Spodoptera eridania and S. ornithogalli (Lepidoptera: Noctuidae), which are polyphagous pests that damage various crops such as tomatoes and beans are regulated quarantine species that are highly likely to invade South Korea. Therefore, it is crucial to promptly and accurately identify the presence of S. eridania and S. ornithogalli in crop fields to effectively eradicate as a regulated quarantine species. In this study, we developed a loop-mediated isothermal amplification (LAMP) assay, which allows for rapid in-field identification. To develop the LAMP　assay, we selected target species-specific genomic regions from the whole-genome sequences of one target and 13 other lepidopteran species. We validated each five and six primer sets that consistently produced positive reactions in S. eridania and S. ornithogalli, respectively. To test the sensitivity of the each locus, LAMP reactions were performed using various reaction times using crude DNA, which was extracted from various types of adult tissues. All sensitivity tests were also successful.

국내 유입 가능성이 높은 검역 관리해충인 Spodoptera eridania 및 S. ornithogalli는 전 세계적으로 토마토, 콩 등 여러 종의 작물을 가해하는 광식성 해충이다. 이에 따라 국내 유입 시 해당 작물에 높은 경제적 피해를 입힐 가능성 이 있으므로 신속 정확한 진단이 필요한 실정이다. 따라서 본 연구에서는 상기 두 종을 대상으로 현장 활용이 가능한 LAMP 진단법 개발을 수행하였다. 표적종 두 종 및 비표적종 11종(국내 발생 Spodoptera 종 및 동일 기주 가해종 등)의 전장유전체 정보를 확보한 후 비교 분석을 통해 각 표적종 별 특이적 영역을 확보한 후 해당 영역을 대상으로 LAMP 프라이머를 제작하였다. DNA 농도 10 ng/μL, 반응시간 40분을 기준으로 LAMP 진단을 수행한 결과, Spodoptera eridania는 5개의 LAMP 진단 마커를 개발하였고, S. ornithogalli는 3개의 LAMP 진단 마커를 개발하였다.

The differences in the immune response between body lice, Pediculus humanus humanus, and head lice, Pediculus humanus capitis, were regarded as primary factors determining their differential vector competence. To find any differences in genetic components in immune system between body and head lice, whole genome sequences of head lice were determined by both SBS [sequencing by synthesis, Illumina Genome Analyzer (Illumina-GA)] and pyrosequencing (Roche GS FLX), and compared with the reference genome sequences of body lice. The short DNA reads from Illumina-GA (an average mapping depth of 50-fold) were aligned first to the body louse reference genome, to which Roche GS FLX DNA reads (an average depth of 2.5-fold) were subsequently assembled to make up gaps between mapped consensus. Total consensus showed a size of 114 Mb and a coverage of 96% of the published body louse genome sequences. From this head louse genome sequences, a total of 12,651 genes were predicted and used for comparing with the 10,775 genes previously reported from the body louse genome. The homolog analysis identified 873 head louse-specific genes and 422 body lice-specific genes. Comparison of immune response genes between both louse species showed head lice have more number of immune-related genes than body lice. Head lice were determined to possess all of the 107 immune-related genes reported in the previous study (Kim et al., 2011), suggesting that there is no difference in genetic make-up in terms of the 107 immune-related genes between body and head lice.