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Disruption of Spodoptera exigua larval development by silencing chitin synthase gene A with RNA interference

Published online by Cambridge University Press:  29 July 2008

X. Chen ,
H. Tian ,
L. Zou ,
B. Tang ,
J. Hu  and
W. Zhang
X. Chen
Affiliation:
State Key Laboratory of Biocontrol and Institute of Entomology, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China
H. Tian
Affiliation:
State Key Laboratory of Biocontrol and Institute of Entomology, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China
L. Zou
Affiliation:
State Key Laboratory of Biocontrol and Institute of Entomology, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China
B. Tang
Affiliation:
State Key Laboratory of Biocontrol and Institute of Entomology, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China
J. Hu
Affiliation:
State Key Laboratory of Biocontrol and Institute of Entomology, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China
W. Zhang*
Affiliation:
State Key Laboratory of Biocontrol and Institute of Entomology, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China
*
*Author for correspondence Fax: +86 20 84112297 E-mail: [email protected]
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Abstract

RNA interference (RNAi) is a powerful tool for rapidly analyzing gene functions. However, little is known about the possible use of dsRNA/siRNA as a pest control method. Here, we demonstrate that dsRNA/siRNA can induce the silence of chitin synthase gene A (CHSA), which is an important gene for the growth and development of cuticles and trachea in beet armyworm, Spodoptera exigua. Based on the in vitro RNAi experiments in an insect cell line (Trichoplusia ni High 5), in vivo RNAi was performed by injecting synthesized dsRNA/siRNA into the 4th instar larvae of S. exigua. Significantly lower levels of CHSA transcripts were detected. In addition, the cuticle of these insects was disordered and the epithelial walls of larval trachea did not expand uniformly in injected individuals. Moreover, Injections significantly increased abnormalities relative to control larvae. These results highlighted the possibility of dsRNA/siRNA for gene function studies in lepidopteran insects and future pest control.

Keywords

chitin synthase geneRNAifunctionSpodoptera exiguapest control
Type
Research Paper
Information
Bulletin of Entomological Research , Volume 98 , Issue 6 , December 2008 , pp. 613 - 619
Copyright
Copyright © 2008 Cambridge University Press

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References

Agrawal, N., Malhotra, P. & Bhatnagar, R.K. (2004) siRNA-directed silencing of transgene expressed in cultured insect cells. Biochemical Biophysical Research Communications 320, 428–434.Google Scholar
Arakane, Y., Muthukrishnan, S., Kramer, K.J., Specht, C.A., Tomoyasu, Y., Lorenzen, M.D., Kanost, M. & Beeman, R.W. (2005) The Tribolium chitin synthase genes TcCHS1 and TcCHS2 are specialized for synthesis of epidermal cuticle and midgut peritrophic matrix. Insect Molecular Biology 14, 453–463.Google Scholar
Araujo, S.J., Aslam, H., Tear, G. & Casanova, J. (2005) Mummy/cystic encodes an enzyme required for chitin and glycan synthesis, involved in trachea, embryonic cuticle and CNS development-analysis of its role in Drosophila tracheal morphogenesis. Developmental Biology 288, 179–193.CrossRefGoogle ScholarPubMed
Baum, J., Bogaert, T., Clinton, W., Heck, G., Feldmann, P., Ilagan, O., Johnson, S., Plaetinck, G., Munyikwa, T., Pleau, M., Vaughn, T. & Roberts, J. (2007) Control of coleopteran insect pests through RNA interference. Nature Biotechnology 25, 1322–1326.Google Scholar
Bettencourt, R., Terenius, O. & Faye, I. (2002) Hemolin gene silencing by ds-RNA injected into Cecropia pupae is lethal to next generation embryos. Insect Molecular Biology 11, 267–271.CrossRefGoogle ScholarPubMed
Caplen, N.J., Parrish, S., Imani, F., Fire, A. & Morgan, R.A. (2001) Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems. Proceedings of the National Academy Sciences USA 98, 9742–9747.CrossRefGoogle ScholarPubMed
Chen, X., Yang, X., Senthil Kumar, N., Tang, B., Sun, X., Qiu, X., Hu, J. & Zhang, W. (2007) The class A chitin synthase gene of Spodoptera exigua: molecular cloning and expression patterns. Insect Biochemistry and Molecular Biology 37, 409–417.CrossRefGoogle Scholar
Clemens, J., Worby, C., Simonson-Leff, N., Muda, M., Maehama, T., Hemmings, B. & Dixon, J. (2001) Use of double-stranded RNA interference in Drosophila cell lines to dissect signal transduction pathways. Proceedings of the National Academy Sciences USA 97, 6499–6503.CrossRefGoogle Scholar
Coutinho, P.M. & Henrissat, B. (1999) Life with no sugars? Journal of Molecular Microbiology and Biotechnology 1, 307–308.Google ScholarPubMed
Cruz, J., Mane-Padros, D., Belles, X. & Martin, D. (2006) Functions of the ecdysone receptor isoform-A in the hemimetabolous insect Blattella germanica revealed by systemic RNAi in vivo. Developmental Biology 297, 158–171.CrossRefGoogle ScholarPubMed
Eleftherianos, I., Marokhazi, J., Millichap, P.J., Hodgkinson, A.J., Sriboonlert, A., ffrench-Constant, R.H. & Reynolds, S.E. (2006a) Prior infection of Manduca sexta with non-pathogenic Escherichia coli elicits immunity to pathogenic Photorhabdus luminescens: roles of immune-related proteins shown by RNA interference. Insect Biochemistry and Molecular Biology 36, 517–525.Google Scholar
Eleftherianos, I., Millichap, P.J., ffrench-Constant, R.H. & Reynolds, S.E. (2006b) RNAi suppression of recognition protein mediated immune responses in the tobacco hornworm Manduca sexta causes increased susceptibility to the insect pathogen Photorhabdus. Developmental and Comparative Immunology 30, 1099–1107.CrossRefGoogle Scholar
Eleftherianos, I., Gokcen, F., Felfoldi, G., Millichap, P.J., Trenczek, T.E., ffrench-Constant, R.H. & Reynolds, S.E. (2007) The immunoglobulin family protein Hemolin mediates cellular immune responses to bacteria in the insect Manduca sexta. Cell Microbiology 9, 1137–1147.CrossRefGoogle ScholarPubMed
Hammond, S.M., Bernstein, E., Beach, D. & Hannon, G.J. (2000) An RNA-directed nuclease mediates post-transcriptional gene silencing in Drosophila cells. Nature 404, 293–296.Google Scholar
Lum, L., Yao, S., Mozer, B., Rovescalli, A., Von Kessler, D., Nirenberg, M. & Beachy, P.A. (2003) Identification of Hedgehog pathway components by RNAi in Drosophila cultured cells. Science 299, 2039–2045.Google Scholar
Mao, Y., Cai, W., Wang, J., Hong, G., Tao, X., Wang, L., Huang, Y. & Chen, X. (2007) Silencing a cotton bollworm P450 monooxy genase gene by plant-mediated RNAi impairs larval tolerance of gossypol. Nature Biotechnology 25, 1307–1313.Google Scholar
Meyering-Vos, M., Merz, S., Sertkol, M. & Hoffmann, K.H. (2006) Functional analysis of the allatostatin-A type gene in the cricket Gryllus bimaculatus and the armyworm Spodoptera frugiperda. Insect Biochemistry and Molecular Biology 36, 492–504.CrossRefGoogle ScholarPubMed
Quan, G.X., Kanda, T. & Tamura, T. (2002) Induction of the white egg 3 mutant phenotype by injection of the double-stranded RNA of the silkworm white gene. Insect Molecular Biology 11, 217–222.CrossRefGoogle ScholarPubMed
Rajagopal, R., Sivakumar, S., Agrawal, N., Malhotra, P. & Bhatnagar, R.K. (2002) Silencing of midgut aminopeptidase N of Spodoptera litura by double-stranded RNA establishes its role as Bacillus thuringiensis toxin receptor. Journal of Biological Chemistry 277, 46849–46851.CrossRefGoogle ScholarPubMed
Rajagopal, R., Thamilarasi, K., Venkatesh, G.R., Srinivas, P. & Bhatnagar, R.K. (2005) Immune cascade of Spodoptera litura: cloning, expression, and characterization of inducible prophenol oxidase. Biochemical Biophysical Research Communications 337, 394–400.CrossRefGoogle ScholarPubMed
Tomoyasu, Y. & Denell, R.E. (2004) Larval RNAi in Tribolium (Coleoptera) for analyzing adult development. Development Genes and Evolution 214, 575–578.Google Scholar
Vermehren, A., Qazi, S. & Trimmer, B.A. (2001) The nicotinic alpha subunit MARA1 is necessary for cholinergic evoked calcium transients in Manduca neurons. Neuroscience Letters 313, 113–116.Google Scholar
Zhou, X., Oi, F.M. & Scharf, M.E. (2006) Social exploitation of hexamerin: RNAi reveals a major caste-regulatory factor in termites. Proceedings of the National Academy of Sciences of the United States of America 103, 4499–4504.Google Scholar
Zimoch, L. & Merzendorfer, H. (2002) Immunolocalization of chitin synthase in the tobacco hornworm. Cell Tissue Research 308, 287–297.CrossRefGoogle ScholarPubMed