• Title/Summary/Keyword: internal transcribed spacer(ITS)

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Inhibitory Effects of Aureobasidium pullulans MHAU2101 Isolated from Domestic Pear Blossom Against Fire Blight (국내 배나무 꽃에서 분리한 Aureobasidium pullulans MHAU2101의 화상병 발생 억제 효과)

  • Hyeonseok Oh;Hyo-Won Choi;Yong Hwan Lee;Seung Yeup Lee;Mi-Hyun Lee;Sang-Keun Oh
    • The Korean Journal of Mycology
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    • v.50 no.4
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    • pp.331-341
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    • 2022
  • This study was conducted to identify yeast species isolated from domestic pear blossom through gene sequencing and analysis of morphological characteristics, and to confirm specific yeast species inhibitory effects toward fire blight in immature apples, pears, and crab apple blossoms. Yeast morphological characteristics were consistent with the known characteristics of Aureobasidium pullulans. Nucleotide sequencing of the D1/D2 region of large-subunit (LSU) 26S ribosomal DNA and the internal transcribed spacer (ITS) region confirmed its identity as A. pullulans (MHAU2101). Inoculation of immature fruits with A. pullulans MHAU2101 before exposure to Erwinia amylovora prevented fire blight symptoms in apples and pears. A. pullulans MHAU2101 treated crab apple blossoms had a significantly lower flower infection rate than untreated blossoms, revealing 64% of the potency of streptomycin. The A. pullulans MHAU2101 treated group also displayed lower E. amylovora density in both pistil and hypanthium compared to the untreated group, especially in the hypanthium. This study confirms that A. pullulans MHAU2101 isolated from domestic pear blossom can effectively suppress the onset of fire blight.

Degradation of Malic Acid by Issatchenkia orientalis KMBL 5774, an Acidophilic Yeast Strain Isolated from Korean Grape Wine Pomace

  • Seo, Sung-Hee;Rhee, Chang-Ho;Park, Heui-Dong
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.521-527
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    • 2007
  • Several yeast strains degrading malic acid as a sole carbon and energy source were isolated from Korean wine pomace after enrichment culture in the presence of malic acid. Among them, the strain designated as KMBL 5774 showed the highest malic acid degrading ability. It was identified as Issatchenkia orientalis based on its morphological and physiological characteristics as well as the nucleotide sequences of the internal transcribed spacer (ITS) 1-5.8S rDNA-ITS II region. Phylogenetic analysis of the ITS I-5.8S rDNA-ITS II sequences showed that the KMBL 5774 is the closest to I. orientalis zhuan 192. Identity of the sequences of the KMBL 5774 was 99.5% with those of I. orientalis zhuan 192. The optimal pH of the media for the growth and malic acid degradation by the yeast was between 2.0 and 3.0, suggesting that the strain is an acidophile. Under the optimized conditions, the yeast could degrade 95.5% of the malic acid after 24 h of incubation at $30^{\circ}C$ in YNB media containing 2% malic acid as a sole carbon and energy source.

Taxonomic Position of Korean Isolates of Rhizoctonia solani Based on RAPD and ITS Sequencing of Ribosomal DNA

  • Jeon, Young-Ah;Kim, Wan-Gyu;Kim, Dae-Ho;Kwon, Soon-Wo;Hong, Seung-Beom
    • The Plant Pathology Journal
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    • v.26 no.1
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    • pp.83-89
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    • 2010
  • Taxonomic position of 46 Korean isolates of Rhizoctonia solani which were classified into nine intraspecific groups by anastomosis and cultural characteristics was analyzed by randomly amplified polymorphic DNA (RAPD) and sequence analyses of the internal transcribed spacer (ITS) regions of ribosomal DNA. All the isolates within each group showed highly similar band patterns in RAPD. The ITS regions of the isolates within the same groups showed a high level of sequence similarity above 96.0% whereas similarities among different groups were below 94.4%. When compared with several reference strains of R. solani from foreign countries, all the Korean isolates were clustered with the foreign isolates belonging to the same groups in the phylogenetic tree. All six Korean strains of AG-4 were identified as HG-1 out of 3 subgroup of AG-4. We discussed taxonomic position of Korean isolates of R. solani and showed that sequence analysis with ITS regions could be a rapid and useful method for identification of intraspecific group of R. solani.

Geographical variation and evolutionary relationship of Asparagus cochinchinensis Lour. based on rDNA-ITS sequences and Random Amplified Polymorphic DNA(RAPD) (ITS 부위 염기서열과 RAPD분석을 통한 천문동의 지역별 변이 및 분자진화적 유연관계)

  • Moon, Byeong-Cheol;Choo, Byoung-Kil;Ji, Yun-Ui;Choi, Go-Ya;Yoon, Tae-Sook;Lee, A-Young;Kim, Ho-Kyoung
    • Korean Journal of Oriental Medicine
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    • v.14 no.1
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    • pp.129-135
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    • 2008
  • Phylogenetic relationship and DNA polymorphism among local populations of the Asparagus cochinchinensis have been investigated based on nuclear ribosomal DNA ITS sequences and RAPD analysis in Korea. In result, two genetically distinct groups of local populations except Geoje were recognized by the phylogenetic tree both in rDNA-ITS and RAPD. One was called 'western coast group' that includes the Buan 1, 2 and Taean and the other was 'southern coast group' that includes Haenam, Yeosu and Namhae. Thus, the geographical relationship of Asparagus cochinchinensis was two well-typified clades. These results suggest that the geographical genetic variation of Asparagus cochinchinensis is closely connected with the slow and long period of propagation via the coast in Korean Peninsula.

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Identification of Orchid Mycorrhizal Fungi Isolated from Terrestrial Orchids in Mt. Hambaek, Korea (함백산의 난초과 식물의 뿌리에서 난균근균의 분리 및 동정)

  • Lee, Bong-Hyung;Han, Han-Kyeol;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.43 no.2
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    • pp.129-132
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    • 2015
  • In this study, orchid mycorrhizal fungi (OMF) were isolated from four terrestrial orchids on Mt. Hambeak, Platnathera chlorantha, Platnathera mandarinorum, Cephalanthera falcate, and Cephalanthera longibracteata. OMF were identified using morphological and sequences analysis of fungal internal transcribed spacer (ITS) regions by specific primer of basidiomycetous orchid mycorrhizas; ITS1-OF and ITS4-OF. Four species of orchid mycorrhizal fungi were identified as Ceratobasidium sp, Epulorhiza anaticula, Tulasnella calospora and Tulasnella sp.

Genetic diversity and phylogenetic analysis of genus Paeonia based on nuclear ribosomal DNA ITS sequence

  • Sun, Yan-Lin;Hong, Soon-Kwan
    • Journal of Plant Biotechnology
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    • v.38 no.3
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    • pp.234-240
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    • 2011
  • The genus Paeonia belongs to the family Paeoniaceae having significant medicinal and ornamental importance. The present investigation was undertaken with an aim to understand phylogenetic relationships of three Paeonia species (P. lactiflora, P. obovata, and P. suffruticosa) that are widely distributed in China, Korea, and Japan, using nuclear ribosomal DNA (nrDNA) internal transcribed spacer (ITS) sequence and to compare the phylogeny results with investigations reported earlier using existed sequences of the same species. The size variation obtained among sequenced nrDNA ITS region was narrow and ranged from 722 to 726 bp. The highest interspecific genetic distance (GD) was found between P. lactiflora and P. suffruticosa or P. obovata. The phylogram obtained using our nrDNA ITS sequences showed non-congruence with previous hypothesis of the phylogeny between section Paeonia and section Moutan of genus Paeonia. This result was supported by the phylogenetic relations showed in the phylogram constructed with existed sequences in NCBI. The present study suggested that P. obovata belonging to section Paeonia was phylogenetically closer to P. suffruticosa representing section Moutan of genus Paeonia than P. lactiflora belonging to section Paeonia. The main reason of the paraphyly of section Paeonia is thought to be nucleotide additivity directly caused by origin hybridization. This study provides more sequence sources of genus Paeonia, and will help for further studies in intraspecies population, and their phylogentic analysis and molecular evolution.

Diseases Severity, Genetic Variation, and Pathogenicity of Ceratocystis Wilt on Lansium domesticum in South Sumatra, Indonesia

  • Muslim, Ahmad;Pratama, Rahmat;Suwandi, Suwandi;Hamidson, Harman
    • The Plant Pathology Journal
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    • v.38 no.2
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    • pp.131-145
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    • 2022
  • Ceratocystis wilt disease has caused significant mortality in duku (Lansium domesticum) since 2014 and has now spread to all districts in South Sumatra, Indonesia. Recently, 16 isolates from duku representing populations from various districts in South Sumatra were isolated. Analysis for the morphological characteristic of the isolate showed that the population has a uniform morphology. Genetic analysis based on internal transcribed spacer (ITS) and β-tubulin sequences verified that the population has being dominated by the ITS5 haplotype of Ceratocystis fimbriata and a new ITS group, the ITS7b haplotype that was localized in Musi Banyuasin. Both haplotypes were highly pathogenic to duku. Inoculation tests on various forest and agroforestry plant hosts showed that both haplotypes were highly pathogenic to Acacia mangium, moderately pathogenic to Acacia carsicarpa, Eucalyptus urophylla, and Melaleuca cajuputi, but weakly pathogenic to Dyera costulata, Hevea brasiliensis, and Alstonia scholaris. Therefore, this pathogen becomes a serious threat to Indonesia's biodiversity due to its ability to infect forest and agroforestry plants, especially the indigenous ones.

DNA barcoding analysis of Rosase Multiflorae Fructus and its adulterants (영실(營實)과 그 위품의 유전자 감별)

  • Doh, Eui jeong;Shin, Sangmun;Lee, Guemsan
    • The Korea Journal of Herbology
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    • v.34 no.4
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    • pp.1-8
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    • 2019
  • Objectives : Rosae Multiflorae Fructus is a traditional medicine derived from the fruit of Rosa multiflora Thunb. a member of the Rosaceae family. Even though it has a single origin, the possibility of adulterants has always existed. In fact, we had discovered suspicious commercial samples of Rosae Multiflorae Fructus, imported from China. Methods : To define the taxonomic origin of Rosae Multiflorae Fructus and its adulterants, DNA barcode analysis of the internal transcribed spacer, trnL-F intergenic spacer, and psbA-trnH sequences was carried out. These DNA barcode sequences from the correct origin of Rosae Multiflorae Fructus were analyzed and compared with those of other samples from genus Rosa used as medicinal herbs. Results : The analyses of the three DNA barcode sequences efficiently distinguished Rosae Multiflorae Fructus from six other species in genus Rosa and also separated each species used in this study. According to the DNA barcoding results, none of the suspicious commercial samples were Rosae Multiflorae Fructus. RMF09 was identified as Rosa acicularis, whereas RMF10 and RMF11 were identified as Rosa davurica and Rosa rugosa, respectively. These results corroborated the existence of adulterants of Rosae Multiflorae Fructus. Conclusions : Our research provides useful information that could be used as a criterion for distinguishing between Rosae Multiflorae Fructus and its adulterants. These results will help in the prevention of adulteration and also suggest effective methods for verifying the origin of commercial herbal medicines derived from genus Rosa.

Taxonomic Revision of Notohymena gangwonensis (Protozoa: Ciliophora), with Notes on Its Cortical Granules and Scanning Electron Micrographs

  • Moon, Ji Hye;Kim, Kang-San;Chae, Kyu Seok;Min, Gi-Sik;Jung, Jae-Ho
    • Animal Systematics, Evolution and Diversity
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    • v.36 no.2
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    • pp.113-122
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    • 2020
  • From a moss sample, we isolated and identified Notohymena gangwonensis Kim et al., 2019 based on morphological and molecular data. The moss and type population has completely identical 18S rRNA (nuclear small subunit ribosomal RNA) gene sequences and both are highly similar in morphological and morphometric attributes, except for the diameter and arrangement of the cortical granules. Thus, we reexamined the type materials(i.e., micrographs and gDNA) and resulted in finding mistakes made by the authors of the species. Based on these data and supporting materials newly obtained (i.e., internal transcribed spacer [ITS] 1, ITS2, 5.8S, and partial 28S rDNA sequences, and scanning electron micrographs), we provide improved diagnosis of the species to clarify its identity. In addition, a key for Notohymena species is provided.

Characterization of Colletotrichum Isolates Causing Anthracnose of Pepper in Korea

  • Kim, Joon-Tae;Park, Sook-Young;Choi, Woo-Bong;Lee, Yong-Hwan;Kim, Heung-Tae
    • The Plant Pathology Journal
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    • v.24 no.1
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    • pp.17-23
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    • 2008
  • A total of 33 isolates of Colletotrichum species obtained from pepper, apple, and strawberry in 2001 and 2002 were identified based on mycological characteristics, responses to fungicides carbendazim and the mixture of carbendazim and diethofencarb, and nucleotide sequence analysis of internal transcribed spacer (ITS) regionMost of the Colletotrichum isolates from pepper could be identified as C. acutatum. The pepper isolates produced grey white mycelia that gradually changed to dark gray. The conidia were variable in size, and almost cylindrical in shape with at least one rounded end. They could grow on PDA amended with carbendazim or with the mixture of carbendazim and diethofencarb at $10{\mu}g/ml$, to which the isolates from apple and strawberry were very sensitive. A part of the ITS regions from the Colletotrichum isolates was amplified with the specific primers designed for C. acutatum (Ca1-1) or C. gloeosporioides (Cg1-3). A primer pair of Ca1-1 and a universal primer (ITS4) amplified a 496-bp DNA fragment from all of the pepper isolates examined and one apple isolate. Taken together, it is conclusive that the Colletotrichum isolates causing the typical lesion of anthracnose on pepper fruits are C. acutatum.