새롭게 육성된 낭충봉아부패병 저항성 신품종 토종벌(Apis cerana koreana) 과 기존 농가에서 관행적으로 사육되는 토종벌 사이의 형태학적 차 이를 육안으로 확연하게 구분하는 것은 어렵지만 본 연구에서는 신품종 토종벌(A. c. koreana) 을 기존 토종벌(A. c. koreana) 품종 및 계통 간 형태학적 비교를 통해 신품종 만의 특성을 결정할 수 있는 표현형 정보를 제공하였다. 신품종 토종벌(A. c. koreana)의 외부형질을 이용한 품종 특성은 22가지의 형태학적 특성을 기하학적, 형태학적 분석 방법을 적용하고 토종벌(A. c. koreana)의 로얄젤리 생산량, 일벌, 여왕벌, 수벌의 특성을 비교 분석하였다. 본 연구 결과, 신품종 토종벌(A. c. koreana)은 기존 토종벌과 앞날개의 길이에 차이를 보였으며, 중국의 동양종꿀벌(A. cerana)과 비교한 결과, 일벌은 몸무게, 혀의 길이, 앞날개의 길이 등의 값이 높았다. 또한, 신품종 토종벌(A. c. koreana)은 A. cerana indica 보다 두 가지 부위에서 형태학적인 차이를 보였다. 그리고 신품종 토종벌(A. c. koreana)은 로얄젤리를 다른 품종과 비교하여 많이 분비하여 봉군의 발육에 긍정적인 영향을 끼쳤다. 따라서 본 연구결과는 신규 육성 토종벌(A. cerana)에 대한 형태학적 분석 방법을 이용하여 품종을 분류하는데 도움이 될 것으로 기대한다.

Frankliniella occidentalis is a major pest in agriculture. Following overuse of insecticides, high resistance has developed due to its high reproduction rate and short generation time. To control the resistant strains of the thrips, the ingestion RNAi- based control was established. A total of 67 genes were selected, and their double-stranded RNAs (dsRNA) were delivered to thrips via the leaf disc-feeding method. Among the genes screened, the dsRNA of Toll-like receptor 6 (TLR6) and coatomer protein subunit epsilon (COPE) resulted in the highest mortality (3.8- and 2.8-fold faster LT50 compared to control, respectively) when ingested by thrips. The dsRNA-fed thrips showed 53% and 83% reduced transcription levels of TLR6 and COPE, respectively. This result demonstrates that the observed mortality of thrips following dsRNA ingestion was due to RNAi, and this lethal genes can be employed as a practical tool to control thrips in the field.

A residual contact vial plus water (RCVpW) bioassay method was established to monitor insectiside resistance in field populations of the melon thrips, Thrips palmi. Resistance level against six major insecticides were evalutated in five regions to test applicability of RCVpW as an on-site resistance monitoring tool. Reduced mortality in response to six test insecticides were exhibited compared to the RDA susceptable strain showing 100 % mortality, indicating different degree of resistance. An apparently reduced mortality to emamectin benzoate and chlofenapyr was observed in some field populations, suggesting uneven distribution of resistance to these insecticides in field populations. In addition, spinosad resistance was high and widely distributed in the test regions. Synergistic bioassay revealed that cytochrome P450-mediated metabolic factor is involved in spinosad resistance in the Korean population.

RNA interference (RNAi)-based strategy has been developed to control various phytophagous chewing pests. However, only a few cases of RNAi-based control success have been reported for sucking pests, suggesting that sucking pests likely ingest less amount of transgenic subcellular hairpin RNA (or dsRNA). In this study, as the basic information for the establishment of ingestion RNAi against sucking pests, feeding amount and time course of plant subcellular fractions of the four sucking pest species (Frankliniella occidentalis, Frankliniella intonsa, Tetranychus urticae and Nilaparvata lugans) were determined by quantitative PCR (qPCR). Adults of the four species were starved for 24 h and then fed with kidney bean leaf (F. occidentalis, F. intonsa, T. urticae) or rice leaf (N. lugens) for 48 h. The leaf-fed adults were collected every 6-h interval and their genomic DNA was extracted. The ingested fractions of chloroplast and nuclear were quantified using rubisco and 50s rRNA as marker genes, respectively. The ingested amount of rubisco and 50s rRNA genes in F. occidentalis, F. intonsa and T. urticae showed rapid increasing pattern after feeding and then slightly reduced over time. In contrast, N. lugens neither ingest nuclear nor showed any distinct feeding pattern of chloroplast. These results demonstrate that F. occidentalis, F. intonsa and T. urticae ingest both chloroplasts and nucleus along with cytosol as cell-feeders but N. lugens, a phloem sap feeder, does not ingest nucleus during sucking. Our findings further suggest that ingestion RNAi-based control strategy would work better for cell-feeding sucking pests compared to phloem sap-feeding sucking pests.

Residual contact vial (RCV) method was used to monitor insecticide resistance in field populations of the melon thrips, Thrips palmi. Median lethal doses (LD50) at 8 h post-treatment of six insecticides (chlorfenapyr, cyantraniliprole, cypermethrin, dinotefuran, emamectin benzoate and spinosad), which are commonly used for T. palmi control, were determined at 8 h post-treatment using a susceptible RDA strain. The diagnostic doses for on-site resistance monitoring of the six insecticides, which were determined as two-fold higher doses of LD90 for the RDA strain, were in the range of 0.299 to 164,25 μg-1cm2. To test the applicability of RCV for T. palmi, insecticide resistance levels in three field populations (Gyeonggi; GG_AS, Chungbuk; CB_CJ, Jeonbuk; JB_KJ) were evaluated. Field populations showed reduced mortality (0-50% mortality) to spinosad, cypermethrin, cyantraniliprole and emamectin benzoate, that they have different degree of resistances to these insecticides. In particular, all test field populations exhibited 0% mortality to spinosad, suggesting wide spread of spinosad resistance in the field. Moreover, no detectable mortality to emamectin benzoate was observed in JB_KJ strain, suggesting uneven distribution of emamectin benzoate resistant population of T. palmi. To provide more precise information on resistance profiles and distribution in T. palmi populations, it would be necessary to conduct a large scale resistance mapping for broad geographical regions.

Frankliniella occidentalis is a notorious polyphagous crop pest causing tremendous economic loss. It damages flowers and leaves of host plants and also carries severe plant viruses. During last few decades, it has spread to all continents via transport of plant materials. Following extensive use of insecticides to control F. occidentalis, it has developed high level of resistance due to its short life cycle and high reproductive potential. In this study, RNA interference (RNAi)-based bioassay system was developed to find an alternative control measure for insecticide-resistant population of F. occidentalis. A variety of genes involved in various physiological mechanisms were selected for the test of dsRNA potency (tubulin, v-ATPase, amylase, aquaporin etc.). Each bioassay unit made by 3D printing has a leaf disc placed on 150 ㎕ of 50 ng/ul dsRNA solution and 20 thrips. The mortality was checked, and the dsRNA and leaf disc were replaced every 24 h for 72 h. Of the 20 genes tesetd, tubulin, v-ATPase, and aquaporin showed 31, 38, 38 and 45% of corrected mortality at 72 h post-treatment, respectively. This result suggests the potential of these genes as candidate lethal genes for RNAi-based F. occidentalis control system.

Small hive beetle (Aethina tumida) (SHB) is an invasive species to most northern hemisphere countries, including Korea. In an attempt to obtain basic information for efficient management of SHB, genes encoding conventional insecticide targets [voltage-sensitive sodium channel α-subunit (VSSC) and acetylcholinesterase (AChE)] were annotated and characterized following the analysis of whole transcriptomes of adults and larvae. A single VSSC gene was identified but no apparent mutations associated with pyrethroid resistance were detected. Genes encoding two AChEs (AtAChE1 and AtAChE2) were identified from the SHB transcriptome. AtAChE1 was determined to be the main catalytic enzyme, thereby being a toxicologically more relevant target. No apparent mutations associated with resistance to organophosphorus and carbamate insecticides was identified in the AtAChE1 gene, whereas the S238G mutation, originally identified from the Colorado potato beetle, was detected in the AtAChE2 gene.

To identify genes that commonly respond to the treatment of different insecticides and are responsible for the toleranceenhancement, transcriptomic profiles of larvae treated with sublethal doses of the five insecticides were compared withthat of untreated control. A total of 117,181 transcripts with a mean length of 662 bp were generated by de novo assembly,of which 35,329 transcripts were annotated. Among them, 125, 143, 182, 215 and 149 transcripts were determined tobe up-regulated whereas 67, 45, 60, 60 and 38 genes were down-regulated following treatments with these five insecticides.The most notable examples of commonly responding over-transcribed genes were two cytochrome P450 genes and ninecuticular protein genes. In contrast, several genes composing the mitochondrial energy generation system were significantlydown-regulated in all treated larvae. Considering the distinct structure and mode of action of the five insecticides tested,the differentially expressed genes identified in this study appear to be involved in general chemical defense at the initialstage of intoxication. Their possible roles in the tolerance/resistance development were discussed.

Honey bee swarming is a natural phenomenon that occurs by changes of colony (i.e. population size and queen condition) and environment conditions. As cuticular hydrocarbons (CHCs) are known to be involved in the communication between honey bee nest-mates, we investigated and compared the CHC profiles of worker bees from individual colonies of 9-days before swarming (PPSC), a day before swarming (PSC), swarming (SG) and remaining (non-swarming) (RG). A total of 53 CHCs were identified by GC-MS, among which 11 compounds showed significantly differential expression patterns between swarming states. Before swarming (between PPSC and PSC), detection levels of 4 CHCs were significantly different, suggesting that production of some CHCs changed prior to swarming for swarming preparation. Six CHCs were deferentially produced between PSC and RG. The differential profiles of CHCs with respect to different swarming states are currently under investigation.