목화진딧물의 방제에 RNA interference(RNAi)를 이용하여 새로운 시각으로 새로운 방제를 시도하고자 한다. RNAi를 이용하여 목화진딧물의 방제에 이용할 target유전자들을 선발하기 위하여 gateway system을 이용한 목화진딧물 cDNA library를 제작하였다. 그 결과 RNAi에 적합한 약 100~400bp의 insert를 확인하였으며, blast search 및 EST database 비교분석 결과, 목화진딧물 관련 유전자임을 확인하였고, 최종적으로 8.4x105 titer의 목화진딧물 cDNA library를 완성하였다. 이러한 cDNA library는 att site를 가지는 TRV(Tobacco rattle virus) RNA2 vector에 LR recombination한 다음 Agrobacterium tumefacience(GV2260)에 transformation하였다. Agro-infiltration을 통하여 RNAi기작이 진단된 오이 에 목화진딧물을 접종하여 섭식시켰다. 섭식을 통한 살충 또는 기피효과를 bio-assay함으로써 target유전자를 선발하는 데 이용될 수 있을 것으로 사료된다.
Aphis gossypii was widely distributed throughout the tropical, subtropical and temperate zone. The chemical control of A. gossypii is becoming problem because it was rapidly appeared resistance expression to chemicals. We will attempt to resolve the this problem using RNAi technique. Besides, RNAi technology can be helpful to study the target genes of A. gossypii. In this study we produce cDNA library construction using gateway cloning system for selecting target gene in order to control of A. gossypii using RNAi. As a result, the 100-400bp of insert size, which is appropriate for RNAi was confirmed. Most of insert gene is associated with A. gossypii, after that insert sequence was compared with DNA databases and EST databases using NCBI blast search. Consequentially, A. gossypii of cDNA library with the titer of 3.15x105 clones were completed. And we will perform the LR recombination to transfer cDNA library into TRV2 (tobacco rattle virus) vector with att site. Then, after performing transformation using Agrobacterium tumefaciens (GV 2260), we inoculated to cucumber with A. tumefaciens. An insecticidal effect or a repellent activity against A. gossypii by changing behavior in transgenic cucumber plants were conformed. Also, the selecting target gene in order to control A. gossypii using RNAi may be provided.