• The Korean Journal of Mycology
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• v.28 no.2
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• pp.112-117
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• 2000

A similarity coefficient were analyzed by RFLP of fourteen species of entomopathogenic fungi, isolated from inhabiting pupa and adult insect at forest. Each rDNA ITS I and ITS II with primers of ITS 1 and ITS 4 was amplified by PCR. The amplified products were conserved to 500 bp were not demarcated between genus and species. Four Paeciliomyces tenuipes, two Beauveria bassiana and six Cordyceps militaris were treated by seven restriction enzymes and confirmed in species except JB3 by electrophoresis band. However, the band of C. scarabaeicola showed the identity with B. bassiana. The result of this experiment indicated that the teleomorph of C. scarabaeicola was the same as that of B. bassiana. CfoI and HpaII of seven restricted enzymes were easily discriminating in the genus between Paecilomyees and Cordyceps. Especially, CfoI was more effective to classify the genera of Paecilomyees, Cordyceps and Beauveria than other restriction enzymes. The band patterns of RFLP of P. tenuipes, C. militaris, C. scarabaeicola and B. bassiana were also analyzed by UPGMA program of NTSYS-pc and showed 100% significance. Thus, the similarity coefficient tended to be lower between genera by RFLP analysis, but was higher between species.

• Journal of Veterinary Clinics
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• v.25 no.3
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• pp.207-210
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• 2008

Trichophyton erinacei is a dermatophyte pathogen that infects both humans and hedgehogs. A two-month old female four-toed hedgehog presented to the Chonbuk Animal Medical Center with pruritus, excoriation and crust on her face for ten days. The owner of the hedgehog also exhibited the clinical signs of scaly erythema with fine vesicles on her neck. A presumptive diagnosis of dermatophytosis was made based on the results of an acetate tape preparation in which hyphae and chains of arthroconidia were observed. The crusts from the lesions were then cultured on Sabouraud Dextrose Agar for identification. After 10 days of incubation, downy colored colonies that had a central umbo with a white granular surface and a yellow pigment ring in the reverse were observed. Microscopic analysis revealed the presence of numerous teardrop shaped microconidia singly attached to the sides of the hyphae. In addition, 2-6 roomed macroconidia that were somewhat irregular in shape and size were present, and abundant intermediate sized spores were observed between the micro and macro conidia. To confirm that the culture was T. erinacei, the internal transcribed spacer region of the 5.8S phase of the ribosomal RNA gene (ITS1-5.8S-ITS2 rDNA) was amplified by PCR and then sequenced. A 679-base pair fragment of DNA was then compared with sequences in GenBank and found to be 99% homologous with sequences of T. erinacei (Z97997 and Z97996. The clinical signs were resolved after four weeks of treatment with oral and topical ketoconazole and chlorhexidine. To the best of our knowledge, this represents the first case of T. erinacei isolated from a four-toed hedgehog in Korea.

• Korean Journal of Plant Resources
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• v.19 no.5
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• pp.628-633
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• 2006

ITS DNA sequences from five individuals, representative of five groups designated according to the degree of leaf teeth and lobes from simple to palmate compound leaf in the Viola albida complex, established and further analysed in order to solve the taxonomic difficulty. A total 702 bp was sequenced at the 5.8S ribosomal DNA and internal transcribed spacer 1 and 2. The 5.8S coding region is 163 bp, and has no sequence variations. The ITS1 and ITS2 noncoding regions have a little bit sequence variations, and those were further analysed by the methods of the analysis of variance (ANOVA), the analysis of sequence divergence and the phylogenetic analysis. The result of ANOVA showed no significant differences among individuals investigated. The analysis of sequence divergence with Kimura 2-parameter distance revealed that in-groups showed much less than 0.05 in absolute value among individuals, while two out groups more than 0.05, V. grypoceras and V. orientalis. This result appeared that the sequence divergence among in-groups was not yet occurred in the species level but situated at somewhere below the species level. In the phylogenetic analysis, two outgroups formed the basal clades in order. Five individuals in-groups formed a clade. The clade was, however, not very robust as around 50% in bootstrap value, suggesting that this result was not meaningful in the phylogenetic point of views.

• Korean Journal of Plant Taxonomy
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• v.33 no.4
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• pp.373-386
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• 2003

Internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA were determined for 14 individuals representing eight taxa from Scopolia s. str. and related genera, Anisodus and Atropanthe. We found that the ITS sequences of Korean endemic species, S. parviflora, are significantly different from its allied species, S. japonica. This is contradictory to traditional taxonomic treatments in which those species are regarded as conspecific. S. parviflora exhibited closer relationship to S. carniolica, which is disjunctly distributed in Europe. In spite of substantially high sequence divergence between S. japonica and S. parvlflora/S. carniolica clade, morphological resemblance is evident among the species. Morphological stasis concept (retardation of morphological differentiation or evolution of similar characters among the disjuncts in a similar ecological habitat) was referred to understand this rather unusual evolutionary feature. S. lutescens, another Korean endemic species, shared almost identical ITS sequences with S. parviflora. Lack of diagnostic character distinguishing the taxa suggests that they are conspecific. Anisodus carniolicoides, which was originally described in Scopolia, was grouped with A. luridus and A. tanguticus. The monophyletic Anisodus formed a sister group relationship with a monotypic genus Atropanthe.

• Journal of Life Science
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• v.24 no.11
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• pp.1174-1179
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• 2014

This research was carried out to evaluate whether gamma ray is a useful tool for breeding new strains of mushrooms. For this research, 5 mutant groups, 20 strains of Hypsizigus marmoreus, 2 strains of Lyophyllum decastes, and 1 strain of Lyophyllum shimeji were used. Monokaryon spores from one variety of H. marmoreus were irradiated with 50~2,000 Gy of gamma ray. The propriety dose was 50~200 Gy for mutagenesis. Mutant monokaryon mycelia crossed each order to become dikaryon mycelia. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR, and the products were sequenced. The sequences of the ITS regions (16 partial rDNA, complete ITS1, 5.8 rDNA and partial rDNA) were analyzed by PCR, and strains of H. marmoreus, L. decastes, and L. shimeji were auto-sequenced. The lengths of the sequenced ITSs were 1,052~1,143 nucleotides. Genetic matrices were calculated using Nei-Li's genetic distance coefficient based on ITS sequence. The dissimilarities were 0~3.35% in strains of H. Hypsizigus. In addition, a phylogenetic tree was constructed based on ITS sequences using the neighbor-joining (NJ) method. The phylogenetic tree revealed that 23 strains and 5 mutant groups were divided into 12 clusters; the mutant groups fell into different clusters. These results show that mushroom spores were mutated effectively by gamma ray; therefore, gamma ray could be a useful tool for breeding new strains of mushrooms.

• Korean Journal of Plant Taxonomy
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• v.45 no.3
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• pp.243-253
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• 2015

Series Chinensis, Genus Iris, endemic to the far regions of East Asia, consists of four species and related varieties. This series is divided into two major groups (I. rossii and I. minutiaurea complex). In this study, the ITS region and matK gene sequences within nuclear ribosomal DNA and plastid DNA were analyzed in order to investigate the phylogenetic relationships among the I. minutiaurea complex (I. minutiaurea, I. odaesanensis, and I. koreana) and the taxonomic identities of a putative hybrid in Mt. Palgong. In the internal transcribed spacer (ITS1, 5.8S, and ITS2) region, a total of 106 cloned genomic sequences from three taxa were obtained to study the intragenomic polymorphisms of the ITS regions. Three taxa revealed high levels of intragenomic polymorphisms, indicative of incomplete nrDNA concerted evolution. This incomplete ITS concerted evolution in the series Chinensis may be linked to the recent species divergence and frequent interspecies hybridization of the series Chinensis. In the matK gene, three taxa were fairly separated by eleven variable sites. In eight individuals collected on Mt. Palgong, putative hybrids between I. odaesanensis and I. minutiaurea were clustered in the I. minutiaurea clade in the NJ (neighbor-joining) tree based on the matK gene. However, in the ITS tree, some of them were clustered in the I. odaesanensis clade and others were clustered in the I. minutiaurea clade. Therefore, the individuals on Mt. Palgong were formed by the hybridization between two taxa (I. odaesanensis and I. minutiaurea) and not through the lineage of I. koreana.

• Weed & Turfgrass Science
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• v.4 no.1
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• pp.18-25
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• 2015

A universal DNA barcoding for agricultural noxious weeds is a powerful technique for species identification without morphological knowledge, by using short sections of DNA from a specific region of the genome. Two standard barcode markers, chloroplast rbcL and matK, and a supplementary nuclear ribosomal Internal Transcribed Spacer (ITS) region were used to examine the effectiveness of the markers for Pooideae barcoding using 163 individuals of 29 taxa across 16 genera of Korean Pooideae. The rbcL and ITS revealed a good level of amplification and sequencing success while matK did not. Barcode gaps were 78.6% for rbcL, 96.2% for matK, and 91.7% for ITS, respectively. Resolving powers were 89.3% for rbcL, 92.3% for matK, and 79.1% for ITS. The matK obtained the best both barcode gap and resolving power. However, it should be considered not to employ matK for Pooideae barcode because of low rate of PCR amplification and sequencing success. As a single DNA marker, rbcL and ITS were reasonable for Pooideae barcode. Barcode gap and resolving power were increased when ITS was incorporated into the rbcL. The barcode sequences were deposited to the National Center for Biotechnology Information (NCBI) database for public use.

• Journal of Life Science
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• v.21 no.7
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• pp.992-996
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• 2011

Endophytic fungi were isolated from the roots of plants growing naturally on the island of Dokdo. Plant samples, such as Miscanthus sinensis, Achyranthus japonica and Echinochloa crusgali were isolated from Dongdo, and those such as Honkenya peploides and Artemsia koidzumii were isolated from Seodo. Twenty one strains of endophytic fungi were isolated from these plants. To identify the strains, PCR (polymerase chain reaction) amplification of the partial ITS (Internal Transcribed Spacer) regions was done with universal primers ITS-1 and ITS-4 to determine the nucleotide sequence of the ITS regions. Of the strains isolated from Miscanthus sinensis, 75% were Penicillium sp. and 25% were Aspergillus sp. Fifty five percent of strains isolated from Achyranthus japonica were Penicillium sp., 30% were Aspergillus sp. and 15% were Zygorhynchus sp. Strains isolated from Echinochloa crusgali were Penicillium sp. (50%), Aspergillus sp. (12%), Giberella sp. (13%), Talaromyces sp. (9%) and Umbelopsis sp. (8%). Of the strains isolated from Honkenya peploides, 76% were Penicillium sp. and 24% were Pestalotiopsis sp. Strains isolated from Artemisia koidzumii were Penicillium sp. (81%) and Mucor sp. (19%). As a result of bioassay, Ec-3-1 strain isolated from Echinochloa crusgalli showed plant growth-promotion activity. Of all the endophytic fungi isolated, Penicillium sp. was the most abundantly distributed fungal strain in all plants used in this study.

• Journal of Life Science
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• v.21 no.11
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• pp.1619-1624
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• 2011

Endophytic fungal strains were isolated from the roots of six species plants in the Dokdo islands. Native plant samples, such as Artemisia japonica, Chenopodium album and Solanum nigrum were isolated from Dongdo, and those such as Cyrtomium falcatum, Dianthus longicalyx and Tetragonia tetragonoides were isolated from Seodo. In total, thirty two fungal strains were isolated from these native plants. To identify the fungal strains, polymerase chain reaction (PCR) amplification of internal transcribed spacer (ITS: containing ITS1, 5.8s and ITS2 region) regions was done with universal primers ITS1 and ITS4. Endophytic fungi of four species were isolated from A. japonica, eight species from C. album, three species from S. nigrum, three species from C. falcatum, three species from D. longicalyx and eleven species from T. tetragonoides. Culture filtrates (CF) of isolated endophytic fungi were used to treatwaito-c rice seedlings to test plant growth-promoting activity. As a result of bioassay, Ca-5-2-2 strain isolated from C. album expressed highest plant growth-promotion activity. Of all the endophytic fungi isolated, Penicillium sp., Fusarium sp. and Aspergillus sp. were the most abundantly distributed fungal strains in the six plants used in this study.

• Korean Journal of Plant Taxonomy
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• v.37 no.2
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• pp.115-129
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• 2007

Using internal transcribed spacer (ITS) sequences and inter-simple sequence repeats (ISSRs) data, genetic diversity of a rare species, Hibiscus hamabo Siebold & Zucc. was examined for 3 populations in Jeju Island, Korea. A total of 14 nucleotide (excluding 3 ambiguous nucleotide) site variation in the ITS was observed from 18 individuals (Population 1, Hadori), which differed up to 13 bp in pair-wise comparison. On the contrary, the ITS sequences of all individuals in Populations 2 and 3 were identical. Genetic diversity estimates including Nei's gene diversity (h) generated by ISSR data were substantially high in Population 1 compared to other two populations. Low genetic variation in Populations 1 and 2 is considered due to genetic drift (bottleneck effect) and limited gene flow in these populations. Considering the differences in genetic diversity, protection of the Population 1(Hadori) is very critical for in situ conservation of Hibiscus hamabo in Korea. If ex situ conservation is required, making the full use of Population 1 will be most efficient.