棉铃虫复眼中Clock生物钟基因的昼夜表达模式

doi:10.3864/j.issn.0578-1752.2017.19.010

Abstract

Abstract: 【Objective】The objective of this study is to clone and analyze a circadian clock gene, Clock (Clk) in the compound eyes of Helicoverpa armigera, examine the diurnal changes and determinants of Clk mRNA levels in compound eyes of cotton bollworm, and to determine whether Clk performed circadian functions in compound eyes, which will provide a theoretical reference for understanding the circadian clock machinery in compound eyes. 【Method】The total RNA was isolated from the compound eyes of 2-day-old H. armigera moths, and the Clk was cloned by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). The amino acid sequence of CLK from cotton bollworm was analyzed using bioinformatics softwares. The relative mRNA levels of Clk were determined among various adult organs (head, thorax, abdomen, leg, wing, brain, antennae and compound eye) of H. armigera by quantitative real-time PCR (qRT-PCR). The diurnal change of Clk mRNA levels in compound eyes was measured under different photoperiods. Moths were illuminated by UV, blue and green lights, respectively, for 6 h from the beginning of the scotophase, and the expression levels of Clk in compound eyes were determined after light exposure. Female and male moths were paired for mating, and the expression levels of Clk in compound eyes were determined from the moths that had completed mating 0 h and 3 h. The expression levels of Clk in compound eyes were determined from the moths that were not fed. 【Result】An open reading frame of 1 860 bp was cloned, encoding 619 amino acids, designed as “HeCLK” (GenBank accession number KM233158). The molecular mass of the deduced protein was predicted to be 69.32 kD, and the calculated isoelectric point (pI) was 5.71. Deduced amino acids sequence contained three transmembrane topologies, and several conserved domains of insect CLK (PAS and HLH). HeCLK showed high homology with Spodoptera exigua (97% identity) and Danaus plexippus (74% identity), and low homology with Riptortus pedestris (53% identity) and Leptinotarsa decemlineata (52% identity). qRT-PCR revealed that HeCLK showed the lowest mRNA levels in compound eyes, and the highest mRNA levels in antennas among tested adult organs. The mRNA levels of HeCLK in compound eyes increased during the day, and decreased during the night under 14L﹕10D. The cycling of the circadian clock gene mRNA levels persisted for 1 d under dark condition, but did not persist further under constant darkness. HeCLK was up-regulated in compound eyes after light exposure, but there was no significant difference in mRNA levels of HeCLK among different wavelengths of light. The expression levels of HeCLK were tended to be down-regulated in the compound eyes after copulation, and there was a significant difference in mRNA levels of HeCLK between mated males and virgin males. The expression levels of HeCLK in compound eyes were not influenced by starvation. 【Conclusion】HeCLK from the compound eyes of a nocturnal moth, H. armigera, was cloned. Deduced amino acids sequence contained the conserved domains of CLK proteins, and shared high homology with insect CLK. HeCLK showed the lowest mRNA levels in compound eyes among tested adult organs. The expression levels of HeCLK in peripheral tissues (compound eyes) were regulated by the circadian rhythms, light condition and physiological status of moths, confirming that HeCLK played an important role in circadian rhythms in compound eyes. Whether the circadian clock machinery is similar between compound eyes and the central nervous system need to be further studied.

Key words: Helicoverpa armigera, circadian clock, compound eye, photoreceptor, peripheral tissue, rhythm

YAN Shuo, LIU YanJun, ZHANG XinFang, QIN Meng, LIU Hui, ZHU JiaLin, LI Zhen, ZHANG QingWen, LIU XiaoXia. Daily Expression of Clock Gene in Compound Eye of Helicoverpa armigera[J].Scientia Agricultura Sinica, 2017, 50(19): 3733-3744.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.chinaagrisci.com/EN/10.3864/j.issn.0578-1752.2017.19.010

https://www.chinaagrisci.com/EN/Y2017/V50/I19/3733

References

[1] Tanoue S, Nishioka T. A receptor-type guanylyl cyclase expression is regulated under circadian clock in peripheral tissues of the silk moth. Light-induced shifting of the expression rhythm and correlation with eclosion. The Journal of Biological Chemistry, 2001,276(50): 46765-46769.

[2] Zhu H, Sauman I, Yuan Q, Casselman A, Emery-Le M, Emery P, Reppert S M. Cryptochromes define a novel circadian clock mechanism in monarch butterflies that may underlie sun compass navigation. PLoS Biology, 2008, 6(1): e4.

[3] 张达燕, 梁辉, 司马杨虎, 徐世清. 温度与光照节律对家蚕成虫生物钟基因Cry1和Cry2表达的影响. 蚕业科学, 2013, 39(3): 453-459.

Zhang D Y, Liang H, Sima Y H, Xu S Q. Effects of temperature and light rhythm on expression of clock genes cry1 and cry2 in Bombyx mori adult. Science of Sericulture, 2013, 39(3): 453-459. (in Chinese)

[4] 王文栋, 束梅影, 张达艳, 徐世清. 家蚕昼夜节律生物钟基因的生物信息学分析. 四川动物, 2016, 35(2): 275-282.

Wang W D, Shu M Y, Zhang D Y, Xu S Q. Bioinformatics analysis of circadian rhythm biological clock genes in Bombyx mori. Sichuan Journal of Zoology, 2016, 35(2): 275-282. (in Chinese)

[5] Zhu L, Liu W, Tan Q Q, Lei C L, Wang X P. Differential expression of circadian clock genes in two strains of beetles reveals candidates related to photoperiodic induction of summer diapause. Gene, 2017, 603: 9-14.

[6] Rubin E B, Shemesh Y, Cohen M, Elgavish S, Robertson H M, Bloch G. Molecular and phylogenetic analyses reveal mammalian-like clockwork in the honey bee (Apis mellifera) and shed new light on the molecular evolution of the circadian clock. Genome Research, 2006, 16(11): 1352-1365.

[7] Gegear R J, Casselman A, Waddell S, Reppert S M. Cryptochrome mediates light-dependent magnetosensitivity in Drosophila. Nature, 2008, 454(7207): 1014-1018.

[8] Barberà M, Collantes-Alegre J M, Martínez-Torres D. Characterisation, analysis of expression and localization of circadian clock genes from the perspective of photoperiodism in the aphid Acyrthosiphon pisum. Insect Biochemistry and Molecular Biology, 2017, 83: 54-67.

[9] Kontogiannatos D, Gkouvitsas T, Kourti A. The expression patterns of the clock genes period and timeless are affected by photoperiod in the Mediterranean corn stalk borer, Sesamia nonagrioides. Archives of Insect Biochemistry and Physiology, 2017, 94(1): e21366.

[10] Chang D C, McWatters H G, Williams J A, Gotter A L, Levine J D, Reppert S M. Constructing a feedback loop with circadian clock molecules from the silkmoth, Antheraea pernyi. The Journal of Biological Chemistry, 2003, 278(40): 38149-38158.

[11] Shirasu N, Shimohigashi Y, Tominaga Y, Shimohigashi M. Molecular cogs of the insect circadian clock. Zoological Science, 2003, 20(8): 947-955.

[12] Sandrelli F, Costa R, Kyriacou C P, Rosato E. Comparative analysis of circadian clock genes in insects. Insect Molecular Biology, 2008, 17(5): 447-463.

[13] Tomioka K, Matsumoto A. A comparative view of insect circadian clock systems. Cellular and Molecular Life Sciences, 2010, 67(9): 1397-1406.

[14] 任爽, 魏慧敏, 郝友进, 陈斌. 昆虫钟基因研究进展. 昆虫学报, 2016, 59(3): 353-364.

Ren S, Wei H M, Hao Y J, Chen B. Research progress in circadian clock genes in insects. Acta Entomologica Sinica, 2016, 59(3): 353-364. (in Chinese)

[15] Cashmore A R. Cryptochromes: enabling plants and animals to determine circadian time. Cell, 2003, 114(5): 537-543.

[16] Busza A, Emery-Le M, Rosbash M, Emery P. Roles of the two Drosophila cryptochrome structure domains in circadian photoreception. Science, 2004, 304(5676): 1503-1506.

[17] Lin C, Todo T. The cryptochromes. Genome Biology, 2005, 6(5): 220.

[18] Zhu H, Yuan Q, Froy O, Casselman A, Reppert S M. The two CRYs of the butterfly. Current Biology, 2005, 15(23): R953-R954.

[19] Boothroyd C E, Wijnen H, Naef F, Saez L, Young M W. Integration of light and temperature in the regulation of circadian gene expression in Drosophila. PLoS Genetics, 2007, 3(4): e54.

[20] Fan J Y, Muskus M J, Price J L. Entrainment of the Drosophila circadian clock: more heat than light. Sciences Stke, 2007, 413: pe65.

[21] Ni H, Yan S, Liu X X, Zhang Q W. Cryptochromes mRNA expression under mating and black light treatment on Helicoverpa armigera. Plant Diseases and Pests, 2011, 2(3): 20-23, 30.

[22] Yan S, Ni H, Li H T, Zhang J, Liu X X, Zhang Q W. Molecular cloning, characterization, and mRNA expression of two cryptochrome genes in Helicoverpa armigera (Lepidoptera: Noctuidae). Journal of Economic Entomology, 2013, 106(1): 450-462.

[23] 刘孝明, 张松斗, 马木提·赛丽蔓, 邹驰, 李贞, 张青文, 刘小侠. 光周期和温度对生物钟基因cwo在棉铃虫幼虫节律表达的影响. 应用昆虫学报, 2016, 53(5): 942-952.

Liu X M, Zhang S D, Mamuti S, Zou C, Li Z, Zhang Q W, Liu X X. The effect of photoperiod and temperature on the diurnal expression of the circadian clock gene cwo in larvae of cotton bollworm, Helicoverpa armigera (Hübner). Chinese Journal of Applied Entomology, 2016, 53(5): 942-952. (in Chinese)

[24] Uryu O, Karpova S G, Tomioka K. The clock gene cycle plays an important role in the circadian clock of the cricket Gryllus bimaculatus. Journal of Insect Physiology, 2013, 59(7): 697-704.

[25] Wu K J, Gong P Y. A new and practical artificial diet for the cotton bollworm. Entomologia Sinica, 1997, 4(3): 277-282.

[26] Yan S, Li H T, Zhang J, Zhu J L, Zhang Q W, Liu X X. Sperm storage and sperm competition in the Helicoverpa armigera (Lepidoptera: Noctuidae). Journal of Economic Entomology, 2013, 106(2): 708-715.

[27] Yan S, Zhu J L, Zhu W L, Zhang X F, Li Z, Liu X X, Zhang Q W. The expression of three opsin genes from the compound eye of Helicoverpa armigera (Lepidoptera: Noctuidae) is regulated by a circadian clock, light conditions and nutritional status. PLoS ONE, 2014, 9(10): e111683.

[28] Bairoch A, Bucher P, Hofmann K. The PROSITE database, its status in 1997. Nucleic Acids Research, 1997, 25(1): 217-221.

[29] Combet C, Blanchet C, Geourjon C, Deléage G. NPS@: network protein sequence analysis. Trends Biochemical Sciences, 2000, 25(3): 147-150.

[30] 魏国树, 张青文, 周明牂, 吴卫国. 棉铃虫[Helicoverpa armigera (Hübner)]蛾复眼视网膜电位研究. 生物物理学报, 1999, 15(4): 682-688.

Wei G S, Zhang Q W, Zhou M Z, Wu W G. Studies on the electroretinogram of the compound eyes of Helicoverpa armigera (Hübner) moth. Acta Biophysica Sinica, 1999, 15(4): 682-688. (in Chinese)

[31] Fuller R C, Claricoates K M. Rapid light-induced shifts in opsin expression: finding new opsins, discerning mechanisms of change, and implications for visual sensitivity. Molecular Ecology, 2011, 20(16): 3321-3335.

[32] 闫硕, 朱家林, 朱威龙, 潘李隆, 张青文, 刘小侠. 棉铃虫α-微管蛋白基因的克隆、序列分析及表达模式检测. 中国农业科学, 2013, 46(9): 1808-1817.

Yan S, Zhu J L, Zhu W L, Pan L L, Zhang Q W, Liu X X. Molecular cloning, sequence analysis and expression pattern detection of a-tubulin gene from Helicoverpa armigera (Hübner). Scientia Agricultura Sinica, 2013, 46(9): 1808-1817. (in Chinese)

[33] Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔCt method. Methods, 2001,25(4): 402-408.

[34] Sancar A. Cryptochrome: the second photoactive pigment in the eye and its role in circadian photoreception. Annual Review of Biochemistry, 2000, 69: 31-67.

[35] Saunders D S. Insect photoperiodism: seeing the light. Physiological Entomology, 2012, 37(3): 207-218.

[36] Ruan G X, Gamble K L, Risner M L, Young L A, McMahon D G. Divergent roles of clock genes in retinal and suprachiasmatic nucleus circadian oscillators. PLoS ONE, 2012, 7(6): e38985.

[37] Bobu C, Sandu C, Laurent V, Felder-Schmittbuhl M P, Hicks D. Prolonged light exposure induced widespread phase shifting in the circadian clock and visual pigment gene expression of the Arvicanthis ansorgei retina. Molecular Vision, 2013, 19(1): 1060-1073.

[38] Buonfiglio D C, Malan A, Sandu C, Jaeger C, Cipolla-Neto J, Hicks D, Felder-Schmittbuhl M P. Rat retina shows robust circadian expression of clock and clock output genes in explant culture. Molecular Vision, 2014, 20(6): 742-752.

[39] Huang N, Chelliah Y, Shan Y, Taylor C A, Yoo S H, Partch C, Green C B, Zhang H, Takahashi J S. Crystal structure of the heterodimeric CLOCK: BMAL1 transcriptional activator complex. Science, 2012, 337(6091): 189-194.

[40] Merlin C, François M C, Queguiner I, Maïbèche- Coisné M, Jacquin-Joly E. Evidence for a putative antennal clock in Mamestra brassicae: molecular cloning and characterization of two clock genes-period and cryptochrome-in antennae. Insect Molecular Biology, 2006,15(2): 137-145.

[41] 鲁艳辉, 赵燕燕, 张发成, 郑许松, 朱平阳, 吕仲贤. 二化螟滞育生物钟蛋白TIME-EA4基因的克隆及时空和温度诱导表达分析. 昆虫学报, 2016, 59(4): 392-401.

Lu Y H, Zhao Y Y, Zhang F C, Zheng X S, Zhu P Y, Lü Z X. Cloning and spatiotemporal and temperature-induced expression profiling of diapause bioclock protein TIME-EA4 gene in the rice stem borer, Chilo suppressalis (Lepidoptera: Pyralidae). Acta Entomologica Sinica, 2016, 59(4): 392-401. (in Chinese)

[42] Chong N W, Chaurasia S S, Haque R, Klein D C, Iuvone P M. Temporal-spatial characterization of chicken clock genes: circadian expression in retina, pineal gland, peripheral tissues. Journal of Neurochemistry, 2003, 85(4): 851-860.

[43] Liu S, Cai Y N, Sothern R B, Guan Y Q, Chan P. Chronobiological analysis of circadian patterns in transcription of seven key clock genes in six peripheral tissues in mice. Chronobiology International, 2007, 24(5): 793-820.

[44] Singh D, Rani S, Kumar V. Daily expression of six clock genes in central and peripheral tissues of a night-migratory songbird: evidence for tissue-specific circadian timing. Chronobiology International, 2013, 30(10): 1208-1217.

[45] Brady A K, Snyder K A, Vize P D. Circadian cycles of gene expression in the coral, Acropora millepora. PLoS ONE, 2011, 6(9): e25072.

[46] Merlin C, Lucas P, Rochat D, François M C, Maïbèche-Coisne M, Jacquin-Joly E. An antennal circadian clock and circadian rhythms in peripheral pheromone reception in the moth Spodoptera littoralis. Journal of Biological Rhythms, 2007, 22(6): 502-514.

[47] Schuckel J, Siwicki K K, Stengl M. Putative circadian pacemaker cells in the antennae of the hawkmoth Manuca sexta. Cell and Tissue Research, 2007, 330(2): 271-278.

[48] Suri V, Qian Z, Hall J C, Rosbash M. Evidence that the TIM light response is relevant to light-induced phase shifts in Drosophila melanogaster. Neuron, 1998, 21(1): 225-234.

[49] Ceriani M F, Darlington T K, Staknis D, Más P, Petti A A, Weitz C J, Kay S A. Light-dependent sequestration of TIMELESS by CRYPTOCHROME. Science, 1999, 285(5427): 553-556.

[50] Hollander A, Yin C M. Neurological in?uences on pheromone release and calling behaviour in the gypsy moth, Lymantria dispar (L.). Physiological Entomology, 1982, 7(2): 163-166.

[51] Raina A K, Klun J A. Brain factor control of sex pheromone production in the female corn earworm moth. Science, 1984, 225(4661): 531-533.

[52] Delisle J, McNeil J N. Calling behaviour and pheromone titre of the true armyworm Pseudaletia unipuncta (Haw.) (Lepidoptera: Noctuidae) under different temperature and photoperiodic conditions. Journal of Insect Physiology, 1987, 33(5): 315-324.

[53] Raina A K. Neuroendocrine control of sex pheromone biosynthesis in Lepidoptera. Annual Review of Entomology, 1993, 38: 329-349.

[54] 闫硕, 李慧婷, 朱威龙, 朱家林, 张青文, 刘小侠. 光强度对棉铃虫交配行为的影响. 昆虫学报, 2014, 57(9): 1045-1050.

Yan S, Li H T, Zhu W L, Zhu J L, Zhang Q W, Liu X X. Effects of light intensity on the sexual behavior of the cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae). Acta Entomologica Sinica, 2014, 57(9): 1045-1050. (in Chinese)

[55] Li H T, Yan S, Li Z, Zhang Q W, Liu X X. Dim light during scotophase enhances sexual behavior of the oriental tobacco budworm Helicoverpa assulta (Lepidoptera: Noctuidae). Florida Entomologist, 2015, 98(2): 690-696.

[56] Schal C, Cardé R T. Effects of temperature and light on calling in the tiger moth Holomelina lamae (Freeman) (Lepidoptera: Arctiidae). Physiological Entomology, 1986, 11(1): 75-87.

[57] Kamimura M, Tatsuki S. Effects of photoperiodic changes on calling behavior and pheromone production in the oriental tobacco budworm moth, Helicoverpa assulta (Lepidoptera: Noctuidae). Journal of Insect Physiology, 1994, 40(8): 731-734.

[58] Burks C S, Brandl D G, Higbee B S. Effect of natural and artificial photoperiods and fluctuating temperature on age of first mating and mating frequency in the navel orangeworm, Amyelois transitella. Journal of Insect Science, 2011,11(1): 48.

[59] Kawazu K, Adati T, Tatsuki S. The effect of photoregime on the calling behavior of the rice leaf folder moth, Cnaphalocrocis medinalis (Lepidoptera: Crambidae). Japan Agricultural Research Quarterly, 2011, 45(2): 197-202.

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

Comments

Recommended 0

No Suggested Reading articles found!

Daily Expression of Clock Gene in Compound Eye of Helicoverpa armigera

RichHTML

PDF

Abstract

Cite this article

share this article

References

Related Articles 9

Metrics

Comments

Recommended 0

[1]	YANG GaiQing, WANG LinFeng, LI WenQing, ZHU HeShui, FU Tong, LIAN HongXia, ZHANG LiYang, TENG ZhanWei, ZHANG LiJie, REN Hong, XU XinYing, LIU XinHe, WEI YuXuan, GAO TengYun. Study on Milk Quality Based on Circadian Rhythm [J]. Scientia Agricultura Sinica, 2023, 56(2): 379-390.
[2]	ChunTao ZHANG,Tao MA,Yan TU,QiYu DIAO. Effects of Circadian Rhythm on Rumen Fermentation and Nutrient Digestion of Mutton Sheep [J]. Scientia Agricultura Sinica, 2022, 55(18): 3664-3674.
[3]	ZHANG LinLin,ZHI Hui,TANG Sha,ZHANG RenLiang,ZHANG Wei,JIA GuanQing,DIAO XianMin. Characterizations of Transcriptional and Haplotypic Variations of SiTOC1 in Foxtail Millet [J]. Scientia Agricultura Sinica, 2021, 54(11): 2273-2286.
[4]	DING Lan,GU Bao,LI PeiYing,SHU Xin,ZHANG JianXia. Genome-Wide Identification and Expression Analysis of SAP Family in Grape [J]. Scientia Agricultura Sinica, 2019, 52(14): 2500-2514.
[5]	ZHAO Jie, REN SuWei, LIU Ning, AI XinYu, MA Ji, LIU XiaoNing. Cloning and Expression of Phosphatidylethanolamine Binding Protein in Helicoverpa armigera [J]. Scientia Agricultura Sinica, 2018, 51(8): 1493-1503.
[6]	YAN Shuo, XIONG XiaoFei, CHU YanNa, LI Zhen, WU PengXiang, YANG QingPo, CUI WeiNa, XU JinTao5, XU LiXia, ZHANG QingWen, LIU XiaoXia. Effects of Predator-Induced Stress from Harmonia axyridis on the Development and Stress Protein Gene Expression of Helicoverpa armigera [J]. Scientia Agricultura Sinica, 2017, 50(21): 4118-4128.
[7]	LI Wei-zheng, HU Jing-jing, LI Hui-ling, GUO Xian-ru, YAN Feng-ming, YUAN Guo-hui. Effect of Piperine- and Sanshool-Experiences on Larval Feeding of Helicoverpa armigera [J]. Scientia Agricultura Sinica, 2015, 48(10): 2076-2084.
[8]	QI Hui-hui, ZHANG Yun-hui, WANG Jian, PENG He, ZHANG Zhi, CHENG Deng-fa. Rhythm of Rice Leaf Folder (Cnaphalocrocis medinalis) to the Searchlight Trap [J]. Scientia Agricultura Sinica, 2014, 47(22): 4436-4444.
[9]	SUN Xia, WANG Xiu-Feng, ZHENG Cheng-Shu, XING Shi-Yan, SHU Huai-Rui. The cDNA Cloning and Analysis of Sequence Information and Quantitative Express of Chrysanthemum Rhythms Clock Output Gene CmGI (GIGANTEA) [J]. Scientia Agricultura Sinica, 2012, 45(13): 2690-2703.