Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (5): 877-884.doi: 10.3864/j.issn.0578-1752.2012.05.007

• PLANT PROTECTION • Previous Articles     Next Articles

mRNA Expression and Physiological Function of Chitin Synthase1 from Oxya chinensis (Thunberg)

 YU  Zhi-Tao, LIU  Xiao-Jian, MA  恩Bo, GUO  Ya-Ping, ZHANG  Jian-Zhen   

  1. 1.山西大学应用生物学研究所，太原 030006
  • Received:2011-06-20 Online:2012-03-01 Published:2011-10-02

PDF

961

Cited

2

Abstract: 【Objective】The objective of this study is to analyze the mRNA expression characteristics and physiological function of chitin synthase1 gene (OcCHS1) from the Chinese rice grasshopper, Oxya chinensis (Thunberg), and to provide a theoretical basis for effective pests control based on RNAi.【Method】Based on the known partial conservative cDNA fragment of OcCHS1 (GenBank accession number: HM214491), primers were designed for mRNA expression analysis. The developmental and tissue-specific expression patterns of OcCHS1 were determined by RT-PCR and qPCR. RNAi experiment was performed to explore the physiological function. 【Result】 OcCHS1 was expressed at all the developmental stages. Tissue-specific analysis showed that OcCHS1 was highly expressed in the integument, followed in the trachea, and lowest in other tissues. RNAi-based gene silencing experiment showed that the fourth day of the 4th instar nymphs injected by OcCHS1-speci?c dsRNA could repress the transcript levels of OcCHS1 by 70.8% compared to those in the control nymphs. Moreover, dsRNA injections signi?cantly increased abnormalities, such as stunted development, uncompleted molting, crimpy abdomen and eventually died. Consequentially, 85.2% of mortalities were observed, while the mortality in the control nymphs was only 7.4%. 【Conclusion】OcCHS1 is essential for development and mainly responsible for chitin synthesis in cuticle and trachea of O. chinensis. Moreover, silencing of chitin synthase 1 gene by RNAi is lethal to O. chinensis.

Key words: Oxya chinensis (Thunberg), chitin synthase 1 gene, mRNA expression, RNA interference

[1]郑哲民. 蝗虫分类学. 西安: 陕西师范大学出版社, 1993.

Zheng Z M. Taxonomy of Acridoidea. Xi’an: Shanxi Normal University Press, 1993. (in Chinese)

[2]Cohen E. Chitin synthesis and inhibition, a revisit. Pest Management Science, 2001, 57(10): 946-950.

[3]Tellam R L, Vuocolo T, Johnson S E, Jarmey J, Pearson R D. Insect chitin synthase cDNA sequence, gene organization and expression. European Journal of Biochemistry, 2000, 267(19): 6025-6042.

[4]Ibrahim G H, Smartt C T, Kiley L M, Christensen B M. Cloning and characterization of a chitin synthase cDNA from the mosquito Aedes aegypti. Insect Biochemistry and Molecular Biology, 2000, 30(12): 1213-1222.

[5]Gagou M E, Kapsetaki M, Turberg A, Kafetzopoulos D. Stage-specific expression of the chitin synthase DmeChSA and DmeChSB genes during the onset of Drosophila metamorphosis. Insect Biochemistry and Molecular Biology, 2002, 32(2): 141-146.

[6]Zhang J Z, Zhu K Y. Characterization of a chitin synthase cDNA and its increased mRNA level associated with decreased chitin synthesis in Anopheles quadrimaculatus exposed to diflubenzuron. Insect Biochemistry and Molecular Biology, 2006, 36(9): 712-725.

[7]Zhu Y C, Specht C A, Dittmer N T, Muthukrishnan S, Kanost M R, Kramer K J. Sequence of a cDNA and expression of the gene encoding a putative epidermal chitin synthase of Manduca sexta. Insect Biochemistry and Molecular Biology, 2002, 32(11): 1497-1506.

[8] Hogenkamp D G, Arakane Y, Zimoch L, Merzendorfer H, Kramer K J, Beeman R W, Kanost M R, Specht C A, Muthukrishnan S. Chitin synthase genes in Manduca sexta: characterization of a gut-specific transcript and differential tissue expression of alternately spliced mRNAs during development. Insect Biochemistry and Molecular Biology, 2005, 35(6): 529-540.

[9]Bolognesi R, Arakane Y, Muthukrishnan S, Kramer K J, Terra W R, Ferreira C. Sequences of cDNAs and expression of genes encoding chitin synthase and chitinase in the midgut of Spodoptera frugiperda. Insect Biochemistry and Molecular Biology, 2005, 35(11): 1249-1259.

[10]Ashfaq M, Sonoda S, Tsumuki H. Developmental and tissue-specific expression of CHS1 from Plutella xylostella and its response to chlorfluazuron. Pesticide Biochemistry and Physiology, 2007, 89(1): 20-30.

[11]Chen X, Yang X, Kumar N S, Tang B, Sun X J, Qiu X M, Hu J, Zhang W Q. The class A chitin synthase gene of Spodoptera exigua: molecular cloning and expression patterns. Insect Biochemistry and Molecular Biology, 2007, 37(5): 409-417.

[12]Kumar N S, Tang B, Chen X F, Tian H G, Zhang W Q. Molecular cloning, expression pattern and comparative analysis of chitin synthase gene B in Spodoptera exigua. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 2008, 149(3): 447-453.

[13]Qu M B, Liu T, Yang J, Yang Q. The gene, expression pattern and subcellular localization of chitin synthase B from the insect Ostrinia furnacalis. Biochemical and Biophysical Research Communications, 2011, 404(1): 302-307.

[14]Qu M B, Yang Q. A novel alternative splicing site of class A chitin synthase from the insect Ostrinia furnacalis - Gene organization, expression pattern and physiological signi?cance. Insect Biochemistry and Molecular Biology, 2011, 41(12): 923-931.

[15]Arakane Y, Hogenkamp D G, Zhu Y C, Kramer K J, Specht C A, Beeman R W, Kanost M R, Muthukrishnan S. Characterization of two chitin synthase genes of the red flour beetle, Tribolium castaneum, and alternate exon usage in one of the genes during development. Insect Biochemistry and Molecular Biology, 2004, 34(3): 291-304.

[16]Zhang J Z, Liu X J, Zhang J Q, Li D Q, Sun Y, Guo Y P, Ma E B, Zhu K Y. Silencing of two alternative splicing-derived mRNA variants of chitin synthase 1 gene by RNAi is lethal to the oriental migratory locust, Locusta migratoria manilensis (Meyen). Insect Biochemistry and Molecular Biology, 2010, 40(11): 824-833.

[17]Merzendorfer H, Zimoch L. Chitin metabolism in insects: structure, function and regulation of chitin synthases and chitinases. The Journal of Experimental Biology, 2003, 206: 4393-4412.

[18]Fire A, Xu S Q, Montgomery M K, Kostas S A, Driver S E, Mello C C. Potent and specific genetic interference by double-stranded RNA in Caenorhabtidis elegans. Nature, 1998, 391: 806-811.

[19]Mello C C, Jr D C. Revealing the world of RNA interference. Nature, 2004, 431: 338-342.

[20]杨中侠, 文礼章, 吴青君, 王少丽, 徐宝云, 张友军. RNAi技术在昆虫功能基因研究中的应用进展. 昆虫学报, 2008, 51(10): 1077-1082.

Yang Z X, Wen L Z, Wu Q J, Wang S L, Xu B Y, Zhang Y J. Application of RNA interference in studying gene functions in insects. Acta Entomologica Sinica, 2008, 51(10): 1077-1082. (in Chinese)

[21]Arakane Y, Muthukrishnan S, Kramer K J, Specht C A, Tomoyasu Y, Lorenzen M D, Kanost M, Beeman R W. The Tribolium chitin synthase genes TcCHS1 and TcCHS2 are specialized for synthesis of epidermal cuticle and midgut peritrophic matrix. Insect Molecular Biology, 2005, 14(5): 453-463.

[22]Chen X, Tian H, Zou L, Tang B, Hu J, Zhang W. Disruption of Spodoptera exigua larval development by silencing chitin synthase gene A with RNA interference. Bulletin of Entomological Research, 2008, 98(6): 613-619.

[23]Baum J A, Bogaert T, Clinton W, Heck G R, Feldmann P, Ilagan O, Johnson S, Plaetinck G, Munyikwa T, Pleau M, Vaughn T, Roberts J. Control of coleopteran insect pests through RNA interference. Nature Biotechnology, 2007, 25(11): 1322-1326.

[24]Mao Y B, Cai W J, Wang J W, Hong G J, Tao X Y, Wang L J, Huang Y P, Chen X Y. Silencing a cotton bollworm P450 monooxygenase gene by plant-mediated RNAi impairs larval tolerance of gossypol. Nature Biotechnology, 2007, 25(11): 1307-1313.

[25]Wilson T G, Cryan J R. Lufenuron, a chitin-synthesis inhibitor, interrupts development of Drosophila melanogaster. The Journal of Experimental Zoology, 1997, 278(1): 37-44.

[26]Candy D J, Kilby B A. Studies on chitin synthesis in the desert locust. The Journal of Experimental Biology, 1962, 39: 129-140.

[27]Ampasala D R, Zheng S C, Zhang D Y, Ladd T, Doucet D, Krell P J, Retnakaran A, Feng Q L. An epidermis-specific chitin synthase cDNA in Choristoneura fumiferana: cloning, characterization, developmental and hormonal-regulated expression. Archives of Insect Biochemistry and Physiology, 2011, 76(2): 83-96.

[28]Tian H G, Peng H, Yao Q, Chen H X, Xie Q, Tang B, Zhang W Q. Developmental control of a lepidopteran pest Spodoptera exigua by ingestion of bacteria expressing dsRNA of a non-midgut gene. PLoS ONE, 2009, 4(7): e6225.

[29]李大琪, 杜建中, 张建琴, 郝耀山, 刘晓健, 王亦学, 马恩波, 张建珍, 孙  毅. 东亚飞蝗几丁质酶家族基因的表达特性与功能研究. 中国农业科学, 2011, 44(3): 485-492.

Li D Q, Du J Z, Zhang J Q, Hao Y S, Liu X J, Wang Y X, Ma E B, Zhang J Z, Sun Y. Study on expression characteristics and functions of chitinase family genes from Locusta migratoria manilensis (Meyen). Scientia Agricultura Sinica, 2011, 44(3): 485-492. (in Chinese)
[1] ZHANG Qi, CHEN ErHu, SUN DeHong, TANG PeiAn. Relationship Between Glutathione S-Transferase Genes CfGSTe1 and CfGSTd1 and Ethyl Formate Tolerance in Cryptolestes ferrugineus [J]. Scientia Agricultura Sinica, 2026, 59(5): 1008-1019.
[2] CHEN ErHu, TANG JingJie, HU ShunJie, TANG PeiAn. The Roles of Heat Shock Protein Genes CfHsp70-1 and CfHsp70-2 in Enhancing the High-Temperature Tolerance after Heat Acclimation in Cryptolestes ferrugineus [J]. Scientia Agricultura Sinica, 2025, 58(5): 918-928.
[3] XIAO ZhuoDan, QIAO JiaZheng, GAO YuLan, SHANG ZhangYin, LIU Huai, WANG Jia. Silencing of Cytochrome P450 Genes CYP6CY53 and CYP302A1 in Aphis craccivora Enhances the Sensitivity to Flonicamid [J]. Scientia Agricultura Sinica, 2025, 58(18): 3664-3675.
[4] CHEH ErHu, YUAN GuoQing, CHEN Yan, CHEN MengQiu, SUN ShengYuan, TANG PeiAn. Mitochondrial Protein-Coding Genes Nad5, Nad6 and Atp6 are Involved in Phosphine Resistance of Cryptolestes ferrugineus [J]. Scientia Agricultura Sinica, 2024, 57(9): 1722-1733.
[5] LUO LiDan, CHEN JiaMing, AN Qi, LIU Lei, SUN QinZhe, LIU Huan, WANG SenShan, SONG LiWen. Effects of Extreme High Temperature on Trehalose Content and Trehalose Transporter Gene in Tetranychus truncatus [J]. Scientia Agricultura Sinica, 2024, 57(6): 1091-1101.
[6] ZHAO YiYan, GUO HongFang, LIU WeiMin, ZHAO XiaoMing, ZHANG JianZhen. Effects of Apolipophorin on Ovarian Development and Lipid Deposition in Locusta migratoria [J]. Scientia Agricultura Sinica, 2024, 57(4): 711-720.
[7] YUAN GuoQing, CHEN ErHu, TANG PeiAn. The Mechanisms of Mitochondrial Protein-Coding Genes ND6 and ATP6 in Regulating Cold Tolerance of Cryptolestes ferrugineus [J]. Scientia Agricultura Sinica, 2024, 57(22): 4483-4494.
[8] LI ChuXin, SONG ChenHu, ZHOU JinHuan, LI JiaXin, WANG XinLiang, TIAN XuBin, SONG Zhen. Research on Prevention and Control Technology of Citrus Yellow Vein Clearing Virus Based on VIGS [J]. Scientia Agricultura Sinica, 2024, 57(22): 4473-4482.
[9] WANG Ni, SHI ZheYi, YOU YuanZheng, ZHANG Chao, ZHOU WenWu, ZHOU Ying, ZHU ZengRong. Effects of miRNA on Gene Expression of Sphingolipids Metabolism and Small RNA Analysis of Silencing NlSPT1 and NlSMase4 in Nilaparvata lugens [J]. Scientia Agricultura Sinica, 2024, 57(20): 4022-4034.
[10] CHEH ErHu, SHEN DanRong, DU WenWei, MENG HongJie, TANG PeiAn. Cuticle Protein Genes are Involved in Phosphine Resistance of Cryptolestes ferrugineus [J]. Scientia Agricultura Sinica, 2023, 56(9): 1696-1707.
[11] SHAO HongYang, MENG Xiang, ZHANG Tao, CHEN Min. Analysis of Cytochrome P450 Genes in Response to Quercetin and Function of CYP6ZB2 in Hyphantria cunea [J]. Scientia Agricultura Sinica, 2023, 56(7): 1322-1332.
[12] GUAN RuoBing,LI HaiChao,MIAO XueXia. Commercialization Status and Existing Problems of RNA Biopesticides [J]. Scientia Agricultura Sinica, 2022, 55(15): 2949-2960.
[13] YIN Fei,LI ZhenYu,SAMINA Shabbir,LIN QingSheng. Expression and Function Analysis of Cytochrome P450 Genes in Plutella xylostella with Different Chlorantraniliprole Resistance [J]. Scientia Agricultura Sinica, 2022, 55(13): 2562-2571.
[14] WU Wei,XU HuiLi,WANG ZhengLiang,YU XiaoPing. Cloning and Function Analysis of a Serine Protease Inhibitor Gene Nlserpin2 in Nilaparvata lugens [J]. Scientia Agricultura Sinica, 2022, 55(12): 2338-2346.
[15] CHEN ErHu,MENG HongJie,CHEN Yan,TANG PeiAn. Cuticle Protein Genes TcCP14.6 and TcLCPA3A are Involved in Phosphine Resistance of Tribolium castaneum [J]. Scientia Agricultura Sinica, 2022, 55(11): 2150-2160.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd