• Title/Summary/Keyword: Neighbor-Joining

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Forensically Important Blow Flies Chrysomya pinguis, C. villeneuvi, and Lucilia porphyrina (Diptera: Calliphoridae) in a Case of Human Remains in Thailand

  • Monum, Tawatchai;Sukontason, Kabkaew L.;Sribanditmongkol, Pongruk;Sukontason, Kom;Samerjai, Chutharat;Limsopatham, Kwankamol;Suwannayod, Suttida;Klong-klaew, Tunwadee;Wannasan, Anchalee
    • Parasites, Hosts and Diseases
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    • v.55 no.1
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    • pp.71-76
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    • 2017
  • This is the first study to report Chrysomya pinguis (Walker) and Lucilia porphyrina (Walker) (Diptera: Calliphoridae) as forensically important blow fly species from human cadavers in Thailand, in addition to Chrysomya villeneuvi (Patton) already known in Thailand. In 2016, a fully decomposed body of an unknown adult male was discovered in a high mountainous forest during winter in Chiang Mai province. The remains were infested heavily with thousands of blow fly larvae feeding simultaneously on them. Morphological identification of adults reared from the larvae, and molecular analysis based on sequencing of 1,247 bp partial mitochondrial cytochrome c oxidase subunit 1 gene (CO1) of the larvae and puparia, confirmed the above mentioned 3 species. The approving forensic fly evidence by molecular approach was described for the first time in Thailand. Moreover, neighbor-joining phylogenetic analysis of the CO1 was performed to compare the relatedness of the species, thereby affirming the accuracy of identification. As species of entomofauna varies among cases in different geographic and climatic circumstances, C. pinguis and L. porphyrina were added to the list of Thai forensic entomology caseworks, including colonizers of human remains in open, high mountainous areas during winter. Further research should focus on these 3 species, for which no developmental data are currently available.

Genotypic Diversity of the Complete Open-Reading Frame 7 Sequences of Porcine Reproductive and Respiratory Syndrome Viruses in Korea and Coexistence of Two Genotypes

  • Chu, Jia-Qi;Kim, Myung-Cheol;Park, Chang-Sik;You, Myung-Jo;Jun, Moo-Hyung
    • Journal of Veterinary Clinics
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    • v.25 no.3
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    • pp.139-145
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    • 2008
  • To investigate the genotypic diversity of the porcine reproductive and respiratory syndrome viruses (PRRSV) in Korea, we examined 92 clinical samples from three provinces by RT-PCR and a nested PCR, and the complete open-reading frame 7 (ORF 7) sequences of 15 samples selected from 72 PCR-positive specimens were analyzed. When we compared nucleotide (amino acid) sequences of 80 isolates from Korea and overseas countries, the sequences of 7 samples belonged to North American (NA)-genotype, and those of 8 samples, to European (EU)-genotype. The nucleotide (amino acid) identities between two genotypes were 63.7% (59.8%) to 65.1% (63.1%). When compared with NA prototype VR-2332, the 7 strains of NA-genotype shared 89.8% (93.6%) to 91.2% (96.0%) identity of nucleotide (amino acid) sequence. The 8 strains of EU-type shared 93.6% (92.3%) to 94.3% (93.8%) identity of nucleotide (amino acid) sequence as compared to EU prototype Lelystad. In phylogenetic tree analysis by neighbor-joining method, all of the 8 EU-type strains were clustered into group 4 distinct from ED-prototype Lelystad (group 1). In NA-genotype, 24 domestic isolates reported previously and the 7 strains of NA-type determined in this study were clustered into group 1, while US prototype VR 2332 was classified into different group (group 2). These results suggest that emergence of EU-genotype and the dual-infection of NA- and EU-genotypes may be prevalent in the pig farms in Korea. The high degree of genetic diversity of field PRRSVs should be taken into consideration for control and preventive measures.

Phylogenetic Relationships among Groupers (Genus Epinephelus) Based on Mitochondrial Cytochrome b DNA Sequences

  • KANG Geo Young;SONG Choon Bok
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.37 no.5
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    • pp.414-422
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    • 2004
  • To infer phylogenetic relationships among Epinephelus species inhabiting coastal regions of Korean peninsula, mitochondrial cytochrome b genes from 9 species belonging to the subfamily Epinephelinae were PCR-amplified, cloned and sequenced. Aligned cytochrome b sequences of 10 species containing one additional sequence from GenBank were 1,140 base pairs in length, including 439 variable and 330 parsimony informative sites. The cytochrome b genes of 10 species, as other vertebrates studied to date, exhibit unequal base compositions: an entirely low G content ($15.2{\pm}0.3{\%}$on average) and almost equal T, C and A contents ($29.3{\pm}0.8{\%},\;30.7{\pm}1.0{\%},\;and\;24.8{\pm}0.5{\%}$ on average, respectively).In third codon positions, transitional substitutions especially between Epinephelus species and outgroup species are almost certainly saturated or near saturation. Phylogenetic analyses were performed with sequence data from 8 Epinephelus species and 2 outgroup species (Cephalopholis urodela and Vaviola louti) by using distance-based (neighbor-joining and minimum evolution) and parsimony-based (maximum parsimony) methods. The results showed that the monophyly of the genus Epinephelus was supported by relatively high bootstrap values. However, phylogenetic relationships among E. areolatus, E. moara, E. septemfasciatus, and Epinephelus sp were poorly resolved. Within the genus Epinephelus, three resolved monophyletic groups were found: clade 1 included E. akaara and E. awoara;, clade 2 included E. fasciatus and E. merra; and clade 3 included E. akaara, E. awoara, E. fasciatus, E. merra, E. areolatus, E. moara, E. septemfasciatus and Epinephelus Sp.

ISAG-recommended Microsatellite Marker Analysis Among Five Korean Native Chicken Lines

  • Choi, Nu-Ri;Hoque, Md. Rashedul;Seo, Dong-Won;Sultana, Hasina;Park, Hee-Bok;Lim, Hyun-Tae;Heo, Kang-Nyeong;Kang, Bo-Seok;Jo, Cheorun;Lee, Jun-Heon
    • Journal of Animal Science and Technology
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    • v.54 no.6
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    • pp.401-409
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    • 2012
  • The objective of this study was to determine genetic variation of five Korean native chicken lines using 30 microsatellite (MS) markers, which were previously recommended by ISAG (International Society for Animal Genetics). The initial study indicated that two microsatellite markers, MCW0284 and LEI0192, were not amplified in these lines and excluded for further analysis. Twenty eight microsatellite markers were investigated in 83 birds from five Korean native chicken lines. The identified mean number of alleles was 4.57. Also, the expected, observed heterozygosity (He, Ho) and polymorphism information content (PIC) values were estimated in these markers and they ranged from 0.31~0.868, 0.145~0.699, and 0.268~0.847, respectively. The results were used for the discrimination of five chicken lines using genetic distance values and also neighbor-joining phylogenetic tree was constructed. Based on the He and PIC values, eighteen markers are enough for the discrimination of these Korean native chicken lines for the expected probability of identity values among genotypes of random individuals (PI), random half sibs ($PI_{half-sibs}$) and random sibs ($PI_{sibs}$). Taken together, these results will help the decision of conservation strategies and establishment of traceability system in this native chicken breed. Also, the use of ISAG-recommended microsatellite markers may indicate that the global comparison with other chicken breeds is possible.

Investigation of KIT Gene Polymorphisms in Korean Cattle

  • Hoque, Md. Rashedul;Lee, Seung-Hwan;Lim, Da-Jeong;Cho, In-Cheol;Choi, Nu-Ri;Seo, Dong-Won;Lee, Jun-Heon
    • Journal of Animal Science and Technology
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    • v.54 no.6
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    • pp.411-418
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    • 2012
  • KIT gene is the major causative gene for coat color variation in diverse animal species. This gene regulates melanocyte migration from the neural crest to target tissues and the mutation of this gene can affect dominant white phenotypes in animals. Because this gene has a major influence for the coat color variation, single nucleotide polymorphisms (SNPs) in 14 Korean cattle (Hanwoo) and 5 Holstein individuals were investigated. The Hanwoo DNA samples included three different colored (5 Black, 5 Yellow and 4 Stripe) animals. Total 126 polymorphisms have been identified and 23 of them are located in the exon region. Also, 5 bp (TTCTC) and 3 bp (TCT) intronic indels in intron 3 and intron 5, respectively, were identified. Out of 23 exonic polymorphisms, 15 SNPs are the missense mutations and the rest of the SNPs are silence mutations. The neighbor-joining phylogenetic tree was constructed for the different colored animals using the obtained KIT gene sequences. Holstein breed showed a clear breed-specific cluster in the phylogenetic tree which is differed from Hanwoo. Also, three colored Hanwoo animals were not discriminated among the breeds. The KIT gene polymorphisms identified in this study will possibly give some solutions for the color variations in cattle with further verifications.

Evaluation of the genetic structure of indigenous Okinawa Agu pigs using microsatellite markers

  • Touma, Shihei;Arakawa, Aisaku;Oikawa, Takuro
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.2
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    • pp.212-218
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    • 2020
  • Objective: Agu pigs are indigenous to the Okinawa prefecture, which is the southernmost region of Japan. Agu pigs were exposed to a genetic bottleneck during the 20th century, due to the introduction of European pig breeds. The objective of this study was to elucidate the genetic structure of Agu pigs and to determine their relationships with those of five European breeds, two Chinese breeds and Ryukyu wild boar using microsatellite markers. Methods: A total of 203 DNA samples from 8 pig breeds were used in this study. Genotyping was performed using 21 microsatellite markers distributed across 17 chromosomes. Results: Numbers of effective alleles in Agu pigs were fewer than in European breeds and Ryukyu wild boar. Among domestic pigs, Agu pigs had the lowest heterozygosity (0.423) and highest inbreeding coefficient (FIS = 0.202), indicating a severe loss of heterozygosity in Agu pigs possibly due to inbreeding. Neighbor-joining tree analysis was performed based on Reynolds' genetic distances, which clustered Agu pigs with Duroc pigs. However, principal component analysis revealed a unique genetic position of the Agu pig, and the second principal component separated Agu pigs from all other breeds. Structure analysis with the optimal assumption of seven groups (K = 7) indicated that Agu pigs form an independent cluster from the other breeds. In addition, high and significant FST values (0.235 to 0.413) were identified between Agu pigs and the other breeds. Conclusion: This study revealed a substantial loss of genetic diversity among Agu pigs due to inbreeding. Our data also suggest that Agu pigs have a distinctive genetic structure, although gene flows from European breeds were observed.

Acinetobacter marinus sp. novo and Acinetobacter seohaensis sp. nov., Isolated from Sea Water of the Yellow Sea in Korea

  • Yoon, Jung-Hoon;Kim, In-Gi;Oh, Tae-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1743-1750
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    • 2007
  • Two Gram-negative, nonmotile, coccobacilli, SW-$3^T$ and SW-$100^T$, were isolated from sea water of the Yellow Sea in Korea. Strains SW-$3^T$ and SW-$100^T$ contained ubiquinone-9 (Q-9) as the predominant respiratory lipoquinone and $C_{18:1}\;{\omega}9c$ and $C_{16:0}$ as the major fatty acids. The DNA G+C contents of strains SW-$3^T$ and SW- $100^T$ were 44.1 mol% and 41.9 mol%, respectively. A neighbor-joining tree based on l6S rRNA gene sequences showed that the two isolates fell within the evolutionary radiation enclosed by the genus Acinetobacter. Strains SW-$3^T$ and SW-$100^T$ exhibited a l6S rRNA gene similarity value of 95.7% and a mean DNA-DNA relatedness level of 9.2%. Strain SW-$3^T$ exhibited l6S rRNA gene sequence similarity levels of 93.5-96.9% to the validly described Acinetobacter species and fifteen Acinetobacter genomic species. Strain SW-$100^T$ exhibited l6S rRNA gene sequence similarity levels of less than 97.0% to the other Acinetobacter species except Acinetobacter towneri DSM $14962^T$ (98.0% similarity). Strains SW-$3^T$ and SW-$100^T$ exhibited mean levels of DNA-DNA relatedness of 7.3-l6.7% to the type strains of some phylogenetically related Acinetobacter species. On the basis of phenotypic, phylogenetic, and genetic data, strains SW-$3^T$ and SW-$100^T$ were classified in the genus Acinetobacter as two distinct novel species, for which the names Acinetobacter marinus sp. novo (type strain SW-$3^T$=KCTC $12259^T$=DSM $16312^T$) and Acinetobacter seohaensis sp. novo (type strain SW-$100^T$=KCTC $12260^T$=DSM $16313^T$) are proposed, respectively.

Identification of Lacrymaria velutina (Pers. Ex Fr.) Konrad & Maubl. from Micheon-myeon, Jinju-city, Korea

  • Han, Ki-Soo;Volk, Thomas J.;Kim, Hee-Kyu
    • Mycobiology
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    • v.38 no.4
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    • pp.249-255
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    • 2010
  • We identified Lacrymaria velutina of the Coprinaceae in Korea. The unusually large and sturdy fruiting body, fibrillose to fibrillose-scaly cap and stalk without a volva with an obscure superior hairy ring zone or hairy annulus, and blackish brown, warted spores distinguished this species from closely related Psathyrella species. An illustrated account of the microscopic traits is presented. Fruiting bodies with obtusely hemispherical caps, 2.5~6 cm, becoming convex with age; surface dry, densely fibrillose-scaly with split margin; stipe, 4.5~6 cm, equal, hollow, fibrillose, dry, whitish above the superior ring zone, light brown below; crowded gills, adnexed, dark black at maturity. Pileipellis typically cellular with the gill edge appearing white and beaded. Blackish brown basidiospores that discolor in concentrated sulfuric acid. Spores elliptical, warted, $9\sim11{\times}6\sim8{\mu}m$, with prominent snout-like germpores. Cheilocystidia abundant, $57\sim68{\times}19\sim25{\mu}m$, and narrowly elongated clavate, often clustered in threes or fours. Pleurocystidia rarely present, $45\sim47.5{\times}12\sim13{\mu}m$, and clavate to utriform. This trait distinguishes our sample as L. velutina from other Psathyrella spp. of the Coprinaceae, which have smooth spores. This taxon was clarified by the observation that Psathyrella spores fade in concentrated sulfuric acid. A molecular phylogenetic study revealed that our specimen was Lacrymria velutipes, which is closely related to Lacrymaria lacrymabunda. Moreover, those two species are clearly distinguishable from other Psathyrella species, which agreed with the morphologically distinctive traits described above. We believe that this is the first report of this taxon, which has not been described in Korea.

Development of EST-SSR markers for genetic diversity analysis in little millet (Panicum sumatrense) genetic resources

  • Lee, Myung-Chul;Choi, Yu-Mi;Lee, Sukyeung;Yoon, Hyemyeong;Oh, Sejong
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.74-74
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    • 2018
  • Little millet (Panicum sumatrense) is well known for its salt and drought stress tolerance and high nutritional value, but very limited knowledge of genetic variation and genomic information is available. This study was to develop highly polymorphic EST-SSR markers based on cross-species transferability of derived SSRs from switchgrass EST databases and characterize newly developed EST - SSRs to better understand the genetic diversity of collected 37 germplasm accessions of little millet. A total of 779 primer pairs were designed from the 22,961 EST sequences of switchgrass (Pancium virgatum), of which 48 EST - SSR markers were developed based on the trials of transferability of these primers in little millet. The EST - SSR amplicons showed reproducible single band polymorphism and produced a total of 160 alleles with an average of 3.3 alleles per locus in 37 accessions of little millet. T he average values of expected and observed heterozygosities were 0.266 and 0.123, respectively. T he polymorphic information content (PIC) values were observed in range of 0.026 to 0.549 with an average of 0.240. The genetic relatedness among the little millet accessions was evaluated by neighbor-joining dendrogram, which grouped all accessions into two distinct groups. The validation thus demonstrated the utility of the switchgrass EST - SSR markers in assessing genomic relationships in little millet. T he findings from this study could be useful for designing strategies for the identification of diverse germplasm for conservation and future molecular breeding programs for little millet.

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Classification of Viruses Based on the Amino Acid Sequences of Viral Polymerases (바이러스 핵산중합효소의 아미노산 서열에 의한 바이러스 분류)

  • Nam, Ji-Hyun;Lee, Dong-Hun;Lee, Keon-Myung;Lee, Chan-Hee
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.285-291
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    • 2007
  • According to the Baltimore Scheme, viruses are classified into 6 main classes based on their replication and coding strategies. Except for some small DNA viruses, most viruses code for their own polymerases: DNA-dependent DNA, RNA-dependent RNA and RNA-dependent DNA polymerases, all of which contain 4 common motifs. We undertook a phylogenetic study to establish the relationship between the Baltimore Scheme and viral polymerases. Amino acid sequence data sets of viral polymerases were taken from NCBI GenBank, and a multiple alignment was performed with CLUSTAL X program. Phylogenetic trees of viral polymerases constructed from the distance matrices were generally consistent with Baltimore Scheme with some minor exceptions. Interestingly, negative RNA viruses (Class V) could be further divided into 2 subgroups with segmented and non-segmented genomes. Thus, Baltimore Scheme for viral taxonomy could be supported by phylogenetic analysis based on the amino acid sequences of viral polymerases.