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

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Cellulose degrading basidiomycetes yeast isolated from the gut of grasshopper in Korea (한국의 메뚜기의 장에서 분리된 Cellulose를 분해하는 담자균 효모)

  • Kim, Ju-Young;Jang, Jun Hwee;Park, Ji-Hyun;Jung, Hee-Young;Park, Jong-Seok;Cho, Sung-Jin;Lee, Hoon Bok;Limtong, Savitree;Subramani, Gayathri;Sung, Gi-Ho;Kim, Myung Kyum
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.362-368
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    • 2018
  • Grasshoppers play vital role in the digestion of photosynthetically fixed carbons. With the aid of intestinal microflora, the grasshopper can degrade leaves constituents such as cellulose and hemicellulose. The purpose of this study was to examine cellulolytic yeast isolates from the gut of grasshoppers collected in Gyeonggi Province, South Korea. Among the yeast isolates, ON2, ON17 (two strains), and ON6 (one strain) showed positive cellulolytic activity in the CMC-plate assay. The sequence analyses of D1/D2 domains of the large subunit rDNA gene and the internal transcribed spacer (ITS) regions revealed that the strains ON2 and ON17 were most closely related to Papiliotrema aspenensis CBS $13867^T$ (100%, sequence similarity in D1/D2 domains; 99.4% sequence similarity in ITS) and strain ON6 related to Saitozyma flava (100% in D1/D2 domains; 99.0% in ITS). All these three yeast strains are capable of degrading cellulose; therefore, the members of endosymbiotic yeasts may produce their own enzymes for carbohydrate degradation and convert mobilized sugar monomers to volatile fatty acids. Thus, the endosymbiotic yeast strains ON2, ON17 (represents the genus Papilioterma) and ON6 (Saitozyma) belonging to the family Tremellomycetes, are unreported strains in Korea.

The complete plastid genome and nuclear ribosomal transcription unit sequences of Spiraea prunifolia f. simpliciflora (Rosaceae)

  • Jeongjin CHOI;Wonhee KIM;Jee Young PARK;Jong-Soo KANG;Tae-Jin YANG
    • Korean Journal of Plant Taxonomy
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    • v.53 no.1
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    • pp.32-37
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    • 2023
  • Spiraea prunifolia f. simpliciflora Nakai is a perennial shrub widely used for horticultural and medicinal purposes. We simultaneously obtained the complete plastid genome (plastome) and nuclear ribosomal gene transcription units, 45S nuclear ribosomal DNA (nrDNA) and 5S nrDNA of S. prunifolia f. simpliciflora, using Illumina short-read data. The plastome is 155,984 bp in length with a canonical quadripartite structure consisting of 84,417 bp of a large single-copy region, 18,887 bp of a short single-copy region, and 26,340 bp of two inverted repeat regions. Overall, a total of 113 genes (79 protein-coding genes, 30 tRNAs, and four rRNAs) were annotated in the plastome. The 45S nrDNA transcription unit is 5,848 bp in length: 1,809 bp, 161 bp, and 3,397 bp for 18S, 5.8S, and 26S, respectively, and 261 bp and 220 bp for internal transcribed spacer (ITS) 1 and ITS 2 regions, respectively. The 5S nrDNA unit is 512 bp, including 121 bp of 5S rRNA and 391 bp of intergenic spacer regions. Phylogenetic analyses showed that the genus Spiraea was monophyletic and sister to the clade of Sibiraea angustata, Petrophytum caespitosum and Kelseya uniflora. Within the genus Spiraea, the sections Calospira and Spiraea were monophyletic, but the sect. Glomerati was nested within the sect. Chamaedryon. In the sect. Glomerati, S. prunifolia f. simpliciflora formed a subclade with S. media, and the subclade was sister to S. thunbergii and S. mongolica. The close relationship between S. prunifolia f. simpliciflora and S. media was also supported by the nrDNA phylogeny, indicating that the plastome and nrDNA sequences assembled in this study belong to the genus Spiraea. The newly reported complete plastome and nrDNA transcription unit sequences of S. prunifolia f. simpliciflora provide useful information for further phylogenetic and evolutionary studies of the genus Spiraea, as well as the family Rosaceae.

A phylogenetic analysis of Korean Artemisia L. based on ITS sequences (ITS 염기서열에 의한 한국산 쑥속(Artemisia L.)의 계통분류학적 연구)

  • Lee, Jeong-Hoon;Park, Chung-Berm;Park, Chun-Geon;Moon, Sung-Gi
    • Korean Journal of Plant Resources
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    • v.23 no.4
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    • pp.293-302
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    • 2010
  • Taxa of Artemisia collected in Korea were constructed by molecular phylogenetic analysis based on the internal transcribed spacer(ITS) regions of nrDNA. The length of the ITS sequences aligned using the clustal X program was 636~643 bp, and the lengths of the ITS1 and ITS2 regions were 251~255 bp and 217~222 bp, respectively. The total number of variable sites was 95 for the entire sequence, and a parsimony- informative site represented an efficacious site in ITS1 rather than in ITS2. The maximum parsimony tree as calculated by the MEGA 4 program was clustered into five clades. The taxa(A. capillaris, A. japonica var. japonica, A. japonica var. hallaisanensis, A. japonica subsp. littoricora) degenerated ovary of clade 1 was supported as the subgenus Dracunculus by Ling's classification system. The results show that A. nakaii and A. fukudo were quite similar genetically(Boostrap 99%) and that the scientific name of Korean A. dubia should be reconsidered. A. sp. distributed in Ganghwa province was grouped with A. argyi(Boostrap 89%). These results suggest that the molecular techniques used in this study could be useful for the phylogenetic analysis of Korean Artemisia herbs having variations in their morphological characteristics.

Diversity Analysis of Endophytic Fungi Isolated from the Roots of Coastal Plants in Taean Peninsula (태안반도에 자생하는 해안식물 뿌리에서 분리한 내생진균의 다양성 분석)

  • You, Young-Hyun;Yoon, Nam-Gyeong;Yoon, Hyeokjun;Kim, Hyun;Lim, Sung Hwan;Choo, Yeon-Sik;Kim, Jong-Guk
    • The Korean Journal of Mycology
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    • v.42 no.1
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    • pp.79-85
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    • 2014
  • Forty-two endophytic fungi were isolated from the roots of coastal plants, such as Aster sphathulifolius Maxim., Aster tripolium L., Phragmites australis, and Puccinellia nipponica Ohwi, naturally growing in Taean peninsula. The identity of the endophytic fungal strains was analyzed using the sequence of internal transcribed spacer regions. All fungi belonged to the phylum Ascomycota, and they were classified into eight orders (Botryosphaeriales, Capnodiales, Diaporthales, Dothideales, Eurotiales, Helotiales, Hypocreales, and Pleosporales) and thirteen genera (Alternaria, Aureobasidium, Cadophora, Cladosporium, Davidiella, Diaporthe, Fusarium, Gibberella, Macrophomina, Metarhizium, Neosartorya, Penicillium, and Phoma). Among the analyzed fungi, the fungi belonging to the genus Penicillium in Eurotiales were the most widely distributed. The host plant Aster tripolium L. was found to contain the most diverse endophytic fungal species among the coastal plants.

Gibberellins Production and Identification of Endophytic Fungi Isolated from Aquatic Plant in Fresh Water (담수에 자생하는 수생식물에서 분리된 내생균류의 지베렐린 생산과 동정)

  • You, Young-Hyun;Kang, Sang-Mo;Choi, Yu-Mi;Lee, Myung-Chul;Kim, Jong-Guk
    • The Korean Journal of Mycology
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    • v.43 no.1
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    • pp.71-76
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    • 2015
  • Aquatic plant Hydrocharis dubia (Blume) Backer was collected from the Dalsung wetland in Daegu. Sixteen endophytic fungi with different colony morphologies were isolated from the roots of aquatic plants. Waito-c rice (WR) seedlings were treated with fungal culture filtrates (FCF) for screening plant growth-promoting activity. In the results, HD1008 strain isolated from aquatic plant showed highest plant growth-promoting activity. The FCF of HD1008 strain was analyzed using gas chromatography mass spectrometry (GC/MS) with selected ion monitoring (SIM). Analysis of the FCF of HD1008 strain found that it contained gibberellins (GA) ($GA_1$, 1.2 ng/100 mL; $GA_4$, 5 ng/100 mL). Phylogenetic tree of HD1008 strain was constructed by partial internal transcribed spacer (ITS) region and partial beta-tubulin gene sequences. Therefore, we describe HD1008 strain as a new gibberellin-producing Penicillium trzebinskii based on morphological and molecular characteristics.

Effect of Trichoderma sp. GL02 on alleviating Drought Stress in Pepper Plants (Trichoderma sp. GL02에 의한 고추 식물의 건조 스트레스 완화 효과)

  • Kim, Sang Tae;Yoo, Sung-Je;Song, Jaekyeong;Weon, Hang-Yeon;Sang, Mee Kyung
    • Korean Journal of Organic Agriculture
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    • v.28 no.3
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    • pp.417-430
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    • 2020
  • Drought stress is one of major environmental stresses in plants; this leads to reduce plant growth and crop yield. In this study, we selected fungal isolate for mitigating drought stress in pepper plants. To do this, 41 fungi were isolated from rhizosphere or bulk soils of various plants in Jeju, Gangneung, Hampyeong in Korea. Out of 41 isolates, we screened two isolates without phytotoxicity through seed germination of tomato, pepper, and cabbage treated with fungal spores; through following plant assay, we selected GL02 as a candidate for alleviating drought stress in pepper plants. As a result of greenhouse test of pepper plants in drought condition, the stomatal conductance on leaves of pepper plants treated with GL02 was increased, whereras the malondialdehyde (MDA) and electrolyte leakage were decreased compared to that in control plants. When stressed plants were rewatered, stomatal conductance of the plants treated with GL02 was increased; the electrolyte leakage was decreased. Based on internal transcribed spacer (ITS) sequencing analysis, isolate GL02 was belonging to genus Trichoderma. Taken together, drought stress in pepper plants treated with GL02 was alleviated, when it was rewatered after drought-stressed, the plants could be recovered effectively. Therefore, Trichoderma sp. GL02 could be used as a bio-fertilizer to alleviate drought stress in pepper plants.

Development and Evaluation of Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Tylenchulus semipenetrans Using DNA Extracted from Soil

  • Song, Zhi-Qiang;Cheng, Ju-E;Cheng, Fei-Xue;Zhang, De-Yong;Liu, Yong
    • The Plant Pathology Journal
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    • v.33 no.2
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    • pp.184-192
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    • 2017
  • Tylenchulus semipenetrans is an important and widespread plant-parasitic nematode of citrus worldwide and can cause citrus slow decline disease leading to significant reduction in tree growth and yield. Rapid and accurate detection of T. semipenetrans in soil is important for the disease forecasting and management. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed to detect T. semipenetrans using DNA extracted from soil. A set of five primers was designed from the internal transcribed spacer region (ITS1) of rDNA, and was highly specific to T. semipenetrans. The LAMP reaction was performed at $63^{\circ}C$ for 60 min. The LAMP product was visualized directly in one reaction tube by adding SYBR Green I. The detection limit of the LAMP assay was $10^{-2}J2/0.5g$ of soil, which was 10 times more sensitive than conventional PCR ($10^{-1}J2/0.5g$ of soil). Examination of 24 field soil samples revealed that the LAMP assay was applicable to a range of soils infested naturally with T. semipenetrans, and the total assay time was less than 2.5 h. These results indicated that the developed LAMP assay is a simple, rapid, sensitive, specific and accurate technique for detection of T. semipenetrans in field soil, and contributes to the effective management of citrus slow decline disease.

First Report of Colletotrichum fioriniae Causing Anthracnose on Fruit of Pumpkin (Cucurbita moschata) in Korea (Colletotrichum fioriniae에 의한 호박 과실 탄저병의 발생 보고)

  • Kim, Joon-Young;Kim, Byung-Sup
    • Research in Plant Disease
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    • v.26 no.3
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    • pp.190-193
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    • 2020
  • In 2014, Anthracnose symptoms were observed on fruit of pumpkin in Gangneung, Gangwon in Korea. The typical symptoms on fruit surfaces were circular lesions, brown, shriveled with salmon-colored spore masses on the spots. A fungal isolate was obtained from symptomatic on fruit and cultured on potato dextrose agar at 25℃. The color of the upper surface of the colony varied from white to gray with occurrence of salmon-colored conidial masses and was red to violet on the underside. The causal fungus of anthracnose isolated from the diseased fruits was identified as Colletotrichum fioriniae based on mycological characteristics and sequences of the internal transcribed spacer, glyceraldehyde-3-phosphate dehydrogenase, chitin synthase-1 gene, histone H3, actin gene, beta-tubulin gene region. Pathogenicity tests of the isolate were performed on pumpkin fruit by both wound and non-wound inoculation with conidial suspension (1×106 conidia/ml) and incubated at 25℃. Lesions were observed on both wounded and non-wounded fruit of pumpkin after 7 days inoculation in only C. fioriniae. The pathogen was re-isolated from the lesions of inoculated fruits. To our knowledge, this is the first report of anthracnose on fruit of pumpkin caused by C. fioriniae in Korea.

Morphological and Molecular Characterization of Lymnaeid Snails and Their Potential Role in Transmission of Fasciola spp. in Vietnam

  • Dung, Bui Thi;Doanh, Pham Ngoc;The, Dang Tat;Loan, Ho Thi;Losson, Bertrand;Caron, Yannick
    • Parasites, Hosts and Diseases
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    • v.51 no.6
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    • pp.657-662
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    • 2013
  • Freshwater snails of the family Lymnaeidae play an important role in the transmission of fascioliasis worldwide. In Vietnam, 2 common lymnaeid species, Lymnaea swinhoei and Lymnaea viridis, can be recognized on the basis of morphology, and a third species, Lymnaea sp., is known to exist. Recent studies have raised controversy about their role in transmission of Fasciola spp. because of confusion in identification of the snail hosts. The aim of this study is, therefore, to clarify the identities of lymnaeid snails in Vietnam by a combination of morphological and molecular approaches. The molecular analyses using the second internal transcribed spacer (ITS2) of the nuclear ribosomal DNA clearly showed that lymnaeids in Vietnam include 3 species, Austropeplea viridis (morphologically identified as L. viridis), Radix auricularia (morphologically identified as L. swinhoei) and Radix rubiginosa (morphologically identified as Lymnaea sp.). R. rubiginosa is a new record for Vietnam. Among them, only A. viridis was found to be infected with Fasciola spp. These results provide a new insight into lymnaeid snails in Vietnam. Identification of lymnaeid snails in Vietnam and their role in the liver fluke transmission should be further investigated.

Isolation and Characteristics of Endolichenic Fungi Producing Antifungal Compound (항진균성 물질을 생산하는 지의류 내생 곰팡이의 선별 및 특성)

  • Hwang, Hyun-Gook;Kim, Yi-Na;Baik, Keun-Sik;Choi, Sang-Ki
    • Korean Journal of Microbiology
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    • v.47 no.1
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    • pp.97-101
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    • 2011
  • To isolate a novel antifungal compound, we obtained 107 kinds of endolichenic fungi from Lichen Bioresources Center and examined their antifungal capability. Two fungi EL123 and EL156 showed high antifungal activity against Candida albicans in both MYA and EMM media. Nucleotide sequence analysis and NCBI Blast analysis in ITS region including 5.8S rRNA revealed that EL123 has 95% homology with Thielavia microspora and EL156, 99% with Cryptosporiopsis diversispora which belong to Ascomycetes. It observed that EL156 formed a branched mycelium whereas EL123 formed a straight one. EL156 also produced the antifungal substance faster than EL123 when they grew on MY liquid medium.