In this study, we developed ten microsatellite markers specific to L. angelina using the Illumina NextSeq 500 platform. Forty-three individuals of L. angelina collected from three localities in South Korea were genotyped to validate these markers and to preliminarily assess population genetic characteristics. The observed number of alleles, observed heterozygosity (HO), and expected heterozygosity (HE) at a locus ranged from 4–13, 0.211–0.950, and 0.659–0.871 in the population with the largest sample size (20 individuals), respectively, thereby validating the suitability of the markers for population analyses. Our preliminarily assessment of the population genetic characteristics indicates the presence of inbreeding in all populations, an isolation of the most geographically distant population (Seocheon), and lower HO than HE. The microsatellite markers developed in this study will be useful for studying the population genetics of L. angelina collected from additional sites in South Korea and from other regions. †These authors contributed equally to this paper.

We sequenced 15,803 bp of the leaf-rolling-weevil, Apoderus jekelii (Coleoptera: Attelabidae) mitochondrial genome (mitogenome) that lacked ~8,000 bp of the A+T-rich region for the completion of the genomic sequence. The A. jekelii mitogenome, which includes 1,169 bp of A+T-rich region, possesses typical sets of genes [13 protein-coding genes (PCGs), 22 tRNA genes, and 2 rRNA genes]. Phylogenetic analyses using the eight concatenated PCG sequences, which are commonly available for the mitogenome sequences of Curculionoidea, revealed Attelabidae as monophyletic, as well as the sister relationship between current A. jekelii and congeneric species A. coryli in Attelabidae, with the highest nodal supports both in Bayesian inference and maximum likelihood methods. In order to gain a more comprehensive picture of the phylogenetic relationships among the lineages of Attelabidae, an extended analysis with more taxonomic sampling will be necessary. †These authors contributed equally to this paper.

The complete mitochondrial genome of Ostrinia palustralis memnialis Walker, 1859 (Lepidoptera: Crambidae) was determined to be 15,246 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 an arrangement identical to that observed in most lepidopteran genomes. The A+T content of the whole genome, PCGs, srRNA, lrRNA, tRNAs, and the A + T-rich region all are well within the range found in other Pyraloidea. Phylogenetic analyses of the concatenated sequences of the 13 PCGs using Bayesian inference and Maximum-likelihood methods placed O. palustralis as a sister group to O. furnacalis and O. nubilalis, with the highest nodal support. The subfamilies within Crambidae, such as Nymphulinae, Spilomelinae, and Pyraustinae, all formed monophyletic groups with the highest nodal support.

In this study, we sequenced two complete mitogenomes, belonging to the families Scythrididae (Scythris sinensis Felder & Rogenhofer, 1775) and Coleophoridae (Coleophora therinella Tengström, 1848) firstly in each family. Gelechioidea is one of the controversial lineages of Lepidoptera in its phylogenetic position and number of families. Phylogenetic analyses with concatenated sequences of the 13 PCGs and two RNA genes using the maximum likelihood method, placed Coleophoridae, represented only by C. therinella, as a sister group to the families Depressariidae and Autostichidae, with very low nodal support (7%). Scythrididae represented only by S. sinensis was placed as the sister to the family Stathmopodidae, with relatively high nodal support (86%). As more mitogenome sequences from the extended taxonomic groups are obtained further robust phylogenetic inference will be possible.

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.

Drosophila suzukii Matsumura (Diptera: Drosophilidae) is known as a pest that has a wide host range. This study was carried out to investigate the occurrence of D. suzukii by trapping D. suzukii in Korean strawberry greenhouse and its surrounding woodland from November, 2017 to April, 2019. As a result of trap monitoring, the greatest capture was recorded in fall and secondary peak was confirmed in summer in a lower capture. It was confirmed that there was a statistically significant difference between the occurrence among inside and outside of the greenhouse and the woodland. The strawberry greenhouse cultivation area of Korea is mainly located in the plain region, whereas most of the other host plants for D. suzukii are usually grown at the ridge of the mountain or hillside. Therefore, if considering the living environment of host plant, monitoring result that showed more capture in woodland than strawberry greenhouse would be explained.

To control an external parasitic mite, a honey bee line possessing high hygienic behavior (HHB) against an external parasitic mite, Varroa destructor, has been bred in South Korea and an assessment method has been necessitated to diagnose HHB line from the low hygienic behavior (LHB) line. Thus, in this study, we developed single nucleotide polymorphism (SNP) markers from whole genome sequencing of each 20 worker bees from HHB and LHB lines of A. mellifera ligustica (Hymenoptera: Apidae). An average of 319,445,977 sequence reads was mapped to the known A. mellifera reference genome (an average of 87.46%). In 2,316,128 and 3,266,756 SNPs from each HHB and LHB line, an average of 93.6% and was located in the intergenic spacers and introns, whereas, the remaining 6.4% was located in the genic region, respectively. Among them 20 SNPs that were fixed at each line possessing within-individual homozygosity were selected and each four SNPs were used to diagnose the two honey bee lines either by typical PCR-restriction fragment length polymorphism method or allele-specific PCR. The remaining six SNPs had the size difference, enabling relatively easy differentiation between the two honey bee lines on typical agarose gel and another remaining six SNPs only has sequence difference including SNP sites. Thus, these SNP markers can be used to diagnose the honey bee line with HHB from LHB line against V. destructor.

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.

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.

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.

Bumblebee, Bombus ardens ardens (Apidae: Hymenopera), is an important resource for pollination that is most widely distributed in Korea. This study utilized microsatellite markers for investigation of genetic diversity and geographic relevance of the B. a. ardens populations in Korea. Through Next Generation Sequencing analysis, we identified 10 microsatellite markers and genotyped for 107 individuals of B. ardens collected from 10 populations. At each locus the number of alleles ranged from 10 to 23; the observed and expected heterozygosities ranged from 0.8909 to 1.0000 and 0.6641 to 0.8422, respectively; and inbreeding coefficient(FIS) ranged from –0.5053 to –0.0891. Significant deviation from the Hardy–Weinberg equilibrium was not observed at any locus. Population structure analysis indicated that there are three genetic groups in Korea with each Jeongseon and Ulleung-do composed of different gene pool from the remaining other populations. Similarly, Principal coordinates analysis also showed the same pattern. FST and RST analyses showed that each Jeongseon and Ulleung-do population had a significant genetic distance from other populations. Considering these results, genetic isolation of Ulleung-do may be explained by “Oceanic island” status and Jeongseon, which showed the positive FIS (0.069) and genetic isolation may be caused by its location on the east side of Baekdudaegan and by on-going inbreeding with a small population size.

The nerippe fritillary butterfly, Argynnis nerippe , is listed as an endangered species in Korea. Establishment of effective conservation strategies can be aided by the development and application of molecular markers that can be used to investigate the population genetics of the butterfly. Therefore, in this study, we identified ten microsatellite markers specific to A. nerippe using the Next-Seq 500 platform, and applied these markers to investigate the characteristics of five South Korean butterfly populations. Genotyping of 48 A. nerippe individuals from five localities showed that at each locus the number of alleles ranged from 4 to 14, and that the observed and expected heterozygosities were 0.324–0.863 and 0.138–0.985, respectively. Significant deviation from the Hardy–Weinberg equilibrium was not observed at any locus. Population structure analysis indicated that there are two genetic groups in Korea, but no population-based gene pool assignments were found. Analysis of FST, RST, and a principal coordinates analysis suggested that the Gureopdo and Yaecheon populations were isolated from other populations. Genetic isolation of the Gureopdo population may be a consequence of unequal population change between Gureopdo and inland populations and to the offshore habitat of Gureopdo. Genetic isolation of the Yaecheon population may be a consequence either of the southernmost location of the population or of the limited sample size available. Further studies with increased sample sizes will be necessary to draw robust conclusions on population isolation and to devise conservation strategies.

Bombyx mandarina (Lepidoptera: Bombycidae) is generally regarded as the ancestor of the domesticated B. mori. Recently, over 40 mitochondrial genomes (mitogenomes) mainly from B. mori strains preserved in China and wild individuals of B. mandarina were sequenced to verify the progenitor of B. mori. At this point, we also were curious about the origin and the relationships of Korean silkworms to foreign B. mandarina and B. mori. As a first step, we sequenced the complete mitogenome of the B. mandarina collected in Korea and compared it to pre-exsiting data (37 strains of B. mori and 14 individuals of B. mandarina). The complete mitogenome of B. mandarina was 15,694-bp long, consisted of 13 protein-coding genes, two rRNA genes, 22 tRNA genes and one non-coding region. The 494-bp long A+T-rich region possessed the highest A/T content (95.3%) than any other region of the genome. Overall, the general mitogenome characteristics of the genus Bombyx species have an identical gene arrangement, similar A/ T content (average 82.3%) and so on. Phylogenetic analysis, however, showed that B. mori and B. mandarina formed a distant group each with the highest nodal support. For more findings of mitogenome characteristics of Bombyx including the Korean B. mandarina and those preserved in Korea more mitogenomes, particularly from Korea, might be needed.

The yellow-legged hornet, Vespa velutina nigrithorax (Hymenoptera: Vespidae), invaded into South Korea in 2003. Currently, V. velutina is distributed throughout South Korean regions,. A genetic trait of invasive species provides important data for environmental risk assessment. In this study, we analysed the genetic variation of the V. velutina among the populations of South Korea and other foreign countries using partial sequences of mitochondrial COI, CytB and 16S rRNA. We further developed intergenic spacers (IGS2 and IGS3) to detect genetic variation among Korean populations. Each single haplotype was recovered from the domestic V. velutina from each coding gene sequence. Korean individuals shared the haplotype with that of Japan and China. Phylogenetic analysis (excluding 16S rRNA) using available sequences and ours showed the presence of two groups: the Indonesia and Malaysia group and another group (Korea, Japan, China, France, Vietnam and Thailand). The newly developed IGS2 and IGS3 markers resulted in each seven and four haplotypes, providing better resolution than coding gene sequences. Analysis of Molecular Variance using the combined sequences of IGS2 and IGS3 showed that the majority of variance was allocated to each population and only minority of variance is allocated to among-populations. This result may indicate that the V. v. nigrithorax invaded in Korea shows a single panmictic population and this possibly suggest that Korean population may have originated from small founder individuals.

The dung beetle, Copris tripartitus, has been listed as an endangered insect in Korea. In contrast to typical genomic characteristics, the 15,457-bp long, the complete mitochondrial genome (mitogenome) of C. tripartitus has a few unusual characteristics. The genome has the trnP-trnT arrangement at the ND4L-ND6 junction, which is found only in two species of Scarabaeoidea, including C. tripartitus, in 53 available species. Twelve protein coding genes (PCGs) have typical ATT and ATG start codons, whereas COII had the start codon GTA, which is unprecedented in Scarabaeidae. The 297-bp A + T-rich region, which is the shortest available in Scarabaeidae, harbors three repeat units (26, 29, and 21 bp), each consisting of duplicated identical sequences interspersed in the A + T-rich region. Phylogenetic analyses were performed using the concatenated sequences of the 13 PCGs and two rRNA genes using both the Bayesian inference (BI) and maximum likelihood (ML) methods for the families Scarabaeidae and Lucanidae that are available for their mitogenomes in Scarabaeoidea. Our consensus phylogeny yielded monophylies of each family and the subfamily Scarabaeinae in Scarabaeidae with the high nodal supports (0.99-1 by BI and 79-100 by ML). Among the three tribes, represented by multiple genera in Scarabaeinae each Oniticellini and Onitini was supported as monophyletic groups with the highest nodal supports (1 by BI and 100 by ML), whereas Onthophagini was non-monophyletic group, presenting the tribe relationships: ((((Oniticellini + Onthophagini [Caccobius, Milichus, Onthophagus]) + Onthophagini [Digitonthophagus, Phalops]) + Onitini) + (Phanaeini + Eurysternini) + Coprini).

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.

Currently, phylogenetic relationships of Bombycoidea have been reconstructed using molecular data, but generated several hypotheses on among-familial relationships [(especially, “Satruniidae+Bombycidae+Sphingidae” group)] within Bombycoidea. Thus, in this study, we sequenced 39 mitogenomes from both Bombycoidea and Lasiocampoidea. Consequently, one topology was obtained on the basis of ML and BI methods. It suggested the monophyly of the two superfamilies (Bombycoidea and Lasiocampoidea) consistently with a high nodal support; the sister relationship between Saturniidae and Sphingidae in “SBS”; and the sister relationships between Bombycidae and the Saturniidae + Sphingidae group.

The effects of cold storage temperature and exposure duration on immature stages of spotted-wing drosophila (SWD) Drosophila suzukii (Diptera: Drosophilidae) on ‘Campbell Early’ grapes were examined to establish a phytosanitary control method. The immature stages (eggs, larvae and pupae) of SWD were all dead after a 6-day cold treatment at 1°C and 8-day cold treatment at 1.5 and 2°C. Small-scale tests using pupae, which were the most-cold tolerant stage, confirmed the validity of the selected temperature and exposure durations. Conversely, the 8- and 10-day at 1°C treatments showed 100% mortality, suggesting that these treatments can guarantee quarantine security against infestations of SWD on exported ‘Campbell Early’ grapes.

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.