수집한 46개의 균주를 배양하여 rDNA의 ITS 염기서열 분석으로 유전적 계통분류 한 결과, Heiricum속이 아닌 균 주가 2균주가 있었음을 확인하였다. 본 결과를 바탕으로 효능이 있는 것으로 밝혀진 H.erinaceus와 가까운 유연관 계를 지니는 종들을 위주로 계통별로 균사 생장실험, 자실 체 발생조사 등을 연구할 예정이다. 또한 H.erinaceus은 아니지만, 생리활성효능이 뛰어난 종의 유무를 테스트하여 기존의 품종보다 효능 면에서 뛰어난 균주를 선발 교잡할 예정이다. 이와 같은 연구를 통해 우수품종을 선발하여 최 적 환경조건 테스트까지 거친다면 노루궁뎅이 버섯 소비 시장 구축에 도움이 되리라 생각된다.

The genetic relationships among five genera, seven species of Theaceae were examined through a random amplified polymorphic DNA (RAPD) analysis and internal transcribed spacer (ITS). In RAPD analysis, five of 15 arbitrary primers showed polymorphic bands, which were able to classify different genera and species of Theaceae. The genetic variations of Theaceae were from 0.031 to 0.484. In ITS analysis, the ITS sequences were analyzed using BLAST and showed high identities with sequences of Theaceae, seven species published in NCBI GenBank database, which ranged from 98 to 100%. Sequence alignment of seven species showed 34.9% identities for ITS 1 region and 43.7% for ITS 2 region. Pairwise sequence divergences among seven species ranged from 0 to 0.330%. In phylogenetic tree, they were divided into three groups. In conclusion, the molecular data generated in the present investigation will help to understand the genetic relationships of Theaceae and also might be useful for further studies in intra-species, inter-species, and molecular evolution researches.

The bumblebee, Bombus ignitus (Hymenoptera: Apidae), is a valuable natural resource that is widely utilized for greenhouse pollination in South Korea. Understanding the magnitude of genetic diversity and geographic relationships is of fundamental importance for long term preservation and utilization. As a first step, we sequenced a partial COI gene of mitochondrial DNA (mtDNA) corresponding to the “DNA barcode” region and the complete internal transcribed spacer 2 (ITS2) of nuclear ribosomal DNA from 88 individuals collected in nine South Korean localities. The complete ITS2 sequences were longest among known insects, ranging in size from 2,034 bp ~ 2,052 bp, harboring two duplicated 112-bp long repeats. The 658-bp long mtDNA sequences provided only six haplotypes with a maximum sequence divergence of 0.61% (4 bp), whereas the ITS sequences provided 84 sequence types with a maximum sequence divergence of 1.02% (21 sites). The combination of the current COI data with those of published data suggest that the B. ignitus in South Korea and China are genetically a large group, but those in Japan can be roughly separated into another group. Overall, a very high per generation migration ratio, a very low level of genetic fixation, and no discernable hierarchical population were found to exist among the South Korean populations of B. ignitus, which suggests panmixia. This finding is consistent with our understanding of the dispersal capability of the species.

수집한 83개의 뽕나무버섯속 균주의 rDNA의 ITS 영역을 증폭하여 염기서열을 분석한 결과 수집한 균주목록과 종이 다르게 동정된 균주가 52%였으며. 같은 종으로 분류한 균주들 간에도 균사 및 균사체의 배양 특성에 많은 차이를 보였다. 수집 균주간의 유전적인 유연관계 분석 결과 A. tabescens, A. mellea, A. novae-zelandia, A. gallica, A. ostoyae 등으로 분류되었고, A. gallica, A, cepistipes, A, gemina는 매우 가까운 유연관계를 보여 ITS 유전자 수준에서 종을 동정하기는 어려웠다. ASI10104 등 12균주는 A. gallica, A, cepistipes, A, gemina와 매우 가까운 유연관계를 보였으며, ASI10017과 ASI10114는 A. sinapina, ASI10045는 A. borealis, ASI10002와 ASI10025는 A . ostoyae와 같은 그룹으로 분류되었다. 따라서 뽕나무버섯속 균주에 대한 보다 정확한 동정을 위해서는 보다 많은 종류의 유전적인 분석이 필요할 것으로 판단된다.

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.

This work was performed to confirm the molecular discrimination through the nrITS1 sequences among 3 taxa of Scirpus L. sensu lato (s.l.) species. S. planiculmis represented only 2 base sequence variations with S. maritimus in spite that they showed different morphological features. The nucleotide sequences of the ITS1 region from S. planiculmis were shown to have 99.1% homology with S. maritimus and 60.4% homology with S. triqueter. Although the morphology of S. planiculmis is similar with S. triqueter, molecular basis of the size and sequences on ITS1 region were shown to have distinctive differences. For divergency investigation on same sites and metapopulation, sequencing was conducted on ITS1 region with partial 5.8S and 18S regions. All plants of each species collected at the same site had identical band size pattern and sequences. Intraspecific molecular divergency was not identified in spite that these species live in different wetland sites. The ITS1 sequences described here provided a powerful genetic tool for phylogenetic studies which was difficult by morphological identification as high rate of morphological plasticity

The leaf beetle, Chrysolina aurichalcea (Coleoptera: Chysomelidae), is a pest damaging plants of Compositae. In order to understand the genetic diversity and geographic variation we sequenced a portion of mitochondrial COI gene (658 bp) and complete nuclear internal transcribed spacer 2 (ITS2) of the species collected from seven Korean localities. A total of 17 haplotypes (CACOI01 ~ CACOI17), with the maximum sequence divergence of 3.04% (20 bp) were obtained from COI gene sequence, whereas 16 sequence types (ITS2CA01 ~ ITS2CA16), with the maximum sequence divergence of 2.013% (9 bp) were obtained from ITS2, indicating substantially larger sequence divergence in COI gene sequence. Phylogenetically, the COI gene provided two haplotype groups with a high nodal support (≥ 87%), whereas ITS2 provided one sequence type group with a high nodal support (≥ 92%). The result of COI gene may suggest the presence of historical biogeographic barriers that bolster genetic subdivision in the species. Different grouping pattern between COI gene and ITS2 sequences were interpreted in terms of recent dispersal, reflected in the ITS2 sequence. Finally, finding of unique haplotypes and sequence types only from Beakryeng-Islet population was interpreted as an intact remnant of ancient polymorphism. As more samples are analyzed using further hyper-variable marker, further fruitful inference on the geographic contour of the species might be available.

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.

The seven-spotted lady beetle, Coccinella septempunctata (Coleoptera: Coccinellidae), known also as the seven-spot ladybird, is natural enemy for aphids and has a broad ecological range, living almost anywhere there are aphids for it to eat. 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 nine Korean localities. A total of 21 haplotypes (CSCOI01 ~ CSCOI21), with the maximum sequence divergence of 4.56% (30 bp) were obtained from COI gene sequence (from 78 individuals), whereas 65 sequence types (CSITS201 ~ CSITS265), with the maximum sequence divergence of 2.06% (11 positions) were obtained from ITS2 (from 79 individuals), indicating substantially larger sequence divergence in mitochondrial gene sequence. Both COI gene and ITS2 shows the distribution pattern that only a few haplotypes or sequence types are widely distributed, whereas majority of them are highly restricted in one geographic location, even represented as a single individual. Unlikely the ITS2 sequence types the mitochondrial COI haplotypes evidenced the presence of two main phylogenetic groups, reciprocally monophyletic to each other. Geographically, these two groups are spread in all localities surveyed. Considering both COI gene and ITS2 sequence together, current our data may suggest the presence of ancestral polymorphism, rather than on-going speciation, but more scrutinized analysis will be performed soon. Due partially by the presence of both COI groups in all surveyed localities, the genetic diversity estimates of all localities are similar from the perspective of COI gene, but ITS data showed extremely lower genetic diversity of one islet locality, Anmyeon-do (locality 2; 0.002530 vs. 0.008054 ~ 0.012060). Analysis of gene flew estimates between localities indicates that most populations are highly interconnected to each other. However, one islet locality, Anmyeon-do (locality) has shown statistically significant distance from the remaining localities on the basis of only ITS2 data (FST = 0.19 ~ 0.34), requiring scrutinized phylogeographic inference on this population with expanded sampling. As more scrutinized analysis is performed, further fruitful inference on the geographic contour of the species might be available.

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.

Ionizing treatments were applied at 5Gy, 10Gy, 50Gy, 100Gy and 500Gy to mushroom mycelium (Lentinula edodes) in order to assesss the effect of the gamma-ray radiation. We mutated by gamma irradiation, 12 strains were isolated. 9 strains of 12 strains were appeared antagonistic interaction on solid medium. Growth rate of mycelium in JMIR-3 and JMIR-6 strains were similar to their control. But, the growth rate of strain JMIR-1, JMIR-2, JMIR-4, JMIR-5, JMIR-7, JMIR-8, JMIR-9, JMIR-10, JMIR-11 and JMIR-12 had generally different compared to control. In the sequence of ITS region of selected strains, it was revealed that the total length ranged from 696 to 780bp. The reciprocal homologies of the ITS region sequences in 5Gy irradiation group (JMIR-5, JMIR-6, JMIR-7 and JMIR-8) and 10Gy irradiation group (JMIR-9, JMIR-10, JMIR-1 and JMIR-12) indicated 99%. The reciprocal homologies of the ITS region sequences in 500Gy irradiation group (JMIR-1, JMIR-2, JMIR-3 and JMIR-4) indicated from 95 to 99%. It seemed that mycelium growth and ITS sequence could be changed the irradiation dose of the gamma-ray radiation.

The bumblebee, Bombus ignitus (Hymenoptera: Apidae), is a valuable natural resource that is one of the most notably utilized for greenhouse pollination in Korea. In order to understand the nature of genetic relationships, gene flow, and population structure of the species we sequenced a partial COI gene of mitochondrial DNA (mtDNA) corresponding to “animal barcode” region and the complete internal transcribed spacer 2 (ITS2) of the nuclear ribosomal DNA (nrDNA) collected from Korean localities. Although the 658-bp long mtDNA sequence provided only six haplotypes with the maximum sequence divergence of 0.61% (4 bp), the ITS sequences provided 84 sequence types with the maximum sequence divergence of 1.02% (21 sites), confirming better applicability of the ITS sequences to the study of intraspecific variation. The complete ITS2 sequences of B. ignitus were shown to be longest among known insects, ranging in size from 2,034 bp ~ 2,052 bp, harboring two duplicated repeats. Overall, a very high per generation migration ratio, a very low level of genetic fixation, and no discernable hierarchical population/ population group were noted to exist among populations of B. ignitus on the basis of both molecules, thus suggesting that the B. ignitus populations on the Korean peninsula are panmictic, which is consistent with our understanding of the dispersal capability of the species

In an effort to gain a better understanding of the nature of the population genetic structure of the pest insect, Spodoptera litura (Lepidoptera: Noctuidae), tobacco cutworms were collected from six Korean and five Chinese localities and their mitochondrial A+T-rich region and nuclear internal transcribed spacer 2 (ITS2) regions were cloned and sequenced. A total of 106 A+T-rich region haplotypes and 92 ITS2 sequence types were obtained from 158 individuals. Compared to the preliminary mitochondrial COI gene sequence data that provided all identical sequence types in all samples, the A+T-rich region and nuclear ITS2 are highly variable. Overall, a low level of genetic fixation in the A+T-rich region (FST = 0 ~ 0.02965) and nuclear ITS2 (FST = 0 ~ 0.34491), and no discernable isolated population was noted to exist among most S. litura populations. The presence of a majority of within-population variations, rather than variations among populations or between China and Korea, indicates that the S. litura populations are composed of heterogeneous individuals. The fixation index at hierarchical level of among regions shows no significance, suggesting that the S. litura populations in both Korea and China are profoundly interrelated with one another. This finding is in accord with the current knowledge that S. litura has sufficient flight capacity for dispersal.

최근들어 수입량이 급증되고 있는 차가버섯의 분류체계를 확립하고, 이들 종 및 계통간의 유연관계 확립과 품종 구분을 위해 계통학적 정보를 지닌 ITS 영역의 염기서열, PCRRFLP 및 STS 프라이머를 사용하여 종 특이적인 마커를 개발하였다. 시베리아 캄차카 반도에서 수집된 74008 균주의 염기서열을 이용하여 Inonotus spp.의 유연관계를 분석한 결과 2개의 그룹으로 나뉘어 졌고, I. obliquus DSM 856P균주와 약 98%의 가장 높은 유사성을 나타내어 차가버섯임이 확인되었다. 또한 ITS 증폭산물을 제한효소로 처리하여 밴드패턴을 비교하였을 때 종 및 계통에 따라 밴드가 다르게 나타났으며, STS primer를 이용하여 증폭산물을 비교하였을 때 종간에는 밴드패턴이 다르나 계통내에서는 동일한 밴드패턴을 보였다. 따라서 차가버섯의 품종 구분을 위해서는 STS 마커와 PCR-RFLP를 동시에 사용함으로서 품종 구분이 좀 더 명확하리라 사료된다.

인공재배된 만가닥버섯(Hypsizygus mamoreus )과 잿빛만가닥버섯(Lyophyllum decastes )을 ITSⅠ^Ⅳ 부위의 염기서열에 의해 종속간 유연관계 및 RAPD 다형성을 분석하였다. ITSⅠ^Ⅳ영역부위 종속간 유연관계에서 Group1(SPA 100, 101, 102)은 만가닥버섯 에 속하였으며, Group2(11균주) 잿빛만가닥버섯 의 대조 분리군 11균주는 동일한 종으로 동정되었다. ITS결과 14개 균주 시 4개 그룹으로 분류되었으며, ClusterⅠ과 ClusterⅡ는 58%의 유사도를 ClusterⅢ과 ClusterⅣ는 41%의 유사성을 보였다. 또한 인공재배한 잿빛만가닥버섯의 종 다양성을 분석하기위해 RAPD를 수행한 결과 가장 수량이 양호하며 우량계통인 SPA 202는 잿빛만가닥버섯인 Lyophyllum decastes SPA 203과 그룹화 되었으며 75%의 유사성을 보여주었고, Lyophyllum decastes 공시균주인 SPA 103과 SPA 104의 유사성은 65%로 나타났다.

지렁이과 (family Megascolecidae) 왕지렁이속 (genus Amynthas) 그룹을 rDNA ITS 유전자를 이용하여 계통 유연관계를 알아보고자 하였다. 2과 10속에서 26종의 DNA 염기서열을 이용하여 종간 계통적 유연관계를 MP (Maximum Parsimony), NJ (Neighbor Joining), QP (Quartet Puzzling)로 계통도를 작성하였다. 염기서열이 확보된 Megascolecidae (지렁이과)의 국내외