The bluetongue is a serious vector-borne viral disease that affect wild and domestic ruminants. It is transmitted by midges belonging to the genus of Culicoides which count more than 1350 species. Since 1998, the disease has been spread in Europe and North Africa in 2000 including Algeria, while Korea reported serotype 1 of the virus on 2015. To know further about the existence and distribution of Culicoides species in Algeria, adult midges were collected from 17 different regions in Algeria from 2009 to 2015. The Interactive Identification Key of Culicoides (IIKC) has been used, followed by molecular identification by the sequencing of partial COI gene. At least 21 genetically different species have been identified. Among them, at least 5 potential virus vector species are present in Algeria. This study provides important insights for the understanding of genetic diversity of Culicoides and the existing potential of bluetongue spread in Algeria.

This study is the identification of root-knot nematode (RKN), Meloidogyne hapla, from strawberry in Korea using molecular analyses. Strawberry plants showed localized stunting and galled roots. Molecular analyses of COⅡ/lrRNA, 28S rDNA D2-D3 segment and internal transcribed spacer (ITS) region were employed for the identification of Meloidogyne spp. Polymerase Chain Reaction (PCR) amplification of COⅡ/lrRNA region produced a single fragment ca 528bp. Restriction digestion of the amplified PCR products with Dra1 enzyme produced two fragments at 200 and 250bp indication M. hapla. 28S rDNA D2-D3 segment and ITS were cloned and sequenced. 28S rDNA D2-D3 segment and ITS region produced a single fragment of 1004bp and 560bp, respectively. In BLAST search in Genbank, all sequences accord with other known M. hapla. As a result, all of the RKN samples were M. hapla. This study suggests that the dominant species of RKN on strawberry is M. hapla in Korea.

We developed a polymerase chain reaction (PCR)-based molecular method for sexing and identification using sexual dimorphism between the Zinc Finger-X and -Y (ZFX-ZFY) gene and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for mitochondrial DNA (mtDNA) cytochrome B (CYTB) gene in meat pieces and commercial sausages from animals of different origins. Sexual dimorphism based on the presence or absence of SINE-like sequence between ZFX and ZFY genes showed distinguishable band patterns between male and female DNA samples and were easily detected by PCR analyses. Male DNA had two PCR products appearing as distinct two bands (ZFX and ZFY), and female DNA had a single band (ZFX). Molecular identification was carried out using PCR-RFLP of CYTB gene, and showed clear species classification results. The results yielded identical information on the sexes and the species of the meat samples collected from providers without any records. The analyses for DNA isolated from commercial sausage showed that pig was the major source but several sausages originated from chicken and Atlantic cod. Applying this PCR-based molecular method was useful and yielded clear sex information and identified the species of various tissue samples originating from livestock.

From August to October 2013, a powdery mildew was found on Japanese artichoke(Stachys sieboldii) in Jinju, Korea. White colonies were present on leaves and petioles. Severely infected lesions were discolored, being brown. In the present study, the morphological features of anamorphic and teleomorphic Korean specimens were studied. To complete the identification, the sequence of the internal transcribed spacer region of the ribosomal RNA gene(ITS rRNA) was analyzed. Based on morphological characteristics, including anamorphic and teleomorphic features, as well as analysis of the ITS rRNA gene region, the fungus associated with the symptoms was identified as Neoerysiphe galeopsidis. Although the host ranges of powdery mildew caused by N. galeopsidis has been previously recorded, no full description or illustration of fungal symptoms and signs on Japanese artichoke has yet appeared. To the best of our knowledge, this is the first full description of powdery mildew disease on Japanese artichoke featuring molecular identification, morphological features, symptoms, and signs in Korea.

동절기에 제주도 지역에서 도래하는 떼까마귀의 유전적 특성과 집단 간 유연관계를 구명하기 위해, 미토콘드리아 COI 유전자 서열의 다형성에 기반한 모계 계통 구조와 계통 유연관계를 분석하였다. 떼까마귀 DNA는 우도와 제주도 내에서 발견된 깃털과 사체 시료에서 분리하였다. 결정된 COI 서열들(n=41)은 떼까마귀(Corvus frugilegus)에서 기존 에 보고된 서열들과 97.0% 이상 일치하였다. 제주도 떼까마귀 COI 서열들은 3가지 haplotype(J01-J03)으로 구분되었으 나 지역-특이적인 양상을 보이지 않아, 이들이 하나의 모계 기원에서 유래한 집단임을 알 수 있었다. 떼까마귀 전체 COI 서열에서 8개의 COI haplotype들이 발견되었다. 이 중 3가지 haplotype들은 러시아 동부, 몽골, 한국 등 동북아시 아의 COI 서열들을 포함하였고, 나머지 5가지는 중앙아시아, 중동아시아, 러시아 서부, 유럽국가의 떼까마귀에서 발견 되었다. 계통수 상에서 떼까마귀의 COI 서열들은 측소적 종분화 단계인 2아종, C. f. frugilegus와 C. f. pastinator인 2개의 모계 계통으로 뚜렷하게 구분되었다. DNA barcoding 분석을 통한 연구결과는 모계 계통의 구조, 계통 유연관계 및 분자생태를 이해하는 데 중요한 정보를 제공할 것이다.

Glucose-regulated protein 78 (GRP78) is a member of the heat shock protein 70 (HSP70) family that is specific to endoplasmic reticulum (ER). It is known as chaperones and signaling regulators that respond to ER stresses in vertebrates. However, its function in invertebrates, including insects, is uncertain. Here we determined a full cDNA sequence and the expression patterns of grp78 of Aphis gossypii, which is a major pest of numerous crop plants worldwide. Its cDNA had highly conserved motifs of the HSP70 family and the C-terminal motif of KDEL characteristic of ER-specific HSPs. It showed 98.5% identity with the GRP78 of the pea aphid Acyrthosiphon pisum. Real-time RT-PCR analysis showed that the grp78 level was higher in the fourth instar nymph than in the younger instar and adult stages. Its level was not affected by thermal stress of 10 to 40°C for 1 h. The grp78 level was proportional to the ingestion of a sucrose solution ranging in concentration from 0% to 30% in a Parafilm feeding chamber. In addition, the grp78 level varied among aphids feeding on leaves from 14 different host plants for 24 h; it was higher with eggplant and pepper but lower with pigweed and tobacco than any other plants. Our study suggests that the grp78 level is regulated by nutritional condition of A. gossypii.

Cotton aphid infests more than 700 plants and a major pest of various horticultural crops worldwide. The glucose regulated protein 78 (GRP78) is a member of heat shock protein 70. Its expression is associated with the nutritional changes as well as environmental stresses. The full sequences of grp78 cDNA of Aphis gossypii was determined. It had conserved motifs of hsp genes and terminated in KDEL which is common to GRP78. Quantitative realtime PCR showed that its level was changed during development and also upregulated by starvation. However, its level was not much changed by heat stress. The level of grp78 can be use to understand nutritional physiology on insects.

Root-lesion nematodes, Pratylenchus sp., are recognized as one of the most important plant parasitic nematodes. They infest various crop plants and cause severe root damages which lead to the yield loss of agricultural crops. There are 72 species recorded in the world, but 14 species in Korea. Soil samples were collected from chrysanthemum cultivars in various regions. Morphological and molecular diagnosis firstly recognized two species as Pratylenchus kumamotoensis and P. pseudocoffeae in Korea. Morphologically, P. kumamotoensis characterized by pharyngeal gland lobes overlapping with intestine dorsally. P. pseudocoffeae, which is similar with P. coffeae, but pharyngeal gland lobes are longer than P. coffeae. In molecular diagnosis, nucleotide sequences of D2-D3 region (700-800 bp) within 28S rDNA and ITS region including ITS1, 5.8S and ITS2 (700-1,000 bp) between 18S and 28S rDNA were determined from collected samples. NCBI database comparison of D2-D3 of P. kumamotoensis and P. pseudocoffeae were 99% similar with those of corresponding species, respectively. ITS of P. pseudocoffeae was 99% similar within species. ITS sequence of P. kumamotoensis was firstly determined in this study. Distribution of P. kumamotoensis was only reported in Japan but P. pseudocoffeae is present in Japan and USA. Here we firstly report the presence of these two species in Korea.

Toothbrushes play an essential role in oral hygiene. However, they can be significant in microbial transmission and can increase the risk of infection, since they can serve as a reservoir for microorganisms in healthy, oral-diseased and medically ill adults. This study was conducted to evaluate toothbrush contamination in six toothbrushes donated from four people. Two participants each supplied two toothbrushes - one used in the bathroom and one used in the workplace. The other two people each donated two toothbrushes used in the workplace. Polymerase chain reaction was used to construct a 16S rRNA clone library. Sequences of cloned DNA were compared with those from the reference organisms provided by GenBank. A total 120 clones, representing 20 clones for each toothbrush, were analyzed. They are composed of six pylum, 46 genera and 79 species. The most dominant species were Streptococcus oralis, Streptococcus parasanguinis and Haemophilus parainfluenzae. Enterobacter and Escherichia were recovered from toothbrushes used domestically. Toothbrushes used in the workplace did not contain Enterobacteria.

옥수수 포장에서 발생하는 Trichogramma속 조명나방 알기생벌의 종 분포를 조사하기 위해 채집된 알기생벌로부터 핵내 ITS2 DNA 전체 염기서열 정보를 해독하였다. 그리고 종 구별을 위한 참고정보로 NCBI GenBank에 등록된 Trichogramma속 60종의 ITS2 전체 염기서열을 확보하여 비교하였다. 국내 채집 알기생벌은 ITS2 DNA 길이와 3′ 말단 염기서열 패턴에 따라 3개 그룹(K-1, -2, -3)으로 구분되었다. 국내 채집 그룹 내 염기서열 차이 추정값(Evolutionary distance, d)은 0.005 이하로 그룹 간 비교 시 d 값(≥0.080)보다 낮았다. 그룹 및 GenBank 등록 종 간 비교 시 K-1은 T. ostriniae, K-2는 T. dendrolimi, K-3은 T. confusum과 d 값이 각각 0.016, 0.001, 0.002로 가장 작았다. 추론된 분자계통수에서 K-1은T. ostriniae, K-2는 T. dendrolimi와 각각 분지되었으나 K-3는 T. confusum, T. chilonis, T. bilingensis와 함께 분지되었다. NCBI BLAST 결과에서도 K-1은 T. ostriniae와 K-2는 T. dendrolimi와 99% identity를 보였다. 그러나 K-3의 홍천 채집 기생벌들은 T. confusum, T. chilonis와 99-100% identity를 보였지만, 고창 채집 기생벌은 T. bilingensis, T. confusum, T. chilonis와 98% identity를 보였다. 이상의 분석 결과 본 연구에서 채집된 알기생벌 K-1과 K-2는 각각 T. ostriniae와 T. dendrolimi, 단일한 종으로 추정되었으나 K-3는 ITS2 정보만으로 종을 추정하기 어려웠다.

국내 과채류 시설재배지 토양 내 분포하는 뿌리혹선충의 계통학적 특성을 조사하였다. 토마토, 오이, 수박, 참외를 재배하는 12개 과채류 시설재배지 토양 내 뿌리혹선충의 밀도를 조사한 결과, 모든 시설재배지 토양에 광범위하게 분포하였으며 토양 300 g 당 평균 72±6~2,898±468마리로 검출되었다. 시설재배지 토양에서 수집한 뿌리혹선충 2령 유충을 대상으로 PCR-RFLP 계통분석을 수행하였다. 시설재배지 토양에서 분리한 12개 뿌리혹선충의 mtDNA PCR 증폭산물을 대상으로 제한효소 HinfI을 처리한 결과 900, 410, 290 및 170 bp의 DNA 절편 양상을 나타내는 Group A와 900, 700 및 170 bp의 DNA 절편 양상을 나타내는 Group B로 분류되었다. 각 그룹에 속하는 뿌리혹선충의 mtDNA 유전자 염기서열(1,483~1,521 bp)을 결정하여 계통분석한 결과, Group A에 속하는 9개의 뿌리혹선충은 고구마뿌리혹선충(Meloidogyne incognita)과 99.73~99.93%의 상동성을 나타내었고 그리고 Group B에 속하는 3개의 뿌리혹선충은 땅콩뿌리혹선충(Meloidogyne arenaria)과 99.54~ 99.73%의 상동성을 나타내어 유사한 종으로 확인되었다.

표고버섯 재배지에서 버섯파리는 가장 문제가 되는 주요해충이다. 버섯파리에 의한 피해를 입은 농가는 표고버섯의 생산뿐만 아니라 상품성이 현저히 떨어져 많은 피해를 입고 있다. 국내 표고버섯 재배양식은 크게 원목과 톱밥배지에 의한 방법으로 나뉘는데 재배양식의 차이 및 지역 간 주요 피해원인이 되는 버섯파리의 종과 분포양상을 조사하기 위해 봄부터가을까지 경기도와 충청도 지역의 주요 표고버섯 재배지에서 버섯파리를 채집하였다. 채집은 황색 끈끈이트랩(참고, Yellow sticky trap)을 이용하였고 버섯파리의 종 동정은 cytochrome c oxidase subunit I(COI)의 염기서열 분석을 통해 버섯파리류의 다양한종을 분류하였다. 그 결과, 원목을 이용하는 표고버섯 재배지에서는 Bradysia difformis, B. alpicola가우점종인것으로 나타났고, 톱밥배지로 표고버섯을 재배하는 농가의 경우, B. difformi와 Scatopsidae sp.의종이 다량 동정되었다. 그 외에도 B. trispinifera, B.longimentura, B. chlorocornea, B. protohilaris 및Lycoriella ingenua가 동정되었다.

Trombiculid mites are known to be the vector of tsutsugamushi disease by transmitting Orientia tsutsugamushi to human. Although the classification of trombiculid mites is necessary for vector surveillance, their classification by morphological observation is only possible at the larval stage and not easy because of similar shapes as well as tiny body sizes. Further the classification need the specimen production process, it takes much time and the accuracy of classification is changed according to the technology of the researcher. The internal transcribed spacers (ITS) regions of 8 trombiculid mite species were analyzed by amplification using tick common ITS primer sets. We designed molecular marker sets for the identification of five Leptotrombidium species, the lengths of marker L. orientale (1078 bp), L. pallidum (820 bp), L. palpale (1202 bp), L. scutellare (447 bp) and for L. zetum (621 bp) and three Neotrombicula species, the lengths of marker N. gardellai (264 bp), N. japonica (460 bp) and N. kwangneungensis (309 bp) based on alignment of ITS sequences. The markers will be helpful for exact classification of trombiculid mites. This study is the first report on molecular marker of ITS regions of trombiculid mites.

2012년 북한강수계 전역에 걸쳐 대발생을 일으킨 남조 Anabaena의 분자생물학적 특성을 검토하였다. 시료는 2012년 7월 13일에 남조 Anabaena 밀도가 높았던 3개 호소- 의암호, 청평호, 팔당호에서 각각 채집하였다. 분석은 선행연구 문헌을 근거로 하는 형태학적 분석, 16S rRNA 염기서열 분석을 통한 분자계통학적 분석을 동시에 실시 하였다. 결과를 종합하면 북한강수계 3개 호소에서 조류 대발생을 일으킨 남조 Anabaena는 모두 동일종 Anabaena crassa (Lemmermann) Komark.-Legn. & Cronberg 이며, 동시에 출현하고 본 종과 형태 및 분자생물학적으 로 매우 유사한 A. circinalis는 환경요인에 따라 형태변 이가 쉽게 일으키는 동일종으로 밝혀졌다.

Anginosus group streptococci (AGS) were classified based on the nucleotide sequences of the 16S rRNA gene (16S rDNA) and comprised Streptococcus anginosus, Streptococcus intermedius, and Streptococcus constellatus. It is known that AGS is a causative factor of oral and systematic diseases. The purpose of this study was to discriminate the 56 clinical strains of AGS isolated from Korean oral cavities using phylogenetic analysis of 16S rDNA and species‐specific PCR at the species-level. The 16S rDNA of clinical strains of AGS was sequenced using the dideoxy chain termination method and analyzed using MEGA version 5 software. PCR was performed to identify the clinical strains using species‐specific primers described in previous studies and S. intermedius‐specific PCR primers eveloped in our laboratory. The resulting phylogenetic data showed that the 16S rDNA sequences can delineate the S. anginosus, S. intermedius, and S. constellatus strains even though the 16S rDNA sequence similarity between S. intermedius and S. constellatus is above 98%. The PCR data showed that each speciesspecific PCR primer pair could iscriminate between clinical strains at the species‐level through phylogenetic analysis of 16S rDNA nucleotide sequences. These results suggest that phylogenetic analysis of 16S rDNA and PCR are useful tools for discriminating between AGS strains at the species-level.

To identify subspecies of the sika deer, Cervus nippon, and to select C. n. hortulorum individuals for restoration of this subspecies in Korea, we obtained cytochrome b partial sequences (375 bp) of sika deer (C. nippon) from deer farms in northeastern China, North Korea, mainland Korea, and Jeju Island. Based on noninvasive samples, these cytochrome b sequences were compared with the corresponding haplotypes of sika deer, which were obtained from GenBank. We identified five individuals of sika deer from deer farms in northeastern China and North Korea as C. n. hortulorum, and found that interbreeding between individuals belonging to different subspecies is common on deer farms in far-eastern Asia. We concluded that the five C. n. hortulorum individuals on deer farms from northeastern China and North Korea, detected in this study, cannot be considered as pure descendants of C. n. hortulorum in the wild.

To identify subspecies and stocks of minke whale meats purchased from Korean markets during 2005-2007, we first obtained their complete sequences of mitochondrial DNA cytochrome b and control region sequences, and compared these sequences to the corresponding sequences of the common minke whale (Balaenoptera acutorostrata), obtained from GenBank. From analyses with partial cytochrome b sequences (383 bp) and non-coding, partial control region sequences (463 bp), Korean mink whale meats are identified as products from the North Pacific minke whale (B. a. scammoni). In addition, the sequences of the partial control region from these meats showed G at site no. 298 and G or A at site no. 463, and the meats appeared to originate from the J stock within this subspecies. Thus, because the J stock has been protected since 1986, implementation of strict regulation measures to reduce their accidental fisheries by catch seems urgent. In addition, B. a. scammoni is distinct from B. a. acutorostrata, with an average Jukes-Cantor distance of 2.21% in the complete control region sequence analysis (935 bp) and 1.31% in the complete cytochrome b gene sequence analysis; the current results support the current subspecies classification, although further sequencing analyses with nuclear genes are necessary.