• Journal of Plant Biotechnology
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• v.45 no.3
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• pp.236-241
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• 2018

Lilium is a perennial bulbous plant belonging to the liriotypes genus. Our aim was to study the phylogenetic relationships of the Lilium family. Two varieties of Lilium ledebourii, 44 varieties of the gene bank, and one variety from the Tulipa family served as the out group. In order to study the diversity between lilium masses, ITS regions were used to design the marker. The results showed that the guanine base is the most abundant nucleotide. Relatively high conservation was observed in the ITS regions of the populations (0.653). Phylogenetic analysis showed that sargentiae and hybrid varieties are older than other varieties of the Lilium family. Also, the location of L. ledebourii varieties (Damash and Namin) was identified in a phylogenetic tree by using the ITS marker. Overall, our research showed that ITS molecular markers are very suitable for phylogenetic studies in the Lilium family.

• 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.

• The Korea Journal of Herbology
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• v.28 no.3
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• pp.75-84
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• 2013

Objectives : The origin of Breeae Herba (So-gye) and Cirsii Herba (Dae-gye) is differently prescribed in Korean and Chinese modern pharmacopoeia. Since the similar morphological characteristics and chaotic plant names, moreover, the aerial part of Carduus crispus have been used as the Cirsii Herba. To develop a reliable method for correct identification of these herbal medicines and to evaluate the genetic relationship of these closely related plant species, we analyzed sequences of DNA barcode regions. Methods : Thirty-one samples of 6 medicinal plants (B. segeta, B. setosa, C. japonicum var. maackii, C. setidens, C. chanroenicum, and C. crispus) were collected from different habitate and nucleotide sequences of DNA barcode regions (rDNA-ITS, matK, and rbcL) were analyzed after amplification using appropriate primers reported in previous studies. The nucleotides of species-specific authentic marker and phylogenetic relations were estimated based on the entire sequences of DNA barcodes by the analysis of ClastalW and UPGMA, respectively. Results : In comparative analysis of DNA barcode sequences, we obtained specific nucleotides to discriminate the medicinal plant of Breeae/Cirsii Herba in species level and evaluated the phylogenetic relationship of these species. Futhermore, we identified distinct marker nucleotides enough to authenticate respective species. These sequence differences at corresponding positions were avaliable genetic markers to determine the botanical origins of Breeae Herbal as well as Cirsii Herba. Conclusions : These marker nucleotides would be useful to identify the official herbal medicines by providing of definitive information that can identify each plant species and distinguish from unauthentic adulterants and substitutes.

• Journal of Life Science
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• v.21 no.3
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• pp.385-392
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• 2011

The otter (Lutra lutra) in Korea is classified as a first grade endangered species and is managed under state control. We performed a phylogenetic analysis of the otter that inhabits the Changnyeong, Jinju, and Geoje areas in Gyeongsangnamdo, Korea using mtDNA and microsatellite (MS) markers. As a result of the analysis using the 676-bp D-loop sequence of mtDNA, six haplotypes were estimated from five single nucleotide polymorphisms. The genetic distance between the Jinju and Geoje areas was greater than distances within the areas, and the distance between Jinju and Geoje was especially clear. From the phylogenetic tree estimated using the Bayesian Markov chain Monte Carlo analysis by the MrBays program, two subgroups, one containing samples from Jinju and the other containing samples from the Changnyeong and Geoje areas were clearly identified. The result of a parsimonious median-joining network analysis also showed two clear subgroups, supporting the result of the phylogenetic analysis. On the other hand, in the consensus tree estimated using the genetic distances estimated from the genotypes of 13 MS markers, there were clear two subgroups, one containing samples from the Jinju, Geoje and Changnyeong areas and the other containing samples from only the Jinju area. The samples were not identically classified into each subgroup defined by mtDNA and MS markers. It could be inferred that the differential classification of samples by the two different marker systems was because of the different characteristics of the marker systems used, that is, the mtDNA was for detecting maternal lineage and the MS markers were for estimating autosomal genetic distances. Nonetheless, the results from the two marker systems showed that there has been a progressive genetic fixation according to the habitats of the otters. Further analyses using not only newly developed MS markers that will possess more analytical power but also the whole mtDNA are needed. Expansion of the phylogenetic analysis using otter samples collected from the major habitats in Korea should be helpful in scientifically and efficiently maintaining and preserving them.

• The Korea Journal of Herbology
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• v.31 no.5
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• pp.15-23
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• 2016

Objectives : Araliae Continentalis Radix and Angelicae Pubescentis Radix have been used as the same medicinal name Korean and Chinese traditional medicines, respectively. The authentic Araliae Continentalis Radix is described only the root of Aralia continentalis in the Korean Pharmarcopoeia. However, the dried root of Angelica biserrata, Levisticum officinale, or Heracleum moellendorffii also has been distributed adulterants of Araliae Continentalis Radix. To develop a reliable method for identifying Araliae Continentalis Radix from adulterants, we carried out the analyses of universal DNA barcode sequences.Methods : Four plants species were collected from different habitate and nucleotide sequences of matK and rbcL were analyzed. The species-specific sequences and phylogenetic relationship were estimated using entire sequences of two DNA barcodes, respectively.Results : In comparative analysis of matK sequences, we were identified 104 positions of marker nucleotide for Ar. continentalis, 3 for An. biserrata, 4 for L. officinale and 8 for H. moellendorffii enough to distinguish individual species, respectively. Furthermore, we obtained marker nucleotides in rbcL at 42 positions for Ar. continentalis, 5 for An. biserrata and 2 for H. moellendorffii, but not for L. officinale. The phylogenetic tree of matK and rbcL were showed that all samples were clustered into four groups constituting homogeneous clades within the species.Conclusions : We confirmed that species-specific marker nucleotides of matK sequence provides distinct genetic information enough to identify four species. Therefore, we suggest that matK gene is useful DNA barcode for discriminating authentic Araliae Continentalis Radix from inauthentic adulterants.

• ALGAE
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• v.31 no.2
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• pp.91-103
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• 2016

A goal of phylogeography is to relate phylogenetic patterns to potential historic and contemporary geographic isolating events. Genetic breaks found in phylogeographic studies may denote boundaries between more generally applicable biogeographic regions. We investigated the distribution of Lessonia variegata, an important habitat forming alga, found on most rocky shores around New Zealand’s main islands, plus related species from surrounding waters. L. variegata has been shown to consist of four distinct cryptic species. Our aim was to compare the distribution of L. variegata with proposed bioregions; and to develop phylogeographic hypotheses to explain its present day distribution. Both a mitochondrial (atp8-sp) and plastid (RuBisCo spacer) marker, with different mutation rates, were used to gain information of the phylogenetic history of Lessonia. The data revealed high phylogeographic structuring and reciprocal endemism for all L. variegata cryptic species. One species (L. variegata / N) is confined to the northern part of the North Island of New Zealand; L. variegata / W is found at the southeast of the North Island and the northern South Island; L. variegata / K is endemic to the northeast South Island; and L. variegata / S is restricted to the southern part of the South Island. No overlapping areas of L. variegata species distribution were found. The data showed that genetic breaks in Lessonia do mostly correlate to bioregions, and highlight the importance of Cape Campbell at the northeast of the South Island and East Cape in the North Island, well known phylogeographic breaks, as a barrier between adjacent species.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.1046-1054
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• 2001

Bioleaching is the process in which insoluble metal sulfide is oxidized by specialized iron- and/or sulfur-oxidizing lithotrophic bacteria in acidic, metal-rich environments. Most of these processes are carried out by the genus Thiobacillus. Three novel Thiobacillus strains (Thiobacillus thiooxidans AZ11, Thiobacillus thiooxidans MET, and thiobacillus thiooxidans TAS) associated with bioleaching have been isolated from soil and sludge (Korean patent No. 1999-0073060 for T. thiooxidans AZ11, Korean patent No. 1999-0005798 for T. thiooxidans MET, and Korean patent No. 1999-0073059 for T. thiooxidans TAS). A partial sequence of 16S ribosomal RNA gene (16S rDNA) and the entire sequence of 16S/23S intergenic spacer region (ISR) were determined in the three above novel strains and in Thiobacillus ferrooxidans ATCC19859 as a reference strain. When phylogenetic analysis was performed based on G+C contents and sequence alignments, T. ferroxidans ATCC19859 was found to be closely related to previously registered T. ferrooxidans strains in a monophyletic manner, while the three novel T. thiooxidans strains were classified in a paraphyletic manner. Close examination on the base composition of 16S/23S ISR revealed that the 5\` part (nucleotide residues 21-200) was specific for the genus Thiobacillus. On the other end, the 3\` part (nucleotide residues 201-520) showed specificity in T. ferrooxidans strains, but not in T. thiooxidans strains. These results suggest that the proximal and distal halves of 16S/23S could be used as a genetic marker for the identification of the genus Thiobacillus and the species T. ferrooxidans, respectively.

• Parasites, Hosts and Diseases
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• v.37 no.4
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• pp.215-228
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• 1999

To choose one or more appropriate molecular markers or gene regions for resolving a particular systematic question among the organisms at a certain categorical level is still a very difficult process. The primary goal of this review, therefore, is to provide a theoretical information in choosing one or more molecular markers or gene regions by illustrating general properties and phylogenetic utilities of nuclear ribosomal DNA (rDNA) and mitochondrial DNA (mtDNA) that have been most commonly used for phylogenetic researches. The highly conserved molecular markers and/or gene regions are useful for investigating phylogenetic relationships at higher categorical levels (deep branches of evolutionary history). On the other hand, the hypervariable molecular markers and/or gene regions are useful for elucidating phylogenetic relationships at lower categorical levels (recently diverged branches). In summary, different selective forces have led to the evolution of various molecular markers or gene regions with varying degrees of sequence conservation. Thus, appropriate molecular markers or gene regions should be chosen with even greater caution to deduce true phylogenetic relationships over a broad taxonomic spectrum.

• Korean Journal of Poultry Science
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• v.42 no.1
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• pp.15-26
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• 2015

In this study, genotyping was executed by using 27 microsatellite markers for genetic diversity of 469 Korean Native Chickens [20 population, each population is 24 samples but Hanhyup A line is 13 samples). in total 469 samples were collected from National Institute of Animal Science (Korean Native Chicken (NR, NY, NG, NL and NW), Ogye (NO), Leghorn F,K (NF and NK), Black and Brown cormish (NH and NS), Rhode Island Red C, D (NC and ND), Total is 12 populations] and Hanhyup [H line (HH), F line (HF), G line (HG), V line (HV), S line (HS), W line (HW), Y line (HY), A line (HA), total is 8 populations]. [The allele number were observed 5 (ADL0268) to 20 (MCW0127) each markers. Observed heterozygostiy ($H_{obs}$), expected heterozygosity ($H_{exp}$), polymorphism Information Content (PIC) were observed 0.359 to 0.677, 0.668 to 0.881 and 0.646 to 0.869, respectively. Using these markers, the calculated the heterozygote deficit within chicken line ($F_{is}$) value each population from mean 0.117. Phylogenetic tree showing the genetic relationship among 20 population using standard genetic distance calculated from 27 microsatellite markers. genetic distances revealed the closest (0.175) between NC and ND. on the other hand, Farthest genetic distances (0.710) revealed between NF and HV. STRUCTURE analysis and Principal Components Analysis (PCA) showed that results of similar phylogenetic tree. The expected probability of identity values on random individuals (Total population and only Hanhyup line) was estimated at $8.80{\times}10^{-83}$ and $3.87{\times}10^{-117}$, respectively. In conclusion, This study shows the useful data that be utilized as a basic data of Korean Native Chicken breeding and development for commercial chicken industry to meet the consumer's demand.

• ALGAE
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• v.28 no.1
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• pp.31-43
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• 2013

This review provides a comprehensive summary of the PCR primers and profiles currently in use in our laboratory for red algal DNA barcoding and phylogenetic research. While work focuses on florideophyte taxa, many of the markers have been applied successfully to the Bangiales, as well as other lineages previously assigned to the Bangiophyceae sensu lato. All of the primers currently in use with their respective amplification profiles and strategies are provided, which can include full fragment, overlapping fragments and what might best be called "informed overlapping fragments", i.e., a fragment for a marker is amplified and sequenced for a taxon and those sequence data are then used to identify the best primers to amplify the remaining fragment(s) for that marker. We extend this strategy for the more variable markers with sequence from the external PCR primers used to "inform" the selection of internal sequencing primers. This summary will hopefully serve as a useful resource to systematists in the red algal community.