Construction of a cDNA library of Aphis gossypii Glover for use in RNAi

다국어 초록 (Multilingual 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 λ. 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 × 106 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.

참고문헌 (Reference)

1 김용헌, "오이재배 시설하우스에서 천적유지식물을 이용한 진딧물의 생물적 방제" 한국응용곤충학회 42 (42): 81-84, 2003

2 최용석, "시설오이에서 azoxystrobin, 미끌애꽃노린재, 콜레마니진디벌을 이용한꽃노랑총채벌레와 목화진딧물 종합관리 사례" 한국응용곤충학회 52 (52): 379-386, 2013

3 Ko NY, "cDNA library construction of Bemisia tabaci (Hemiptera: Aleyrodidae) for using RNAi" Chungnam National University 2015

4 Leshkowitz D, "Whitefly (Bemisia tabaci) genome project: analysis of sequenced clones from egg, instar, and adult (viruliferous and non-viruliferous) cDNA libraries" 7 : 79-, 2006

5 Scott JG, "Towards the elements of successful insect RNAi" 59 : 1212-1221, 2013

6 Pitino M, "Silencing of aphid genes by dsRNA feeding from plants" 6 : e25709-, 2011

7 Price DR, "RNAi-mediated crop protection against insects" 26 : 393-400, 2008

8 Mutti NS, "RNAi knockdown of a salivary transcript leading to lethality in the pea aphid, Acyrthosiphon pisum" 6 : 1-7, 2006

9 Upadhyay SK, "RNA interference for the control of whiteflies (Bemisia tabaci) by oral route" 36 : 153-161, 2011

10 Xu R, "Protocol: Streamline cloning of genes into binary vectors in Agrobacterium via the Gateway TOPO vector system" 4 : 4-, 2008

11 Liu E, "Optimized cDNA libraries for virus-induced gene silencing (VIGS) using tobacco rattle virus" 4 : 1-13, 2008

12 Sapountzis P, "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" 51 : 20-32, 2014

13 Huvenne H, "Mechanisms of dsRNA uptake in insects and potential of RNAi for pest control: A review" 56 : 227-235, 2010

14 Al-Ayedh H, "Insecticidal potency of RNAi-based catalase knockdown in Rhynchophorus ferrugineus(Oliver)(Coleoptera: Curculionidae)" 72 : 2118-2127, 2016

15 Leclant F, "Insect Pests of Cotton" CAB International 285-323, 1994

16 Whyard S, "Ingested double-stranded RNAs can act as species-specific insecticides" 39 : 824-832, 2009

17 Paik WH, "Illustrated Flora and Fauna of Korea 13 (Insecta V)"

18 CoyMR, "Gene silencing in adult Aedes aegypti mosquitoes through oral delivery of double-stranded RNA" 136 : 741-748, 2012

19 Jaubert-Possamai S, "Gene knockdown by RNAi in the pea aphid Acyrthosiphon pisum" 7 : 63-, 2007

20 Sasaki Y, "Evidence for high specificity and efficiency of multiple recombination signals in mixed DNA cloning by the Multisite Gateway system" 107 : 233-243, 2004

21 Baum JA, "Control of coleopteran insect pests through RNA interference" 25 : 1322-1326, 2007

22 Bolongnesi R, "Characterizing the mechanism of action of double-stranded RNA activity against Western corn rootworm (Diabrotica virgifera virgifera LeConte)" 7 : e47534-, 2012

23 Shakesby AJ, "A water-specific aquaporin involved in aphid osmoregulation" 39 : 1-10, 2009