• Title/Summary/Keyword: rDNA internal transcribed spacer sequence

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Westerdykella reniformis: A New Record from Field Soils in Korea

  • Adhikari, Mahesh;Kim, Sang Woo;Gwon, Byeong Heon;Ju, Han Jun;Lee, Hyang Burm;Lee, Youn Su
    • The Korean Journal of Mycology
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    • v.48 no.1
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    • pp.47-53
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    • 2020
  • During a survey of fungal diversity in different provinces of South Korea in 2017, a new fungal isolate was discovered. This fungal isolate was identified as Westerdykella reniformis, based on its morphological characteristics and phylogenetic analysis, using internal transcribed spacer (ITS) and 28S ribosomal DNA (28S rDNA) sequence data. To our knowledge, W. reniformis has not previously been reported in South Korea. Thus, in this study, we report a new record of a species from the Dothideomycetes class in Korea, and provide a detailed description with morphological illustrations.

Phylogenetic Analysis of Phellinus linteus and Related Species Comparing the Sequences of rDNA Internal Transcribed Spacers

  • Lee, Jae-Dong;Kim, Gi-Young;Park, Joung-Eon;Park, Hyung-Sik;Nam, Byung-Hyouk;An, Won-Gun;Lee, Tae-Ho
    • 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.

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Identification of Aspergillus Strain with Antifungal Activity Against Phytophthora Species

  • KANG SUNG WOO;HONG SUK IN;KIM SEUNG WOOK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.227-233
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    • 2005
  • Fungal strain CGF was isolated from the soil of ChungNam Province, South Korea. Based on the 28S rDNA sequence analysis and the sequence of the internal transcribed spacer (ITS) region of ribosomal DNA, together with morphological and cultural characteristics, this strain was identified as Aspergillus sclerotiorum and renamed Aspergillus sclerotiorum CGF. This is the first strain of Aspergillus sclerotiorum identified in Korea. When the antifungal activity of A. sclerotiorum CGF was evaluated, among the phytopathogenic fungi, mycelial growth of only Phytophthora species was inhibited. Oermination of P. capsid zoospore was also inhibited. The bioactive compound of A. sclerotiorum CGF was highly thermo- and pH-stable.

A Duplex PCR for Detection of Phytophthora katsurae Causing Chestnut Ink Disease (밤나무 잉크병균, Phytophthora katsurae의 검출을 위한 Duplex PCR)

  • Lee, Dong-Hyeon;Lee, Sun-Keun;Kim, Hye-Jeong;Lee, Sang-Hyun;Lee, Sang-Yong;Lee, Jong-Kyu
    • Research in Plant Disease
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    • v.18 no.2
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    • pp.73-79
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    • 2012
  • Phytophthora katsurae is a fungal pathogen responsible for chestnut ink disease. We designed two duplex primer sets (SOPC 1F/1R+KatI 3F/5R, SOPC 1-1F/1-1R+KatI 3F/5R) to detect P. katsurae. SOPC 1F/1R and SOPC 1-1F/1-1R primer pairs were designed for sequence characteristic amplification regions (SCAR) marker, and KatI 3F/5R primer pair was used for P. katsurae-specific primer designed from internal transcribed spacer (ITS) region. To assess the sensitivity of duplex PCR, genomic DNA was serially diluted 10-fold to make the final concentrations from 1 mg/ml to 1 ng/ml. The sensitivity for two primer sets were 1 ${\mu}g/ml$ and 100 ng/ml, respectively. To find detection limits for zoospores of P. katsurae, each zoospore suspension was serially diluted 10-fold to make the final concentrations from $1{\times}10^6$ to $1{\times}10^2$ cells/ml, and then DNA was extracted. The limits of detection for all of two primer sets were $1{\times}10^5$ cells/ml. All of two primer sets were specific to P. katsurae in PCR detection and did not produce any P. katsurae-specific PCR amplicons from other 16 Phytophthora species used as the control. This study shows that duplex PCR using two primer sets might be a useful tool for rapid and efficient detection of P. katsurae.

Development of Molecular Markers for the authentication of Zanthoxyli Pericarpium by the analysis of rDNA-ITS DNA barcode regions (rDNA-ITS DNA 바코드 부위 분석을 통한 산초(山椒) 기원종 감별용 유전자 마커 개발)

  • Kim, Wook Jin;Ji, Yunui;Lee, Young Mi;Kang, Young Min;Choi, Goya;Moon, Byeong Cheol
    • 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.

Phylogenetic Analysis of Caterpillar Fungi by Comparing ITS 1-5.8S-ITS 2 Ribosomal DNA Sequences

  • Park, Joung-Eon;Kim, Gi-Young;Park, Hyung-Sik;Nam, Byung-Hyouk;An, Won-Gun;Cha, Jae-Ho;Lee, Tae-Ho;Lee, Jae-Dong
    • Mycobiology
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    • v.29 no.3
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    • pp.121-131
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    • 2001
  • This study was carried out to identify the phylogenetic relationships among several caterpillar fungi by comparing the sequences of internal transcribed spacer regions(ITS1 and ITS2) and 5.8S ribosomal DNA(rDNA) repeat unit. The sequences of ITS1, ITS2, and the 5.8S rDNA from 10 strains of Cordyceps species, 12 strains of Paecilomyces, 3 strains of Beauveria, 2 strains of Metarhizium and 1 strains of Hirsutella were amplified, determined and compared with the previously known Cordyceps species. The sequences of 5.8S rDNA were more conserved in length and variation than those of ITS regions. Although the variable ITS sequences were often ambiguously aligned, the conserved sites could be found. In the phylogenetic tree, the species generally divided into three clusters, supported by their morphology and/or host ranges. The 5.8S rDNA and TTS1 sequences among 10 species of Cordyceps militaris were identical and only one base pair in ITS2 sequence was different. Cordyceps sinensis and Cordyceps ophioglossoides were also clearly different, although they belonged to the same cluster. The Geniank database search of species revealed sister taxa of an entomogenous fungus. Metarhizium was used as an putgroup in all taxa.

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Limits of Direct PCR Amplification from Seaweeds Using Arbitrary and ITS Primers (해조류로부터 Arbitrary 및 ITS Primer들을 사용한 직접 PCR 유전자 증폭반응의 한계)

  • 김용국;진형주;박선미;진덕희;홍용기
    • Journal of Life Science
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    • v.9 no.1
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    • pp.15-21
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    • 1999
  • The random amplified ploymorphic DNAs (RAPD) assay is a simple and useful tool in identification of appropriate genetic markers, that requires no knowledge of target DNA sequence. RAPD products were generated directly from seaweed tissues, without prior nucleic acid extraction, of Porphyra yezoensis, Ulva pertusa and Undaria pinnatifida. The nuclear rDNA internal transcribed spacer (ITS) fragment however was not amplified directly from the seaweed tissues. Using DNA extracted by the LiCl method, both the ITS and RAPD's have been amplified by the polymerase chain reaction. RAPD of P yezoensis, thallus (n) and conchocelis (2n) produced lots of different polymorphic bands (36-50$\%$) depending on the arbitrary primers used. Difference was also observed between direct tissues amplification and DNA extracts amplification (53-57$\%$). Thus it is important to use the same ploidy of tissue for DNA extraction and as a RAPD template.

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First Report of Chrysanthemum (Chrysanthemum morifolium) Crown Rot Caused by Fusarium solani in Korea

  • Mun, Hye-Yeon;Jeong, Je-Yong;Kim, Chang-Jeon;Lee, Hyang-Burm
    • The Plant Pathology Journal
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    • v.28 no.1
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    • pp.49-54
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    • 2012
  • In August 2010, a severe crown rot was observed on chrysanthemum ($Chrysanthemum$ $morifolium$ Ramat., variety Sinro) in several greenhouses located at Damyang and Muan, Jeonnam province, Korea. Three isolates (EML-CHS1, -CHS2, and -CHS3) of $Fusarium$ were isolated from the affected plants and identified based on morphological characteristics and rDNA internal transcribed spacer (ITS) sequence analysis. Sequence analysis by BLAST indicated that EMLCHS1, -CHS2 and CHS3 were closest to a $Fusarium$ species, $F.$ $solani$ with > 99% sequence similarity. Pathogenicity tests were performed on chrysanthemum with spore suspensions containing $3.4{\times}10^6$ spores/ml using the dipping method. Ten days after inoculation, similar symptoms to those observed in the greenhouses were seen on the inoculated plants. The causal fungus was reisolated from the artificially inoculated basal stems, fulfilling Koch's postulates. To our knowledge, this is the first report of crown rot by $Fusarium$ $solani$ on chrysanthemum ($Chrysanthemum$ $morifolium$) in Korea.

Two Unreported Glomeromycota Fungi Isolated from Rhizospheres of Smilax china (청미래덩굴의 근권에서 분리된 2종의 Glomeromycota 미기록종)

  • Park, Hyeok;Ka, Kang-Hyeon;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.47 no.3
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    • pp.275-280
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    • 2019
  • We isolated fungal spores belonging to the phylum Glomeromycota from the rhizospheres of Smilax china, cultured in a greenhouse. We identified the isolated spores using sequence analysis of 18S partial rDNA region, internal transcribed spacer and 28S rDNA regions. We confirmed 2 unreported spores of Glomeromycota fungal species, Diversispora eburnea and Paraglomus laccatum. Here, we described the morphological characteristics and results of phylogenetic analysis of the confirmed species.

Occurrence of Stem Canker and Tuber Rot on Yam Caused by Rhizoctonia solani AG 2-2IIIB in Korea (Rhizoctonia solani AG 2-2IIIB에 의한 마 뿌리썩음병의 한국 내 발생)

  • Hong, Sung Kee;Lee, Jae Kook;Lee, Young Kee;Lee, Sang Yeob;Kim, Wan Gyu;Shim, Hong Sik
    • The Korean Journal of Mycology
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    • v.40 no.4
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    • pp.266-270
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    • 2012
  • Stem canker and tuber rot symptoms were observed on yam grown in Andong and Jinju, Korea in 2011. A total of 20 isolates of Rhizoctonia and allied fungi were obtained from the symptomatic plants. Among the isolates, 8 isolates were identified as Rhizoctonia solani and 12 isolates as Ceratobasidium sp. based on rDNA-internal transcribed spacer (ITS) sequence similarity. In the cluster analysis of rDNA-ITS sequences, 7 isolates of R. solani belonged to AG 2-2IIIB and remaining one to AG 1-1A. In addition, among the 12 isolates of Ceratobasidium sp., 7 isolates belonged to AG-Fa, three isolates to AG-A and the other two isolates to AG-Fb and AG-O, respectively. Pathogenicity tests showed that all the R. solani AG 2-2IIIB isolates are pathogenic on stem and tuber of yam but R. solani AG 1-1A and all the Ceratobasidium isolates are non-pathogenic. The results indicate that R. solani AG 2-2IIIB is an important pathogen causing stem canker and tuber rot on yams grown in the study areas. This is the first report of R. solani AG 2-2IIIB causing stem canker and tuber rot of yam in Korea.