transitions: Lessons from Drosophila research. Annual Review of Entomology, 58, 497–516.
Yamazaki Y, Kiuchi M, Takeuchi H, Kubo T. 2011. Ecdysteroid biosynthesis in workers of the European honeybee Apis mellifera L. Insect Biochemistry and Molecular Biology, 41, 283–293.
Yang D L, Hao J, Wang Y N, Xi G S. 2014. Research progress of ecdysone in adult insects. Chinese Bulletin of Life Sciences, 26, 874–881. (in Chinese)
Zhao M T, Wang Y, Zhou Z S, Wang R, Guo J Y, Wan F H. 2016. Effects of periodically repeated heat events on reproduction and ovary development of Agasicles hygrophila (Coleoptera: Chrysomelidae). Journal of Economic Entomology, 109, 1586–1594.
Zhao X. 2009. Thermal adaptation of Agasicles hygrophila (Coleoptera: Chrysomelidae) in responses to temperature stress. MSc thesis, Southwest University, China. pp. 10–19. (in Chinese)

JIA-2022-0121 Halloween基因AhCYP307A2和AhCYP314A1调控莲草直胸跳甲(鞘翅目:叶甲科)末龄幼虫-蛹-成虫转变，卵巢发育和卵子发生

Halloween genes AhCYP307A2 and AhCYP314A1 modulate last instar larva–pupa–adult transition, ovarian development and oogenesis in Agasicles hygrophila (Coleoptera: Chrysomelidae)

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