• Title/Summary/Keyword: Sequence-based molecular analysis

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Revisiting Rhytisma lonicericola: Morphological Characterization and Molecular Phylogenetic Analysis

  • Jung, Bok-Nam;Park, Ji-Hyun;Shin, Hyeon-Dong
    • Mycobiology
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    • v.50 no.2
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    • pp.150-154
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    • 2022
  • Rhytisma lonicericola was identified as a tar spot fungus on Lonicera sp. in 1902, and has since been recorded on several species of Lonicera in China, Japan, and Korea. Most of the previous records of R. lonicericola have been based on a list of disease occurrences in the absence of any formal morphological identification or molecular analyses. Using six newly obtained specimens collected in the past 2 years, we confirmed the tar spot fungus found on L. japonica in Korea as R. lonicericola based on morphological examinations and molecular phylogenetic analyses. This fungus was distinguished from R. xylostei, another tar spot fungus on Lonicera, by ascospore size and geographical distributions. We present detailed mycological information and, for the first time, DNA sequence data useful for the identification of R. lonicericola.

Expression of dehydration responsive element-binding protein-3 (DREB3) under different abiotic stresses in tomato

  • Islam, Mohammad Saiful;Wang, Myeong-Hyeon
    • BMB Reports
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    • v.42 no.9
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    • pp.611-616
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    • 2009
  • We investigated the expression pattern of dehydration responsive element-binding protein-3 in tomato under different abiotic stresses. Full length LeDREB3 cDNA was isolated from tomato plant, followed by phylogenetic analysis based on deduced amino acid sequences that revealed significant sequence similarity to DREB proteins belonging to diverse families of plant species. Southern blot analysis showed duplicate copies of LeDREB3 in the tomato genome while organ-specific expression profiling indicated constitutive expression of LeDREB3 in all tested organs, which was particularly strong in flower. LeDREB3 expression was significantly induced by Nacl, drought, low temperature and $H_2O_2$. Moreover, LeDREB3 was slightly regulated by treatment with ABA and MV. These observations suggest that the LeDREB3 gene may be involved in the response of the tomato plant to stress.

Identification and molecular characterization of a rabbit hemorrhagic disease virus variant (KV0801) isolated in Korea

  • Yang, Dong-Kun;Kim, Byoung-Han;Lee, Kyung-Woo;Kim, Ji-Yeon;Kim, Hee-Jin;Choi, Sung-Suk;Chun, Ji-Eun;Son, Seong-Wan
    • Korean Journal of Veterinary Research
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    • v.49 no.3
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    • pp.207-213
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    • 2009
  • Rabbit hemorrhagic disease (RHD) is caused by RHD virus (RHDV) and is one of the most fatal diseases of rabbits. Acute death of rabbits occurred in a farm located in the Gyeonggi province of South Korea. The virus was isolated and confirmed as RHDV based on reverse transcription polymerase chain reaction and hemagglutination assay (HA), and the isolate was designated as KV0801. The nucleotide sequence of the complete VP60 gene of KV0801 was determined and the corresponding amino acid sequence was deduced. Molecular analysis showed that the KV0801 isolate can be classified as a pandemic antigenic variant strain, RHDVa. The VP60 nucleotide sequence and deduced amino acid homology between KV0801 and other Korean isolate, RHF89, which was isolated in 1988, were 92.1 and 94.3%, respectively. The pathogenicity of the KV0801 isolate at an HA titer ranging from 16,384 to 0.16 HA units was evaluated in five-month-old SFP rabbits. The rabbits inoculated with KV0801 isolate containing more than 1.63 HA units died within six days of inoculation. These results suggest that a highly pathogenic RHDVa is circulating in the rabbit populations of Korea.

Biodiversity of Bacterial lipase genes

  • Kim, Hyung-Kwoun
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2001.06a
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    • pp.163-164
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    • 2001
  • A number of bacterial species produce extracellular lipases. Among them, many lipase genes have been cloned and sequenced. A comparison of primary sequences revealed only very limited sequence homology among them. Based on the sequence homologies and molecular sizes (Mr), bacterial lipases were classified into four discrete groups. From soil samples taken around Taejon, five different lipase-producing bacteria were isolated; Proteus vulgaris K80, Bacillus stearothermophilus Ll, B. pumilus B26, Staphylococcus haemolyticus L62, S. aureus B56. Nucleotide sequence analysis showed that Staphylococcus lipase genes (L62 and B56) composed of pre-pro-mature parts, Bacillus lipase genes (Ll and B26) pre-mature parts, and Proteus lipase gene (K80) mature part only. In addition, the molecular sizes of their mature parts were quite different from 19,000 to 45,000. Finally, they had very little homology (less than 20%) in their amino acid sequences. Judging from the above results, lipase K80 belonged to bacterial lipase Group I, lipase L1 and lipase B26 Group III, and lipase L62 and lipase B56 Group IV. This diversity in their primary structures was also reflected in their enzymatic properties; temperature effects, pH effects, substrate specificity, detergent effects, and so on.

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Molecular Identification and Expression of Myosin Light Chain in Shortspine Spurdog (Squalus mitsukurii)

  • Kim, Soo Cheol;Sumi, Kanij Rukshana;Sharker, Md Rajib;Kho, Kang Hee
    • Journal of Marine Life Science
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    • v.3 no.1
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    • pp.1-8
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    • 2018
  • Myosin is considered as the vital motor protein in vertebrates and invertebrates. Our present study was conducted to decipher the occurrence of myosin in dog fish (Squalus mitsukurii). We isolated one clone containing 979 bp cDNA sequence, which consisted of a complete coding sequence of 453 bp and a deduced amino acid sequence of 150 amino acids from the open reading frame with molecular weight, isoelectric point and aliphatic index are 16.72 Kda, 4.49 and 78.00, respectively. It contained 428 bp long 3' UTR with single potential polyadenylation signals (AATAAA). The predicted EF CA2+ binding domains were identified in residue 6-41, 83-118 and 133-150. A BLAST search indicates this protein exhibits a strong similarity to whale shark (Rhincodon typus) MLC3 (91% identical) and also house mouse (Mus musculus) MLC isoform 3f (81% identical). Phylogenetic analysis revealed that this protein is a MLC 3 isoform like protein. This protein also demonstrates highly conserved region with other myosin proteins. Homology modeling of S. mitsukuri was performed using crystal structure of Gallus gallus skeletal muscle myosin II based on high similarity. Reverse transcription-polymerase chain reaction (PCR), quantitative PCR results exhibits dogfish myosin protein is highly expressed in muscle tissue.

First Molecular Characterization of Hypoderma actaeon in Cattle and Red Deer (Cervus elaphus) in Portugal

  • Ahmed, Haroon;Sousa, Sergio Ramalho;Simsek, Sami;Anastacio, Sofia;Kilinc, Seyma Gunyakti
    • Parasites, Hosts and Diseases
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    • v.55 no.6
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    • pp.653-658
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    • 2017
  • Hypoderma spp. larvae cause subcutaneous myiasis in several animal species. The objective of the present investigation was to identify and characterize morphologically and molecularly the larvae of Hypoderma spp. collected from cattle (Bos taurus taurus) and red deer (Cervus elaphus) in the district of Castelo Branco, Portugal. For this purpose, a total of 8 larvae were collected from cattle (n=2) and red deer (n=6). After morphological identification of Hypoderma spp. larvae, molecular characterization was based on PCR-RFLP and mitochondrial CO1 gene sequence analysis. All larvae were morphologically characterized as the third instar larvae (L3) of H. actaeon. Two restriction enzymes were used for molecular identification of the larvae. TaqI restriction enzyme was not able to cut H. actaeon. However, MboII restriction enzyme differentiated Hypoderma species showing 210 and 450 bp bands in H. actaeon. Furthermore, according to the alignment of the mt-CO1 gene sequences of Hypoderma species and to PCR-RFLP findings, all the identified Hypoderma larvae were confirmed as H. actaeon. This is the first report of identification of Hypoderma spp. (Diptera; Oestridae) from cattle and red deer in Portugal, based on morphological and molecular analyses.

Characterization of Trichoderma spp. Associated with Green Mold of Oyster Mushroom by PCR-RFLP and Sequence Analysis of ITS Regions of rDNA

  • Park, Myung-Soo;Seo, Geon-Sik;Bae, Kyung-Sook;Yu, Seung-Hun
    • The Plant Pathology Journal
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    • v.21 no.3
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    • pp.229-236
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    • 2005
  • Molecular profIles of PCR-RFLP and sequence analysis of internal transcribed spacer (ITS) regions of rDNA were compared between morphologically distinguishable species of Trichoderma isolated from substrates of oyster mushroom in Korea, T. atroviride, T. citrinoviride, T. harzianum, T. longibrachiatum, T. virens, and two unidentified species, Trichoderma sp. 1 and 2. PCR­RFLP analysis divided the Trichoderma spp. into six RFLP groups, A, B, C, D, E, and F. The RFLP groups were generally agreed with described morphological species, except that the RFLP group A containing the two unidentified species. A neighbor-joining tree based on ITS sequences well supported RFLP groups observed by RFLP analysis of ITS regions of rDNA. Additionally, the two unidentified species, Trichoderma sp. 1 and 2, which could not be distinguished by PCR­RFLP analysis, were separated in sequence analysis of ITS regions of rDNA.

A Phylogenetic Relationships of Araliaceae Based on PCR-RAPD and ITS Sequences (PCR-RAPD와 ITS 서열 분석에 의한 두릅나무과 (Araliaceae) 의 유연관계 분석)

  • 김남희;양덕춘;엄안흠
    • Korean Journal of Plant Resources
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    • v.17 no.2
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    • pp.82-93
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    • 2004
  • Phylogenetic relationships among species in Araliaceae were analyzed using PCR-RAPD and sequence of ITS region of nuclear ribosomal DNA based on samples collected in Korea. RAPD analysis showed various polymorphic bands which were able to differentiate species and genus, and specific bands showing variations among individuals within species. Cluster analysis using gel images revealed high molecular variability within species of Aralia eleta. No significant variation was found among cultivated species of Panax ginseng, but they showed high genetic differences with wild type of the species. In ITS analysis, specific sequences for each genus and species were observed and these were allowed to differentiate species and genus. Phylogenetic analysis using ITS sequences showed that Acanthopanax and Kalopanax had a close relationship, and Aralia and Panax are monophyletic, but genus Hedera is different species from other species in family Araliaceae in this study. The results showing close relationship between genera Aralia and Panax were also observed in RAPD analysis. Contrary to the results of RAPD analysis of Panax ginseng, sequence analysis of ITS showed no significant difference between wild mountain ginseng and cultivated species of P. ginseng. Also, both RAPD and ITS analysis of P. ginseng showed no significant genetic variability among cultivation sites. Results indicate that P. ginseng cultivating in Korea is monophyletic. The molecular analysis used in this study agreed on classification using morphological feature. These results suggest that molecular techniques used in this study could be useful for phylogenetic analysis of Araliaceae.

Taxonomic Re-evaluation of Korean Gyromitra Species Based on Morphological and Phylogenetical Studies

  • Cho, Sung-Eun;Kwag, Young-Nam;Han, Sang-Kuk;Kim, Chang Sun
    • The Korean Journal of Mycology
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    • v.49 no.3
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    • pp.363-371
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    • 2021
  • Several species of Gyromitra are highly poisonous when consumed raw the presence of gyromitrin. In this study, Gyromitra specimens deposited in the Korea National Arboretum were re-examined and subjected to molecular sequence analysis. G. gigas, G. aff. perlata, and G. tianshanensis were identified based on their morphological features and molecular phylogenetic analysis for the first time in Korea. Furthermore, internal transcribed spacer and 28S rRNA sequences of G. esculenta and G. infula were analyzed to confirm their phylogeny.

Inter Simple Sequence Repeat (ISSR) Polymorphism and Its Application in Mulberry Genome Analysis

  • Vijayan Kunjupillai
    • International Journal of Industrial Entomology and Biomaterials
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    • v.10 no.2
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    • pp.79-86
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    • 2005
  • Molecular markers have increasingly been used in plant genetic analysis, due to their obvious advantages over conventional phenotypic markers, as they are highly polymorphic, more in number, stable across different developmental stages, neutral to selection and least influenced by environmental factors. Among the PCR based marker techniques, ISSR is one of the simplest and widely used techniques, which involves amplification of DNA segment present at an amplifiable distance in between two identical microsatellite repeat regions oriented in opposite direction. Though ISSR markers are dominant like RAPD, they are more stable and reproducible. Because of these properties ISSR markers have recently been found using extensively for finger printing, pohylogenetic analysis, population structure analysis, varietal/line identification, genetic mapping, marker-assisted selection, etc. In mulberry (Morus spp.), ISSR markers were used for analyzing phylogenetic relationship among cultivated varieties, between tropical and temperate mulberry, for solving the vexed problem of identifying taxonomic positions of genotypes, for identifying markers associated with leaf yield attributing characters. As ISSR markers are one of the cheapest and easiest marker systems with high efficiency in generating polymorphism among closely related varieties, they would play a major role in mulberry genome analysis in the future.