Browse > Article
http://dx.doi.org/10.5338/KJEA.2017.36.4.36

Acute Oral Toxicity of dsRNA to Honey Bee, Apis mellifera  

Lim, Hye Song (Division of Ecological Conservatioin, Bureau of Ecological Research, National Institute of Ecology)
Jung, Young Jun (Division of Ecological Conservatioin, Bureau of Ecological Research, National Institute of Ecology)
Kim, Il Ryong (Division of Ecological Conservatioin, Bureau of Ecological Research, National Institute of Ecology)
Kim, Jin (Central Research Institute)
Ryu, Sungmin (Central Research Institute)
Kim, Banni (Graduate School of Future Convergence Technology, Hankyong National University)
Lee, Jung Ro (Division of Ecological Conservatioin, Bureau of Ecological Research, National Institute of Ecology)
Choi, Wonkyun (Division of Ecological Conservatioin, Bureau of Ecological Research, National Institute of Ecology)
Publication Information
Korean Journal of Environmental Agriculture / v.36, no.4, 2017 , pp. 241-248 More about this Journal
Abstract
BACKGROUND: RNA interference (RNAi) eliminates or decreases gene expression by disrupting the target mRNA or by interfering with translation. Recently, RNAi technique was applied to generate new crop traits which provide protection against pests. To establish the environmental risk assessment protocol of RNAi LMO in lab scale, we developed dsRNA expression system using E. coli and tested acute oral toxicity assay to honey. METHOD AND RESULTS: The dsRNA expression vector, L4440, was chosen and cloned 240 bp of Snf7 and GFP gene fragment. To develop the maximum dsRNA induction condition in E. coli, we tested induction time, temperature and IPTG concentration in media. To estimate the risk assessment of dsRNA to honey bee, it has been selected and cultured with dsRNA supplement for 48 hours according to OECD guideline. As a result, the optimum condition of dsRNA induction was $37^{\circ}C$, 4 hours and 0.4 mM IPTG concentration and the difference between Snf7 and GFP dsRNA molecules from E. coli was not significant in survival and behavior to honey bee. Furthermore, blast search results indicated that effective match of predicted dsRNA fragments were not existed in honey bee genome. CONCLUSION: In this study, we developed and tested the acute oral toxicity of dsRNA using E. coli expression system to honey bee.
Keywords
Acute oral toxicity; Apis mellifera; dsRNA; Living modified organisms;
Citations & Related Records
Times Cited By KSCI : 2  (Citation Analysis)
연도 인용수 순위
1 Bettencourt, R., Terenius, O., & Faye, I. (2002). Hemolin gene silencing by dsRNA injected into Cecropia pupae is lethal to next generation embryos. Insect Molecular Biology, 11(3), 267-271.   DOI
2 Bolognesi, R., Ramaseshadri, P., Anderson, J., Bachman, P., Clinton, W., Flannagan, R., Ilagan, O., Lawrence, C., Levine, S., Moar, W., Mueller, G., Tan, J., Uffman, J., Wiggins, E., Heck, G., & Segers, G. (2012). Characterizing the mechanism of action of doublestranded RNA activity against western corn rootworm (Diabrotica virgifera virgifera LeConte), PLoS One, 7(10), e47534.   DOI
3 Clifford, R. J., Milillo, M., Prestwood, J., Quintero, R., Zurawski, D. V., Kwak, Y. I., Waterman, P. E., Lesho, E. P., & Mc Gann, P. (2012). Detection of bacterial 16S rRNA and identification of four clinically important bacteria by real-time PCR, PLoS One, 7(11), e48558.   DOI
4 Coy, M. R., Sanscrainte, N. D., Chalaire, K. C., Inberg, A., Maayan, I., Glick, E., Paldi, N., & Becnel, J. J. (2012). Gene silencing in adult Aedes aegypti mosquitoes through oral delivery of double-stranded RNA. Journal of Applied Entomology, 136(10), 741-748.   DOI
5 Elbashir, S. M., Lendeckel, W., & Tuschl, T. (2000). RNA interference is mediated by 21- and 22- nucleotide RNAs. Genes & Development, 15, 188-200.
6 Fire, A., Xu, S., Montgomery, M. K., Kostas, S. A., Driver, S. E., & Mello, C. C. (1998). Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans, Nature, 391(6669), 806-811.   DOI
7 Henry, M., Beguin, M., Requier, F., Rollin, O., Odoux, J.- F., Aupinel, P., Aptel, J., Tchamitchian, S. & Decourtye, A. (2012). A common pesticide decreases foraging success and survival in honey bees. Science, 336(6079), 348-350.   DOI
8 Henne, W. M., Buchkovich, N. J., & Emr, S. D. (2011). The ESCRT pathway. Developmental Cell, 21(1), 77-91.   DOI
9 Jeon, M. J., Seo, M. J., Youn, N. Y., & Yu, Y. M. (2014). RNA Interference of Chitinase gene in Spodoptera litura. The Korean Journal of Pesticide Science, 18(3), 202-209.   DOI
10 Jaubert-Possamai, S., Trionnaire, G. L., Bonhomme, J., Christophides, G. K., Rispe, C., & Tagu, D. (2007). Gene knockdown by RNAi in the pea aphid Acyrthosiphon pisum. BMC Biotechnology, 7(1), 63-70.   DOI
11 Kim, J. H., Seo, E. Y., Kim, J. K., Lim, H. S., Yu, Y. M., & Youn, Y. N. (2015). Gene expression in plant according to RNAi treatment of the tobacco whitefly. Korean Journal of Agricultural Science, 42(2), 81-86.   DOI
12 Koi, J., Ramaseshadri, P., Bolognesi, R., Segers, G., Flannagan, R., & Park, Y. (2014). Ultrastructural changes caused by Snf7 RNAi in larval enterocytes of western corn rootworm Diabrotica virgifera virgifera Le Conte. PLoS One, 9(1), e83985.   DOI
13 Lima, P. C., Botwright, N. A., Harris, J. O. & Cook, M. (2014). Development of an in vitro Model System for Studying Bacterially Expressed dsRNA-Mediated Knockdown inNeoparamoeba genus. Marine Biotechnology, 16(4), 447-455.   DOI
14 Niimi, T., Kuwayama, H., & Yaginuma, T. (2005). Larval RNAi Applied to the Analysis of Postembryonic Development to the Analysis of Postembryonic Development in the Ladybird Beetle, Harmonia axyridis. Journal of Insect Biotechnology Sericology, 74(3), 95-102.
15 Timmons, L., Court, D. L., & Fire, A. (2001). Ingestion of bacterially expressed dsRNAs can produce species and potent genetic interference in Caenorhabditis elegans. Gene, 263(1), 103-112.   DOI
16 Quan, G., Ladd, T., Duan, J., Wen, F., Doucet, D., Cusson, M., & Krell, P. J. (2013). Characterization of a spruce budworm chitin deacetylase gene: Stage and tissue specific expression, and inhibition using RNA interference. Insect Biochemistry Molecular Biology, 43(8), 683-691.   DOI
17 Rajagopal, R., Sivakumar, S., Agrawal, N., Malhotra, P., & Bhatnagar, R. K. (2002). Silencing of midgut Aminopeptidase N of Spodoptera litura by doublestranded RNA establishes its role as Bacillus thuringiensis toxin receptor. Journal of Biological Chemistry, 277(49), 46849-46851.   DOI
18 Ramaseshadri, P., Segers, G., Flannagan, R., Wiggins, E., Clinton, W., Ilagan, O., McNulty, B., Clark, T., & Bolognesi, R. (2013). Physiological and cellular responses caused by RNAi-mediated suppression of Snf7 orthologue in western corn rootworm (Diabrotica virgifera virgifera) larvae. PLoS One, 8(1), e54270.   DOI
19 Rodriguez-Cabrera, L., Trujillo-Bacallao, D., Borras-Hidalgo, O., Wright, D. J., & Ayra-Pardo, C. (2010). RNAi mediated knockdown of a Spodoptera frugiperda trypsin like serine protease gene reduces susceptibility to a Bacillus thuringiensis Cry1Ca1 protoxin. Environmental Microbiology, 12(11), 2894- 2903.   DOI
20 Tan, J., Levine, S. L., Bachman, P. M., Jenson, P. D., Mueller, G. M. Uffman, J. P., Meng, C., Song, Z., Richards, K. B., & Beevers M. H. (2015) No impact of DvSnf7 RNA on honey bee (Apis mellifera L.) adults and larvae in dietary feeding tests. Environmental Toxicology and Chemistry, 35(2), 287-294.
21 Turner, C. T., Davy, M.W., MacDiarmid, R. M., Plummer, K. M., Birch, N. P., & Newcomb, R. D. (2006). RNA interference in the light brown apple moth, Epiphyas postvittana (Walker) induced by double- stranded RNA feeding. Insect Molecular Biology, 15(3), 383-391.   DOI
22 Velez, A. M., Jurzenski, J., Matz, N., Zhou, X., Wang, H., Ellis, M., & Siegfried, B. D. (2016). Developing an in vivo toxicity assay for RNAi risk assessment in honey bees, Apis mellifera L. Chemosphere, 144, 1083-1090.   DOI
23 Wegner, C. S., Rodahl, L. M. W., & Stenmark, H. (2011). ESCRT proteins and cell signaling. Traffic, 12(10), 1291-1297.   DOI
24 Bachman, P. M., Bolognesi, R., Moar, W. J., Mueller, G. M., Paradise, M. S., Ramaseshadri, P., Tan, J., Uffman, J. P., Warren, J., Wiggins, B. E., & Levin, S. L. (2013). Characterization of the spectrum of insecticidal activity of a double-stranded RNA with targeted activity against Western Corn Rootworm (Diabrotica vrigifera virgifera LeConte). Transgenic Research, 22(6), 1207- 1222.   DOI
25 Zhu, J. Q., Liu, S., Ma, Y., Zhang, J. Q., Qi, H. S., Wei, Z. J., Yao, Q., Zhang W. Q., & Li, Z. (2012). Improvement of Pest Resistance in Transgenic Tobacco Plants Expressing dsRNA of an Insect-Associated Gene EcR. PLoS One, 7(6), e38572.   DOI
26 Bautista, M. A. M., Miyata, T., Miura, K., & Tanaka, T. (2009). RNA interference mediated knockdown of a cytochrome P450, CYP6BG1, from the diamonback moth, Plutella xylostella, reduces larval resisitance to permethrin. Insect Biochemistry and Molecular Biology, 39(1), 38-46.   DOI