• Title/Summary/Keyword: Molecular phylogenetic tree

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Development of molecular markers for varietal identification of Brassica juncea on the basis of the polymorphic sequence of ITS regions and MITE families (갓 (Brassica juncea) 품종구분을 위한 ITS 영역 및 MITE Family 정보를 이용한 분자표지 개발)

  • Yang, Kiwoung;Yi, Go-eun;Robin, Arif Hasan Khan;Jeong, Namhee;Lee, Yong-Hyuk;Park, Jongin;Kim, Hoyteak;Chung, Mi-Young;Nou, Ill-Sup
    • Horticultural Science & Technology
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    • v.34 no.2
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    • pp.305-313
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    • 2016
  • Brassica juncea (2n = 4x = 36, AABB genome, 1,068 Mb) is a U's triangle species and an amphidiploid derivative of B. rapa and B. nigra. Fifteen varieties were used to study the ITS (internal transcribed spacer) regions of ribosomal DNA and MITEs (miniature inverted-repeat transposable elements) with a view of developing specific molecular markers. ITSs and MITEs are an excellent resource for developing DNA markers for genomics and evolutionary studies because most of them are stably inherited and present in high copy numbers. The ITS (ITS1 and ITS2) sequence was compared with the consensus sequence of B. rapa and B. nigra. Variation in ITS1 created two separate groups among 15 varieties, with 10 varieties in one group and 5 in the other. Phylogenetic analysis revealed two major clusters for those 10 and 5 varieties. Among the 160 different MITE primers used to evaluate the selected 15 varieties of B. juncea, 70 were related to the Stowaway, 79 to the Tourist, 6 to the hAT, and 5 to the Mutator super-families of MITEs. Of 160 markers examined, 32 were found to be polymorphic when fifteen different varieties of B. juncea were evaluated. The variety 'Blackgat' was different from the other mustard varieties with respect to both phenotype and genotype. The diversity of 47 additional accessions could be verified using eight selected molecular markers derived from MITE family sequences. The polymorphic markers identified in this study can be used for varietal classification, variety protection, and other breeding purposes.

Monitoring of genetically close Tsaiya duck populations using novel microsatellite markers with high polymorphism

  • Lai, Fang-Yu;Chang, Yi-Ying;Chen, Yi-Chen;Lin, En-Chung;Liu, Hsiu-Chou;Huang, Jeng-Fang;Ding, Shih-Torng;Wang, Pei-Hwa
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.6
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    • pp.888-901
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    • 2020
  • Objective: A set of microsatellite markers with high polymorphism from Tsaiya duck were used for the genetic monitoring and genetic structure analysis of Brown and White Tsaiya duck populations in Taiwan. Methods: The synthetic short tandem repeated probes were used to isolate new microsatellite markers from the genomic DNA of Tsaiya ducks. Eight populations, a total of 566 samples, sourced from Ilan Branch, Livestock Research Institute were genotyped through novel and known markers. The population genetic variables were calculated using optional programs in order to describe and monitor the genetic variability and the genetic structures of these Tsaiya duck populations. Results: In total 24 primer pairs, including 17 novel microsatellite loci from this study and seven previously known loci, were constructed for the detection of genetic variations in duck populations. The average values for the allele number, the effective number of alleles, the observed heterozygosity, the expected heterozygosity, and the polymorphism information content were 11.29, 5.370, 0.591, 0.746, and 0.708, respectively. The results of analysis of molecular variance and principal component analysis indicated a contracting Brown Tsaiya duck cluster and a spreading White Tsaiya duck cluster. The Brown Tsaiya ducks and the White Tsaiya ducks with Pekin ducks were just split to six clusters and three clusters when K was set equal to 6 and 3 in the Bayesian cluster analysis. The individual phylogenetic tree revealed eight taxa, and each individual was assigned to its own population. Conclusion: According to our study, the 24 novel microsatellite markers exhibited a high capacity to analyze relationships of inter- and intra-population in those populations with a relatively limited degree of genetic diversity. We suggest that duck farms in Taiwan could use the new (novel) microsatellite set to monitor the genetic characteristics and structures of their Tsaiya duck populations at various intervals in order to ensure quality breeding and conservation strategies.

Survey of genetic structure of geese using novel microsatellite markers

  • Lai, Fang-Yu;Tu, Po-An;Ding, Shih-Torng;Lin, Min-Jung;Chang, Shen-Chang;Lin, En-Chung;Lo, Ling-Ling;Wang, Pei-Hwa
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.2
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    • pp.167-179
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    • 2018
  • Objective: The aim of this study was to create a set of microsatellite markers with high polymorphism for the genetic monitoring and genetic structure analysis of local goose populations. Methods: Novel microsatellite markers were isolated from the genomic DNA of white Roman geese using short tandem repeated probes. The DNA segments, including short tandem repeats, were tested for their variability among four populations of geese from the Changhua Animal Propagation Station (CAPS). The selected microsatellite markers could then be used to monitor genetic variability and study the genetic structures of geese from local geese farms. Results: 14 novel microsatellite loci were isolated. In addition to seven known loci, two multiplex sets were constructed for the detection of genetic variations in geese populations. The average of allele number, the effective number of alleles, the observed heterozygosity, the expected heterozygosity, and the polymorphism information content were 11.09, 5.145, 0.499, 0.745, and 0.705, respectively. The results of analysis of molecular variance and principal component analysis indicated a contracting white Roman cluster and a spreading Chinese cluster. In white Roman populations, the CAPS populations were depleted to roughly two clusters when K was set equal to 6 in the Bayesian cluster analysis. The founders of private farm populations had a similar genetic structure. Among the Chinese geese populations, the CAPS populations and private populations represented different clads of the phylogenetic tree and individuals from the private populations had uneven genetic characteristics according to various analyses. Conclusion: Based on this study's analyses, we suggest that the CAPS should institute a proper breeding strategy for white Roman geese to avoid further clustering. In addition, for preservation and stable quality, the Chinese geese in the CAPS and the aforementioned proper breeding scheme should be introduced to geese breeders.

Molecular and Epidemiological Characterization of Enteroviruses Isolated in Chungnam, Korea from 2005 to 2006

  • Baek, Kyung-Ah;Park, Kwi-Sung;Jung, Eun-Hye;Chung, Eun-Hee;Park, Joon-Soo;Choi, Hwa-Jung;Baek, Seung-Hwa;Jee, Young-Mee;Cheon, Doo-Sung;Ahn, Gwang-Sook
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.1055-1064
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    • 2009
  • Enteroviruses were identified and characterized from patients with aseptic meningitis and other enterovirus-related diseases in Chungnam, Korea from 2005 to 2006. Enteroviruses were isolated from 79 of 519 cases (15.2%) in 2005, and 37 of 386 cases (9.6%) in 2006. Based on partial VP1 sequencing, a total of 116 enterovirus isolates were resolved into 13 types. Prevalent among the Chungnam isolates were echovirus 18 and coxsackievirus B5 in 2005, and echoviruses 5 and 25 in 2006. This is the first time echoviruses 5 and 18 have been identified in Korea since enterovirus surveillance began there in 1993. The temporal distribution of enterovirus epidemics in Chungnam showed a remarkable seasonal pattern, with cases occurring during most of the three months of the summer from June to August. The highest rate of enterovirus-positive cases occurred in patients less than 1 year of age. The ratio of male to female enterovirus-positive patients was approximately 1.8:1. Comparison of the VP1 amino acid sequences of the 15 coxsackievirus B5 isolates with reference strains revealed that all Chungnam isolates are substituted at positions 23 (V231), 19 (S19G), 75 (Y75F), and 95 (N95S). Upon comparing the nine ECV5 isolates with foreign strains, it was found that only the Chungnam isolates, with the exception of Kor06-ECV5-239cn, have P at position 153 and F at position 146. The three ECV9 isolates from 2006 show alterations at amino acids 36, 148, and 154 outside of the BC-loop and at position 84 in the BC-loop, whereas the seven isolates from 2005 and the other ECV9 strains in the database only show the alteration at position 84 (D, I, N, S). The five ECV25 isolates have an S residue at position 134, whereas most of the foreign strains have an N residue.

Morphological and Biochemical Characteristics in the Korean Landraces of Mulberry Tree, Morus (한국 재래뽕의 생태 및 품종간 특이성에 관한 연구)

  • Kim, Hyeon-Bok;Kim, Nam-Su;Park, Gwang-Jun
    • Journal of Sericultural and Entomological Science
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    • v.38 no.2
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    • pp.81-92
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    • 1996
  • This study was conducted to characterize mulberry varieties by the analysis of morphological characters as well as biochemical and molecular biological markers. As for the budding stage Geumsang, Chosang i and Yeongbyeonchuwoo were early, but Dangsang 6, Hwangchuwoo were late. The lowest varietiy in rate of death atop was Dangsang 8(0.0%). Suncheonppong was the highest leaf yields in spring and autumn rearing season. In biochemical isozyme analysis, peroxidase gave good zymogram patterns in isoelectric focusing electrophoresis. There were high variations in RAPD analysis among the mulberry trees. From the obtained peroxidase and RAPD variations, cluster phylogenetic analysis was carried out using NT-SYS PC program. There were no clear grouping patterns between native varieties and leading varieties. The highest similarity was observed between Suwonsang 1 and Suwonsang 2 at about 90% similarity level.

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Molecular Identification of the Bloom-forming Cyanobacterium Anabaena from North Han River System in Summer 2012 (북한강 수계 조류대발생 원인종 남조 Anabaena의 분자계통학적 검토)

  • Li, Zhun;Han, Myung-Soo;Hwang, Su-Ok;Byeon, Myeong-Seop;Hwang, Soon-Jin;Kim, Baik-Ho
    • Korean Journal of Ecology and Environment
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    • v.46 no.2
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    • pp.301-309
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    • 2013
  • Between May and August 2012, a massive cyanobacterial bloom with Anabaena has been occurred throughout the North Han River. Sampling was conducted at one station on each lake, L. Uham, L. Cheongpyung, and L. Paldang, where occurred a dense bloom, in 13 July. According to the microscopic examination, the blooms was dominated by one specific filamentous cyanobacterium Anabaena and other phytoplankton. Morphologically, previous literature proven that this Anabaena species is A. crassa (Lemmermann) Komark.-Legn. & Cronberg. However, identification of species in a mixed population is complicated due to limited morphological differences. Therefore, with live sample including trichome, akinete and heterocyst, the sequences of 16S rRNA gene of Anabaena isolates were cloned and analyzed, and three 16S rRNA gene sequences of 1188~1520 bp in length were obtained. It was shown from the homologous analysis results that the obtained 16S rRNA sequences were highly homologous to the relevant sequences of A. crassa in GenBank. The 16S rRNA sequences of 63 species were retrieved from GenBank, and the phylogenetic tree was constructed by using these sequences.

Molecular Biological Studies on the Stress Protein HSP90$\beta$ Gene from Flounder (Paralichthys olivaceus) (넙치(Paralichthys olivaceus) HSP90$\beta$ 유전자의 분자생물학적 연구)

  • Lee, Jae-Hyeong;Kim, Yeong-Tae
    • Microbiology and Biotechnology Letters
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    • v.32 no.4
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    • pp.297-306
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    • 2004
  • Heat shock proteins (HSPs) were induced in cells in the thermal stress, and the HSP90 family is one of the major classes of HSPs. Gene encoding HSPs have been characterized from various mammals and piscine. We have cloned and sequenced the HSP90 cDNA from a brain cDNA library constructed from flounder (Paralichthys oliThe result of sequence analysis shows it to be the HSP90~. The nucleotide sequence of the HSP90$\beta$ was composed of 2791 long, encoding 726 amino acid residues. The flounder hsp90$\beta$ gene showed very high sequence homology with hsp90f3 of European sea bass (96.6%), zebrafish (92.9%), Atlantic salmon (92.0%) and human (89.5%). We also constructed a phylogenetic tree based on HSP90 amino acid sequences from vertebrate species. Gene-specific primers were selected and used in RT-PCR reactions to measure the basal hsp90$\beta$ mRNA. The hsp90f3 gene is constitutively expressed at a fairly high level in all examined tissues (brain, liver, kidney, muscle, and spleen). In order to express protein of flounder hsp90$\beta$ in E. coli, we used the His-tagged pETvector. Then, the expression of flounder HSP90$\beta$ was confirmed by Western blot analysis.

Molecular Identification and Sequence Analysis of Coat Protein Gene of Ornithogalum mosaic virus Isolated from Iris Plant

  • Yoon, Hye-In;Ryu, Ki-Hyun
    • The Plant Pathology Journal
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    • v.18 no.5
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    • pp.251-258
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    • 2002
  • A potyvirus was isolated from cultivated Iris plants showing leaf streak mosaic symptom. Reverse transcription and polymerase chain reaction (RT-PCR) product of 1 kb long which encoded partial nuclear inclusion B and N-terminal region of viral coat protein (CP) genes for potyviruses was successfully amplified with a set of potyvirus-specific degenerate primers with viral RNA samples from the infected leaves: The RT-PCR product was cloned into the plasmid vector and its nucleotide sequences were determined. The nucleotide sequence of a CDNA clone revealed that the virus was an isolate of Ornithogalum moseic virus (OrMV) based on BLAST search analysis and was denoted as OrMV Korean isolate (OrMV-Ky). To further characterize the CP gene of the virus, a pair of OrMV-specific primers was designed and used for amplification of the entire CP gene of OrMV-Kr, The virus was easily and reliably detected from virus-infected Iris leaves by using the RT-PCR with the set of virus-specific primers. The RT-PCR product of the CP gene of the virus was cloned and its sequences were determined from selected recombinant CDNA clones. Sequence analysis revealed that the CP of OrMV-Kr consisted of 762 nucleotides, which encoded 253 amino acid residues. The CP of OrMV-Ky has 94.1-98.0% amino acid sequence identities (20 amino acid alterations) with that of other three isolates of OrMV, Two NT rich potential N-glycosylation motif sequences, NCTS and NWTM, and a DAC triple box responsible for aphid transmission were conserved in CPs of all the strains of OrMV. The virus has 58.5-86.2% amino acid sequence identities with that of other 16 potyviruses, indicating OrMV to be a distinct species of the genus. OrMV-Ky was the most related with Pterostylia virus Yin the phylogenetic tree analysis of CP at the amino acid level. This is the first report on the occurrence of OrMV in Iris plants in Korea. Data in this study indicate that OrMV is found in cultivated Iris plants, and may have mixed infection of OrMV and Iris severe mosaic virus in Korea.

Maternal Origins of the Jeju Native Pig Inferred from PCR-RFLP Haplotypes and Molecular Phylogeny for Mitochondrial DNA CYTB Gene Sequences (미토콘드리아 DNA CYTB 유전자 서열에 대한 분자 계통과 PCR-RFLP 반수체형에 근거한 제주재래돼지의 모계 기원)

  • Han, Sang-Hyun;Ko, Moon-Suck;Jeong, Ha-Yeon;Lee, Sung-Soo;Oh, Hong-Shik;Cho, In-Cheol
    • Journal of Life Science
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    • v.21 no.3
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    • pp.341-348
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    • 2011
  • In an effort to gain greater understanding of the maternal lineages of the Jeju native pig (JNP), we analyzed the mitochondrial DNA (mtDNA) CYTB gene and compared it with those of other pig breeds. PCR-RFLP analysis was conducted with six pig breeds including JNP, and then the RFLP patterns allowed for the separation of the pig breeds into two distinct haplotypes (mtCYTB1 and mtCYTB2). The JNP CYTB sequences were detected in both the European and Asian breed clusters on the phylogenetic tree. The J2 group was sorted with the indigenous cluster of Asian pig lineages and was related closely to Chinese native pig breeds, but a second group, J1, was sorted with the European pig lineages and appeared to be related to Spanish Iberian native pigs, rather than to Asian breeds. These results indicate that the JNP currently raised on Jeju Island have two major maternal origins estimated in Asian and European pigs. We concluded that the JNP that share a common lineage with indigenous Asian pigs were domesticated in the distant past, originating from pigs that were already being raised elsewhere at that time, and that the European pig breeds introduced in the twentieth century have also contributed to the formation of this pig population.

The Identity of Eggplant Powdery Mildews Collected in Korea (한국에서 채집된 가지 흰가루병균의 실체)

  • Cho, Sung-Eun;Choi, In-Young;Shin, Hyeon-Dong
    • The Korean Journal of Mycology
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    • v.45 no.2
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    • pp.91-101
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    • 2017
  • Three species of powdery mildew (Erysiphales) on eggplant (Solanum melongena L.) have been listed in Korea, namely Erysiphe cichoracearum (now genus Golovinomyces), Leveillula taurica, and Sphaerotheca fusca (now genus Podosphaera; syn. Podosphaera xanthii). Since E. cichoracearum was recorded on eggplant for the first time in Korea in 1969, it has been regarded as a major powdery mildew agent on that plant. In 1998, the causal agent of powdery mildew on eggplant was recorded as L. taurica, then as S. fusca in 2002. During our extensive field surveys in Korea, we collected 22 samples of eggplant powdery mildews. Our microscopic observations and molecular sequence analyses showed that all of our samples belonged to the genus Podosphaera, in the absence of either E. cichoracearum or L. taurica, suggesting that P. xanthii is the dominant agent of powdery mildew disease on eggplants in Korea. As there have been no additional findings on L. taurica after the first report on the species, it seems to be a minor species that is rarely found in greenhouses. The presence of E. cichoracearum (syn. Golovinomyces cichoracearum s. lat.) on eggplants is questionable, as the morphological characteristics of E. cichoracearum in the original description of the Korean collection deviate from the morphological variations of this species. In addition, no herbarium material of E. cichoracearum remains. Consequently, it seems that P. xanthii is the main species of powdery mildew on eggplants, whereas L. taurica occurs rarely on eggplants, in Korea. This review provides the historical and recent taxonomy of eggplant powdery mildews in detail.