Mycoplasma hyorhinis (M. hyorhinis) is considered an etiological agent of arthritis in suckling pigs. Recently, some M. hyorhinis strains were shown to produce pneumonia that is indistinguishable from the mycoplasmosis caused by M. hyopneumoniae. In this study, we developed a sensitive and specific PCR assay to detect M. hyorhinis and applied the developed PCR assay for detection of Mycoplasma infection in clinical piglets infected with M. hyorhinis. We developed a new PCR assay using a M. hyorhinis-specific primer pair, Mrhin-F and Mrhin-R, designed from the Mycoplasma 16S-23S rRNA internal transcribed spacer (ITS) region. The primers and probe for the assay were designed from regions in the Mycoplasma 16S-23S rRNA ITS unique to M. hyorhinis. The developed PCR assay was very specific and sensitive for the detection of M. hyorhinis. The assay could detect the equivalent of 1 pg of target template DNA, which indicates that the assay was very sensitive. In addition, M. hyorhinis PCR assay detected only M. hyorhinis and not any other Mycoplasma or bacterial spp. of other genera. The new developed PCR assay effectively detected M. hyorhinis infection in pigs. We suggest that this PCR assay using a M. hyorhinis-specific primer pair, Mrhin-F and Mrhin-R, could be useful and effective for monitoring M. hyorhinis infection in pigs.

A rust on Asiatic dayflower plant (Commelina communis L.) was found in Geoje, Korea, in August 2010. Uredia are mostly produced on abaxial leaf surface or elongated on stem, early naked, surrounded by the ruptured epidermis, cinnamon-brown. Uredospores are globose, ellipsoid or ovate, echinulate, yellowish brown or brown, 20-30 × 20-25 ㎛. On the basis of mycological characteristics and molecular data, the fungus was identified as Uromyces commelinae Cooke. The phylogenetic position of U. commelinae is separate from the other rusts where the economically important rusts of the Poaceae are situated. Although host ranges of the rust caused by U. commelinae were previously recorded, full descriptions and illustrations, including symptoms and signs have not been described. This is the first description of rust disease on C. communis plant with molecular identification, symptoms, and signs.

Nuclear ribosomal DNA (rDNA) was analyzed to identify inter-specific genetic relationships among 8 Cymbidium species (Cymbidium insigne, C. ensifolium, C. marginatum, C. faberi, C. gyokuchin, C. kanran, C. forrestii, and C. goeringii). Nuclear rDNA including 2 internal transcribed spacer (ITS) regions and 5.8S, was amplified using polymerase chain reaction and sequenced. The sequences were compared via pair-wise multiple alignment to determine the genetic relationships among the studied species. The lengths of the ITS1, ITS2, and 5.8S regions were 235 bp, from 255 bp to 257 bp, and from 153 bp to 165 bp, respectively. Sequence similarities in the ITS region ranged from 78.7% between C. gyokuchin and C. kanran to 96.8% between C. ensifolium and C. kanran. A phylogenetic tree was constructed from nuclear rDNA nucleotide sequence data of the 8 cymbidiums and 1 outgroup species to estimate genetic relationships. The tree revealed that cymbidiums could be classified by their ecological traits, such as their temperature preference or inflorescence pattern. The phylogenetic data is applicable for identification, classification, and breeding of cymbidiums.

보존중인 잎새버섯속 균주를 선발하여 배양 및 형태적 특성을 조사하여 비슷한 균주별로 그룹화하여 rDNA의 ITS영역을 증폭하여 염기서열을 결정한 결과 보존시 균주의 학명과 많은 차이를 보였으며, 균사의 모양 및 색깔에도 많은 차이를 보였다. 보존 당시 동정한 결과와 rDNA의 ITS 영역의 염기서열 분석을 통한 결과를 비교한 결과 학명이 다른 균주가 4균주로 전체의 40%를 차지하였다. 국내에서 수집한 잎새버섯속은 모두 G. frondosa로 동정되었고, 일본에서 수집한 3균주 중 1균주는 완전히 다른 속으로 동정되었으며 2균주는 G. frondosa로 동정되었다. 그리고 중국에서 수집한 3균주중 2균주는 완전히 다른 속으로 동정되었고, 1균주만 G. frondosa로 동정되었다. RAPD 분석을 통한 유전적인 다형성 조사에서 같은 종내에서도 분포지역에 따라 서로 상이한 밴드패턴을 보였다. 유전적인 유연관계 분석에서는 G. frondosa 1개의 분류군으로 이루어졌으며, ITS부위 유전자수준의 상동성 분석에서도 비슷한 경향을 보였다. 따라서 기존 목록과 완전히 다른 속으로 동정된 균주들에 대해서는 계통분류학적인 유연관계 분석과 보존중인 자실체 유전자와의 상동성을 비교하여 기존 목록의 학명을 재분류해야 할 것으로 판단된다.

보존중인 Polyporus속 균주를 선발하여 배양 및 형태적 특성을 조사하여 비슷한 균주별로 그룹화하여 rDNA의 ITS 영역을 증폭하여 염기서열을 결정한 결과 보존시 균주의 학명과 많은 차이를 보였으며, 균사의 모양 및 색깔에도 많은 차이를 보였다. 보존 당시 동정한 결과와 rDNA의 ITS 영역의 염기서열 분석을 통한 결과를 비교한 결과 종이 다른 균주가 3균주와 학명이 다른 균주가 4균주로 전체의 53.8%를 차지하였다. 국내에서 수집한 Polyporus속은 경우 P. alveolarius, P. brumalis, P. squamosus, P. tuberaster, P. arcularius 등 5개 종으로 동정되었고, 미국에서 수집한 균주는 P. alveolarius와 P. arcularius로 동정되었다. 그리고 일본에서 수집한 균주는 P. arcularius로 동정되었다. RAPD분석을 통한 유전적인 다형성 조사에서 Polyporus속간에는 완전히 다른 밴드 패턴을 보였지만 같은 종내에서는 비슷한 밴드 패턴을 보였다. 유전적인 유연관계 분석에서는 P. alveolarius 등 5개의 분류군으로 나누어졌으며, ITS부위 유전자수준의 상동성 분석에서도 비슷한 경향을 보였다. 따라서 기존 목록과 완전히 다른 속으로 동정된 균주들에 대해서는 계통분류학적인 유연관계 분석과 보존중인 자실체 유전자와의 상동성을 비교하여 기존 목록의 학명을 재분류해야 할 것으로 판단된다.

보존중인 구름버섯속 균주를 선발하여 배양 및 형태적 특성을 조사하여 비슷한 균주별로 그룹화하여 rDNA의 ITS 영역을 증폭하여 염기서열을 결정한 결과 보존시 균주의 학명과 많은 차이를 보였으며, 균사의 모양 및 색깔에도 많은 차이를 보였다. rDNA의 ITS 영역의 염기서열을 바탕으로 보존 당시 동정된 결과와 염기서열 분석을 통한 결과를 비교한 결과 종이 다른 균주가 5균주와 학명이 다른 균주가 6균주로 전체의 61%를 차지하였다. 국내에서 수집한 구름버섯의 경우 T. versicolor, T. elegans, T. gibbosa 등 3개 속으로만 동정되었고, 미국에서 수집한 균주는 T. junipericola로 동정되었다. Trametes spp의 RAPD 분석을 통한 유전적인 다형성 조사에서 T. versicolor와 T. gibbosa는 아주 다른 밴드 패턴을 보였다. 또한 같은 종내에서서 분포지역에 따라 상이한 밴드 패턴을 보였다. 유전적인 유연관계 분석에서는 T. vericolor 등 4개의 분류군으로 나누어졌으며, ITS부위 유전자수준의 상동성 비교에서도 비슷한 경향을 보였다. 따라서 기존 목록과 완전히 다른 속으로 동정된 균주들에 대해서는 계통분류학적인 유연관계 분석과 보존중인 자실체 유전자와의 상동성을 비교하여 보존진균의 오염 여부를 판단하여 기존 목록의 학명을 재분류해야 할 것으로 판단된다.

Recently, ambrosia beetles have become very important pest of 2~5 year old apple trees with M9 dwarf rootstocks in South Korea. The beetles have killed the branches and stems of the young trees, especially, frozen damage trees in winter or drought stressed tree in spring. By the increase in planting area and weaken property in winter of M9 dwarf rootstock, ambrosia beetles are becoming a key pest in Korean apple orchards using M9 rootstock. According to the survey of damaged apple trees by ambrosia beetles, Xylosandrus germanus Blandford, Xyleborus apicalis Blandford and Xyleborinus saxeseni (Ratzeburg). These insects are hosts of the ambrosia fungi. ITS region of rDNA has shown to be a useful source for phylogenetic studies and identifying speices in previous published articles. We analyzed the nucleotide sequences of ITS1, 5.8S and ITS2 region of ambrosia fungi isolated from three ambrosia beetles, in order to observe molecular variation among the fungi strains and to reveal phylogenetic relationships.

Background : Ixeris dentata is the perennial herbaceous plant in the Ixeris genera within the Compositae family. The whole plant has been used traditionally as herbal medicine. It is widely distributed in South Korea and the genetic difference among the plants harvested from different regions may differ due to the disparity in cultivation climate. Therefore, this research was performed to discriminate the I. dentata that are collected from four locations in South Korea based on sequence analysis of nrDNA-ITS region. Methods and Results : Genomic DNA was extracted from I. dentata obtained from Goesan, Dangjin, Yangpyeong, and Chuncheon. I. stonolifera was included for a comparison of genetic distance with I. dentata. PCR amplification was performed by using an universal barcode nrDNA-ITS primer for DNA barcoding. After sequencing, the data was aligned using ClustalW multiple alignment tool in BioEdit version 7.2.5 software. SNP and phylogenetic analysis were conducted with the MEGA7 program. Phylogenetic analysis was presented with Neighbor-joining method using the K2P model. Statistical significance are evaluated using bootstrap (1,000 replicates). PCR products were amplified with about 800bp length for the ITS1-4 sequences of all samples. Nineteen SNPs were detected within a 578bp fragment of the aligned sequences. The mean GC content was 51.88% for ITS1-4 sequences of them. The interspecific genetic distance between I. dentata and I. stonolifera was 0.029%. The highest region-specific distance was confirmed to 0.010% between the plants from Dangjin with Goesan and Chuncheon group. Meanwhile, the mean intraspecific distance among the plants from Yangpyeong was 0.002%. Conclusion : In the phylogenetic analysis, the plants from Goesan and Chuncheon were placed within the same clade, while the plants from Dangjin formed independent clade. On the other hand, the plants from Yangpyeong were not grouped into one clade followed by the intraspecies variation. In conclusion, the data from the research will be useful for the regional identification of Dangjin, even if it is required to perform some additional researches.

The 12 cultivars of the Jeju native Citrus are considered to have originated from China. However, the origin of the cultivar ‘Byungkyool’ (Citrus platymamma Hort. ex Tanaka) is not clearly known. We performed PCR analysis by using three primer sets designed from the internal transcribed spacer (ITS) region of nuclear ribosomal DNA (nrDNA) to analyze the phylogenetic relationship between the traditional citrus cultivars and the Byungkyool cultivar. Sequence length of the nrDNA ITS1 region of JNCPCRI (Jeju Native Citrus platymamma Citrus Research Institute) cultivar was 247 bp, 8the ITS2 region was 228 bp and the total ITS region (ITS1-5.8S-ITS2) was 638 bp. Analysis of the genetic relationship based on the sequence analysis at the ITS region of the JNCPCRI cultivar revealed that the ITS1 region of the cultivar was genetically the same as that of the Byungkyool (JQ990189) cultivar, and the ITS2 region was genetically similar to the Binkyool (JQ990180), Hongkyool (JQ990178), Dangyooja (JQ990179), and Pyunkyool (JQ990181) cultivars. Moreover, the total ITS region in the 5.8S rDNA region was genetically similar to the Hongkyool (JQ990178) cultivar. In addition, the total ITS region of the JNCPCRI cultivar was the most closely related to the Cheongkyool (JQ990183) cultivar and has been reported to originate from the Binkyool (JQ990180) and Pyunkyool (JQ990181) cultivars. Although the JNCPCRI cultivar was morphologically the same as the Byungkyool (JQ990189) cultivar, the ITS region showed genetic heterogeneity. Taken together, we conclude that the genetic variation in the ITS region of JNCPCRI cultivar suggests that it was propagated through fertilization with the surrounding citrus cultivars.

Background : Correct identification of Panax species is important to ensure food quality, safety, authenticity and health for consumers. This paper describes a high resolution melting (HRM) analysis based method using internal transcribed spacer (ITS) and 5.8S ribosomal DNA barcoding regions as target (Bar-HRM) to obtain barcoding information for the major Panax species and to identify the origin of ginseng plant. Methods and Results : A PCR-based approach, Bar-HRM was developed to discriminate among Panax species. In this study, the ITS1, ITS2, and 5.8S rDNA genes were targeted for testing, since these have been identified as suitable genes for use in the identification of Panax species. The HRM analysis generated cluster patterns that were specific and sensitive enough to detect small sequence differences among the tested Panax species. Conclusion : The results of this study show that the HRM curve analysis of the ITS regions and 5.8S rDNA sequences is a simple, quick, and reproducible method. It can simultaneously identify three Panax species and screen for variants. Thus, ITS1HRM and 5.8SHRM primer sets can be used to distinguish among Panax species.

제주도에 자생하는 부채 선인장인 백년초의 기원 규명을 목적으로 ITS primer를 이용하여 685 bp의 ITS 영역을 분리하였다. ITS 영역의 염기서열을 분석한 결과 18S rRNA의 길이는 54 bp, 26S rRNA는 55 bp, ITS1은 193 bp, ITS2는 220 bp로 구성되어 있었다. 백년초 ITS 영역은 기존에 보고된 Cucurbitoideae 식물들의 ITS 영역에 비하여 ITS2 스페이서 영역의 239-254 bp보다는 다소 짧았다. 그러나 이들 스페이서 영역의 GC 함량은 백년초의 경우 ITS1은 66.8%, ITS2의 경우에는 67.7%로 Cucurbitoideae 식물들에서 보다 높은 GC 함량을 나타내었다. 백년초 선인장의 rDNA 영역에 가장 높은 상동성을 나타낸 것은 같은 Opuntioideae에 속하는 Pereskiopsis porteri(L78037)로 95%의 유사도를 나타내었다. 백년초 rDNA Clustal W 프로그램을 이용하여 유연관계를 조사한 결과 같은 Opuntioideae에 속하는 Pereskiopsis porteri(L78037)와 같은 cluster로 분리되었다.