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A New Distinct Clade for Iranian Tomato spotted wilt virus Isolates Based on the Polymerase, Nucleocapsid, and Non-structural Genes

  • Abadkhah, Mahsa;Koolivand, Davoud;Eini, Omid
    • The Plant Pathology Journal
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    • v.34 no.6
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    • pp.514-531
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    • 2018
  • Tomato spotted wilt virus (TSWV; Genus Orthotospovirus: Family Tospoviridae) is one of the most destructive viruses affecting a wide range of horticultural crops on a worldwide basis. In 2015 and 2016, 171 leaf and fruit samples from tomato (Solanum lycopersicum) plants with viral symptoms were collected from the fields in various regions of Iran. ELISA test revealed that the samples were infected by TSWV. The results of RT-PCR showed that the expected DNA fragments of about 819 bp in length were amplified using a pair of universal primer corresponding to the RNA polymerase gene and DNA fragments of ca 777 bp and 724 bp in length were amplified using specific primers that have been designed based on the nucleocapsid (N) and non-structural (NSs) genes, respectively. The amplified fragments were cloned into pTG19-T and sequenced. Sequence comparisons with those available in the GenBank showed that the sequences belong to TSWV. The high nucleotide identity and similarities of new sequences based on the L, N, and NSs genes showed that minor evolutionary differences exist amongst the isolates. The phylogenetic tree grouped all isolates six clades based on N and NSs genes. Phylogenetic analysis showed that the Iranian isolates were composed a new distinct clade based on a part of polymerase, N and NSs genes. To our knowledge, this is the first detailed study on molecular characterization and genetic diversity of TSWV isolates from tomato in Iran that could be known as new clade of TSWV isolates.

Molecular Detection and Phylogenetic Analysis of Anaplasma phagocytophilum in Horses in Korea

  • Seo, Min-Goo;Ouh, In-Ohk;Choi, Eunsang;Kwon, Oh-Deog;Kwak, Dongmi
    • Parasites, Hosts and Diseases
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    • v.56 no.6
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    • pp.559-565
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    • 2018
  • The identification and characterization of pathogenic and zoonotic tick-borne diseases like granulocytic anaplasmosis are essential for developing effective control programs. The differential diagnosis of pathogenic Anaplasma phagocytophilum and non-pathogenic A. phagocytophilum-like Anaplasma spp. is important for implementing effective treatment from control programs. The objective of the present study was to investigate the prevalence of Anaplasma spp. in horses in Korea by nucleotide sequencing and restriction enzyme fragment length polymorphism assay. Of the 627 horses included in the study, only 1 (0.2%) was infected with A. phagocytophilum. Co-infection with A. phagocytophilumlike Anaplasma spp. was not detected in the study. The 16S rRNA sequence of A. phagocytophilum was similar (99.5-100%) to A. phagocytophilum 16S rRNA isolated from horses in other countries. PCR adapted to amplify A. phagocytophilum groEL and msp2 genes failed to generate amplicons, suggesting genetic diversity in these genes. This study is the first molecular detection of A. phagocytophilum in horses in Korea. Human granulocytic anaplasmosis and animal infection of A. phagocytophilum have been reported in Korea recently. Because of vector tick distribution, global warming, and the increase of the horse industry, horses should be considered as a potential reservoir for A. phagocytophilum, and cross infectivity should be evaluated even though a low prevalence of infection was detected in this study. Furthermore, continuous surveillance and effective control measures for A. phagocytophilum should be established to prevent disease distribution and possible transmission to humans.

Performance of Korean spontaneous speech recognizers based on an extended phone set derived from acoustic data (음향 데이터로부터 얻은 확장된 음소 단위를 이용한 한국어 자유발화 음성인식기의 성능)

  • Bang, Jeong-Uk;Kim, Sang-Hun;Kwon, Oh-Wook
    • Phonetics and Speech Sciences
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    • v.11 no.3
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    • pp.39-47
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    • 2019
  • We propose a method to improve the performance of spontaneous speech recognizers by extending their phone set using speech data. In the proposed method, we first extract variable-length phoneme-level segments from broadcast speech signals, and convert them to fixed-length latent vectors using an long short-term memory (LSTM) classifier. We then cluster acoustically similar latent vectors and build a new phone set by choosing the number of clusters with the lowest Davies-Bouldin index. We also update the lexicon of the speech recognizer by choosing the pronunciation sequence of each word with the highest conditional probability. In order to analyze the acoustic characteristics of the new phone set, we visualize its spectral patterns and segment duration. Through speech recognition experiments using a larger training data set than our own previous work, we confirm that the new phone set yields better performance than the conventional phoneme-based and grapheme-based units in both spontaneous speech recognition and read speech recognition.

Characterization of an alkaline esterase from an enriched metagenomic library derived from an oil-spill area

  • Baek, Seung Cheol;Jo, Jeong Min;Jeong, Soo-Mi;Lee, Jae Pil;Lee, Hyun Woo;Kim, Jungho;Kim, Hoon
    • Journal of Applied Biological Chemistry
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    • v.62 no.1
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    • pp.73-79
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    • 2019
  • A novel esterase gene (est7S) was cloned from an enriched metagenomic library derived from an oil-spill area. The gene encoded a protein of 505 amino acids, and the molecular mass of the Est7S was estimated to be 54,512 Da with no signal peptide. Est7S showed the highest identity of 40% to an esterase from a sludge metagenome compared to the characterized enzymes with their properties, although it showed 99% identity to a carboxylesterase in the genome sequence of Alcanivorax borkumensis SK2. Est7S had catalytic triad residues, Ser183, Glu312, and His420, and the GESAG motif in most family VII lipolytic enzymes. Est7S was purified from the crude extract of clone SM7 using Sephacryl S-200 HR and HiTrap Q column chromatographies. The purified Est7S was optimally active at $50^{\circ}C$ and pH 10.0. Est7S showed a high specific activity of 366.7 U/mg protein. It preferred short length esters, particularly p-nitrophenyl acetate, efficiently hydrolyzed R- and S-enantiomers of methyl-3-hydroxy-2-methylpropionate, and glyceryl tributyrate. These properties of Est7S may provide potential merits in biotechnological applications such as detergent and paper processing under alkaline conditions.

Identification and structural analysis of novel laccase genes in Flammulina elastica genome (Flammulina elastica의 유전체 정보기반 신규 laccase 유전자 동정 및 구조 분석)

  • Yu, Hye-Won;Park, Young-Jin
    • Journal of Mushroom
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    • v.19 no.1
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    • pp.33-40
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    • 2021
  • The genome sequence of various Flammulina species has recently been reported, thereby revealing a diverse genetic repertoire. In this study, we identified laccase genes and analyzed their structural characteristics in Flammulina elastica (F. elastica) genome. Through genome analysis and bioinformatics approaches, three laccase genes (Fe-lac1, -lac2, and -lac3) were identified, ranging from 1,548 to 1,602 bp in length. These genes contained a copper ion-binding region with ten histidine residues and one cysteine residue and a disulfide bond-forming region with four cysteine residues. Full-length cDNA sequencing analysis revealed that laccase genes contain 12 to 16 introns and signal peptides between 17 and 22 bp in the N-terminus. Structural characterization of the laccase genes identified in this study should help in better understanding the biomass decomposition of F. elastica.

Genetic Identification Monitoring of Cobitidae Distribution in Korea (국내에서 유통되는 미꾸리과(Cobitidae) 어종의 분자동정 모니터링)

  • Kim, Hyunsuk;Shin, Jiyoung;Yang, Junho;Cha, Eunji;Yang, Ji-young
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.55 no.5
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    • pp.742-750
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    • 2022
  • This study aimed to monitor the distribution of Cobitidae in Korea by the identification of species using genetic analysis. Based on the genetic analysis, Cobitidae species in four of five domestic fish farms consisted of only Chinese muddy loach Misgurnus mizolepis, but muddy loach Misgurnus anguillicaudatus was also present it in one fish farm. In the case of imported Cobitidae species, in addition to Chinese muddy loach and muddy loach, the harmful species Paramisgurnus dabryanus, was also present. Chinese muddy loach accounted for 20%, 67%, and 60% of the S6, S7, and S8 samples, respectively. An analysis of the total length, body length, and weight showed that domestic Chinese muddy loach showed higher values than imported muddy loach, and imported Chinese muddy loach showed similar values to P. dabryanus. There were no significant differences in the country of origin of the three species. Thus, the mitochondrial cytochrome c oxidase subunit I gene sequence was analyzed and compared the verification of species identification. The three species of Cobitidae were genetically divided into three groups and determined to have genetic differences. These results indicate that it is necessary to reduce the heterogeneous mixing rate through discriminating species by genetic analysis.

Investigation on Support Mechanism of Geogrid-Encased Stone Columns in Soft Ground (연약지반에 시공되는 지오그리드 감쌈 스톤컬럼의 하중 지지 메카니즘에 관한 연구)

  • Yoo, Chung-Sik;Kim, Sun-Bin
    • Journal of the Korean Geotechnical Society
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    • v.24 no.12
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    • pp.93-101
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    • 2008
  • This paper presents the results of numerical investigation on support mechanism of geogrid-encased stone columns for use in soft ground improvement. A number of cases were analyzed using a 3D stress-pore pressure coupled model that can effectively model construction sequence and drainage as well as reinforcing effects of geogrid-encased stone columns. The results indicated that the geogrid encasement provides additional confinement effect that reduces vertical stress in the soft ground, thus resulting in less excess pore water pressures and associated settlement. Also revealed was that such a confinement effect depends on encasement length and stiffness of geogrid. It is also shown that there exist critical encasement length and stiffness of geogrid for a given condition.

ITS Sequence Variations in Populations of Ilex cornuta (Aquifoliaceae) (호랑가시나무(Ilex cornuta) 개체군의 ITS 염기서열 변이 분석)

  • Son, Sung-won;Kim, Joo-Hwan;Kim, Yong-Shik;Park, Seon-Joo
    • Korean Journal of Plant Taxonomy
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    • v.37 no.2
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    • pp.131-141
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    • 2007
  • Genetic variation of Ilex cornuta Lindley et Paxton was examined by sequence analyses of ITS for 65 individuals from Korea and China. The length of ITS 1 ranged from 253 to 259 bp. The 5.8S was 159 bp and ITS2 was observed to be 231 bp. A total of 8 different ITS types (Single Nucleotide Polymorphism haplotypes), which showed the difference of 1 - 6 bp, were detected from 65 individuals. The sequence polymorphisms of ITS appeared at 9 different sites. All of four individuals collected at Daejeong-eup in Jeju-do exhibited different types, but all individuals from Naju-si and Muan-gun in Jeollanam-do were identical. The variation of ITS was higher in Jeju-do population than in inland population. Since I. cornuta contains various types of ITS sequences, ITS analyses will provide important information on genetic diversity and conservation of this species.

Sequential Longest Section Color Winning Algorithm for Car Paint Sequencing Problem (자동차 페인트 순서 문제의 연속된 최장 구간 색 승리 알고리즘)

  • Lee, Sang-Un
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.20 no.1
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    • pp.177-186
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    • 2020
  • This paper deals with the car paint sequencing problem (CPSP) that the entrance sequence is to same colored group with maximum sequenced cars for the buffer arriving cars from the body shop. This problem classified by NP-complete problem because of the exact solution has not obtained within polynomial time. CPSP is aim to minimum pugging number that each pugging must be performs at color changing time in order to entirely cleaning the remaining previous color. To be obtain the minimum number of moving distance with window concept and minimum number of pugging, this paper sorts same color and arriving sequence. Then we basically decide the maximum length section color time to winner team using stage race method. For the case of the loser team with no more racing or yield to loser team and more longer stage in upcoming racing, the winner team give way to loser team. As a result, all cars(runners) are winner in any stage without fail. For n cars, the proposed algorithm has a advantage of simple and fast with O(nlogn) polynomial time complexity, this algorithm can be get the minimum number of moving distance and purging for all of experimental data.

Characterization of a Novel cry1-Type Gene from Bacillus thuringiensis subsp. alesti Strain LY-99

  • Qi, Xu Feng;Li, Ming Shun;Choi, Jae-Young;Roh, Jong-Yul;Song, Ji Zhen;Wang, Yong;Jin, Byung-Rae;Je, Yeon-Ho;Li, Jian Hong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.18 no.1
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    • pp.18-27
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    • 2009
  • B. thuringiensis strain LY-99 belonging to subsp. alesti (H3a3c), was isolated from Chinese tobacco warehouse and showed significantly high toxicity to Plutella xylostella. For the identification of the cry1-type genes from B. thuringiensis LY-99, an extended multiplex PCRrestriction fragment length polymorphism (PCRRFLP) method was established by using two pairs of universal primers based on the conserved regions of the cry1-type genes to amplify around 2.4 kb cry1-type gene fragments. Then the DNA fragment was cloned into pGEM-T Easy vector and digested with EcoRI and EcoRV enzymes. Through this method, a known cry1-type gene was successfully identified from the reference strain, B. thuringiensis subsp. alesti. In addition, the RFLP patterns revealed that B. thuringiensis LY-99 included a novel cry1A-type gene in addition to cry1Aa, cry1Ac, cry1Be and cry1Ea genes. The novel cry1A-type gene was designated cry1Ah2 (Genbank accession No DQ269474). An inverse PCR method was used to amplify the flank regions of cry1Ah2 gene. Finally, 3143 bp HindIII fragment from B. thuringiensis LY-99 plasmid DNA including 5' region and partial ORF was amplified, and sequence analysis revealed that cry1Ah2 gene from LY-99 showed 89.31% of maximum sequence similarity with cry1Ac1 crystal protein gene. In addition, the deduced amino acid sequence of Cry1Ah2 protein shared 87.80% of maximum identity with that of Cry1Ac2. This protein therefore belongs to a new class of B. thuringiensis crystal proteins.