Entomological Research

The Entomological Society of Korea (한국곤충학회)
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Construction of a cDNA library of Aphis gossypii Glover for use in RNAi

  • KWON, HyeRi (Department of Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • KIM, JungGyu (Department of Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • LIM, HyounSub (Department of Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • YU, YongMan (Department of Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • YOUN, YoungNam (Department of Applied Biology, College of Agriculture and Life Sciences, Chungnam National University)
  • Received : 2017.09.14
  • Accepted : 2018.02.24
  • Published : 2018.09.29
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Abstract

Aphis gossypii Glover is an important insect pest that functions as a viral vector and mediates approximately 45 different viral diseases. As part of a strategy for control of A. gossypii, we investigated the functions of genes using RNAi. To this end, a cDNA library was constructed for various genes and for selecting appropriate targets for RNAi mediated silencing. The cDNA library was constructed using the Gateway cloning system with site-specific recombination of bacteriophage ${\lambda}$. It was used to carry out single step cloning of A. gossypii cDNAs. As a result, a cDNA library with a titer of $8.4{\times}10^6$ was constructed. Since the sequences in this library carry att sites, they can be cloned into various binary vectors. This library will be of value for various studies. For later screening of selected genes, it is planned to clone the library into virus-induced gene silencing (VIGS) vectors, which makes it possible to analyze gene function and allow subsequent transfection of plants. Such transfection experiments will allow testing of RNAi-induced insecticidal activity or repellent activity to A. gossypii, and result in the identification of target genes. It is also expected that the constructed cDNA library will be useful for analysis of gene functions in A. gossypii.

Keywords

References

  1. Al-Ayedh H, Rizwan-ul-Haq M, Hussain A et al. (2016) Insecticidal potency of RNAi-based catalase knockdown in Rhynchophorus ferrugineus(Oliver)(Coleoptera: Curculionidae). Pest Management Science 72: 2118-2127. https://doi.org/10.1002/ps.4242
  2. Baum JA, Bogaert T, Clinton W et al. (2007) Control of coleopteran insect pests through RNA interference. Nature Biotechnology 25: 1322-1326. https://doi.org/10.1038/nbt1359
  3. Bolongnesi R, Ramaseshadri P, Anderson J et al. (2012) Characterizing the mechanism of action of double-stranded RNA activity against Western corn rootworm (Diabrotica virgifera virgifera LeConte). PLoS One e47534: 7. https://doi.org/10.1371/journal.pone.0047534
  4. Choi YS, Whang IS, Han IS et al. (2013) A case study for intergrated pest management of Franklinella occidentalis and Aphis gossypii by simultaneously using Orius laevigatus and Aphidius colemani with azoxystrobin in cucumber plants. Korean Journal of Applied entomology 52: 379-386. https://doi.org/10.5656/KSAE.2013.11.0.077
  5. Coy MR, Sanscrainte ND, Chalaire KC et al. (2012) Gene silencing in adult Aedes aegypti mosquitoes through oral delivery of double-stranded RNA. Journal of Applied Entomology 136: 741-748. https://doi.org/10.1111/j.1439-0418.2012.01713.x
  6. Huvenne H, Smagghe G (2010) Mechanisms of dsRNA uptake in insects and potential of RNAi for pest control: A review. Journal of Insect Physiology 56: 227-235. https://doi.org/10.1016/j.jinsphys.2009.10.004
  7. Jaubert-Possamai S, Le Trionnaire G, Bonhomme J et al. (2007) Gene knockdown by RNAi in the pea aphid Acyrthosiphon pisum. BMC Biotechnology 7: 63. https://doi.org/10.1186/1472-6750-7-63
  8. Kim YH, Kim JH (2003) Biological control of aphids on cucumber in plastic greenhouses using banker plants. Korean Journal of Applied Entomology 42: 81-84.
  9. Ko NY (2015) cDNA library construction of Bemisia tabaci (Hemiptera: Aleyrodidae) for using RNAi. Master thesis, Chungnam National University, pp 63.
  10. Leclant F, Deguine JP (1994) Aphids (Hemiptera: Aphididaae). In: Matthew GA, Tunstal JP (eds) Insect Pests of Cotton, pp 285-323. CAB International, Wallingford, UK.
  11. Leshkowitz D, Gazit S, Reuveni E et al. (2006) Whitefly (Bemisia tabaci) genome project: analysis of sequenced clones from egg, instar, and adult (viruliferous and non-viruliferous) cDNA libraries. BMC Genomics 7: 79. https://doi.org/10.1186/1471-2164-7-79
  12. Liu E, Page JE (2008) Optimized cDNA libraries for virus-induced gene silencing (VIGS) using tobacco rattle virus. Plant Methods 4: 1-13. https://doi.org/10.1186/1746-4811-4-1
  13. Mutti NS, Park Y, Reese JG et al. (2006) RNAi knockdown of a salivary transcript leading to lethality in the pea aphid, Acyrthosiphon pisum. Journal of Insect Science 6: 1-7.
  14. Paik WH (1972) Illustrated Flora and Fauna of Korea 13 (Insecta V), p 751. Ministry of Education, Korea.
  15. Pitino M, Coleman AD, Maffei ME et al. (2011) Silencing of aphid genes by dsRNA feeding from plants. PLoS One 6: e25709. https://doi.org/10.1371/journal.pone.0025709
  16. Price DR, Gatehouse JA (2008) RNAi-mediated crop protection against insects. Trends in Biotechnology 26: 393-400. https://doi.org/10.1016/j.tibtech.2008.04.004
  17. Sapountzis P, Duport G, Balmand S et al. (2014) New insight into the RNA interference response against cathepsin-L gene in the pea aphid, Acyrthosiphon pisum: Molting or gut phenotypes specially induced by injection or feeding treatments. Insect Biochemistry and Molecular Biology 51: 20-32. https://doi.org/10.1016/j.ibmb.2014.05.005
  18. Sasaki Y, Sone T, Yoshida S et al. (2004) Evidence for high specificity and efficiency of multiple recombination signals in mixed DNA cloning by the Multisite Gateway system. Journal of biotechnology 107: 233-243. https://doi.org/10.1016/j.jbiotec.2003.10.001
  19. Scott JG, Michel K, Bartholomay LC et al. (2013) Towards the elements of successful insect RNAi. Journal of Insect Physiology 59: 1212-1221. https://doi.org/10.1016/j.jinsphys.2013.08.014
  20. Shakesby AJ, Wallace IS, Isaacs HV et al. (2009) A water-specific aquaporin involved in aphid osmoregulation. Insect Biochemistry and Molecular Biology 39: 1-10. https://doi.org/10.1016/j.ibmb.2008.08.008
  21. Upadhyay SK, Chandrashekar K, Thakur N et al. (2011) RNA interference for the control of whiteflies (Bemisia tabaci) by oral route. Journal of Biosciences 36: 153-161. https://doi.org/10.1007/s12038-011-9009-1
  22. Whyard S, Singh AD, Wong S (2009) Ingested double-stranded RNAs can act as species-specific insecticides. Insect Biochemistry and Molecular Biology 39: 824-832. https://doi.org/10.1016/j.ibmb.2009.09.007
  23. Xu R, Li QQ (2008) Protocol: Streamline cloning of genes into binary vectors in Agrobacterium via the Gateway TOPO vector system. Plant Methods 4: 4. https://doi.org/10.1186/1746-4811-4-4