• BMB Reports
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• 제35권6호
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• pp.595-603
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• 2002

A gene that encodes a homologue to baculoviral ODVP-6E/ODV-E56, a baculoviral envelope-associated viral structural protein, has been identified and sequenced on the genome of Choristoneura fumiferana granulovirus (ChfuGV). The ChfuGV odvp-6e/odv-e56 gene was located on an 11-kb BamHI subgenomic fragment using different sets of degenerated primers, which were designed using the results of the protein sequencing of a major 39 kDa structural protein that is associated with the occlusion-derived virus (ODV). The gene has a 1062 nucleotide (nt) open-reading frame (ORF) that encodes a protein with 353 amino acids with a predicated molecular mass of 38.5 kDa. The amino acid sequence data that was derived from the nucleotide sequence in ChfuGV was compared to those of other baculoviruses. ChfuGV ODVP-6E/ODV-E56, along with othe baculoviral ODVP-6E/ODV-E56 proteins, all contained two putative transmembrane domains at their C-terminus. Several putative N-and O-glycosylation, N-myristoylation, and phosphorylation sites were detected in the ChfuGV ODVP-6E/ODV-E56 protein. A similar pattern was detected when a hydrophobicity-plots comparison was performed on ChfuGV ODVP-6E/ODV-E56 with other baculoviral homologue proteins. At the nucleotide level, a late promoter motif (GTAAG) was located at -14 nt upstream to the start codon of the GhfuGV odvp-6e/odv-e56 gene. a slight variant of the polyadenylation signal, AATAAT, was detected at the position +10 nt that is downstream from the termination signal. A phylogenetic tree for baculoviral ODVP-6E/ODV-E56 was constructed using a maximum parsimony analysis. The phylogenetic estimation demonstrated that ChfuGV ODVP-6E/ODV-E56 is most closely related to those of Cydia pomonella granulovirus (CpGV) and Plutella xylostella granulovirus (PxGV).

• BMB Reports
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• 제35권6호
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• pp.553-561
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• 2002

A gene that encodes a protein homologue to baculoviral IE-1 was identified and sequenced in the genome of the Choristoneura fumiferana granulovirus (ChfuGV). The gene has an 1278 nucleotide (nt) open-reading frame (ORF) that encodes 426 amino acids with an estimated molecular weight of 50.33 kDa. At the nucleotide level, several cis-acting regulatory elements were detected within the promoter region of the ie-1 gene of ChfuGV along with other studied granuloviruses (GVs). Two putative CCAAT elements were detected within the noncoding leader region of this gene; one was located on the opposite strand at -92 and the other at -420 nt from the putative start triplet. Two baculoviral late promoter motifs (TAAG) were also detected within the promoter region of the ie-1 gene of ChfuGV. A single polyadenylation signal, AATAAA, was located 18nt downstream of the putative translational stop codon of ie-1 from ChfuGV. At the protein level, the amino acid sequence data that was derived from the nucleotide sequence in ChfuGV IE-1 was compared to those of the Cydia pomonella granulovirus (CpGV), Xestia c-nigrum granulovirus (XcGV) and Plutella xylostella granulovirus (PxGV). The C-terminal regions of the granuloviral IE-1 sequences appeared to be more conserved when compared to the N-terminal regions. A domain, similar to the basic helix-loop-helix like (bHLH-like) domain in NPVs, was detected at the C-terminal region of IE-1 from ChfuGV (residues 387 to 414). A phylogenetic tree for baculoviral IE-1 was constructed using a maximum parsimony analysis. A phylogenetic estimation demonstrates that ChfuGV IE-1 is most closely related to that of CpGV.

• Parasites, Hosts and Diseases
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• 제54권4호
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• pp.503-507
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• 2016

The genus Spirometra belongs to the family Diphyllobothriidae and order Pseudophyllidea, and includes intestinal parasites of cats and dogs. In this study, a plerocercoid labeled as Spirometra mansonoides from the USA was examined for species identification and phylogenetic analysis using 2 complete mitochondrial genes, cytochrome c oxidase I (cox1) and NADH dehydrogenase subunit 3 (nad3). The cox1 sequences (1,566 bp) of the plerocercoid specimen (USA) showed 99.2% similarity to the reference sequences of the plerocercoid of Korean Spirometra decipiens (GenBank no. KJ599679), and 99.1% similarity in regard to nad3 (346 bp). Phylogenetic tree topologies generated using 4 analytical methods were identical and showed high confidence levels with bootstrap values of 1.00, 100%, 100%, and 100% for Bayesian inference (BI), maximum-likelihood (ML), neighbor-joining (NJ), and maximum parsimony (MP) methods, respectively. Representatives of Diphyllobothrium and Spirometra species formed a monophyletic group, and the sister-genera status between these species was well supported. Trapezoic proglottids in the posterior 1/5 region of an adult worm obtained from an experimentally infected cat were morphologically examined. The outer uterine loop of the uterus coiling characteristically consisted of 2 complete turns. The results clearly indicated that the examined Spirometra specimen from the USA matched to S. decipiens very well, and indicated possible presence of the life cycle of this species in this region.

• 한국해양학회지:바다
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• 제3권2호
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• pp.90-93
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• 1998

국내 남해안의 적조발생지역에서 채집한 Prorocentrum minimum, P. micans, P. triestinum, P. balticum, Gymnodinium sanguineum, Alexandrium catenella, Scrippsiella trochoidea, Heterosigma akashiwo를 순수배양하고 이로부터 PCR을 이용해 약 700 bp에 해당되는 24S rRNA 유전자의 D1과 D2 변이부위를 증폭하고 클로닝한 후 염기서열을 분석하였다. 분석된 염기서열들을 이미 밝혀진 적조원인종들의 것과 ClustalW 프로그램을 사용하여 배열하고 계통수를 구성한 결과 Prorocentrum과 Alexandrium 속의 것들은 형태적인 분류결과와 종수준까지 거의 일치함을 보여주었다. 이러한 연구결과는 그 24S rRNA 유전자의 염기서열분석이 국내에서 발생하는 적조원인종들에 대한 종 수준에서의 신속한 확인에 이용될 수 있음을 제시한다.

• Animal Systematics, Evolution and Diversity
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• 제29권2호
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• pp.136-143
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• 2013

The collared scops owl that occurs in Korea is a protected species but its exact specific status has been questioned. To resolve the species status, a molecular phylogenetic analysis was conducted using two fragments of mitochondrial DNA, cytochrome b (cyt b, 891 bp) and NADH dehydrogenase subunit 2 (ND2, 627 bp) genes. Phylogenetic trees of cyt b revealed that all Korean specimens formed a monophyletic group with Japanese scops owl Otus semitorques with very low sequence divergence (d=0.008). We obtained a similar ND2 tree as well (d=0.003); however, the genetic distance between Korean individuals and O. lempiji from GenBank (AJ004026-7, EU348987, and EU601036) was very high and sufficient enough to separate them as species (cyt b, d=0.118; ND2, d=0.113). We also found that Korean species showed high differentiation from O. bakkamoena (AJ004018-20 and EU601034; cyt b, d=0.106; ND2, d=0.113) and O. lettia (EU601109 and EU601033, cyt b, d=0.110; ND2, d=0.117) as well. Therefore, we suggest that the Korean collared scops owl should be designated as Otus semitorques.

• Journal of Plant Biotechnology
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• 제47권1호
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• pp.15-25
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• 2020

Allium L. is one of the largest genera of the Amaryllidaceae family, with more than 920 species including many economically important species used as vegetables, spices, medicines, or ornamental plants. Currently, DNA barcoding tools are being successfully used for the molecular taxonomy of Allium. A total of 46 Allium species were collected from their native areas, and DNA was extracted using the IBRC DNA extraction kit. We used specific primers to PCR amplify matK. DNA sequences were edited and aligned for homology, and a phylogenetic tree was constructed using the neighbor-joining method. The results show thymine (38.5%) was the most frequent and guanine (13.9%) the least frequent nucleotide. The matK regions of the populations were quite highly conserved, and the amount of C and CT was calculated at 0.162 and 0.26, respectively. Analysis of the nucleotide substitution showed C-T (26.22%) and A-G (8.08%) to have the highest and lowest percent, respectively. The natural selection process dN/dS was 1.16, and the naturality test results were -1.5 for Tajima's D and -1.19 for Fu's Fs. The NJ dendrogram generated three distinct clades: the first contained Allium austroiranicum and A. ampeloprasum; the second contained A. iranshahrii, A. bisotunense, and A. cf assadi; and the third contained A. rubellum and other species. In this study, we tested the utility of the matK region as a DNA barcode for discriminating Allium. species.

• Asian-Australasian Journal of Animal Sciences
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• 제32권10호
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• pp.1491-1500
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• 2019

Objective: European pigs have been imported to improve the economically important traits of Thai pigs by crossbreeding and was finally completely replaced. Currently Thai indigenous pigs are particularly kept in a small population. Therefore, indigenous pigs risk losing their genetic diversity and identity. Thus, this study was conducted to perform large-scale genetic diversity and phylogenetic analyses on the many pig breeds available in Thailand. Methods: Genetic diversity and phylogenetics analyses of 222 pigs belonging to Thai native pigs (TNP), Thai wild boars (TWB), European commercial pigs, commercial crossbred pigs, and Chinese indigenous pigs were investigated by genotyping using 26 microsatellite markers. Results: The results showed that Thai pig populations had a high genetic diversity with mean total and effective ($N_e$) number of alleles of 14.59 and 3.71, respectively, and expected heterozygosity ($H_e$) across loci (0.710). The polymorphic information content per locus ranged between 0.651 and 0.914 leading to an average value above all loci of 0.789, and private alleles were found in six populations. The higher $H_e$ compared to observed heterozygosity ($H_o$) in TNP, TWB, and the commercial pigs indicated some inbreeding within a population. The Nei's genetic distance, mean $F_{ST}$ estimates, neighbour-joining tree of populations and individual, as well as multidimensional analysis indicated close genetic relationship between Thai indigenous pigs and some Chinese pigs, and they are distinctly different from European pigs. Conclusion: Our study reveals a close genetic relationship between TNP and Chinese pigs. The genetic introgression from European breeds is found in some TNP populations, and signs of genetic erosion are shown. Private alleles found in this study should be taken into consideration for the breeding program. The genetic information from this study will be a benefit for both conservation and utilization of Thai pig genetic resources.

• Journal of Microbiology and Biotechnology
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• 제33권8호
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• pp.1013-1022
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• 2023

Arbuscular mycorrhizal fungi (AMF) are widespread soil endophytic fungi, forming mutualistic relationships with the vast majority of land plants. Biochar (BC) has been reported to improve soil fertility and promote plant growth. However, limited studies are available concerning the combined effects of AMF and BC on soil community structure and plant growth. In this work, a pot experiment was designed to investigate the effects of AMF and BC on the rhizosphere microbial community of Allium fistulosum L. Using Illumina high-throughput sequencing, we showed that inoculation of AMF and BC had a significant impact on soil microbial community composition, diversity, and versatility. Increases were observed in both plant growth (the plant height by 8.6%, shoot fresh weight by 12.1%) and root morphological traits (average diameter by 20.5%). The phylogenetic tree also showed differences in the fungal community composition in A. fistulosum. In addition, Linear discriminant analysis (LDA) effect size (LEfSe) analysis revealed that 16 biomarkers were detected in the control (CK) and AMF treatment, while only 3 were detected in the AMF + BC treatment. Molecular ecological network analysis showed that the AMF + BC treatment group had a more complex network of fungal communities, as evidenced by higher average connectivity. The functional composition spectrum showed significant differences in the functional distribution of soil microbial communities among different fungal genera. The structural equation model (SEM) confirmed that AMF could improve the microbial multifunctionality by regulating the rhizosphere fungal diversity and soil properties. Our findings provide new information on the effects of AMF and biochar on plants and soil microbial communities.

• 한국자원식물학회지
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• 제23권4호
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• pp.293-302
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• 2010

한국산 쑥속 분류군의 계통분류학적 연구를 위해 Nuclear ribosome DNA의 ITS 염기서열 분석을 실시하였다. 정렬된 염기의 총 길이는 635~643 bp이며, ITS1과 ITS2 부위의 길이는 각각 251~255 bp와 217~222 bp로 나타났다. 염기서열 변이를 보이는 site는 95개로 확인되었다. 그 중 ITS1이 35개, ITS2가 26개로 총 72개의 site가 계통학적으로 유효한 것으로 나타남으로써 ITS1이 ITS2보다 종 분화의 변이가 다양하게 발생하는 것으로 확인되었다. ITS 염기서열을 기초한 계통학적 분석은 쑥속 내에 5개의 Clade를 형성하였다. 그 결과 자방이 퇴화된 분류군들(사철쑥, 제비쑥, 섬쑥, 갯제비쑥)이 하나의 분계조(Clade 1)를 형성함으로써 아속 수준(Subgen. Dracunculus)으로 취급되는 결과를 뒷받침 하였다. 애기비쑥과 큰비쑥은 거의 동일한 유전적 정보를 보였으며(Boostrap 99%), 한국산 참쑥의 학명은 재고 되어야할 것으로 사료된다. 또한, 강화약쑥(A. sp.)은 황해쑥과 매우 가까운 상동성을 보였다(Boostrap 89%). 따라서, 형태적 형질의 변이가 다소 연속적인 쑥속은 DNA 염기서열에 기초한 분자계통학적 연구가 유용한 방법으로 판단되며, 본 ITS 연구결과는 한국산 쑥속의 계통분류를 이해하는데 유용한 형질로 기여할 것으로 기대된다.

• Mycobiology
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• 제37권3호
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• pp.183-188
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• 2009

The molecular phylogeny in nine different commercial cultivated strains of Pleurotus nebrodensis was studied based on their internal transcribed spacer (ITS) region and RAPD. In the sequence of ITS region of selected strains, it was revealed that the total length ranged from 592 to 614 bp. The size of ITS1 and ITS2 regions varied among the strains from 219 to 228 bp and 211 to 229 bp, respectively. The sequence of ITS2 was more variable than ITS1 and the region of 5.8S sequences were identical. Phylogenetic tree of the ITS region sequences indicated that selected strains were classified into five clusters. The reciprocal homologies of the ITS region sequences ranged from 99 to 100%. The strains were also analyzed by RAPD with 20 arbitrary primers. Twelve primers were efficient to applying amplification of the genomic DNA. The sizes of the polymorphic fragments obtained were in the range of 200 to 2000 bp. RAPD and ITS analysis techniques were able to detect genetic variation among the tested strains. Experimental results suggested that IUM-1381, IUM-3914, IUM-1495 and AY-581431 strains were genetically very similar. Therefore, all IUM and NCBI gene bank strains of P. nebrodensis were genetically same with some variations.