Culex orientalis is belonging to the mimeticus group of the genus Culex and shows various patterns of white pale spots on the wing and wing venation which cause an ambiguity to identify as this species. To confirm whether or not these variations are limited within the species, we observed 230 Cx. orientalis specimens collected in Korea and divided them into 51 variations according to their wing spots patterns. To compare a molecular similarity between the variations, the ITS2 regions of five major variations were analyzed. The results showed that there are more than 97% nucleotide sequence similarity between the variations as well as within a variation. This results suggest that the wing variations of Cx. orientalis are limited a within-species divergence. To further confirm, ITS2 regions of other species (Cx. mimeticus and Cx. jacksoni) in the mimetiucs group will be analyzed and compared with those of Cx. orientalis variations.

Larval stages of Callipogon relictus (Semenov) (Coleoptera: Cerambycidae), a gigantic longhorn beetle designated as a natural monument of Korea, has never been studied as it is hardly discovered in nature. The DNA barcoding gene, mt-COI, was used to identify a dead larva found in the Gwangneung forest of the Korea National Arboretum. Based on the result, we provide the morphology of the immature stage, with the illustrations of diagnostic characteristics.

프로테오믹스 기법을 이용하여 벼 고온 스트레스 관련 단백질을 분리 동정하기 위하여 에서 고온처리한 벼의 줄기로부터 단백질을 분리하였다. 분리한 단백질로부터 Rubisco 단백질을 제거하기 위해 15% PEG fractionation을 실시한 후 상등액 분획의 단백질을 이차원전기 영동한 후, CBB 염색을 통해 차별적 발현을 보이는 단백질을 분석하였다. 총 46개의 단백질 spot이 발현양에 변화를 보였으며, 그 중 24개의 단백질이 고온 스트레스에 의해

Cecidomyiidae and Sciaridae (Diptera: Sciaroidea) are mostly mycophagous, feeding on decaying plant materials and fungi. On the shiitake bedlogs, great number of larvae of the species cause serious damage, feeding on the mycelium of the shiitake mushroom. We confirmed five species emerged from the shiitake bedlogs, which are two cecidomyiids and three sciarids. Using the DNA barcording, the mitochondrial cytochrome c oxidase subunit I (COI) region (658 bp), the larvae and adult flies were identified, and Camptomyia corticalis was confirmed as a major pest on shiitake mushroom.

Tetranychid mites are one of the most diverse group including at least 1,200 species in the world. Species identification is difficult due to the small size and similar morphology within the group. We collected 17 species of spider mites from various host plants in different regions of Korea and determined species identity by the comparison of morphological characters and nucleotide sequences of internal transcribed spacer 2 (ITS2) and the cytochrome oxidase subunit I (COI). In addition, we report three new species that were firstly identified in Korea. Amphitetranychus quercivorus (Ehara and Gotoh) was collected from Mongolian oak plant in Daegu, Schizotetranychus miscanthi Saito was from the common reed plant in Ulleungdo, and S. cercidiphylli Ehara was from Bamboo plant in Jeju. Morphological identification of three species were similar with those of Japanese samples, but the ITS2 and COI sequences of A. quercivorus and the COI sequence of S. miscanthi were different with Japanese species at the rates of 1/419, 2/331 and 3/332 nucleotides, repsectively. S. cercidiphylli can be identified by the aedeagus shape of males but we firstly sequenced ITS2 and COI of this species. Our results can be used for the identification of spider mites which are important in plant quarantine.

The sweetpotato whitefly, Bemisia tabaci, is a vector insect of more than 100 plant-diseased viruses as well as a serious pest of various horticultural crops. B. tabaci is a species-complex that consists of at least 24 biotypes, which show different biological characteristics including host range, fecundity, insecticide resistance and virus transmission. Here we identified biotype, endosymbiotic bacteria, and tomato yellow leaf curl virus (TYLCV) acquisition of various B. tabaci populations collected in Korea. In addition, we compared those profiles with B. tabaci collected from Bangladesh and Myanmar, and the greenhouse whitefly, Trialeurodes vaporariorum. PCR diagnosis of cytochrome oxidase I (COI) showed that all B. tabaci populations of Korea were Q-biotype and closely related with a subgroup I (MedBasin 1), which is indigenous to the Western Mediterranean area. Ribosomal DNA analysis of 5 endosymbionts showed that both Cardinium and Hamiltonella were detected in most tested populations while the presence of Arsenophonus, Fritschea and Wolbachia dependent on populations. Our results suggest that the acquisition of TYLCV do not related with the endosymbiont profile of B. tabaci.

In the family Sciaridae, only 3 species have been recorded in Korea, as the pests on various crops and mushrooms in glasshouse (Bradysia difformis Frey, 1948, Bradysia procera (Winnertz, 1868), Lycoriella ingenua (Dufour, 1839)), even though Sciaridae is one of the species-rich families in the order Diptera. There are plenty of species unknown in Korea which are very hard to identify by morphological characters in the larval, pupal stages and also female adults as well. Therefore, using the sequences of cytochrome oxidase I gene (COI) from adult male, we tested the utilities of DNA barcode to identify the species of sciarid flies.

Because mealybugs are one of the most economically damaging groups of insects on food crops and ornamental plants, some are regulated-species in quarantine with the foreign trade of agricultural products. However, the absence of morphological characteristics enabling the discrimination of early life stages often causes a significant delay or the rejection of a shipment when fruit is discovered containing them, causing much economic loss. A PCR-based method for species identification was developed for six mealybug species known from Korean pears including two regulated insects, Planococcus kraunhiae (Kuwana) and Crisicoccus matsumotoi (Siraiwa). Six sets of species-specific primers were designed based on the sequence comparison of the internal transcribed spacer 1 and 2 regions. Efficiency tests against many mealybug samples showed that this method could effectively discriminate different mealybug species regardless of their developmental stages. Blind tests against 11 field collected mealybug nymph samples indicated that a single PCR is enough to discriminate unidentified mealybugs collected on Korean pears. This new method will be useful in quarantine as well as pest monitoring by providing an easy, accurate way of identifying any life stage of these mealybugs.

Acquisition of a reference Tetranychusstrains that are completely susceptible to acaricides and retain identical genetic backgrounds to acaricide-resistant strains is an essential step in elucidating mechanisms of resistance. To establish both completely susceptible and acaricide-resistant strains for this purpose, I collected Tetranychus mite populations from various regions in South Korea including both heavily cultivated and remote regions. Suitability as a susceptible or resistant reference strain was tested by determining species identity as Tetranychus urticae along with baseline susceptibility to common acaricides. The majority of mite populations collected from cultivated areas belonged to a monophyletic group of the previously reported green-type T. urticae as determined by mtCOI- and ITS2-based phylogenetic analysis. These strains showed relatively reduced levels of susceptibility to the acaricides tested, suggestive of the development of resistance. Among them, the AbaR strain was classified as a minor group in the T. urticae complex. The UD strain, originally collected from a remote island region, was found to be susceptible to the acaricides tested and generated an independent phylogenetic branch. The UD strain was also considered a minor group in the T. urticae complex. Phenotypic analysis based on morphological characters confirmed that both the AbaR and UD strains were statistically undistinguishable from the major green-type T. urticae. Taken together, I propose that the UD strain be used as a susceptible reference strain as it provides baseline susceptibility to acaricides and a wild-type genetic background for the resistance studies.

The soybean aphid, Aphis glycines Matsumura 1917, is well known as a soybean pest in the world. Recently, it has been introduced to North America causing serious damage in U.S. As a cooperative research with USDA-ARS, we have investigated A. glycines in soybean fields, and also examined the colonies on the overwintering host Rhamnus davurica in order to find its natural enemies. It was generally reported that A. glycines has host alternation between the soybean, Glycine max (summer host) and the Dahurian buckthorn, Rhamnus davurica (winter host) in East Asia. However, it was very difficult to identify the soybean aphid, A. glycines, from R. davurica due to the co-existance of at least three Aphis species and the seasonal polymorphisms of each species (e.g, gynopara, ovipara, and male). For species identification, we tested 3 molecular markers, mitochondrial COI, COII, and nuclear EF1α, for 14 collected samples (7 samples from G. max and 7 samples from R. davurica). As a result, we found two different species, A. gossypii and other Aphis sp., are mixed together with A. glycines on R. davurica. We report the biology of A. glycines in Korea, and present species identification using molecular phylogenetic approach.

Protease from various sources have been studied biotecnologically. For biotechnological applications, one highly preferred enzyme is protease. There have been no reports of cloned genes encoding digestive proteases in the Laccotrephes japonenis, Ranatra unicolor, Muljarus japonicus. These insects are considered to be a predator of aquatic insects. RT-PCR was used to amplify cDNA fragments for digestive proteases from total RNA the hole body of the insects. The flanking sequences of the 5'- and 3'- end of the these genes were characterized by RACE-PCR. Sequence analysis showed that these genes contained complete ORF. The deduced amino acid sequences of these protease showed 62% identity to the serine protease of Creontiades dilutus, 58% to Lygus loneolaris trypsin-like serine proteinase , 54% to Triatonatoma infestans salivary trypsin. In the further study, to generate digestive protease, the DNA fragment coding for serine protease, trypsin-like serine protease were cloning into suttle vector pBACⅠ, and infected to Spodoptera frugiperda (sf9) insect cell. After that, we expect to carry out the proteolytic activity of these recombinant proteases. This is intended as a basis for future studies on the digestive protease in the insects.

Only two species of Eurydema occur in Korea and Japan: E. dominulus (Scopoli, 1763) [= E. pulchra (Westwood, 1837), syn. n.] and E. gebleri Kolenati, 1846 [= E. rugosa Motschulsky, 1861, syn. n.]. In order to prove the above taxonomical change, we focused on three major works (1)morphology, (2)molecular identification and (3)biological experiment (inter-specific copulation, confirmation of fertilization and interbreeding). All the results of these three major works support to confirm new taxonomical change amongst these four species. The inter-specific copulation occurs (E. pulchra_male X E. gebleri_female) in nature, but the eggs from inter-specific copulation are confirmed not to be fertilized as a result of the biological experiment. A new identification key to species occurring in Korea and Japan is presented for the all stages (from eggs to adult).

In order to obtain novel genes related to the human craniofacial development, molecular cloning and sequencing, and in situ hybridization using craniofacial tissue sections were performed and followed by protein structure simulation. Totally 231 clones were obtained from the subtracted craniofacial tissue cDNA library of human embryo. Random cloning using the non-redundant clones from the craniofacial tissue of human embryo was done and obtained 398 clones from the premade human chondrocyte cDNA library. Their partial sequence data showed that 214 clones of subtracted cDNA library of craniofacial tissue were still non-redundant in Genebank search. And 20 clones among 498 clones of premade chondrocyte cDNA library were known to be undefined genes. Through in situ hybridization screening in the craniofacial tissue sections of 10 weeks old human embryo 36 clones were found to be positive in specific tissues. Depending on the cell types of sirnilar developmental origin, the positive reactions could be divided into five groups. Among the 20 clones of undefined genes from human chondrocyte cDNA library, 7 clones showed characteristic positive reaction in human cartilage tissue by in situ hybridization. From the simulated protein structure, motif analysis and in situ hybridization studies for the 7 undefined clones, Ch89, Ch96, Ch129, Ch285 clones may function in the outer space of the cell constituting a part of matrix protein complex, and Ch276 as a transmembrane protein which might partic ipate in matrix calcification around chondrocytes. Ch153 is a kind of antirnicrobial protein also acting as an inflammation mediator, and Ch334 clone is a zinc finger protein, of which expression increases in human adult tissues We presume these novel genes from human chondrocytes may provide a new path of chondrocyte development and functions of human craniofacial tissues

과피와 과육의 다양한 색은 복숭아 분류에 가장 널리 사용되는 상업적 기준 중 하나이다. 새로운 적색 과육 품종을 육성하기 위해서는 많은 교배 조합과 세대가 진전되어야 한다. 따라서 육종 효율을 높이기 위해서는 목적 형질을 가진 개체에 적용할 조기 선발 분자표지를 개발할 필요가 있다. 과육색이 다르게 발현되는 복숭아 품종의 유전자 발현을 비교하기 위해 2개의 cDNA library를 제작하였다. 적색 과육 품종인 ‘조생혈도’와 백색 과육 품종인 ‘미백도’의 유전자 발현 차이를 보기 위해 차세대 염기서열 분석(NGS) 기술을 사용하였고, 두 품종으로부터 얻은 EST의 염기서열을 결정하고 기존에 보고된 유전자와의 상동성을 분석하였다. ‘조생혈도’와 ‘미백도’의 EST database로부터 72쌍의 SNP 분자표지를 선발하였고, 적색 과육 품종 8개와 백색 과육 품종 24개를 구분할 수 있는 SNP 분자표지를 HRM 방법으로 분석하였다. 본 연구에서는 복숭아 EST database를 기반으로 HRM 분석 방법을 이용하여 복숭아 품종의 적육계와 백육계를 구분할 수 있는 효율적인 SNP 분자표지를 개발하였다. 이러한 SNP 분자표지는 복숭아 육종에 유용하게 사용할 수 있으며, 복숭아 품종의 다양한 색 변화에 관한 분자 기작 연구에 좋은 참고자료가 될 수 있을 것이다.

Background: Cnidium officinale Makino and Ligusticum chuanxiong Hort. are important medicinal crops in Korea. However, there is insufficient information on the identification of thrips, which attack these plants. Until now, one species of thrips has been recorded as a main pest. Methods and Results: To identify the thrips emerging in C. officinale Makino and L. chuanxiong Hort., these plants were independently cultivated in two local areas. Thirty individuals of each plant species were selected randomly and surveyed for the presence of thrips. After confirming the existence of thrips, 100 thrips individuals were collected from each crop using the beating method. To identify thrips species, we performed PCR-restriction fragment length polymorphism (RFLP)-based analysis using ITS2 primer sets. Six thrips species were identified: western flower thrips (Frankliniella occidentalis), flower thrips (F. intonsa), onion thrips (Thrips tabaci), chrysanthemum thrips (T. nigropilosus), chilli thrips (Scirtothrips dorsalis), and grass thrips (Anaphothrips obscurus). The proportion of these species differed between the host plant species. Conclusions: Six thrips species were major pests of two medicinal crops. Integrated pest management is required to control these thrips species, and will enhance the yield and quality of C. officinale and L. chuanxiong.

Background : Cudrania tricuspidata Bureau is a widely used medicinal perennial woody plant. Obtaining information about the genetic diversity of plant populations is highly important for conservation and germplasm utilization. In this study, we developed single nucleotide polymorphism (SNP) markers derived from chloroplast genomic sequences to identify distinct Korean-specific ecotypes of C. tricuspidata via amplification refractory mutation system (ARMS)-PCR analyses. We performed molecular authentication of twelve C. tricuspidata ecotypes from different regions using DNA sequences in the chloroplast TrnL-F intergenic region. Methods and Results : SNPs were identified based on the results of nucleotide sequence for the intergenic region of TrnL-TrnF gene (chloroplast). Molecular markers were designed for those SNPs with additional mutations on the second base from SNPs for amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). HRM pattern analyses were performed using the Mx3005P QPCR System (Agilent Technologies, CA, USA). Conclusion : We collected 12 individual lines of C. tricuspidata from various region in South Korea and China. Based on the nucleotide sequence in the trnL-trnF intergenic region of these lines, six SNPs and a deletion of 12 bps were identified and 12 individual lines were able to be grouped in one Korean ecotype and two different ecotypes of chinese lines, chinese line 1 and 2. The SNP markers developed in this study are useful for rapidly identifying these specific C. tricuspidata ecotypes collected from different regions.