• Title/Summary/Keyword: ITS region sequence

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Genetic Diversity and Phylogenetic Relationship in Korean Strains of Lentinus lepideus Based on PCR Polymorphism (PCR 다형성 분석에 의한 한국산 잣버섯의 유전적 다양성 및 유연관계)

  • Lee, Jae-Seong;Cho, Hae-Jin;Yoon, Ki-Nam;Alam, Nuhu;Lee, Kyung-Lim;Shim, Mi-Ja;Lee, Min-Woong;Lee, Yun-Hae;Jang, Myoung-Jun;Ju, Young-Chul;Cheong, Jong-Chun;Shin, Pyung-Gyun;Yoo, Young-Bok;Lee, U-Youn;Lee, Tae-Soo
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.105-111
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    • 2010
  • Lentinus lepideus, known as train wrecker fungus, has been used for nutritional and medicinal purposes. Recently, commercial cultivation technique and a new cultivar of the mushroom were developed. To investigate the genetic diversity and phylogenetic relationship for identifying the mushroom strains and cultivar, one commercial and 13 strains of Lentinus lepideus from different geographical regions of Korea were analyzed by ITS regions of rDNA and RAPD of genomic DNA. Three strains of Lentinus edodes were also used for the analysis. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 173 to 179 bp and 203 to 205 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical with 156 base pairs. A phylogenetic tree based on the ITS region sequences indicated that selected strains could be classified into four clusters, while 3 strains of L. edodes was divided into a new cluster. Ten primers out of 20 arbitrary primers used in the RAPD-PCR efficiently amplified the genomic DNA. The numbers of amplified DNA bands varied with the primers and strains, with polymorphic DNA fragments in the range from 0.2 to 2.6 kb. The results showed that phylogenetic relationship among Korean strains of Lentnus lepideus is high, but genetic diversity is low.

Comparative Molecular Phylogenetic Relationships in Different Strains of Pleurotus spp. (느타리속 버섯 계통의 분자생물학적 유연관계의 비교연구)

  • Cho, Hae-Jin;Lee, Jae-Seong;Yoon, Ki-Nam;Alam, Nuhu;Lee, Kyung-Lim;Shim, Mi-Ja;Lee, Min-Woong;Cheong, Jong-Chun;Shin, Pyung-Gyun;Yoo, Young-Bok;Lee, U-Youn;Lee, Tae-Soo
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.112-119
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    • 2010
  • Pleurotus spp. have been used for edible and medicinal purposes in Asian countries for a long time. The fruiting bodies of the Pleurotus ostreatus, Pleurotus citrinopileatus and Pleurotus salmoneostramineus contained many physiologically beneficial substances for human health. Therefore, it is necessary to study the genetic diversity of Pleurotus mushroom cultivars commercially cultivated in Korea. Eleven strains of Pleurotus spp. were collected from different geographical regions in South-East Asia and ITS regions of rDNA and RAPD of genomic DNA were analyzed. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 167 to 254 bp and 156 to 213 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, and the 5.8S sequences were identical. A phylogenetic tree based on the ITS region sequences indicated that selected strains could be classified into 4 clusters. Eleven Pleurotus species were also analyzed by RAPD with 20 arbitrary primers. Ten of these primers were efficiently amplified the genomic DNA. The number of amplified bands varied with the primers and strains, with polymorphic fragments in the range from 0.1 to 2.0kb. The results revealed that genetic diversity of selected strains of P. ostreatus, P. citrinopileatus and P. salmoneostramineus is low.

Molecular Analysis of Pathogenic Molds Isolated from Clinical Specimen (임상검체에서 분리된 병원성 사상균의 분자생물학적 분석)

  • Lee, Jang Ho;Kwon, Kye Chul;Koo, Sun Hoe
    • Korean Journal of Clinical Laboratory Science
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    • v.52 no.3
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    • pp.229-236
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    • 2020
  • Sixty-five molds isolated from clinical specimens were included in this study. All the isolates were molds that could be identified morphologically, strains that are difficult to identify because of morphological similarities, and strains that require species-level identification. PCR and direct sequencing were performed to target the internal transcribed spacer (ITS) region, the D1/D2 region, and the β-tubulin gene. Comparative sequence analysis using the GenBank database was performed using the basic local alignment search tool (BLAST) algorithm. The fungi identified morphologically to the genus level were 67%. Sequencing analysis was performed on 62 genera and species level of the 65 strains. Discrepancies were 14 (21.5%) of the 65 strains between the results of phenotypic and molecular identification. B. dermatitidis, T. marneffei, and G. argillacea were identified for the first time in Korea using the DNA sequencing method. Morphological identification is a very useful method in terms of the reporting time and costs in cases of frequently isolated and rapid growth, such as Aspergillus. When molecular methods are employed, the cost and clinical significance should be considered. On the other hand, the molecular identification of molds can provide fast and accurate results.

Investigation of Fungi in Pesticide Fumigated Oak Wilt-Diseased Logs (훈증방제 처리한 참나무시들음병 감염목의 사상균 조사)

  • Suh, Dong Yeon;Son, Seong-Yeol;Kim, Seong Hwan;Seo, Sang Tae;Kim, Kyung Hee;Ko, Han Kyu
    • The Korean Journal of Mycology
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    • v.40 no.4
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    • pp.288-291
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    • 2012
  • Korean oak wilt disease caused by Raffaelea quercus-mongolicae is vectored by the ambrosia beetle Platypus koryoensis. To prevent the spread of the disease, the beetle infested oak tree had been cut into logs, covered with plastic vinyl, fumigated with a pesticide, and stored for three years on the site where the tree was cut. This study was carried out to get information on the fungi colonizing the fumigated oak wood. Wood disk samples collected from the fumigated oak logs at two locations in the Taejo Mountain, Cheonan city, were used for fungal isolation. A total of 99 filamentous fungal isolates were obtained from the wood disk samples. Hypocrea spp., Trichoderma spp. and Penicillium spp. were identified based on morphological characteristics and nucleotide sequence analysis of translation elongation factor 1-alpha gene and ITS rDNA region. Trichoderma was the major fungal group. R. quercus-mongolicae, and P. koryoensis were not detected from the fumigated oak wood. Our work provided evidence that after three years of storage, the fumigated oak wilt-diseased logs should be no longer harmful source of oak wilt disease transmission.

Plant growth-promoting activity and identification of endophytic fungi isolated from native plant in East coast (동해안 자생식물로부터 분리된 내생균류의 식물생장촉진활성 및 동정)

  • You, Young-Hyun;Jin, Yong Ju;Kang, Sang-Mo;Oh, Sejong;Lee, Myung-Chul;Kim, Jong-Guk
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.14-20
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    • 2015
  • Coastal plant species, Plantago camtschatica Cham. native to the coastal region of the East Sea were sampled and then morphologically different 20 endophytic fungal strains were purely isolated. Phylogenetic analysis of isolates was done by the Bayesian program based on sequenced internal transcribed spacer (ITS-rDNA) region. Culture filtrates of each of 20 isolates were treated to Waito-c rice (WR) seedlings for verifying plant growth-promoting activity, respectively. As the results, E/PC/10/1 strain showed the highest plant growth-promoting activity among them. The culture filtrate of the strain E/PC/10/1 was revealed as containing gibberellins ($GA_1$, $GA_3$, $GA_4$) by using HPLC, and gas GC/MS with selected ion monitoring (SIM). Finally, this strain was identified as novel Penicillium spinulosum species that producing new GAs with microscopic observation and further molecular analysis with beta-tubulin gene sequence.

Occurrence of Leaf Spot Caused by Stemphylium lycopersici on Cirsium setidens in Korea (Stemphylium lycopersici에 의한 고려엉겅퀴 점무늬병의 발생)

  • Choi, Hyo-Won;Kim, Seok Gu;Hong, Sung Kee;Lee, Young Kee;Lee, Jae Guem;Kim, Hyo Won;Lee, Eun Hyeong
    • The Korean Journal of Mycology
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    • v.44 no.3
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    • pp.201-205
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    • 2016
  • In August 2015, leaf spot symptoms were observed on Korean gondre thistle (Cirsium setidens) in Youngwol, Korea. During the early stage, the symptoms appeared as one or more small gray-brown to brown spots on plant leaves. The spots showed extensive enlargement over time and eventually became large dark brown to black lesions on the whole leaf. Stemphylium species were consistently isolated from affected leaves. All isolates were identified as S. lycopersici, S. solani, or S. xanthosomatis based on morphological and cultural characteristics. The isolates were confirmed as S. lycopersici based on a multilocus sequence analysis using the ribosomal internal transcribed spacer (ITS) region, elongation factor 1, GAPDH (glyceraldehyde-3-phosphate dehydrogenase), and the noncoding region between the vacuolar membrane ATPase catalytic subunit A gene and a gene involved in vacuolar biogenesis. Pathogenicity was tested by spore suspension inoculation on wounded or unwounded gondre leaves. The lesions were observed on inoculated leaves within 3 days after inoculation, regardless of wound. To our knowledge, this is the first report of the leaf spot on gondre thistle caused by S. lycopersici in Korea or elsewhere.

Difference of Gray Mold Severity at Roses Caused by Botrytis cinerea Strains (잿빛곰팡이병원균 Botrytis cinerea 균주 종류별 장미 발병 정도의 차이)

  • Hwang, Kyu-Hyon;Hong, Seung-Min;Lee, Young-Soon;Lee, Hyun-Ju;Seo, Myeong-Whoon
    • Research in Plant Disease
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    • v.25 no.1
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    • pp.16-21
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    • 2019
  • Botrytis cinerea is the pathogen for a gray mold generating problems during the cultivation and transportation of roses. But there is little information about the difference of the symptom severity caused by gray mold on rose varieties and pathogen strains. 16 strains were collected from the rose cultivation area to confirm the degree of disease occurrence against strains and each variety. Collected 16 strains were identified based on the sequences analysis of ITS region of ribosomal DNA by using specific primers. The sequence analysis was performed by comparing the sequences to find a difference. To confirm the difference in disease occurrence for each strains, the difference was classified from 0 to 5 stages using charmant variety as a control. The data was confirmed through Kruskal-Wallis ANOVA. The result showed the significant difference in the pathogenicity caused by strains. WNG6_5 showed the lowest pathogenicity with 0.24 and WNG6_3 showed the highest with 3.20. The difference between two strains were almost 3.0. In addition, nine varieties of roses were more investigated with three strains such as the strains of WNG6_5, Hwa_1, and WNG6_3. The result showed that the Love Letter variety showed resistance and the Ice Bear variety was sensitive to three strains. Taken together, this study showed the significant difference by the interactions of rose varieties and gray mold strains.

Isolation and characterization of cellulolytic yeast belonging to Moesziomyces sp. from the gut of Grasshopper (메뚜기의 내장에서 분리한 Moesziomyces 속에 속하는 셀룰로오스 분해 효모의 분리 및 특성)

  • Kim, Ju-Young;Jung, Hee-Young;Park, Jong-Seok;Cho, Sung-Jin;Lee, Hoon Bok;Sung, Gi-Ho;Subramani, Gayathri;Kim, Myung Kyum
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.234-241
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    • 2019
  • An intensive interaction between yeasts and insects has highlighted their relevance for attraction to food and for the insect's development and behavior. Yeast associated in the gut of insects secretes cellulase which aided in the food digestion (cellulose degradation). Three strains of cellulose-degrading yeast were isolated from the gut of adult grasshoppers collected in Gyeonggi Province, South Korea. The strains $ON22^T$, $G10^T$, and $G15^T$, showed positive cellulolytic activity in the carboxymethyl cellulose (CMC)-plate assay. The phylogenetic tree based on sequence analysis of D1/D2 domains of the large subunit rRNA gene and the internal transcribed spacer (ITS) regions revealed that the strains $ON22^T$ (100 and 98.4% sequence similarities in D1/D2 domains and ITS) and $G10^T$ (99.8 and 99.5% in D1/D2 domain and ITS region) were most closely related to the species Moesziomyces aphidis JCM $10318^T$; $G15^T$ (100% in D1/D2 domains and ITS) belongs to the species Moesziomyces antarcticus JCM $10317^T$, respectively. Morphology and biochemical test results are provided in the species description. Cellulase with its massive applicability has been used in various industrial processes such as biofuels like bioethanol productions. Therefore, this is the first report of the cellulolytic yeast strains $ON22^T$, $G10^T$, and $G15^T$ related to the genus Moesziomyces in the family Ustilaginaceae (Ustilaginales), in Korea.

Phylogenetic Analysis of Ruminant Theileria spp. from China Based on 28S Ribosomal RNA Gene

  • Gou, Huitian;Guan, Guiquan;Ma, Miling;Liu, Aihong;Liu, Zhijie;Xu, Zongke;Ren, Qiaoyun;Li, Youquan;Yang, Jifei;Chen, Ze;Yin, Hong;Luo, Jianxun
    • Parasites, Hosts and Diseases
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    • v.51 no.5
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    • pp.511-517
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    • 2013
  • Species identification using DNA sequences is the basis for DNA taxonomy. In this study, we sequenced the ribosomal large-subunit RNA gene sequences (3,037-3,061 bp) in length of 13 Chinese Theileria stocks that were infective to cattle and sheep. The complete 28S rRNA gene is relatively difficult to amplify and its conserved region is not important for phylogenetic study. Therefore, we selected the D2-D3 region from the complete 28S rRNA sequences for phylogenetic analysis. Our analyses of 28S rRNA gene sequences showed that the 28S rRNA was useful as a phylogenetic marker for analyzing the relationships among Theileria spp. in ruminants. In addition, the D2-D3 region was a short segment that could be used instead of the whole 28S rRNA sequence during the phylogenetic analysis of Theileria, and it may be an ideal DNA barcode.

Analysis of Wave Distribution at Nakdong River Estuary Depending on the Incident Wave Directions Based on SWAN Model Simulation (SWAN 모델을 이용한 낙동강 하구역의 입사파향별 파랑분포 특성)

  • Park, Soon;Yoon, Han-Sam;Park, Hyo-Bong;Ryu, Seung-Woo;Ryu, Cheong-Ro
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.12 no.3
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    • pp.188-196
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    • 2009
  • This study conducted numerical simulations to analyze the wave characteristics(distribution) depending on the directional changes of waves in the Nakdong river estuary by using SWAN(Simulating WAves Nearshore) model. The results from the tests are summarized as below. The wave height rates are generally highly distributed with the incident waves from the S, SSE, SSW, SE, SW in sequence. When the waves from the S, SSW, SSE directions are predominant, the bigger waves were observed in front of sandbars. According to the results of the wave steepness against the wave direction, at the east coast of Gadeok island(northwest of Nakdong estuary), where has mild seabed slopes, the wave height rates distribute in the range of 0.4~0.6; the wave height rates over the west coastal region of Dadeapo(southeast of Nakdong estuary) are 0.5~0.6. The wave height rate tends to be rapidly decreased over the east region of Nakdong river estuary rather than its west region.

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