먹노린재의 방제시기 설정을 위하여 서식지 내 발생 양상을 조사하였다. 전라남도를 중심으로 돌발 대량 발생 한 먹노린재의 발생 조사는 2023년도 벼의 모내기가 완료된 시점부터 전남 곡성군 석곡면과 여수시 화양면 일대 의 친환경단지에서 주 1회 실시하였으며, 벼의 수확이 완료되는 시점까지 진행될 예정이다. 금년도 발생 조사 결과는 지난 2021년도 결과와 비교하여 분석하고자 하였다. 현제까지의 결과, 여수시의 친환경단지 내 먹노린재 의 발생은 모내기가 완료된 이 후 2주 경과 시점에서 발견되었으며, 곡성군은 4주 경과 시점에서 발견되었고, 발생 최고점은 여수시의 경우 모내기 후 6주 경과 시점으로, 곡성군의 경우 5주 경과 시점으로 나타났다. 먹노린재 의 대발생이 지속되던 2021년도의 결과와 비교하였을 때, 2023년도 결과도 유사한 것으로 나타났다. 이에 따라, 먹노린재 친환경제재 살포 시기는 모내기 후 2주 경과 시점이 적절한 것으로 나타났으며, 2차 방제 시점에 대해서 는 추가 조사를 통해서 제안하고자 한다.

2018년 7월 13일부터 2019년 10월 29일까지 1년 3개월 동안, 총 306개 국외 발 국내 입항 선박을 대상으로 한국 미분포 편승자 해충 (not-distributed hitchhiker insect pests in Korea)에 대한 모니터링을 실시하였다. 그 결과, 총 805개체의 편승자 해충을 확보하였으며, 이를 통합 분류학적 종동정 방법(integrative identification method)을 이용하여 총 12목 78과 379종으로 동정하였다. 이 중 7목 21과 42종 67개체 의 한국 미분포종이 확인되었는데, 10종이 다중검출된 것으로 나타났으며, 그 중 7종은 2018년도에 이어 2019년도에도 검출된 것으로 확인되었다. 또한, 농림축산검역본부에 관리해충으로 등재되어 있는 Erthesina fullo (Pentatomidae, Hemiptera)와 Tessaratoma papilosa (Tessaratomidae, Hemiptera)가 검출되었다. 이에 따라, 검출된 미분포종의 사전 조사 및 모니터링 방안 마련뿐만 아니라 침입종(invasive species) 내지 침입 가능종(invasive likelihood species)에 대한 위험성 평가를 위한 data sheet를 제공하였다.

2018년 6월 1일부터 9월 17일까지 109일 동안, 총 112개 국외 발 국내 입항 선박을 대상으로 편승자 해충(hitchhiker insect pests)에 대한 모니터링을 실시하였다. 그 결과, 총 336개체의 편승자 해충을 확보하였으며, 이를 통합 분류학적 종동정 방법(integrative identification method)을 이용하여 9목 47과 159종으로 동정하였다. 이 중 총 3목 9과 14종의 한국 미분포종(not-distributed species)이 확인되어 보고하며, 검출된 미분포종에 대해서 침입종(invasive species) 내지 침입 가능종(invasive likelihood species)에 대한 위험성 평가를 위한 data sheet를 제공하였다. 또한, 미기록(unrecorded species), 미보고종(unreported species) 내지 신기록종(new recorded species)과 용어의 혼동을 피하기 위하여 본 연구에서는 미분포종(not-distributed species)을 제안하여 사용하였다.

We conducted quarantine insect species diversity monitoring using DNA barcoding with 517 lepidopteran samples that were obtained from quarantine inspections of foreign vessels entering Korea. The DNA barcode of each sample was treated as a molecular operational taxonomic unit (MOTU). For species delimitation and species identification of the analyzed samples, we applied a 2% cutoff rule and then identified with using BLAST of NCBI GenBank and BOLD System ver. 3.0. Consequently, 517 analyzed samples were delimited as 214 putative species across 20 families. Of these 214 putative species 145 (368 samples) were considered taxonomically identified if the closest BLAST match was no more than 2% different. Therefore the number of samples that were identified to the species level was relatively low, at approximately 71%. 115 of the 145 species were known in Korea. Of the 30 species that were not known in Korea, three, i.e., Noctua pronuba (Noctuidae), Orthosia hibisci (Noctuidae), and Pieris brassicae (Pieridae), were checked as ‘Regulated pests’ in Korea. We suggest that the three regulated pest species could be prevented from being introduced to Korea if monitoring of the vessels that pass the navigation route that contains these three species is performed consistently. Therefore, we suggest that the monitoring of quarantined insect pest enables the prevention of the introduction of alien species.

Tetranychus urticae and Myzus persicae are one of the most serious insect pests in many crops, vegetables, flowers, and fruit trees worldwide. Many insecticides have been developed to control green peach aphid and two spotted spider mite, but resistance to almost all insecticides has reduced their control effect. Particular groups of plant-beneficial microbials are not only root colonizers that provide plant disease suppression, but in addition are able to infect and kill insect larvae. Antimicrobial compounds produced by biocontrol microbes are effective weapons against a vast diversity of organisms such as fungi, nematodes, and viruses. In this study, we evaluated the effectiveness of mixtures plant extracts and improvement of culture process biocontrol microbials on insecticidal activity. Azadirachta indica and Derris elliptica mixed with micorbials, which are nutrient sources of mung bean extract and lecithin, were more effective than other the mixtures. Leaf spraying with the mixture of Pseudomonas fluorescens significantly showed the highest insecticidal power in vivo for 24 hours after treatment. The effect of spraying mixture was more than 50% at 2000 times dilution, and the spraying concentration of 90% or more showed a dilution of up to 500 times. Our results indicated that the nutrient sources of microbe act as a key antimicrobial metabolite in biocontrol of insect pests, and mixing with plant extracts can provide synergistic effects as an optimal usage of the biocontrol agents.

In Korea, species of the genus Ptecticus Loew, 1855 (Family Stratiomyidae) have been known as three species, P. aurifer (Walker, 1854), P. matsumurae Lindner, 1936 and P. japonicus (Thunberg, 1789) (= P. tenebrifer (Walker, 1849)). Additionally, an unrecorded species, P. sinchangensis Ôuchi, 1938, is founded in South Korea. We therefore report the species for the first time in Korea with morphological diagnoses and key for the identification of species of the genus. In total, the members of the genus Ptecticus are officially recognized as four species in Korea.

A male specimen of Lymantria albescens (called as Okinawa gypsy moth) was captured in Busan, by sex-pheromone trap for Asian Gypsy Moth (AGM) (7R,8S)-cis-7,8-epoxy-2- methyloctadecane [(+)-disparlure]. Up to now, this species is distributed only in Ryukyu Islands of Japan including Ishigaki and Okinawa. The male of Okinawa gypsy moth might be attracted to AGM pheromone trap. If L. albescens is occurred in Korea, more many male individuals must be captured in pheromone trap. Therefore, we considered that the individual might be imported from Japan by inanimate pathway. Although it is high probability that L. albescens might be imported from Okinawa, it is important to a survey on an invasive pathway of the species in a view point of quarantine inspection. Through this presentation, we provided a detection method on Lymantria species using DNA barcoding. On the basis of this study, we will conduct on an invasive pathway and inhabitation possibility.

We carried out DNA barcoding of five Korean Lymantria species to establish identification references library for quarantine inspection. Total of 118 samples including 34 samples obtained through quarantine inspection, two from USDA, and one collected from Philiphine were used for this study. And 30 sequences of 10 species from GenBank of NCBI were used as reference sequences. In a result of DNA barcoding of the Korean Lymantria species, sequence divergence of 148 DNA barcodes ranged from null to 17.0%, intraspecific divergence from null to 1.0%, and interspecific divergence from 5.1 to 17.0%. In NJ tree, L. dispar contained three clusters, which were identified as L. dispar asiatica, L. albescens, and L. xylina, respectively. L. xylina was collected through quarantine inspection on a foreign merchant ship in Yeosu port, and L. albescens was obtained by pheromone trap on L. dispar installed in Busan port. And L. monacha known as single species in Korea was revealed as species complex with three species, L. monacha, L. minomonis, and L. sugii. In subspecies level, L. dispar dispar (EGM) built single cluster, but L. d. asiatica (AGM) and L. d. japonica showed as multiple cluster. Therefore, DNA barcoding lead to rapid and accurate identification in species level, but in subspecies level, only a taxon showing geographically far distance was discriminated from the others. And the results could provide a taxonomic outline of the Korean Lymantria fauna and might be used as identification reference for Lymantria species in quarantine inspection.

A taxonomic review on the Korean Lymantria Hübner, 1819 was conducted. In a result, a total of nine species under four subgenera including two new recorded species were detected as followings: L. dispar asiatica Vnukovskij, 1926, L. xylina Swinhoe, 1903, L. monacha (Linnaeus, 1758), L. minomonis Matsumura, 1933, L. sugii Kishida, 1986, L. lucescens (Butler, 1881), L. mathura Moore, 1865, L. fumida Butler, 1877, and L. bantaizana Matsumura, 1933. Of the two unrecorded species, L. minomonis was found only in Is. Bogildo of Jeollanam-do, the southern part of Korea, and the other one, L. sugii was collected in the middle part of Korea. On the two species, L. xylina and L. fumida, the Korean specimens could not be examined through this study. Therefore, we considered that the two species might be excluded from the Korean Lymantria fauna. Each species was identified on the basis of wing pattern and genitalia of male/female adult. We provided diagnosis, male/female adult habitus photos, male genitalia photos, and female ovipositor photos.

Allomyrina dichotoma is a typical pet beetles in Korea. From 2010, similar symptoms of milky spore disease were found collectively in grubs of the species reared in insect farms. They shared a specific symptom that the skin of last instar larvae was changed softer with opaque white and infested grubs eventually died. To clarify the cause of the symptom, we collected the larvae of A. dichotoma from five farms and examined intestinal bacterial florae of them using pyrosequencing technique. From those results, a member of Paenibacillus was found only in the larvae showing the symptom of disease. Through PCR analysis using a Paenibacillus specific primer set, we obtained the partial 16S rRNA gene sequence and confirmed the microbe as Paenibacillus sp. For detailed characterizing, a whole guts was extracted from each larva showing the sign of the disease and incubated at 70℃ for 15 min to isolate spore forming bacteria. After then, each content of guts was cultured on MYPGPNAL agar medium (12.5 μg/ml of nalidixic acid) at 30℃. The 16S rRNA gene sequence analysis for six isolate showed that they were closely related to P. rigui (97.5~98.0% similarity) and to P. chinjuensis (95.9~96.6% similarity). Additional tests including API test and cellular fatty acid composition analysis were performed, but the strain couldn’t be identified at species level, suggesting it may represent novel species of the genus Paenibacillus.

DNA barcoding is a strong species identification tool for all animal taxa, and can easily be conducted when materials are under DNA friendly conditions. In contract, a full-length (659 bp) sequencing has been limited for the degraded DNAs extracted from old museum specimens. The initial challenges to retrieve the authentic DNA fragments from old museum specimens were attempted by obtaining short sequences (<300 bp) with the cloning process after PCR, making it both expensive and time-consuming. In this study, we employed a modified method to analyze the full-length DNA barcoding regions in 31~52 year-old butterfly specimens (301 dried specimens of 39 species) using direct sequencing after PCR with two different methods: 1) the successful PCR rates of 0 to 5.6% using four universal primer sets were too low to obtain authentic sequences and the cross-contamination was detected in almost all successful amplicons; 2) the success rates of PCR using specie-specific overlapping primer sets were distinctly high, reaching up to 75% with 98% authentic and 2% non-specific sequences. Thus, the result showed the method that using species-specific primer set per species yields the most effective success rates of both PCR and sequencing from degraded DNA without incorrect sequences.

In DNA barcoding, the DNA degradation of old museum specimens has been limited full-length (658bp) sequencing. The challenges associated with the retrieval and authentication of degraded DNA extracts from fossil and old museum specimens were principally limited to analyze the relatively short sequences (<300 bp). Furthermore, almost protocols in other to analyzed the degraded DNA contained the cloning process after PCR causing the time-consuming and the rising costs. To overcome these problematic circumstances, we tried a modified method to analyze full-length of DNA barcoding region in 30~60 year-old butterfly specimens (225 samples in 28 species), using direct sequencing after PCR with species-specific overlapping primer sets per each species. As a result, all of 28 species have been successfully analyzed, although 178 samples (79%) are completely generated barcoding sequences ranged from 640 to 658 bp and 47 samples (21%) are partially sequenced ranged from 100 to 500 bp. Thus, the result showed that the direct PCR sequencing using the overlapping primer sets per species appears to have great potential efficiency for analysis of degraded DNA without incorrect sequences.

The leaf beetle, Chrysolina aurichalcea (Coleoptera: Chysomelidae), is a pest damaging plants of Compositae. In order to understand the genetic diversity and geographic variation of the species we sequenced a portion of mitochondrial COI gene (658 bp) and complete nuclear internal transcribed spacer 2 (ITS2) collected from seven Korean localities. A total of 18 haplotypes (BARCA01 ~ BARCA18), with the maximum sequence divergence of 3.04% (20 bp) were obtained from COI gene sequence, whereas 17 sequence types (ITS2CA01 ~ ITS2CA17), with the maximum sequence divergence of 2.013% (9 bp) were obtained from ITS2, indicating substantially larger sequence divergence in mitochondrial gene sequence. Phylogenetically, the mitochondrial DNA has shown several haplotypes formed independent groups with substantially high node support (≥ 90%), whereas no such grouping was evidenced for ITS2, indicating different behaviors of the two molecules. Such difference may reflect a diverse dynamics of the species such as biogeographic history, mating behaviors, and also possibly different mode of inheritance of the two molecules, but requires further scrutinized examination of the dataset. In terms of population genetic perspective, overall no population subdivision was detected from both molecules, except for locality 7 (Eocheong islet) from mitochondrial DNA. As more scrutinized analysis is performed, further fruitful inference on the geographic contour of the species might be available.

In traditional taxonomy on the family Cantharidae, color pattern of the body and the structure of the male genitalia have been often used as diagnostic characters in identification of the specific level. However, these characters caused the difficulty in identifying the female in case a species was described only by male specimens or has the several color types among individuals. In this study, we attempted to evaluate the species reality of Asiopodabrus fragiliformis which was often difficult to be identified due to individual variation in color pattern and lack of information of female, through searching for new morphological diagnostic characters as well as DNA barcoding analysis, including their closely relative species from Russia and Japan. The results showed that A. fragiliformis was represented as three clusters strongly supported by high value of boots trap (>99%) and over 3% branch length. The pairwise distances between species of Asiopodabrus were detected larger, ranged from 3.4–9.5%, than the intragroup distance ranged from 0–2.9% indicating presence of a barcoding gap. And then, the three clusters were respectively determined as A. fragiliformis, A. kurvatovi and a new species through the analysis of morphology and COI gene. Therefore, we suggest that the species delineation on polymorphic species and the female specimens of closely resembling species would be more exactly and effectively determined if DNA barcoding and the traditional taxonomy are used as complementary methods for identification.

The tribe Atimiini LeConte containing two genera (Atimia and Paratimia) and about 12 species is distributed in East Asia and North America. The systematic position of the Atimiini has long puzzled the Coleopterist due to the Lamiid-like aspect of the Atimia. But the Atimiini has regarded as a tribe of the subfamily Aseminae since Webb (1912) associated the Atimia with the Asemum on the basis of larval characters. The tribe is very specialized in food habits, the species of Atimia are all restricted to trees of the Cupressaceae and Taxodiaceae, including Cupressus, Libocedrus, Thuja, Chamaecyparis, Juniperus, Taxodium, Sequoia, and their relatives. Members of the Atimiini are characterized by the combination of following features: head transverse, front short, vertical, mouthparts nearly horizontal; antennae 11-segmented, shorter than the body in both sexes, eyes large, moderately granulated, deeply emarginate, embracing antennal insertion; labrum transverse, ciliated; palpi unequal in length, the maxillary longer; pronotum quadrate, transverse; anterior coxae rounded, cavities usually not angulated, completely closed behind; mesonotum with a large, divided, stridulatory area; scutellum subquadrate; intermediate coxal cavities closed; metasternum deeply emarginate posteriorly, metepisterna narrow, attenuated behind; legs short; femora feebly clavate; tibiae armed with short spurs; wings with a closed cell in the anal sector. Here, we report the tribe Atimiini for the first time in Korea based on provisional identified species, Atimia spec. okayamensis Hayashi. We also provide a diagnosis, habitus photo, and drawings of diagnostic characters.

A new species, Hatchiana n. sp., is confirmed by morphological and molecular data. The genus Hatchiana Fender, 1966 belonging to Podabrini LeConte, 1881 is distinguished by the shape of tarsal claws: all tarsal claws with blunt tooth in both sexes. The genus was recognized as a subgenus of Podabrus Westwood, 1838. Recently, however, Hatchiana was suggested as separated genus. The genus Hatchiana was known as 12 species in Palaeartic region including three species from Korea. In this study, we found Hatchiana n. sp. from several areas of Taean-gun, Chungcheongnam-do, Korea. This new species is different from closely relative species, Hatchiana jirisanensis (Kang & Kim, 2000) by following morphological characters: size of compound eye, length of antennae, shape of pronotum, shape of scutellum, and shape of aedeagus. Also, we compare the DNA barcoding region (the former region of CO1 gene) between these two species as molecular characters. In result, Hatchiana n. sp. is distinct from Hatchiana jirisanensis Kang & Kim by discrepancy of three percents in CO1 sequence. Therefore, the Korean Hatchiana is confirmed as four species, adding Hatchiana n. sp.