• Title/Summary/Keyword: ITS rDNA sequences

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The Incidence of Alternaria Species Associated with Infected Sesamum indicum L. Seeds from Fields of the Punjab, Pakistan

  • Nayyar, Brian Gagosh;Woodward, Steve;Mur, Luis A.J.;Akram, Abida;Arshad, Muhammad;Naqvi, S.M. Saqlan;Akhund, Shaista
    • The Plant Pathology Journal
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    • v.33 no.6
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    • pp.543-553
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    • 2017
  • Sesame (Sesamum indicum) is an important oil seed crop of Asia. Yields can be negatively impacted by various factors, including disease, particularly those caused by fungi which create problems in both production and storage. Foliar diseases of sesame such as Alternaria leaf blight may cause significant yield losses, with reductions in plant health and seed quality. The work reported here determined the incidence of Alternaria species infecting sesame seeds grown in the Punjab, Pakistan. A total of 428 Alternaria isolates were obtained from 105 seed samples and grouped into 36 distinct taxonomic groups based on growth pattern and morphological characters. Isolation frequency and relative density of surface sterilized and non-surface sterilized seeds showed that three isolates (A13, A47 and A215) were the most common morphological groups present. These isolates were further identified using sequencing of the Internal Transcribed Spacer (ITS) region of ribosomal DNA (rDNA) and the Alternaria major allergen gene (Alt a 1). Whilst ITS of rDNA did not resolve the isolates into Alternaria species, the Alt a 1 sequences exhibited > 99% homology with Alternaria alternata (KP123850.1) in GenBank accessions. The pathogenicity and virulence of these isolates of Alternaria alternata was confirmed in inoculations of sesame plants resulting in typical symptoms of leaf blight disease. This work confirms the identity of a major source of sesame leaf blight in Pakistan which will aid in formulating effective disease management strategies.

Detection of DNA Sequence Polymorphism by Polymerase Chain Reaction in Fraxinus mandshurica Rupr Growing in Korea (P.C.R 기법(技法)을 이용(利用)한 들메나무 DNA sequence의 변이조사(變異調査))

  • Na, Chun Soo;Noh, Eun Woon;Kim, Young Joong;Shin, Chang Ho;Song, Won Sup;Kim, Sea Hyun
    • Journal of Korean Society of Forest Science
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    • v.81 no.4
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    • pp.320-324
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    • 1992
  • It has been reported that there are two distinct phenotypes in Fraxinus mandshurica Rupr. growing in Korea. Recently developed polymerase chain reaction(PCR) was used to detect DNA sequence polymorphism in the species. Using a thermostable DNA polymerase and synthetic DNA primers, unknown DNA sequences from the species were randomly amplified. The two types of the species produced different DNA amplification pattern with three different primers tested. Although DNA polymorphism was detected among individuals within types, each type has its own distinct pattern. The two types could be easily differentiated by trier characteristic predominant bands.

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Survey of Perkinsus olseni infection in Manila clam, Ruditapes philippinarum in 2009 on the west and south coast of Korea using PCR technique (PCR 기법을 이용한 2009년 우리나라 서해안과 남해안 바지락, Ruditapes philippinarum의 Perkinsus olseni 감염에 관한 보고)

  • Lee, Nam-Sil;Hwang, Jee-Youn;Choi, Dong-Lim;Park, Myoung-Ae
    • Journal of fish pathology
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    • v.23 no.2
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    • pp.145-153
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    • 2010
  • Prevalence of a protozoan parasite Perkinsus olseni in Manila clam Ruditapes philippinarum was surveyed from July to December 2009 on the west and south coast of Korea. P. olseni infection was diagnosed using two primer sets, P.olseni NTS Forward/P.olseni NTS Reverse set and PolsITS-140F/PolsITS-600R set in polymerase chain reaction(PCR). The results using PolsITS-140F and PolsITS-600R primer set was retained up to 60% at all stations from July to December, except for Padori. Especially, Goheung showed 100% prevalence from October to December. The results about comparison of the 4 station's DNA sequences which were analyzed from PCR products(457bp) using PolsITS-140F and PolsITS-600R primer set, there were only 2base differences at Sunjedo.

Genetic Diversity of Xanthomonas arboricola pv. pruni Isolated in Korea (우리나라에서 분리한 Xanthomonas arboricola pv. pruni의 유전적 다양성)

  • Park, So-Yeon;Lee, Young-Sun;Shin, Jong-Sub;Koh, Young-Jin;Jung, Jae-Sung
    • Journal of Life Science
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    • v.19 no.5
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    • pp.684-687
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    • 2009
  • Xanthomonas arboricola pv. pruni, the causal agent of bacterial shot holes in stone fruits, was known to have a low population diversity. To investigate the genetic characteristics of X. arboricola pv. pruni isolated in Korea, three strains which have identical 16S rDNA sequences - including type strain (LMG852), Japanese isolate (MAFF301420) and Korean isolate (XWD1) - were analysed based on the nucleotide sequences of three DNA regions and RAPD pattern. No sequence diversity among the three strains was found within the ITS, glnA and atpD gene sequences. However, five of 756 nucleotides of the atpD gene determined (accession number FJ429319) were different from those of the French strain available from the Genbank database. RAPD analyses performed with 40 different arbitrary primers revealed that two strains isolated from Korea and Japan showed similarity in their band patterns distinguished by type strain. These results suggest that Korean and Japanese strains are very close and belong to a population with a low genetic diversity, and might have a different origin from strains found in West Europe.

Diversity of Butyrivibrio Group Bacteria in the Rumen of Goats and Its Response to the Supplementation of Garlic Oil

  • Zhu, Zhi;Hang, Suqin;Mao, Shengyong;Zhu, Weiyun
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.2
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    • pp.179-186
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    • 2014
  • This study aimed to investigate the diversity of the Butyrivibrio group bacteria in goat rumen and its response to garlic oil (GO) supplementation as revealed by molecular analysis of cloned 16S rRNA genes. Six wethers fitted with ruminal fistulas were assigned to two groups for a cross-over design with 28-d experimental period and 14-d interval. Goats were fed a basal diet without (control) or with GO ruminal infusion (0.8 g/d). Ruminal contents were used for DNA extraction collected before morning feeding on d 28. A total bacterial clone library was firstly constructed by nearly full-length 16S rRNA gene cloned sequences using universal primers. The resulting plasmids selected by Butyrivibrio-specific primers were used to construct a Butyrivibrio group-specific bacterial clone library. Butyrivibrio group represented 12.98% and 10.95% of total bacteria in control and GO group, respectively. In libraries, clones were classified to the genus Pseudobutyrivibrio, Butyrivibrio and others within the family Lachnospiraceae. Additionally, some specific clones were observed in GO group, being classified to the genus Ruminococcus and others within the family Ruminococcaceae. Based on the criterion that the similarity was 97% or greater with database sequences, there were 29.73% and 18.42% of clones identified as known isolates (i.e. B. proteoclasticus and Ps. ruminis) in control and GO groups, respectively. Further clones identified as B. fibrisolvens (5.41%) and R. flavefaciens (7.89%) were specifically found in control and GO groups, respectively. The majority of clones resembled Ps. ruminis (98% to 99% similarity), except for Lachnospiraceae bacteria (87% to 92% similarity) in the two libraries. The two clone libraries also appeared different in Shannon diversity index (control 2.47 and GO group 2.91). Our results indicated that the Butyrivibrio group bacteria had a complex community with considerable unknown species in the goat rumen.

First Report of Leaf Spot of Datura metel Caused by Alternaria tenuissima in Korea

  • Aktaruzzaman, Md.;Kim, Joon-Young;Afroz, Tania;Kim, Byung-Sup
    • Research in Plant Disease
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    • v.21 no.4
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    • pp.330-333
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    • 2015
  • In June 2013, we collected leaf spot disease samples of Datura metel from Gangneung, Gangwon Province, Korea. The symptoms observed were small circular to oval dark brown spots with irregular in shape or remained circular with concentric rings. We isolated the pathogen from infected leaves and cultured the fungus on potato dextrose agar. We examined the fungus morphologically and confirmed its pathogenicity according to Koch's postulates. The results of morphological examinations, pathogenicity tests, and the rDNA sequences of the internal transcribed spacer regions (ITS1 and ITS4), glycerol-3-phosphate dehydrogenase (G3PDH) and the RNA polymerase II second largest subunit (RPB2) gene sequence revealed that the causal agent was Alternaria tenuissima. To the best of our knowledge, this is the first report of leaf spot of D. metel caused by A. tenuissima in Korea as well as worldwide.

Isolation of a Pestalotiopsis Species Degrading Mucilage from Fruit of Opuntia ficus-indica var. Saboten

  • Huh, Yoon-Hee;Ko, Young-Hwan
    • Journal of Applied Biological Chemistry
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    • v.50 no.4
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    • pp.221-226
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    • 2007
  • The high molecular-weight mucilage extracted and purified from cactus fruit of Opuntia ficus-indica var. Saboten was degraded by the cell-free culture filtrate of a fungus isolated from soil. TLC analysis of the polymeric mucilage after incubation with the fungal culture filtrate confirmed its degradation. When the degradation products were tested for their qualitative reactions with ninhydrin and phenol-sulfuric acid, only phenol-sulfuric acid gave positive development, and ninhydrin did not show any observable color reaction. This coloring reaction suggested the presence of a carbohydrate without an amino group within the mucilage. Analyses by HPLC and liquid gel permeation chromatography on sephadex G-100 also provided additional information on degradation of the mucilage by the fungal culture filtrate. The sequences of ITS-5.8S rDNA from the fungal isolate that was cultivated for the preparation of mucilage-degrading enzyme showed 99% similarity to those of Pestalotiopsis aquatica.

Characterization of a Zygomycete Fungus, Mortierella minutissima from Freshwater of Yeongsan River in Korea

  • Nguyen, Thi Thuong Thuong;Lee, Hyang Burm
    • The Korean Journal of Mycology
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    • v.44 no.4
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    • pp.346-349
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    • 2016
  • During a survey of fungal diversity of the order Mortierellales, a zygomycete strain, EML-YS717-1, was isolated from a freshwater sample collected from the Yeongsan River in Gwangju, Korea. Based on its morphological characteristics and a phylogenetic analysis of the internal transcribed spacer (ITS1 and ITS2) and 5.8S rDNA sequences, the strain was identified as Mortierella minutissima. To the best of our knowledge, M. minutissima, has not previously been authentically reported in Korea.

Preliminary Application of Molecular Monitoring of the Pacific Herring (Clupea pallasii) Based on Real-time PCR Assay Utilization on Environmental Water Samples

  • Kim, Keun-Yong;Heo, Jung Soo;Moon, Seong Yong;Kim, Keun-Sik;Choi, Jung-Hwa;Yoo, Joon-Taek
    • Korean Journal of Ecology and Environment
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    • v.54 no.3
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    • pp.209-220
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    • 2021
  • Pacific herring, Clupea pallasii, a keystone species with significant ecological and commercial importance, is declining globally throughout much of its range. While traditional fishing equipment methods remain limited, new sensitive and rapid detection methods should be developed to monitor fisheries resources. To monitor the presence and quantity of C. pallasii from environmental DNA (eDNA) extracted from seawater samples, a pair of primers and a TaqMan® probe specific to this fish based on mitochondrial cytochrome b (COB) sequences were designed for the real-time PCR (qPCR) assay. The combination of our molecular markers showed high specificity in the qPCR assay, which affirmed the success of presenting a positive signal only in the C. pallasii specimens. The markers also showed a high sensitivity for detecting C. pallasii genomic DNA in the range of 1 pg~100 ng rxn-1 and its DNA plasmid containing COB amplicon in the range of 1~100,000copies rxn-1, which produced linear standard calibration curves (r2=0.99). We performed a qPCR assay for environmental water samples obtained from 29 sampling stations in the southeastern coastal regions of South Korea using molecular markers. The assay successfully detected the C. pallasii eDNA from 14 stations (48.2%), with the highest mean concentration in Jinhae Bay with a value of 76.09±18.39 pg L-1 (246.20±58.58 copies L-1). Our preliminary application of molecular monitoring of C. pallasii will provide essential information for efficient ecological control and management of this valuable fisheries resource.

Isolation and Identification of Wild Yeasts from Soils of Fields in Daejeon Metropolitan City and Chungcheongnam-do, Korea (대전광역시와 충청남도 밭 토양으로부터 야생효모의 분리 및 동정)

  • Han, Sang-Min;Han, Jae-Won;Bae, Sang-Min;Park, Won-Jong;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.44 no.1
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    • pp.1-7
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    • 2016
  • This study focused on isolation and identification of wild yeasts from soils in fields near mountains and elucidation of its yeast distribution. Several kinds of yeasts were isolated from various soils of Daejeon metropolitan city and Chungcheongnam-do in Korea and identified by BLAST search of nucleotide sequences of internal transcribed spacer (ITS) region including 5.8S rRNA and D1/D2 region of 26S rDNA. Ninety-seven strains of 20 species from 61 soil samples were isolated, of which Cryptococcus podzolicus (11 strains), Debaryomyces hansenii (6 strains), and Trichosporon asahii (6 strains) were dominant species.