• Journal of Life Science
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• v.11 no.2
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• pp.126-134
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• 2001

The phylogenetic tree displayed the presence of five groups in the Phellinus genus, which were distinguished based on their morphology. Most of the p. linteus appeared a cluster which was highly significant with the exception of P. linteus KACC 500122 and KACC 500411. They formed the sister taxa of P 1inteus where P. baumii, Phellinus sp. MPNU 7003, MPNU 7007, and MPNU 7010 had similar morphological characteristics. Also, P. nigricans IMSNU 32024 and P. pini var, carniformans IMSNU 32031 were grouped in the same cluster with P. igniarius KCTC 6227, KCTC 6228, and P. chrysoloma KCTC 6225 extracted from the Gen-Bank database. P. torulosus IMSNU 32028 and Phellinus sp. MPNU 7011 formed a closed group, however, these species had a distant taxa when compared with the other Phellinus species. The nucleotide sequences of the internal transcribed spacer (ITS) regions of ribosomal DNA (rDNA) including the 5.85 rDNA were determined from 24 strains of the Phellinus genus in order to analyze their phylogenetic relationship. These fungi were divided into two basic groups based on their ITS length, however, this grouping was different from that based on their morphological characteristics. Although various ITS sequences were ambiguously aligned, conserved sites were also identified. Accordingly, a neighbor-joining tree was constructed using the nucleotide sequence data of the conserved sites of the ITS regions and the 5.8S rDNA.

• Korean Journal of Plant Taxonomy
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• v.41 no.2
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• pp.130-137
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• 2011

Pedicularis growing at Mt. Halla of Jeju Island is known as an endemic species of P. hallaisanensis Hurusawa. On the other hand, the plant is morphologically similar to P. amoena, P. spicata, and P. verticillata in gross morphology, so the taxonomic treatment of the taxon remains controversial. To clarify the taxonomic position of the plants, we examined external morphological characters and nuclear ribosomal DNA ITS sequences for P. hallaisanensis and its related species. The plants of Mt. Halla are clearly different from P. amoena and P. verticillata in the morphology of calyx lobes, the length of galea and lower lip, density of glandular hairs on plants, presences of the radical leaves after anthesis and molecular data. However, P. hallaisanensis is not clearly separated from P. spicata distributed in N. E. Asia on external morphological characters and DNA sequences of internal transcribed spacers. In this study, the morphological and molecular data suggested that P. hallaisanensis should be merged into the former species.

• The Korean Journal of Mycology
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• v.49 no.3
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• pp.417-423
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• 2021

Apple decline symptoms were frequently observed on cv. Fuji apple orchards located in Gyeonggi, Gyeongbuk, and Gangwon provinces during surveys conducted from May until the end of September 2020. Three fungal strains were isolated from the margins of internal lesions of diseased apple trees, and their morphological characteristics were considered similar to Botryosphaeria sinensis. Phylogenetic analysis using internal transcribed spacer (ITS) regions, translation elongation factor 1-alpha (tef1), beta-tubulin (tub2), and the second largest subunit of RNA polymerase II (rpb2) gene sequences confirmed the closest relationship of isolates with B. sinensis at the species level. According to a pathogenicity test, the appearance of dark-brown discolorations and vascular necrosis on apple branches inoculated with the isolated strain KNUF-20-014 was observed. To the best of our knowledge, this is the first report of B. sinensis as the causal agent of apple disease in Korea.

• Mycobiology
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• v.51 no.2
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• pp.109-113
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• 2023

Auricularia is one of the broadly cultivated edible mushrooms in Korea. Most of the Korean Auricularia strains used for cultivation and breeding are known as A. auricula-judae. Recently, this species has been reported to belong to a species complex. Therefore, this study was carried out to genetically clarify the bred and cultivated Korean A. auricula-judae strains. The internal transcribed spacer (ITS) and IGS1 rDNA region sequences were determined from 10 A. auricula-judae strains by PCR and sequencing. Variation in the nucleotide sequence and sequence length of the two rDNA regions were found among the seven A. auricula-judae strains. A maximum-likelihood (ML) phylogenetic tree based on the ITS sequences clearly placed all the 10 Korean A. auricula-judae strains in the A. heimuer clade of the A. auriculajudae complex. A. heimuer is diverged from A. auricula-judae. An ML phylogenetic tree based on the IGS1 sequences revealed the close relationship between Korean A. heimuer strains to Chinese A. heimuer strains. But each strain could be distinguishable by the IGS1 sequence. Furthermore, progeny strains in the seven Korean strains could be differentiated from their parental strains by the IGS1 sequence based phylogenetic tree. Our results are expected to be used to complement the distinction of domestic Auricularia cultivars.

• Journal of Life Science
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• v.22 no.11
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• pp.1470-1476
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• 2012

A genetic variation of Lepista nuda and two genus Lepista species (L. irina and L. sordida) were analyzed by random amplified polymorphic DNA (RAPD) and internal transcribed spacer (ITS) sequence analysis. In the resulting RAPD analysis, 22 out of 40 random primers amplified polymorphic RAPD fragment patterns, the amplified bands were 355, and DNA fragment sizes were 200-400bp. Intraspecific genetic dissimilarity of the 10 L. nuda strains were calculated to range from 0% to 21.60%, L. sordida from 16.93% to 24.82%, L. irina were 20.62% to 25.54%, and intraspecific genetic dissimilarity of L. sordida and L. irina was 23.49%. The 673 base pairs were sequenced during the analysis of the ITS I and II region; six L. nuda strains intraspecific genetic dissimilarities ranged from 1.58% to 11.47%, L. nuda and L. sordida from 3.83% to 12.88%, L. nuda and L. irina from 7.11% to 15.61%, and intra-specific genetic variation between L. sordida and L. irina was 4.79%. The findings showed that RAPD and ITS sequencing could be used for developing molecular genetic markers and screening of unidentified genus Lepista species.

• The Korea Journal of Herbology
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• v.29 no.6
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• pp.45-53
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• 2014

Objectives : Due to the morphological similarity of in the roots of herbal medicine, the official herbal medicine is very difficult to authenticate between the original plants of Patriniae Radix and two adulterant Patrinia species. Therefore, we introduced DNA barcode analysis to establish a powerful tool for the authentication of Patriniae Radix from its adulterants. Methods : To analyze DNA barcode regions, genomic DNA was extracted from twenty-nine specimens of Patrinia scabiosaefolia, Patrinia villosa, Patrinia saniculifolia, and Patrinia rupestris, and internal transcribed spacer 2(ITS2), matK and rbcL genes were amplified. For identification of species specific sequences, a comparative analysis was performed by the ClastalW based on entire sequences of ITS2, matK and rbcL genes, respectively. Results : In comparison of three DNA barcode sequences, we identified 22, 22, and 12 species-specific nucleotides enough to distinguish each four species from ITS2, matK and rbcL gene, respectively. The sequence differences at the corresponding positions were available genetic marker nucleotides to discriminate the correct species among analyzed four species. These results indicated that comparative analysis of ITS2, matK and rbcL genes were useful genetic markers to authenticate Patriniae Radix. Conclusions : The marker nucleotides enough to distinguish P. scabiosaefolia, P. villosa, P. saniculifolia, and P. rupestris, were obtained at 22 SNP marker nucleotides from ITS2 and matK DNA barcode sequences, but they were confirmed at 12 SNP marker nucleotides from rbcL. These differences could be used to authenticate Patriniae Radix from its adulterants as well as discriminating each four species.

• Journal of Mushroom
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• v.10 no.2
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• pp.93-99
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• 2012

This study was carried to identify a correct species and asses genetic diversity within the same species of Grifola spp. preserved in Division of applied Microbiology. Contaminated isolates showed different growth rates, morphology and color of hyphae. We have reconstructed the phylogenetic tree of a select group of Grifola spp. using nucleotide sequences of the internal transcribed spacer region(ITS) region. The phylogenetic tree was constructed by using the neighbor-joining method. PELF primers of 20-mer were used to assess genetic diversity of preserved isolates. Sequence analysis showed that four strains were identified completely different nomenclature. According to the analysis of ITS sequences, the genus Grifola clustered into one group, most of which correlated with species-groups identified by RAPD method. Eight isolates included strain GM01 showed high similarity with Grifola frondosa. All isolates were collected in the Japan(GM01, GM02, GM03) was identified as Grifola frondosa and isolates of the China(GM05, GM06, GM08) was identified as Bjerkandera fumosa, Grifola frondosa and Dichomitus squalens, respectively. RAPD analysis of genetic polymorphisms of genus Grifola showed a very different band patterns on the isolat. As the result of RAPD and ITS region sequences analysis for preserved isolates, it seems likely that 4 isolates of Grifola spp. may be need to reclassify or eliminate from preserved catalogue.

• Journal of Mushroom
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• v.9 no.1
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• pp.27-33
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• 2011

This study was carried to identify a correct species and asses genetic diversity within the same species of Trametes spp. preserved in Division of applied Microbiology The morphological and cultural characteristics of preserved strains were observed through microscope and investigated on PDA, respectively. Contaminated isolates showed different growth rates, morphology and color of hyphae. We have reconstructed the phylogenetic tree of a select group of Trametes spp. using nucleotide sequences of the internal transcribed spacer region(ITS) region. The phylogenetic tree was constructed by using the neighbor-joining method. PELF primers of 20-mer were used to assess genetic diversity of preserved isolates. Sequence analysis showed that five strains were different species and six strains were identified completely different nomenclature. According to the analysis of ITS sequences, the genus Trametes clustered into four distinct group, most of which correlated with species-groups identified by RAPD method. Seven isolates included TM 01 strain showed high similarity with Trametes versicolr, TM 07 and TM 10 high similarity with Trametes gibbosa, and TM 05 high similarity with Trametes elegans. But isolates collected in the United States was identified as T. junipericola. T. gibbosa and T. versicolor by RAPD analysis of genetic polymorphisms showed a very different band patterns and these strains showed different band patterns on areas. As the result of RAPD and ITS region sequences analysis for preserved isolates, it seems likely that 11 isolates of Trametes spp. may be need to reclassify or eliminate from preserved catalogue.

• Journal of Mushroom
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• v.10 no.1
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• pp.37-43
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• 2012

This study was carried to identify a correct species and asses genetic diversity within the same species of Polyporus spp. preserved in Division of applied Microbiology. Contaminated isolates showed different growth rates, morphology and color of hyphae. We have reconstructed the phylogenetic tree of a select group of Polyporus spp. using nucleotide sequences of the internal transcribed spacer region(ITS) region. The phylogenetic tree was constructed by using the neighbor-joining method. PELF primers of 20-mer were used to assess genetic diversity of preserved isolates. Sequence analysis showed that three strains were different species and four strains were identified completely different nomenclature. According to the analysis of ITS sequences, the genus Polyporus clustered into five distinct group, most of which correlated with species-groups identified by RAPD method. Four isolates included strain PM02 showed high similarity with P. arcularius, four isolates included strain PM03 high similarity with P. alveolaris, three isolates included strain PM01 high similarity with P. tuberaster, and PM 06 and PM04 high similarity with P. brumalis and P. squamossus. Isolates were collected in the United States(PM10, PM11) was identified as P. alveolarius and P. arcularius. RAPD analysis of genetic polymorphisms of genus Polyporus showed a very different band patterns. As the result of RAPD and ITS region sequences analysis for preserved isolates, it seems likely that 6 isolates of Polyporus spp. may be need to reclassify or eliminate from preserved catalogue.

• The Korea Journal of Herbology
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• v.30 no.3
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• pp.41-47
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• 2015

Objectives : Due to the morphological similarity of the pericarp and description of multi-species in National Pharmacopoeia of Korea and China, the Zanthoxylum Pericarpium is difficult to authenticate adulterant in species levels. Therefore, we introduced the sequence analysis of DNA barcode and identification of single nucleotide polymorphism(SNP) to establish a reliable tool for the distinction of Zanthoxylum Pericarpium from its adulterants. Methods : To analyze DNA barcode region, genomic DNA was extracted from twenty-four specimens of authentic Zanthoxylum species and inauthentic adulterant and the individual internal transcribed spacer regions (rDNA-ITS and ITS2) of nuclear ribosomal RNA gene were amplified using ITS1, ITS2-S2F, and ITS4 primer. For identification of species-specific sequences, a comparative analysis was performed using entire DNA barcode sequences. Results : In comparison of four Zanthoxylum ITS2 sequences, we identified 16, 4, 6, and 4 distinct species-specific nucleotides enough to distinguish Z. schinifolium, Z. bungeanum, Z. piperitum, and Z. simulans, respectively. The sequence differences were available genetic marker to discriminate four species. Futhermore, phylogenetic relationship revealed a clear classification between different Zanthoxylum species showing 4 different clusters. These results indicated that comparative analysis of ITS2 DNA barcode was an useful genetic marker to authenticate Zanthoxylum Pericarpium in species levels. Conclusions : The marker nucleotides, enough to distinguish Z. schinifolium, Z. piperitum, Z. bungeanum, and Z. simulans, were obtained at 30 SNP marker nucleotides from ITS2 sequences. These differences could be used to authenticate official Zanthoxylum Pericarpium from its adulterants as well as discriminating each four species.