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Journal of Integrative Agriculture  2022, Vol. 21 Issue (11): 3278-3292    DOI: 10.1016/j.jia.2022.08.079
Plant Protection Advanced Online Publication | Current Issue | Archive | Adv Search |
Identification of chorion genes and RNA interference-mediated functional characterization of chorion-1 in Plutella xylostella

DONG Shi-jie1, 2, 3, 4, 5*, LIU Bo6*, ZOU Ming-min1, 2, 3, 4, 5, LIU Li-li1, 2, 3, 4, 5, CAO Min-hui1, 2, 3, 4, 5, HUANG Meng-qi1, 2, 3, 4, 5, LIU Yan1, 2, 3, 4, 5, Liette VASSEUR1, 2, 3, 4, 7, YOU Min-sheng1, 2, 3, 4, 5, PENG Lu1, 2, 3, 4, 5

1State Key Laboratory of Ecological Pest Control for Fujian-Taiwan Crops, Institute of Applied Ecology, Fujian Agriculture and Forestry University, Fuzhou 350002, P.R.China

2International Joint Research Laboratory of Ecological Pest Control, Ministry of Education, Fujian Agriculture and Forestry University, Fuzhou 350002, P.R.China

3Ministerial and Provincial Joint Innovation Centre for Safety Production of Cross-Strait Crops, Fujian Agriculture and Forestry University, Fuzhou 350002, P.R.China

4Key Laboratory of Integrated Pest Management for Fujian-Taiwan Crops, Ministry of Agriculture, Fuzhou 350002, P.R.China

5Fujian Provincial Key Laboratory of Insect Ecology, Fujian Agriculture and Forestry University, Fuzhou 350002, P.R.China

6Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518120, P.R.China 

7Department of Biological Sciences, Brock University, St. Catharines, Ontario L2S 3A1, Canada

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摘要  

绒毛膜形成期是昆虫卵子发生的最后阶段,在这一阶段,滤泡细胞通过合成分泌绒毛膜蛋白,转移至发育的卵母细胞表面沉积形成卵壳,为胚胎发育提供保护屏障。目前对绒毛膜基因家族的研究大多集中在模式昆虫,如家蚕和果蝇在世界性害虫小菜蛾中仍缺少对绒毛膜基因家族的系统鉴定以及其功能分析因此明确绒毛膜基因家族在小菜蛾基因组上的分布情况及其转录特征,解析绒毛膜基因在小菜蛾卵子发生过程,以及胚胎发育过程中的重要作用,可为小菜蛾的遗传调控提供潜在分子靶标。本研究分析鉴定了小菜蛾绒毛膜基因的数量及染色体定位、分子特征、进化关系及其启动子区的序列特性,基于转录组数据以及qPCR实验,分析了绒毛膜基因在不同龄期和不同组织的表达模式,并基于RNAi揭示了PxCho-1生殖功能。在小菜蛾中一共鉴定得到15个绒毛膜基因,分为AB两大类。不同类型绒毛膜基因以成对的方式分布在染色体上。部分绒毛膜基因对,共享一个双向启动子调控区。系统发育分析表明,AB两类绒毛膜基因具有高度的保守性,并且在对应类别中小菜蛾绒毛膜基因均具有物种特异性。不同龄期与组织的表达谱与qPCR分析均显示,绒毛膜基因主要在小菜蛾雌成虫中显著高表达,并且在卵黄完全沉积的卵巢内显著高表达,说明绒毛膜基因在小菜蛾雌成虫的生殖发育中具有重要作用,且主要作用于卵子发生后期。抑制PxCho-1基因转录虽然对卵黄沉积没有影响,但会导致卵子变小,孵化率急剧下降,同时导致卵子内卵壳柱状层的排列松弛以及外卵壳绒毛变短。本研究为探索小菜蛾雌性生殖调控机制奠定了理论基础,有利于筛选潜在的小菜蛾遗传防控分子靶标。



Abstract  

Choriogenesis is the last step of insect oogenesis, a process by which the chorion polypeptides are produced by the follicular cells and deposited on the surface of oocytes in order to provide a highly specialized protective barrier to the embryo.  The essential features of chorion genes have yet to be clearly understood in the diamondback moth, Plutella xylostella, a worldwide Lepidoptera pest attacking cruciferous crops and wild plants.  In this study, complete sequences for 15 putative chorion genes were identified, and grouped into A and B classes.  Phylogenetic analysis revealed that both classes were highly conserved and within each, branches are also species-specific.  Chorion genes from each class were located in pairs on scaffolds of the Pxylostella genome, some of which shared the common promoter regulatory region.  All chorion genes were highly specifically expressed in the Pxylostella adult females, mostly in the ovary with full yolk, which is a crucial period to build the shells of the eggs.  RNAi-based knockdown of chorion-1, which is located on the Px_scaffold 6 alone, although had no effect on yolk deposition, resulted in smaller eggs and sharply reduced hatchability.  Additionally, inhibition of PxCho-1 expression caused a less dense arrangement of the columnar layers, reduced exochorion roughness and shorter microvilli.  Our study provides the foundation for exploring molecular mechanisms of female reproduction in Pxylostella, and for making use of chorion genes as the potential genetic-based molecular target to better control this economically important pest.

Keywords:  Plutella xylostella        chorion genes       RNAi       oogenesis       pest control  
Received: 17 November 2021   Accepted: 18 April 2022
Fund: 

This work was funded by the National Natural Science Foundation of China (32172404), the Natural Science Foundation of Fujian Province, China (2019J01666), the Fujian Agriculture and Forestry University Fund for Distinguished Young Scholars, China (xjq201903), and the “111” Program - Innovation Center for Ecologically Based Pest Management of Subtropical Crops, Fujian Agriculture and Forestry University, China.  

About author:  DONG Shi-jie, E-mail: dongsj01@163.com; LIU Bo, E-mail: liubo03@caas.cn; Correspondence YOU Min-sheng, E-mail: msyou@fafu.edu.cn; PENG Lu, E-mail: pl526520@163.com * These authors contributed equally to this study.

Cite this article: 

DONG Shi-jie, LIU Bo, ZOU Ming-min, LIU Li-li, CAO Min-hui, HUANG Meng-qi, LIU Yan, Liette VASSEUR, YOU Min-sheng, PENG Lu. 2022. Identification of chorion genes and RNA interference-mediated functional characterization of chorion-1 in Plutella xylostella. Journal of Integrative Agriculture, 21(11): 3278-3292.

Benaki D C, Aggeli A, Chryssikos G D, Yiannopoulos Y D, Kamitsos E I, Brumley E, Case S T, Boden N, Hamodrakas S J. 1998. Laser-Raman and FT-IR spectroscopic studies of peptide-analogues of silkmoth chorion protein segments. International Journal of Biological Macromolecules, 23, 49–59.
Bomfim L, Vieira P, Fonseca A, Ramos I. 2017. Eggshell ultrastructure and delivery of pharmacological inhibitors to the early embryo of R. prolixus by ethanol permeabilization of the extraembryonic layers. PLoS ONE, 12, e0185770.
Bouts D M, Melo A C, Andrade A L, Silva-Neto M A, Paiva-Silva Gde O, Sorgine M H, da Cunha Gomes L S, Coelho H S, Furtado A P, Aguiar E C, de Medeiros L N, Kurtenbach E, Rozental S, Cunha-E-Silva N L, de Souza W, Masuda H. 2007. Biochemical properties of the major proteins from Rhodnius prolixus eggshell. Insect Biochemistry Molecular Biology, 37, 1207–1221.
Carter J M, Baker S C, Pink R, Carter D R, Collins A, Tomlin J, Gibbs M, Breuker C J. 2013. Unscrambling butterfly oogenesis. BMC Genomics, 14, 283.
Chen Z, Nohata J, Guo H, Li S, Liu J, Guo Y, Yamamoto K, Kadono-Okuda K, Liu C, Arunkumar K P, Nagaraju J, Zhang Y, Liu S, Labropoulou V, Swevers L, Tsitoura P, Iatrou K, Gopinathan K P, Goldsmith M R, Xia Q, Mita K. 2015. A comprehensive analysis of the chorion locus in silkmoth. Scientific Reports, 5, 16424.
Drevet J R, Skeiky Y A, Iatrou K. 1994. GATA-type zinc finger motif-containing sequences and chorion gene transcription factors of the silkworm Bombyx mori. Journal of Biological Chemistry, 269, 10660–10667.
Drevet J R, Swevers L, Iatrou K. 1995. Developmental regulation of a silkworm gene encoding multiple GATA-type transcription factors by alternative splicing. Journal of Molecular Biology, 246, 43–53.
Epstein L, Lamport D T A. 1984. An intramolecular linkage involving isodityrosine in extensin. Phytochemistry, 23, 1241–1246.
Furlong M J, Wright D J, Dosdall L M. 2013. Diamondback moth ecology and management: Problems, progress, and prospects. Annual Review of Entomology, 58, 517–541.
Goldsmith M R. 1989. Organization and developmental timing of the Bombyx mori chorion gene clusters in strain C108. Developmental Genetics, 10, 16–23.
Hibner B L, Burke W D, Lecanidou R, Rodakis G C, Eickbush T H. 1988. Organization and expression of three genes from the silkmoth early chorion locus. Developmental Biology, 125, 423–431.
Huebner E, Anderson E. 1972. A cytological study of the ovary of Rhodnius prolixus. III. Cytoarchitecture and development of the trophic chamber. Journal Morphology, 138, 1–39.
Iatrou K, Tsitilou S G, Kafatos F C. 1984. DNA sequence transfer between two high-cysteine chorion gene families in the silkmoth Bombyx mori. Proceedings of the National Academy of Sciences of the United States of America, 81, 4452–4456.
Irles P, Bellés X, Piulachs M D. 2009. Brownie, a gene involved in building complex respiratory devices in insect eggshells. PLoS ONE, 4, e8353.
Irles P, Piulachs M D. 2011. Citrus, a key insect eggshell protein. Insect Biochemistry and Molecular Biology, 41, 101–108.
Kafatos F C, Tzertzinis G, Spoerel N A, Nguyen H T. 1995. Choriongenes: An overview of their structure, function and transcriptional regulation. In: Goldsmith M R, Wilkins A S, eds., Molecular Model Systems in the Lepidoptera. Cambridge University Press, UK. pp. 181–215.
Kumar S, Stecher G, Li M, Knyaz C, Tamura K. 2018. MEGA X: Molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution, 35, 1547–1549.
Lecanidou R, Papantonis A. 2010a. Modeling bidirectional transcription using silkmoth chorion gene promoters. Organogenesis, 6, 54–58.
Lecanidou R, Papantonis A. 2010b. Silkmoth chorion gene regulation revisited: promoter architecture as a key player. Insect Molecular Biology, 19, 141–151.
Lecanidou R, Rodakis G C, Eickbush T H, Kafatos F C. 1986. Evolution of the silk moth chorion gene superfamily: gene families CA and CB. Proceedings of the National Academy of Sciences of the United States of America, 83, 6514–6518.
Leclerc R F, Regier J C. 1994. Evolution of chorion gene families in lepidoptera: characterization of 15 cDNAs from the gypsy moth. Journal of Molecular Evolution, 39, 244–254.
Li Z, Feng X, Liu S S, You M, Furlong M J. 2016. Biology, ecology, and management of the diamondback moth in China. Annual Review of Entomology, 61, 277–296.
Lou Y H, Pan P L, Ye Y X, Cheng C, Xu H J, Zhang C X. 2018. Identification and functional analysis of a novel chorion protein essential for egg maturation in the brown planthopper. Insect Molecular Biology, 27, 393–403.
Margaritis L H. 1985. The egg-shell of Drosophila melanogaster III. Covalent crosslinking of the chorion proteins involves endogenous hydrogen peroxide. Tissue Cell, 17, 553–559.
Mitsialis S A, Kafatos F C. 1985. Regulatory elements controlling chorion gene expression are conserved between flies and moths. Nature, 317, 453–456.
Papantonis A, Swevers L, Iatrou K. 2015. Chorion genes: A landscape of their evolution, structure, and regulation. Annual Review of Entomology, 60, 177–194.
Papantonis A, Vanden Broeck J, Lecanidou R. 2008. Architectural factor HMGA induces promoter bending and recruits C/EBP and GATA during silkmoth chorion gene regulation. Biochemical Journal, 416, 85–97.
Parks S, Spradling A. 1987. Spatially regulated expression of chorion genes during Drosophila oogenesis. Genes Development, 1, 497–509.
Peng L, Wang L, Yang Y F, Zou M M, He W Y, Wang Y, Wang Q, Vasseur L, You M S. 2017. Transcriptome profiling of the Plutella xylostella (Lepidoptera: Plutellidae) ovary reveals genes involved in oogenesis. Gene, 637, 90–99.
Peng L, Wang Q, Zou M M, Qin Y D, Vasseur L, Chu L N, Zhai Y L, Dong S J, Liu L L, He W Y, Yang G, You M S. 2020. CRISPR/Cas9-mediated vitellogenin receptor knockout leads to functional deficiency in the reproductive development of Plutella xylostella. Frontiers in Physiology, 10, 1585.
Peng L, Wang L, Zou M M, Vasseur L, Chu L N, Qin Y D, Zhai Y L, You M S. 2019. Identification of halloween genes and RNA interference-mediated functional characterization of a halloween gene shadow in Plutella xylostella. Frontiers in Physiology, 10, 1120.
Petri W H, Wyman A R, Kafatos F C. 1976. Specific protein synthesis in cellular differentiation. III. The eggshell proteins of Drosophila melanogaster and their program of synthesis. Development Biology, 49, 185–199.
Regier J C, Kafatos F C, Kramer K J, Heinrikson R L, Keim P S. 1978. Silkmoth chorion proteins. Their diversity, amino acid composition, and the NH-terminal sequence of one component. Journal of Biology Chemistry, 253, 1305–1314.
Regier J C, Weigmann B M, Leclerc R F , Friedlander T P. 1994. Loss of phylogenetic information in chorion gene families of Bombyx mori gene conversion. Molecular Biology and Evolution, 11, 72–87.
Rørth P. 2009. Collective cell migration. Annual Review of Cell and Development Biology, 25, 407–429.
Rodakis G C, Moschonas N K, Kafatos F C. 1982. Evolution of a multigene family of chorion proteins in silkmoths. Molecular and Cellular Biology, 2, 554–563.
Santos A, Ramos I. 2021. ATG3 is important for the chorion ultrastructure during oogenesis in the insect vector Rhodnius prolixus. Frontiers in Physiology, 12, 638026.
Skeiky Y A, Iatrou K. 1990. Silkmoth chorion antisense RNA. Structural characterization, developmental regulation and evolutionary conservation. Journal of Molecular Biology, 213, 53–66.
Skeiky Y A, Iatrou K. 1991. Synergistic interactions of silkmoth chorion promoter-binding factors. Molecular and Cellular Biology, 11, 1954–1964.
Sourmeli S, Kravariti L, Lecanidou R. 2003. In vitro analysis of Bombyx mori early chorion gene regulation: Stage specific expression involves interactions with C/EBP-like and GATA factors. Insect Biochemistry and Molecular Biology, 33, 525–540.
Spoerel N, Nguyen H T, Kafatos F C. 1986. Gene regulation and evolution in the chorion locus of Bombyx mori. Structural and developmental characterization of four eggshell genes and their flanking DNA regions. Journal of Molecular Biology, 190, 23–35.
Spradling A C. 1993. Developmental genetics of oogenesis. In: Bate M, Martinez Arias A, eds., The Development of Drosophila Melanogaster. Cold Spring Harbor Laboratory Press, NY. pp. 1–70.
Swevers L, Raikhel A S, Sappington T W, Shirk P, Iatrou K. 2005. Vitellogenesis and post-vitellogenic maturation of the insect ovarian follicle. In: Gilbert L I, Gill S, Iatrou K, eds., Reproduction and Development. Elsevier, NY. pp. 87–155.
Telfer W H. 2009. Egg formation in Lepidoptera. Journal of Insect Science, 9, 50.
Tootle T L, Williams D, Hubb A, Frederick R, Spradling A. 2011. Drosophila eggshell production: Identification of new genes and coordination by Pxt. PLoS ONE, 6, e19943.
Tsitilou S G, Rodakis G C, Alexopoulou M, Kafatos F C, Ito K, Iatrou K. 1983. Structural features of B family chorion sequences in the silkmoth Bombyx mori, and their evolutionary implications. EMBO Journal, 2, 1845–1852.
Tsukada J, Yoshida Y, Kominato Y, Auron P E. 2011. The CCAAT/enhancer (C/EBP) family of basic-leucine zipper (bZIP) transcription factors is a multifaceted highly-regulated system for gene regulation. Cytokine, 54, 6–19.
Velentzas A D, Velentzas P D, Katarachia S A, Anagnostopoulos A K, Sagioglou N E, Thanou E V, Tsioka M M, Mpakou V E, Kollia Z, Gavriil V E, Papassideri I S, Tsangaris G T, Cefalas A C, Sarantopoulou E, Stravopodis D J. 2018. The indispensable contribution of s38 protein to ovarian-eggshell morphogenesis in Drosophila melanogaster. Scientific Reports, 8, 16103.
Waring G L. 2000. Morphogenesis of the eggshell in Drosophila. International Review of Cytology, 198, 67–108.
Woods H A. 2010. Water loss and gas exchange by eggs of Manduca sexta: trading off costs and benefits. Journal of Insect Physiology, 56, 480–487.
Woods H A, Bonnecaze R T, Zrubek B. 2005. Oxygen and water flux across eggshells of Manduca sexta. Journal of Experimental Biology, 208, 1297–1308.
Wu X, Tanwar P S, Raftery L A. 2008. Drosophila follicle cells: Morphogenesis in an eggshell. Seminars in Cell and Developmental Biology, 19, 271–282. 
Yamauchi H, Yoshitake N. 1984. Formation and ultrastructure of the micropylar apparatus in Bombyx mori ovarian follicles. Journal of Morphology, 179, 47–58.
You M, Yue Z, He W, Yang X, Yang G, Xie M, Zhan D, Baxter S W, Vasseur L, Gurr G M, Douglas C J, Bai J, Wang P, Cui K, Huang S, Li X, Zhou Q, Wu Z, Chen Q, Liu C, et al. 2013. A heterozygous moth genome provides insights into herbivory and detoxification. Nature Genetics, 45, 220–225.
Zrubek B, Woods H A. 2006. Insect eggs exert rapid control over an oxygen-water tradeoff. Proceeding of the Royal Society Biological Sciences, 273, 831–834.

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