• 제목/요약/키워드: ITS region of rDNA

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Isolation and Identification of Three Newly Reported Ascomycete Fungal Species Isolated from Soil in Korea

  • Mohammad Hamizan Azmi;Seong-Keun Lim;Seok-Jin Park;Min-Gyeong Song;Jun-Soo Cha;Seung-Yeol Lee;Hee-Young Jung
    • The Korean Journal of Mycology
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    • v.52 no.2
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    • pp.97-108
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    • 2024
  • In this study, three fungal isolates belonging to the phylum Ascomycota under classes Leotiomycetes, Eurotiomycetes, and Sordariomycetes were isolated from soil in Korea. These species were designated as KNUF-22-003, KNUF-22-005, and KNUF-20-NI016, respectively, and identified based on their phylogenetic relationships and morphological characteristics. The isolates were confirmed through molecular phylogenetic analyses of their internal transcribed spacer (ITS) region, 28S rDNA large subunit (LSU), and actin (ACT1 ) gene sequences. Cultural and morphological characteristics of strains KNUF-22-003, KNUF-22-005, and KNUF-20-NI016 were matched with Chaetomella oblonga CBS110.78T, Oidiodendron chlamydosporicum CBS403.69T, and Sarocladium subulatum CBS217.35T, respectively. To the best of our knowledge, this is the first report on C. oblonga, O. chlamydosporicum, and S. subulatum in Korea.

Discovery of novel haplotypes from wild populations of Kappaphycus (Gigartinales, Rhodophyta) in the Philippines

  • Roleda, Michael Y.;Aguinaldo, Zae-Zae A.;Crisostomo, Bea A.;Hinaloc, Lourie Ann R.;Projimo, Vicenta Z.;Dumilag, Richard V.;Lluisma, Arturo O.
    • ALGAE
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    • v.36 no.1
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    • pp.1-12
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    • 2021
  • As the global demand for the carrageenophyte Kappaphycus is steadily increasing, its overall productivity, carrageenan quality, and disease resistance are gradually declining. In the face of this dilemma, wild Kappaphycus populations are viewed as sources of new cultivars that could potentially enhance production; therefore, assessment of their diversity is crucial. This study highlights the morphological and genetic diversity of wild Kappaphycus species obtained from two sites in the Philippines. Nucleotide alignments of available 5' region of the mitochondrial cytochrome c oxidase subunit I (COI-5P) and cox2-3 spacer sequences of Kappaphycus confirmed the presence of K. alvarezii in Guiuan, Eastern Samar and K. striatus in Bolinao, Pangasinan. Based on the concatenated sequences of the COI-5P and the cox2-3 spacer, nine novel haplotypes were observed along with other published haplotypes. However, there was no relationship between haplotype and morphology. These newly recognized haplotypes indicate a reservoir of unutilized wild genotypes in the Philippines, which could be taken advantage of in developing new cultivars with superior traits. DNA barcodes generated from this study effectively expand the existing databank of Kappaphycus sequences and can provide insights in elucidating the genetic diversity of Kappaphycus species in the country.

Stem Rot on Adzuki Bean (Vigna angularis) Caused by Rhizoctonia solani AG 4 HGI in China

  • Sun, Suli;Xia, Changjian;Zhang, Jiqing;Duan, Canxing;Wang, Xiaoming;Wu, Xiaofei;Lee, Suk-Ha;Zhu, Zhendong
    • The Plant Pathology Journal
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    • v.31 no.1
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    • pp.67-71
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    • 2015
  • During late August and early September 2011, stem rot symptoms were observed on adzuki bean plants (Vigna angularis) growing in fields located in Beijing and Hebei Province, China, respectively. In this study, four isolates were obtained from infected stems of adzuki bean plants. Based on their morphology, and sequence and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analyses of the ribosomal DNA internal transcribed spacers (rDNA-ITS) region, the four isolates were identified as Rhizoctonia solani in anastomosis group (AG) 4 HGI. Pathogenicity tests showed that all isolates were strongly pathogenic to adzuki bean and resulted in serious wilt symptoms which was similar to observations in the fields. Additionally, the isolates infected several other crops and induced related rot on the roots and basal stems. To our knowledge, this is the first report of Rhizoctonia solani AG 4 HGI causing stem rot on adzuki bean.

Undescribed Fungal Species of Eupenicillium, Mortierella, and Trichoderma Isolated in the Vicinity of Demilitarized Zone in Yeoncheon-gun, Gyeonggi-do, Korea (경기도 연천비무장지대 근역에서 분리한 국내 미기록 Eupenicillium, Mortierella, Trichoderma 진균 종 보고)

  • Ahn, Geum Ran;Kim, Ji Eun;Oh, Yun Seok;Lee, Kyoung Min;Jin, Hyeop;Kim, Min Uk;Kim, Jun Young;Kim, Seong Hwan
    • The Korean Journal of Mycology
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    • v.46 no.4
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    • pp.359-367
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    • 2018
  • As an effort to explore fungal diversity, fungal survey was undertaken in 2017 in the vicinity of demilitarized zone (DMZ) located in Yeoncheon-gun, Gyeonggi-do, Korea. For the survey, wild plants and soils were sampled and subjected to fungal isolation. A total of 18 genera and 23 species including five unrecorded fungal species, Mortierella sclerotiella, M. sossauensis, M. verticillate, Eupenicillium saturniforme, and Trichoderma hispanicum, were obtained from the survey. This study described their morphological characteristics including colony features formed on media, light microscopic images and molecular characteristics of nucleotide sequences of the internal transcribed spacer (ITS) region, 18S and 28S rDNA, ${\beta}-tubulin$ gene, calmodulin, and translation elongation factor $1{\alpha}$ ($tef1{\alpha}$) nDNA genes.

Sequence analysis of partial LSU rDNA of three Alexandrium species (Dinophyceae) hitherto unreported

  • Kim, Keun-Yong;Makoto Yoshida;Kim, Chang-Hoon
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2003.10a
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    • pp.35-35
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    • 2003
  • We, for the first time, reported molecular sequences of large subunit ribosomal DNA Dl-D3 region of A. hiranoi, A. leei and A. satoanum hitherto unreported. In addition, this study presented the full-length sequences of A. affine, A. fraterculus, A. catenella and A. tamarense occurring in Korean coastal waters. In total, 17 Alexandrium morphospecies were subjected to the phylogenetic analysis using the Maximum-likelihood (ML) method. The alignment result of sequences of A. hiranoi and A. pseudogonyaulax showed that there were only two substitutions without length heterogeneity implying their genetic affiliation. In ML tree, A. leei formed a deeply diverging branch probably because of the accelerated evolutionary rate, and its phylogenetic position was so ambiguous to resolve the phylogenetic relationship to the residual taxa. An A. satoanum culture showing morphological variation in the sulcal plate formed an independent divergent branch with consistent sister relationship to A. hiranoi/A. pseudogonyaulax clade supported by the high posterior probability (PP) value. Blast search in GenBank showed the sequence data of A. affine, A. fraterculus, A. catenella and A. tamarense corresponded to their morphological species designation. In ML tree, Alexandrium species were commonly split into four main clades. The inter-clade relationships were not clear and usually supported by the week PP values. In general, the sulcal plate of Alexandrium species seemed to reflect the true phylogeny at the main clade level, and the connection between the 1 and the apical pore complex seemed to reflect the phylogeny at the subclade level.

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Sequencing, Genomic Structure, Chromosomal Mapping and Association Study of the Porcine ADAMTS1 Gene with Litter Size

  • Yue, K.;Peng, J.;Zheng, R.;Li, J.L.;Chen, J.F.;Li, F.E.;Dai, L.H.;Ding, SH.H.;Guo, W.H.;Xu, N.Y.;Xiong, Y.ZH.;Jiang, S.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.7
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    • pp.917-922
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    • 2008
  • A disintegrin-like and metalloprotease (reprolysin type) with thrombospondin type 1 motif (ADAMTS1) plays a critical role in follicular rupture and represents a major advance in the proteolytic events that control ovulation. In this study, a 9,026-bp DNA sequence containing the full coding region, all 8 introns and part of the 5'and 3' untranslated region of the porcine ADAMTS1 gene was obtained. Analysis of the ADAMTS1 gene using the porcine radiation hybrid panel indicated that pig ADAMTS1 is closely linkage with microsatellite marker S0215, located on SSC13q49. The open reading frame of its cDNA covered 2,844 bp and encoded 947 amino acids. The coding region of porcine ADAMTS1 as determined by sequence alignments shared 85% and 81% identity with human and mouse cDNAs, respectively. The deduced protein contained 947 amino acids showing 85% sequence similarity both to the human and mouse proteins, respectively. Comparative sequencing of three pig breeds revealed one single nucleotide polymorphism (SNP) within exon 7 of which a G-C substitution at position 6006 changes a codon for arginine into a codon for proline. The substitution was situated within a PvuII recognition site and developed as a PCR-RFLP marker for further use in population variation investigations and association analysis with litter size. Allele frequencies of this SNP were investigated in seven pig breeds/lines. An association analysis in a new Qingping female line suggested that different ADAMTS1 genotypes have significant differences in litter size (p<0.01).

Cultural characteristics of fungal species associated with deterioration or foxing of paper and chemical removal (종이변색균류의 배양적 특성 및 화학적 방법에 의한 변색제거)

  • Jo, Seong-Eun;Kim, Yong-Tae;Jeong, So-Yeong;Jo, Byeong-Muk;Lee, Jong-Gyu
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 2009.04a
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    • pp.295-303
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    • 2009
  • The annals of Joseon dynasty, especially the volumes of King SeJong(1418-1450 A.D.), were heavily deteriorated by fungi. Investigations on the deteriorating and foxing fungi were carried out. Fungal structures on the beeswax, which were coated on the both side of Han-Ji, were suspected to be involved in the deterioration, and were observed by SEM. Isolation and culturing of these fungi were tried by scrubing swab samples and placing on the artificial media. Culture-independent approaches were used to identify the fungal strains associated with damages of beeswax and foxing of the paper by the analyses based on DNA sequences data from the specific ITS region of rDNA regions. In addition, well-known paper staining fungi(PSF), i.e., Aspergillus terreus var. terreus, Fusarium oxysporum, Chaetomium globosum, Cladosporium cladosporioides, and Alternaria solani, were compared in the mycelial growth and stain on beeswax and papers under different environmental conditions (temperature, light, moisture, etc). Fungal strains isolated from the air samples in the storage room and shelves were identified as Irpex sp., Arthrinium sacchari, Cladosporium tenuissimum, Aspergillus sclerotiorum, Sistotrema brinkmannii, and Hypoxylon bovei var. microsporum The isolated strains were compared in growth and stain patterns on beeswax and papers(Han-Ji, Hwa-Ji, and Yang-Ji) whether these can cause damage or foxing on the annals or not.

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The Rapid Detection of Pathogens in Organically Grown Vegetables Using PCR-DGGE (PCR-DGGE를 이용한 유기농 채소의 유해 미생물 신속 검지)

  • Kwon, Oh Yeoun;Son, Seok Min
    • Food Engineering Progress
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    • v.15 no.4
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    • pp.370-375
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    • 2011
  • In this study the polymerase chain reaction (PCR) combined with denaturing gradient gel electrophoresis (DGGE) was evaluated as a method permitting the rapid detection of pathogens in fresh originally grown vegetables. A universal primer (341GCf/534r) was selected for its ability to amplify the V3 region of 16S-rRNA genes in their target pathogens (Salmonella typhimurium, Pseudomonas fluorescens, Bacillus cereus, Listeria monoytogenes, Staphyloocus aureus, E. coli). The 194 bp fragments in PCR were successfully duplicated as expected. The amplified fragments of the same size from six different pathogens also showed good separation upon DGGE. The detection limit of PCR-DGGE for six pathogens in fresh-cut lettuces were over $10^{5}$ CFU/g when sampled by stomaching. However, when the sampling method was changed from stomaching to shaking, the detection limit of six pathogens in organic vegetables was shown to increase by over $10^{1}$ CFU/g, but only those of B. cereus were over $10^{3}$ CFU/g. Therefore, PCR-DGGE was shown to be a reliable method for the detection of pathogens in fresh-cut vegetables.

Growth Promotion and Induction of Systemic Resistance Against Phytophthora capsici on Red-pepper Plant by Treatment of Trichoderma harzianum MPA167 (근권 Trichoderma harzianum MPA167 처리에 의한 생육촉진과 고추 역병균에 대한 고추의 유도저항성)

  • Yang, Nuri;Lee, Sae Won;Kim, Heung Tae;Park, Kyungseok
    • The Korean Journal of Pesticide Science
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    • v.17 no.4
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    • pp.394-401
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    • 2013
  • Trichoderma harzianum is one of rhizosphere fungus usually lives near the plant root regions in the soil. T. harzianum plays an important role in plant growth promotion and increases disease resistance against various plant pathogens on crops. In this study, the strain T. harzianum MPA167 was isolated from the barley rhizosphere soil in Suwon, Korea. Among 183 isolates, the strain T. harzianum MPA167 was selected as promising strain in which based on hyperparasitical activity against Phytophthora capsici and estimated disease control activity against P. capsici in the greenhouse conditions. The strain T. harzianum MPA167 was identified using 23s rDNA internal transcribed spacer(ITS) region sequences. MPA167 treatment ($1{\times}10^6$ spores/ml) showed greater disease suppression against Phytophthora blight of red-pepper caused by P. capsici in greenhouse compared with the water-treated control. Volatiles derived from T. harzianum MPA167 elicit growth promotion of tobacco and Arabidopsis seedlings in I-plate assay. In addition, T. harzianum MPA167 strain was also found to be effective for the growth promotion and induction of systemic resistance on red-papper plant. These results suggest that MPA167 might be used as one of the potential biocontrol agents.

Screening, Gene Cloning, and Characterizations of an Acid-Stable α-Amylase

  • Liu, Xinyu;Jia, Wei;An, Yi;Cheng, Kun;Wang, Mingdao;Yang, Sen;Chen, Hongge
    • Journal of Microbiology and Biotechnology
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    • v.25 no.6
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    • pp.828-836
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    • 2015
  • Based on its α-amylase activity at pH 5.0 and optimal pH of the crude enzyme, a strain (named B-5) with acid α-amylase production was screened. The B-5 strain was identified as Bacillus amyloliquefaciens through morphological, physiological, and biochemical characteristics analysis, as well as 16S rDNA phylogenetic analysis. Its α-amylase gene of GenBank Accession No. GU318401 was cloned and expressed in Escherichia coli. The purified recombinant α-amylase AMY-Ba showed the optimal pH of 5.0, and was stable at a pH range of 4.0-6.0. When hydrolyzing soluble starch, amylose, and amylopectin, AMY-Ba released glucose and maltose as major end products. The α-amylase AMY-Ba in this work was different from the well-investigated J01542-type α-amylase which also came from B. amyloliquefaciens. AMY-Ba exhibited notable adsorption and hydrolysis ability towards various raw starches. Structure analysis of AMY-Ba suggested the presence of a new starch-binding domain at its C-terminal region.