In this study, we developed 11 microsatellite markers specific to A. crataegi using NGS to investigate the genetic relationships of A. crataegi populations from South Korea to circumferential Asian countries (China, Russia, Mongolia, and Japan). Further, two mitochondrial DNA (mtDNA) gene segments (COI and CytB) were sequenced from the samples. The population- and individual-based Principal Coordinates and STRUCTURE analyses collectively suggested that the South Korean population of A. crataegi is most differentiated from the Japanese population, whereas it was closer to Mongolian and Chinese populations. These results collectively suggest that northern populations, in particular, Mongolian populations can be considered as the most genetically compatible one as donee population, when the reintroduction program is launched. †These authors contributed equally to this paper.

Cymbidium flower is mainly grown for exportation to China and Japan, but detection of a few pests including the cotton aphid, Aphis gossypii (Hemiptera: Aphidae), necessitated post-harvest treatment for casual exportation. Thus, we irradiated electron beam to cotton aphids occurring in cymbidium to establish post-harvest method for casual exportation of the flowers. For cymbidium, six categories of product quality were examined after eight different doses were irradiated (100, 200, 300, 400, 500, 600, 800 and 1,000 Gy). One thousand Gy to cymbidium caused an extreme deterioration only in vase life in both colors compared to control (0 Gy). In the case of cotton aphids, adult longevity decreased from 11.23 (100 Gy) to 4.70 (400 Gy) when four different doses were irradiated (100, 200, 300, and 400 Gy), with control being an average of 20.89 days. The numbers of total first instar nymph produced per adult was not significantly differed among five doses (2.21 ~ 2.74 individuals), but was substantially lower compared to control (an average of 51.46; P < 0.0001). Live F1 nymphs did not develope to adults at all five doses, except for a single nymph at 100 Gy, which was dead right after emergence. The results of probit analysis indicated that majority of adults required 3.33 ~ 7.55 days for 90% mortality at 200 Gy and higher, but at 100 Gy it required 41.56 days. Therefore, higher than 100 Gy might be required for complete control of adult cotton aphids and their F1.

In order to accumulate the ecological information of Drosophila suzukii for export negotiations of domestic strawberry, we weekly monitored occurrence of D. suzukii at three strawberry fields located in Gok-seong, Dam-yang and Hwa-sun using the best combination of trap and attractant from November 2017 to May 2018, the period of strawberry production. Monitoring was carried out at three sites (a hill located 150 meter distant to greenhouse, outside greenhouse and inside greenhouse) in Gok-seong. Drosophil suzukii was trapped at three short periods from November to the beginning of December, from the end of December to the beginning of January, and from April in a hill. At outside greenhouse, D. suzukii was trapped at two short periods from November to the early December and the beginning of May. In inside greenhouse, it occurred once after May 24th. Gok-seong data can be summarized as more occurrence for longer period as the distance to greenhouse increases. Dam-yang occurred once in a short period inside greenhouse and hill, respectively on May. In case of Hwa-sun, D. suzukii did not trapped in whole trapping sites during trapping period, except for the surrounding grape and peach cultivation areas at the end of November. Summarized, no occurrence periods are from November 20th, 2017 to May 14th, 2018 inside greenhouse; from December 11th, 2017 to May 3rd, 2018 outside greenhouse; and from January 8th, 2018 to April 12th, 2018 on a hill. In connection with strawberry culture, the D. suzukii tends to occur at the beginning and the end of the strawberry production period. We expect this result can serve as a helpful data for export negotiation for quarantine.

The black-veined white, Aporia crataegi (Lepidoptera: Pieridae), which is distributed mainly in Eastern Asia is presumed to be extinct in South Korea, only with some numbers of dried specimens left, whereas the species is found casually in circumferential countries. One of the common conservation practices for such species is to launch introduction program, but prior population genetic analysis between donor and donee populations might be essential for long-term conservation. In this study, we developed 11 microsatellite markers specific to A. crataegi using Illumina paired-end sequencing to investigate the genetic relationships of A. crataegi populations from South Korea and circumferential Asian countries (China, Russia, Mongolia, and Japan). Further, two mitochondrial DNA (mtDNA) gene segments (COI and CytB) were sequenced from the samples. The population- and individual-based Principal Coordinates and STRUCTURE analyses collectively suggested that the South Korean population of A. crataegi is most differentiated from the Japanese population, whereas it was closer to Mongolian and Chinese populations. The STRUCTURE analysis based on two concatenated mtDNA gene sequences also supported different genetic composition of Japanese population from the remaining populations including that of South Korea and rather similar genetic composition between the populations of South Korea and Mongolia. These results collectively suggest that northern populations, in particular, Mongolian populations can be considered as the most genetically compatible one as doner population, when reintroduction program is launched.

동남아시아에 발생하던 등검은말벌은 최근 국내 뿐만 아니라 프랑스와 일본, 유럽 전역으로 확산되어 전 세계 양봉 산업에 극심한 피해를 주고 있다. 등검은말벌을 방제하기 위해 다양한 방제 방법들이 연구되고 있으며, 그 중 말벌 유인제에 대해 국제적으로 많은 연구가 진행되어 왔으나 이 역시 지속적인 개발이 필요한 실정이다. 따라서 본 연구에서는 시판 유인제와 최근 (주)다목에코텍에서 새롭게 개발한 신규 개발 유인제에 대한 효능 검정을 수행하였 고 양봉장에 출현하는 국내 발생 말벌속의 발생양상을 조사하였다. 담양 1개소와 곡성 2개소에 기존 유인제과 신규 유인제를 장착한 포획기를 각 3쌍씩 설치하여 2016년 9월부터 11월말까지 주 1~2회 관찰하였다. 그 결과 국내 발생 말벌속 중 등검은말벌(7,787 개체)이 가장 많이 포획되었고, 다음으로 장수말벌, 말벌, 좀말벌, 꼬마장수말벌, 털보말벌 순으로 포획되었다. 두 유인제의 포획력 검정 결과, 신규 개발 유인제가 통계적으로 유의하게(P < 0.01) 등검은말벌 포획력이 높음을 보였다.

The honey bee, Apis mellifera ligustica (Hymenoptera: Apidae), strain with a high hygienic behavior (HHB) has been bred for several years in Korea, and a diagnosis system to distinguish it from low hygienic behavior (LHB) strain has been necessitated. Thus, complete mitogenome of the two strains were sequenced through Next-Generation Sequencing technique to detect SNPs. Comparison of the mitogenome sequences from the two strains of A. m. ligustica have detected 23 SNPs in 11 PCGs and these were further confirmed the presence of SNPs using each 10 individuals selected randomly from each strain, indicating that these SNP markers might be useful to diagnose the honeybee strains with the HHB. Therefore, mitogenome sequences are promising genome source to detect SNP markers, particularly for inbred female iso-lines.

벗초파리는 우리나라에서는 심각한 해충으로 인식되지 않아 발생 기간 및 패턴, 기주 범위 등의 생태적인 연구뿐만 아니라 모니터링을 위한 우수 유인제 및 트랩의 연구도 제한적으로 수행된 실정이다. 본 연구는 기존 연구 결과를 참조하여 사과식초 함유 유인제 2종과 화학적 루어 2종 그리고 트랩 2종에 대한 선발 시험을 수행하였다. 선발 시험은 사육중인 벗초파리를 이용한 choice test와 함께 수목원, 딸기 농장, 블루베리 농장에서의 6반복 유인력 검증 실험을 수행하였다. 유인력 비교 결과 ACV + wine의 포획량이 높았으며, 블루베리 농장 인근 결과값은 전체 조사 항목에서 통계적 유의성을 보였다 (P < 0.05). 트랩 선발 실험 결과 모든 실험 장소에서 Dreves 트랩이 높은 포획량을 보였다. 이상의 결과를 통해 ACV + wine과 Dreves 트랩 조합을 선발하였으며, 이를 이용하여 향후 지속적인 모니터링, 월동 발생 조사 등의 연구에 활용할 수 있을 것으로 생각된다.

In this study, we developed 12 microsatellite markers specific to N. pygmaea using Illumina paired-end sequencing. Forty individuals of N. pygmaea collected from three currently known localities in South Korea were genotyped to validate these markers and to preliminarily assess population genetic characteristics. No locus showed significant deviation from the Hardy–Weinberg equilibrium (HWE). Our preliminary data indicate an absence of inbreeding in all populations and an absence of obvious genetic difference. The microsatellite markers developed in this study will be useful for studying the population genetics of N. pygmaea collected from other regions, including additional sites in South Korea.

The Acoptolabrus changeonleei Ishikawa et Kim, 1983 (Coleoptera: Carabidae), has been listed as an endangered insect in South Korea. The complete mitochondrial genome of the species was 16,831 bp with a typical set of genes (13 protein-coding genes [PCGs], 2 rRNA genes, and 22 tRNA genes) and one non-coding region, with the arrangement identical to that observed in most insect genomes. Phylogenetic analyses with concatenated sequences of the 13 PCGs and 2 rRNA genes, using the Bayesian inference (BI) and maximum-likelihood (ML) methods, placed A. changeonleei as a sister to the within-subfamilial species Damaster mirabilissimus in Carabinae, with the highest nodal support by both analyses.

Previously, a partial mitochondrial COI and CytB gene sequences have been used to infer genetic diversity and gene flow of the species. In this study, we additionally collected C. tripartitus from one previous and two new localities, sequenced the COI and CytB genes. Sequence divergence of current samples showed slightly lower values, but nearly equivalent to previous study. Haplotype diversity was still high and nucleotide diversity indicating that Seogwi-dong showed the highest estimates in both gene sequences. Gene flow among populations is high, but a significance difference was detected between Gulupdo and Anmado; and between Gulupdo and Seogwi-dong only in COI sequences (P < 0.05). Considering the high genetic diversity and gene flow in C. tripartitus populations, one of the major issues regarding conservation seems not to be the recovery of genetic diversity as has been suggested before.

The tiny dragonfly, Nannophya pygmaea (Odonata: Libellulidae) is one the smallest dragonflies in the world and listed as a second-degree endangered wild animal in Korea. We developed microsatellite markers and applied selected markers to South Korean populations to understand population genetic characteristics, along with two mitochondrial DNA (mtDNA) gene sequences (COI and ND5). Two mtDNA-based population genetic analysis indicates substantially reduced genetic diversity in an island population (Muuido) compared to others. On the other hand, population-based FST and RST consistently support that N. pygmaea populations are overall well interconnected with a relatively high gene flow. These results may collectively indicate that N. pygmaea populations in South Korea may have rather larger population size than we previously acknowledged based on a single-locus mtDNA sequence and field observation.

Despite of a substantial effort, the phylogeny of Lepidoptera still has unstable taxa and unresolved relationships particularly form Apoditrysia to Ditrysia (e.g. phylogenetic position and familial relationships of Gelechioidea, the monophylies of Tineoidea and Zygaenoidea and so on). We performed phylogenetic analysis using ~13,000-bp length of mitogenome sequences form 50 species (with seven species in two superfamilies form this study) representing 13 superfamilies to improve the lepidopteran phylogeny from Apoditrysia to Ditrysia. A close relationship of Gelechioidea to Obtectomera than to apoditrysian taxa was noteworthy, although the nodal support is very weak (ML, 23%; BI, BPP = 0.94). Monophyly of Zygaenoidea without Epipyropidae (Epipomponia nawai) was also noteworthy in spite of weak nodal support (ML, 64%; BI, 1.0). Scrutinized analysis is currently underway.

We sequenced complete mitochondrial genomes (mt genomes) of four Apis species, such as A. cerana, A. dorsata, A. laboriosa, and A. mellifera ligustica. The A. m. ligustica used in this study was originated from United States and selected for a high hygienic behavior during several years of breeding program. Considering current phylogenetic hypothesis of Apis the positive relationships between phylogeny and gene rearrangement was not supported. Phylogenetic reconstruction using available Apoidea species has shown the relationships ((Apidae + Colletidae) + Crabronidae). Within Apis, a strong support for three groups was obtained: A. florea and A. andreniformis as a basal group to the other A. cerana and A. mellifera group and A. dorsata and A. laboriosa group. The test to find out suitable single mt gene for the phylogeny at the genus level and below-genus level of Apis provided an identical topology from ND4L and ND6 in the ML analysis and ND3 and NF4L in the BI analysis to that from whole mt genome, with relatively strong support, indicating that these individual genes can potentially be utilized for within-species level phylogeny of Apis.

We newly sequenced mitogenomes of five skippers belonging to Lepidoptera to obtain further insight into characteristics of butterfly mitogenomes and performed phylogenetic reconstruction using all available gene sequences (PCGs, rRNAs, and tRNAs) from 85 species in 19 families in eight superfamilies. The general genomic features found in the butterflies also were found in the five skippers: a high A/T composition (79.3% - 80.9%), dominant usage of TAA stop codon, similar skewness pattern in various levels, consistently long intergenic spacer sequence between tRNAGln - ND2 (64-87 bp), the ATACTAA motif betweent RNASer(UCN) and ND1, and characteristic features of the A+T-rich region (the motif ATAGA, varying length of poly-T stretch, and poly-A stretch). The start codon for COI was CGA in four skippers as typical, but Lobocla bifasciatus evidently possessed canonical ATG as start codon. Phylogenetic analyses mainly yielded the consensus superfamilial relationships ((((((Bombycoidea + Noctuoidea + Geometroidea) + Pyraloidea) + Papilionoidea) + Tortricoidea) + Yponomeutoidea) + Hepialoidea) with a high support for most nodes, confirming the validity of Macroheterocera and its sister relationship to Pyraloidea. Within Rhopalocera the familial relationships (Papilionidae + (Hesperiidae + (Pieridae + ((Lycaenidae + Riodinidae) + Nymphalidae))) were strongly supported, confirming invalidity of the superfamily Hesperioidea. On the other hand, superfamilial relationships among Noctuoidea, Geometroidea, and Bombycoidea and the familial relationships among Saturniidae, Sphingidae, and Bombycidae were dubious, requiring further representative taxon sampling.

Up to now the complete mitochondrial genome (mitogenome) sequences of only three species of clitellate have been available. We have determined the complete mitogenome sequences of the elusive Burmese giant earthworm Tonoscolex birmanicus (Clitellata: Megascolecidae), which is endemic to Myanmar. The 15,170-bp long genome contains the 37 genes typical of metazoan mitogenomes [13 protein-coding genes (PCG), two rRNA genes and 22 tRNA genes] and one major non-coding region. All of the 37 genes are transcribed from the same DNA strand. The arrangement of the T. birmanicus mitogenome is identical to that of two within-ordinal species Lumbricus terrestris and Perionyx excavates. All 13 PCGs start with the ATG. For the stop codon, only six PCGs end with the TAA, whereas the remaining ones ends with the incomplete stop codon, T. Genes overlap in a total of 14 bp in five locations, and harbor a total of 16 bp of intergenic spacer sequences in nine locations.

We sequenced 17,329 bp of mitochondrial genome (mitogenome) of the black dwarf honey bee, Apis andreniformis (Hymenoptera: Apidae), that lacks ~200 bp of the A+T-rich region for the completion of the genomic sequence. The gene arrangement of A. andreniformis mitogenome is identical to that of A. cerana. However, the genome contains 5 additional tRNALeu(CUN) located 4 copies between tRNAMet and tRNAGln, and 1 copy between tRNAGln and tRNAAla, along with the typical sets of genes (13 protein-coding genes, 22 tRNAs, and 2 rRNAs) including regular tRNALeu(CUN) and the A+T-rich region (at least 923 bp). Only 1 copy of tRNALeu(CUN) differed by 1 bp from other 4 copies of tRNALeu(CUN). Each additional tRNALeu(CUN) is followed by nearly identical 68-bp long repeat sequence (95.6% identity). All 13 protein coding genes have typical start codons found in insect mitochondrial PCGs (2 ATA, 9 ATT, and 2 ATG).

Lepidoptera is one of the largest insect orders, but the phylogenetic relationships within this order, have yet to be completely described. One of the unresolved relationships includes the monophyly of Papilionoidea in relationship with the monotypic superfamily Hesperioidea. We newly sequenced five hesperid mitochondrial genomes (mitogenomes), representing four subfamilies: Pyrginae (Daimio tethys and Lobocla bifasciatus), Coeliadinae (Choaspes benjaminii), and Hesperiinae (Potanthus flavus), and Heteropterinae (Carterocephalus silvicola). Along with these newly sequenced hesperid genomes phylogenetic analysis was conducted with all available lepidopteran mitogenomes including three reported species of Hesperiidae that consisted of ~70 species in ten lepidopteran superfamilies. The test for the effect of optimization schemes, such as exclusion and inclusion of third codon position of 13 PCGs, other genes (22 tRNAs and two rRNAs), and with and without partitions also was performed. Majority of datasets consistently placed the monophyletic Hesperiidae the sister to ((Pieridae + Lycaenidae) + Nymphalidae), placing another true butterfly family Papilionidae as the basal lineage of this group, presenting the relationships (Papilionidae + (Hesperiidae + ((Pieridae + Lycaenidae) + Nymphalidae))). Consistent to previous result, Pyraloidea was placed as the sister to ((Bombycoidea + Geometroidea) + Noctuoidea), placing the Macrolepidoptera as non-monophyletic group.

In the present study, the 17,694-bp long complete mitochondrial genome (mitogenome) of the dwarf honey bee, Apis florea (Hymenoptera: Apidae), is described with an emphasis on the noteworthy triplicated tRNAser(AGN) region and an extraordinary long A+T-rich region with repeat regions. The gene arrangement of A. florea mitogenome is identical to that of A. mellifera, but has triplicated tRNASer(AGN), each of which contains the precedent 44 bp-long and following another 64 bp-long repeats plus one complete first repeat abutting to tRNAMet. A total of 1,610-bp long two repeat regions in 1,987 bp-long A+T-rich region is composed of nearly identical 141 ~ 219-bp long five tandem repeats and 50 ~ 52-bp long 12 tandem repeats that are encompassed by three non-repeat sequences. One of the common interpretations for such repeat sequence is slipped-strand mispairing and unequal crossing-over events during DNA replication.

We previously reported Pear Pest Forecasting Management System (PPFMS) for the Improvement of pass ratio of Korean exporting pears. It is consisted of regular field forecasting by pear farmers, meteorological data obtained by automatic weather station (AWS), an internet web page (http://pearpest.jnu.ac.kr/) as information collecting and providing ground, and information providing service. Currently, we are expanding this system to the area, Cheonan and Ansung, where pear orchards are organized into exportation-specific group. Further, the information obtained from field forecasting and AWS were up-loaded to under-constructing upgraded webpage (http://www.kpear.kr), with several pest/disease-related information. We hope this pest forecasting management system increases the pass ratio of Korean exporting pears throughout establishment of farmer-oriented forecasting, inspiring farmers’ effort for the prevention and forecasting of diseases and pests occurring at pear orchards.

Gene arrangement in the mitochondrial genome (mitogenome) has been regarded as an important evolutionary event that is useful as a phylogenetic signal. The mountainous duskywing, Erynnis montanus, belongs to a lepidopteran family Hesperiidae. We sequenced 15,530-bp long complete mitogenome of the species. The genome has the typical gene content of animals (13 protein-coding genes, two rRNA genes, 22 tRNA genes, and one major non-coding A+T-rich region). Further, E. montanus mitogenome also contained a high A/T content in the whole genome (81.7%) and the CGA (arginine) as the start codon for the COI gene, as typical in lepidopteran mitogenome. However, unlike other lepidopteran species, including two sequenced skippers, the E. montanus mitogenome has a unique arrangement tRNASer-tRNAAsn, instead of the tRNAAsn-tRNASer found unanimously in other lepidopteran species, providing a new gene arrangement in Lepidoptera. Such rearrangement probably was likely caused by duplication of gene block tRNASer-tRNAAsn and subsequent random loss of tRNAAsn in the first copy and tRNASer in the second copy, resulting in the arrangement tRNASer-tRNAAsn. Considering current phylogenetic relationships among available lepidopteran groups in connection with lepidopteran gene arrangement the new gene arrangement found in E. montanus seems to be apomorphy, requiring cautious interpretation as a phylogenetic signal.