Chironomid communities are indicators of water pollution because of their ability to thrive under freshwater conditions. However, it is difficult to distinguish between chironomid larvae based on morphology. DNA barcoding, based on nucleotide sequences of marker genes, can be used to identify chironomid larvae. Samples of chironomid larvae were collected from Gwangju Stream and Pungyeongjeong Stream, tributaries of the Yeongsan River in South Korea. We identified 3 subfamilies, 13 genera, 16 species, and 1 cryptic species. There were 7 genera and 10 species from the subfamily Chironominae, 5 genera and 5 species from subfamily Orthocladiinae, 1 genus and 1 species from subfamily Tanipodinae, and the cryptic chironomid species of the family Chironomidae. There were 21 individuals from, 7 species and 1 cryptic species from the Gwangju Stream and 24 individuals, belonging to 10 species from the Pungyeongjeong Stream. The only species detected in both streams was Cricotopus bicinctus. The relationship between water quality and the species detected was difficult to explain, but the number of species showed a tendency to increase at sites where water quality was poor. Additional investigations and studies are needed to understand the relationship between water quality and the chironomid species occurring in these two streams.

해외 수입농산물의 양이 점차적으로 증가하여 다양한 병해충의 유입 가능성이 증가함에 따라, 이들을 신속하고 정확하게 판별하기 위하여 DNA 염기서열을 이용하여 형태학적 동정을 보완하고 있다. 현재 바구미과의 Sternochetus속 에 속하는 관리해충은 S. mangiferae와 S. frigidus, 그리고 S. olivieri가 있으며, 태국에서 수입된 망고에서 2018년 9월부터 2019년 1월까지 7차례에 걸쳐 바구미류가 검출되었고, 형태학적 형질 및 DNA 바코드를 이용하여 S. olivieri로 동정하였다. 추가로, Sternochetus속에 속하는 종들의 분류학적 형질을 이용하여 동정의 수단을 제공하였다. 본 연구결 과는 수입 및 반입되는 망고에서 검출되는 바구미류의 신속하고 정확한 진단에 활용될 수 있을 것으로 판단된다.

During the last couple of decades, molecular techniques are widespread tools for the species diversity. However, their application to taxonomy provoked intense debates between traditional and molecular taxonomists. To prevent every kind of disagreement, it should be required to a threshold for standarizing the species delimitation. Here, we tested three species delimitation methods (ABGD, PTP and bPTP) and compared their results with morphospecies on the widest DNA barcoding dataset. Moreover, a possible threshold for species determination within the superfamily has been suggested. It is believed that it could be an important criterion in detection for hidden or cryptic species, tracing of variation, and can help in identification on immature stages of pests.

Promalactis established by Meyrick in 1908, is one of the highest species richness genera of the family Oecophoridae. Over 240 described species are mainly distributed from Oriental and Palaearctic regions. They are usually a tiny-sized and exhibit subtle morphological differences which make problems on morphological-based identification. In this study, we performed DNA barcoding study for the Promalactis by using mitochondrial cytochrome c oxidase subunit I (COI). From analyzing 155 COI sequences, we observed the usefulness of the COI in species identification with intraspecific genetic variation (range 0.0-2.1%) and interspecific genetic divergence (range 2.8-15.4%).

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.

Pear psyllids belong to the most serious pests of pear. They damage pear trees by excessive removal of phloem sap, by soiling the fruits with honeydew which, in turn, provides a substrate for sooty mold, and by transmission of Candidatus Phytoplasma, the causal agents of the pear decline disease. The morphological similarity, the presence of seasonal dimorphism that affect adult colour, size and wing characters, and uncritical use of species names, led much confusion in the taxonomy of pear psyllid species. As a result, pear psyllids have been frequently misidentified. Here we analysed DNA barcodes of eleven pear psyllid species from eastern Asia, Europe and Iran using four mitochondrial gene fragments. The efficiency of identification was notably high and considerable barcoding gaps were observed in all markers. Our results confirm the synonymies of the seasonal forms. Previous misidentifications are also corrected. There is no evidence for the presence of European pear psyllid species in East Asia.

DNA bacoding is a popular DNA diagnostic technique especially for specific and generic level identification during various quarantine activities. BOLD Systems, an internet accessible DNA barcode data portal, currently includes over 12,000 DNA barcodes for the fruit fly family Tephritidae, of which 8,940 barcodes are open to public. These data represent 655 tephritid species, majority of which are pest species. These figures are rapidly increasing as the tephritid barcoding research is increasing every year. Therefore, BOLD has established as the most important DNA barcode source for tephritid identification. It is, therefore, also important to understand the limitations and problems of the DNA barcoding analysis. We here discussed the following potential problems and their possible solutions: (1) misidentification of species listed in BOLD; (2) sibling species with identical or near-identical barcodes; (3) NUMT (nuclear mitochondrial DNA) or pseudogene; and (4) introgression by hybridization between closely related species.

Genus Trissolcus are considered as most effective natural enemy for control Pentatomid pests. Trissolcus japonicus (Ashmead) is native egg parasitoid of the brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae) in Palearctic region. Nevertheless, identification of Palearctic Trissolcus species are still controversial because of morphological homogeneity such as Trissolcus semistriatus (Nees, 1834) and Trissolcus Kozlovi Ryakhovskii, 1975. This study reassessed Korean Trissolcus species diversity and provided DNA barcoding data using mitochondrial cytochrome c oxidase subunit I (COI) gene for 11 Korean Trissolcus species.

Gelechioidea is the superfamily that contains diverse groups comprising over 16,000 species in worldwide. It was estimated that only 25% of the species diversity of the Gelechioidea has been described. In this study, DNA barcoding was assessed based on cytochrome c oxidase subunit I (COI) for the 154 morphologically identified Korean gelechioids. The results of three molecular species delimitation methods were consistent for 72% with their morphological identifications. Based on molecular operational taxonomic unit (MOTU) estimation, we discussed on discordant groups including barcode sharing pairs and cryptic species.

The objectives of this study are to examine the genetic variation and the origin elucidation in oriental fruit flies (Diptera:Tephritidae) using DNA barcode, in prepartion for their future introductions into Korea. About 1,600 specimens of B.dorsalis sp. complex and B. correcta were collected from 10 countries, the Indochina peninsula, the Philippines, Taiwanand South China. A total of 182 cytochrome c oxidase I (COI) sequences were obtained and aligned from these regionalspecimens. Three chinese sequences from the GenBank reference were also included. Six hundreds base pair fragmentswere aligned and trimmed and used for a barcode. The phylogenetic tree was generated using the neighbor-joining methodwith 1,000 bootstrap replicates. There were two distinct groups in the phylogenetic tree, Bactrocera dorsalis sp. complexand B. correcta. Three specimen, intercepted in the hand-carried mango at the airport inspection and collected in the confiscatedmango, smuggled from Vietnam were included for a test. The DNA sequences from the airport were 100% identical toone of various Vietnam specimens, and that from the confiscated mango was mostly similar to those of Vietnam, suggestingthe usefulness of the barcode for elucidating the origin of oriental fruit fly.

The objectives of this study are to examine the genetic variation in oriental fruit flies (Diptera: Tephritidae) and to use it as a barcode for the origin elucidation, in preparation for the their incursions into Korea. About 1,600 specimens of B. dorsalis sp. complex and B. correcta were collected from 10 countries, the Indochina peninsula, the Philippines, Taiwan and South China. A total of 182 cytochrome c oxidase (COI) sequences were obtained and aligned from these regional specimens. Three sequences from the Genebank reference were also included. Six hundreds base pair fragments were aligned and trimmed and used for a barcode. The phylogenetic tree was generated using the neighbor-joining method with 1,000 bootstrap replicates. There were two distinct groups in the phylogenetic tree, Bactrocera dorsalis sp. complex and B. correcta. Three specimen, intercepted in the hand-carried mango at the airport inspection and collected in the confiscated mango, smuggled from Vietnam were included for a test. The DNA from the airport were 100% identical to that of one of various Vietnam specimens, and that from the confiscated mango was mostly similar to those of Vietnam, suggesting the usefulness of the barcode for the origin elucidation tool of oriental fruit fly.

Two cryptic species of the black-tailed bamboo aphid, Takecallis arundicolens (Clarke, 1903), are revealed based on DNA barcoding and nymph morphology. Three species, T. arundicolens, T. sp.1 nov. and T. sp.2 nov. have significant genetic differences ranged from 2-8% but, only subtly differing in their nymph morphology. In this study, molecular analyses and morphological comparisons are presented including species keys for alatoid nymphs.

Many species of braconids play an important role in controlling the populations of insects in agricultural and natural ecosystems, which parasitize pests. Nevertheless, the family Braconidae was scarcely research and DNA barcoding of Braconidae has not been studied in Korea. DNA barcoding can rapidly perform species identification, and classify difficult species in morphological identification. Because most braconids are very small and cryptic in morphology, DNA barcoding is useful in Braconidae taxonomy. In this study, genomic DNA of 280 individual samples were extracted, and, among them, 208 samples were obtained COI (cytochrome c oxidase I) barcode sequences. As a consequence of blast, 48 samples were identified in genus-level, and 51 sample were identified in species-level. Among them, 4 species were revealed as unrecorded in Korea.

Generally, in aphid groups, taxonomically deficient characters and high level of morphological plasticity induced by environmental factors make difficulties for species identification. To solve these problems, DNA barcoding has been widely used for rapid and reliable species identification in aphids. The subfamily Calaphidinae is the second largest group in the family Aphididae with about 400 species belong to 59 genera. But so far, no trial of DNA barcoding has been conducted for the subfamily Calaphidinae. In this study, a total of 446 Cytochrome oxidase I (COI) sequences of 76 morphospecies collected in Korea and other countries were analyzed to detect cryptic diversity. In addition, 551 sequences of 74 species from the Genbank and BOLD system were compared with our new dataset. The final dataset consisted of 998 sequences of 115 species. As a results, we propose 12 cryptic species with discussion on morphological and ecological comparisons. Our results suggest that DNA barcoding is effective for precise species identification in this group and contributes to reveal hidden diversity.

2011년 6월 28일에 파주시 조리읍 장곡리 인근 도로에서 두부쪽이 손상되고, 해당연도에 태어난 맹금류 어린새를 발견하였고, 이를 DNA 바코드 기법으로 동정하였다. Mitochondrial DNA(mtDNA) cytochrome c oxidase I(COI) 유전자의 695bp 절편을 중합효소연쇄반응(polymerase chain reaction, PCR)으로 증폭하고 염기서열을 결정하였다. 결정된 염기서열을 BOLD systems과 NCBI의 BLAST에서 유사도 분석을 수행한 결과 총 5개체의 왕새매가 검색되었고, 염기서열의 동일성은 100%로 조사되었다. 또한, DNA 분자성판별 결과는 해당 개체가 암컷임을 나타내었다. 이러한 결과는 경기도 파주에서 1968년 이후 43년만에 왕새매의 번식이 확인된 중요한 정보로, 향후 광역야생동물구조센터는 야생동물의 사체 수거 시 인근 기탁등록보존기관과 연계하여 DNA시료를 확보하고 보다 정확한 종동정과 성판별 정보를 기록하는 등의 체계적인 관리시스템이 필요할 것으로 사료된다. 또한 이렇게 확보한 왕새매의 DNA 시료와 DNA 바코드 COI 유전자 서열은 유사종 연구의 참조표본(reference standard)로 이용될 수 있을 것이다.

A partial sequence of the mitochondrial cytochrome oxidase subunit I (COI) gene is widely used as a molecular marker for species identification in animals, also termed a DNA barcode. However, the presence of more than one sequence type in a single individual, also known as heteroplasmy, is one of the shortcomings of barcode identification. In this study, we examined the extent and divergence of COI heteroplasmy, including nuclear-encoded mitochondrial pseudogenes (NUMTs), at the genomic-DNA level from 13 insect species, including four individuals of orthopteran Anapodisma miramae. Furthermore, a long fragment of mitochondrial DNA (~13.5 kb) and cDNA from A. miramae were used as a template for COI PCR to compare the patterns of heteroplasmy between DNA sources and to investigate a possible way to avoid ambiguity in DNA barcoding. When multiple numbers of clones originated from genomic DNA were sequenced, heteroplasmy was prevalent in all species (3~16 heteroplasmic copies), with a varying degree of maximum sequence divergence (<1% in 7 species, <4% in 3 species, <6% in 2 species and 2.15-8.03% in four A. miramae individuals). In five species, NUMTs also were observed when genomic DNA was used as a template. Long fragment DNA also is a source of heteroplasmic amplification, but the divergent haplotypes and NUMTs obtained in the genomic DNA-based PCR were not detected in A. miramae. On the other hand, cDNA was heteroplasmy-free, without NUMTs when multiple numbers of clones were sequenced. Consistently, one dominant haplotype was always obtained from the genomic DNA-origin clones in all species and also from the long fragment- and cDNA-origin clones of A. miramae. Furthermore, the dominant haplotype was identical in sequence, regardless of the DNA source. Thus, one possible solution to avoid the barcoding problem in relationship to heteroplasmy could be the acquisition of multiple numbers of barcoding sequences to determine a dominant haplotype that can be assigned as barcoding sequence for a given species.

This study was carried out to provide accurate identification of Noctuidae, using the DNA barcode. In total, 2,040 adult specimens of 443 species were collected from 13 forest areas in Korea. In this study, we conducted the correct identification using external morphology and genitalic characters. In addition, we observed and compared the voucher specimens, preserved in the major entomological collections in Korea, including Hannam university Natural History Museum (102 species), Korea Forest Research Institute (8 species) and the National Science Museum (27 species) for correct identification. For extracting DNA sequences of the mitochondrial gene COI, the hind legs were detached and sampled with tweezers, providing about 25 mg of tissue sample. We amplified and sequenced the standard DNA barcode fragment of 648 basepairs. As a result, we have complete DNA barcodes for all 331 species. These results will be presented in Forest Pests Information Data Sheets for Noctuidae, containing DNA Barcode information, external morphology, ecological characteristics and phenology.