• Horticultural Science & Technology
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• v.35 no.3
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• pp.344-360
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• 2017

Zoysiagrasses are important turf plants used for school playgrounds, parks, golf courses, and sports fields. The two most popular zoysiagrass species are Zoysia japonica and Zoysia sinica. These are widely distributed across different growing zones and are morphologically distinguishable from each other; however, it is phenotypically difficult to differentiate those that grow along the coastal line from those in beach area habitats. A combination of morphological and molecular approaches is desirable to efficiently identify these two plant cultivars. In this study, we used a rapid identification system based on DNA barcoding of the nrDNA-internal transcribed spacer (ITS) regions. The nrDNA-ITS regions of ITS1, 5.8S nrDNA, and ITS2 from Z. japonica, Z. sinica, Agrostis stolonifera, and Poa pratensis were DNA barcoded to classify these grasses according to their molecular identities. The nrDNA-ITS sequences of these species were found at 686 bp, 687 bp, 683 bp, and 681 bp, respectively. The size of ITS1 ranged from 248 to 249 bp, while ITS2 ranged from 270 to 274 bp. The 5.8S coding region ranged from 163 - 164bp. Between Z. japonica and Z. sinica, nineteen (2.8%) nucleotide sites were variable, and the G+C content of the ITS region ranged from 55.4 to 63.3%. Substitutions and insert/deletion (indel) sites in the nrDNA-ITS sequence of Z. japonica and Z. sinica were converted to cleaved amplified polymorphic sequence (CAPS) markers, and applied to the Zoysia grasses sampled to verify the presence of these markers. Among the 62 control and collected grass samples, we classified three groups: 36 Z. japonica, 22 Z. sinica, and 4 Z. japonica/Z. sinica hybrids. Morphological classification revealed only two groups; Z. japonica and Z. sinica. Our results suggest that used of the nrDNA-ITS barcode region and CAPS markers can be used to distinguish between Z. japonica and Z. sinica at the species level.

• Korean Journal of Plant Taxonomy
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• v.36 no.4
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• pp.263-277
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• 2006

Anemone pendulisepala, recently described as a new species, is distributed in sympatry with A. reflexa, A. amurensis, and A. raddeana at Mt. taebeark and Mt. Baekdu. Anemone pendulisepala was previously proposed to be a hybrid species between A. reflexa and A. amurensis becaue it displavs overlapping features with them in involucre shape, petiole length, sepal apex and xylem shape, To verify the taxonomic status and to examine the hybridity of A. pendulisepala, sequences of ITS region of nuclear ribosomal DNA and the psba-trnH, rps16 and trnLF region of cpDNA from 36 accessions of 5 taxa including outgroup were analyzed. In maximum parsimony tree based on ITS sequences, A. pendulisepala had the same sequences of A. reflexa and was clustered with monophyletic A. amurensis, and then A. raddeana. Anemone pendulisepala was distinguished from the other taxa by having four base insertion in rps16 region, two species-specific bases and insertion in trnLF region. In the phylogenetic trees of combined cpDNA, A. pendulisepala showed monophyly with the bootstrap 100%. Anemone pendulisepala exhibited no polymorphism and shared no sequences with putative parental or related taxa examined in this study. Molecular data suggest that A. pendulisepala should be a distinct species, and no evidence of the hybridization was detectcd.

• Korean Journal of Plant Taxonomy
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• v.42 no.3
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• pp.185-196
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• 2012

Morphological and geographical examinations as well as phylogenetic analyses using ITS sequences were performed for Asparagus L. in Korea. A total of five species of Asparagus were confirmed to be distributed in South Korea. The shape of cladophylls, length of pedicels, and shape of perianth were considered to be important characteristics for the identification of Koran Asparagus species. A monophyly of each species was evident in the ITS phylogenetic trees in which multiple accessions (5 to 24, depending on species) represented each of the five Korean species. A. rigidulus Nakai, once considered conspecific to A. schoberioides Kunth, formed a distinct lineage in the ITS trees. Pedicels of A. rigidulus, which is distributed mainly in coastal areas, were about two times longer than those of A. schoberioides occurring in inland areas, suggesting that they should be treated as distinct taxa.

• Korean Journal of Plant Taxonomy
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• v.43 no.1
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• pp.12-21
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• 2013

To clarify the taxonomic position for Astragalus nakaianus and provide correct scientific name for A. mongholicus cultivar in South Korea, we examined external morphological characters and sequence variations from ITS and five cp non-coding DNA regions. Genetic structure was also analyzed for 61 individuals from three populations using nine microsatellite loci. We found no significant difference between the South Korean cultivar and A. mongholicus var. dahuricus when morphology and ITS sequences were considered. Morphologically, A. nakaianus specimens varied somewhat from A. mongholicus var. mongholicus and var. dahuricus in habit, plant height, and lengths of leaf axis and leaflet. Although sequence data from ITS and cp noncoding DNA regions could not distinguished A. nakaianus from A. mongholicus, microsatellite analysis revealed strong structuring between the cultivar and A. nakaianus. Therefore, we conclude that the South Korean A. mongholicus cultivar should be treated as A. mongholicus var. dahuricus and that A. nakaianus should be merged into A. mongholicus as a variety, i.e., A. mongholicus var. nakaianus.

• Korean Journal of Microbiology
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• v.45 no.4
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• pp.430-434
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• 2009

Bacterial 16S rRNA gene sequences have been widely used for the studies on molecular phylogeny, evolutional history, and molecular detections. Bacterial genomes have multiple rRNA operons, of which gene sequences sometimes are variable. In the present study, heterogeneity of the Vibrio 16S rRNA gene sequences were investigated. Vibrio 16S rRNA sequences were obtained from GenBank databases, considering the completion of gene annotation of Vibrio genome sequences. These included V. cholerae, V. harveyi, V. parahaemolyticus, V. splendidus, and V. vulnificus. Chromosome 1 of the studied Vibrio had 7~10 copies of the 16S rRNA gene, and their intragenomic variations were less than 0.9% dissimilarity (more than 99.1% DNA similarity). Chromosome 2 had none or single 16S rRNA gene. Intragenomic 16S rRNA genotypes were detected at least 5 types (V. vulnificus #CMCP6) to 8 types (V. parahaemolyticus #RIMD 2210633, V. harveyi #ATCC BAA-1116). These suggest that Vibrio has high heterogeneity of the 16S rRNA gene sequences.

• Journal of Sericultural and Entomological Science
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• v.51 no.2
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• pp.137-141
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• 2013

In this study, nucleotide sequence structures of intergenic transcribed spacer (ITS) 1, complete 5.8S ribosomal RNA gene and ITS 2 region were analyzed to identify three Peruvian entomopathogenic fungal isolates. The isolates had highly conserved sequence region in 5.8S rRNA gene and unique sequences in ITS 1 and 2 region among them. 5.8S rRNA gene regions were highly conserved and showed high homoloies among tested isolates. In contrast, ITS region showed species-specific sequence region, resulting in inter-genus differencies. Scanning electron microscopic images of these isolates supported the result of ITS-based identification. From these result, Peruvian entomopathogenic fungal isolate J270, J278, were identified as Beauveria bassiana and J271 was identified as Lecanicillium attenuatum.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2005.11a
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• pp.53-59
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• 2005

Sequences of candidate chicken testis-specific genes were analyzed in order to develop a resource for functional genomic studies of the testis and male germ cells. Tentative consensus sequences (TCs) containing ESTs expressed in testis libraries only were selected from the TIGR Gallus gallus Gene Index, resulting in a total of 292 TCs. The transcriptional expression of these genes were evaluated in a variety of chicken tissues, including testis and ovary, Of the panel of 292 TCs, 110 were expressed in a testis-specific manner. The correlation between the number of ESTs assembled into each TC and the number of testis-specific TCs was not significant. Annotation of the TCs using the Gene Ontology database terms showed that the proportion of testis-specific TCs that were classified as having catalytic activity (within the Molecular Function branch) was larger than the proportion of total chicken TCs classified in the same way. Our results might facilitate the investigation of testis-specific genes and their functional analysis in the chicken as well as in other avian species.

• Korean Journal of Plant Taxonomy
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• v.39 no.4
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• pp.237-246
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• 2009

Inter-specific relationships among six species and two varieties, belonging to Scrophularia (Scrophulariaceae), in Korea were re-examined based on both morphological and molecular data. Scrophularia takesimensis, a Korean endemic species, should be separated from S. grayana as an independent species based upon unique morphological characters such as non-winged stems, thickened leaves and glabrous surfaces of leaves, even though they share a lot of other features. It was found that S. grayana is distributed in Sokcho in Korea and along the coast of the East Sea north to far eastern Russia. Although some morphologies of three taxa, S. cephalantha, S. koraiensis var. velutina and S. kakudensis var. microphylla appeared to differ from each other, their ITS DNA sequence data was similar to that of S. kakudensis, suggesting that they might better be designated with the later species. The taxonomic position of S. koraiensis, only found in Korea, has been ambiguous, but ITS DNA sequence data in this study revealed that this species is distinct from S. kakudensis. Consequently, we conclude that the genus Scrophularia in Korea includes five species.

• Korean Journal of Plant Taxonomy
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• v.44 no.1
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• pp.39-50
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• 2014

Phylogenetic positions of Sedirea and Neofinetia were addressed using the chloroplast matK and the nuclear ITS sequences. We also evaluate the usefulness of the makers for the identification of species and localities. Sedirea and Neofinetia form an independent monophyletic genus, respectively, in both matK and nuclear ITS trees. The sister genus of the Neofinetia was Vanda in both trees. In addition, our trees support the separate recognition of the Neofinetia from Vanda rather than the inclusion of Neofinetia into Vanda. The sister group of the Sedirea was (Dimorphorchis(Pteroceras(Saccolabiun+Phalaeonopsis))) clade. The Dimorphorchis was one of the most probable sister genus to the Sedirea. The sister group relationship between Sedirea and Aerides was suggested by their similar morphology, but not supported in molecular trees. The identification of species and localities of Neofinetia was possible using our two molecular markers. However, several pseudo-gene sequences are discovered from the public data base. In addition, the horizontal gene transfer of chloroplast genomes is frequent events in orchid hybrids. Therefore, we need a careful evaluation for the data prior to systematic use. Generation of sequence data from multiple accessions of a species may helpful to reduce these types of error.

• Korean Journal of Plant Taxonomy
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• v.43 no.1
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• pp.46-55
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• 2013

The purpose of this study is to review the taxonomic position of Taxus cuspidata var. latifolia endemic to Ulleung Island with related taxa T. cuspidata var. cuspidata, T. caespitosa, and T. cuspidata var. nana based on external morphological characters and DNA barcoding study. T. cuspidata var. latifolia was similar to T. cuspidata var. cuspidata in the arbor, straight trunk, and symmetric arrangement of leaf. But the unique differences between T. cuspidata var. latifolia and T. cuspidata var. cuspidata were leaf size and the exposure of seed from aril. Additionally, sequences of four chloroplast DNA regions including matK, rbcL, trnL intron and trnL-trnF spacer regions were analyzed. Korean Taxus species and their related taxon T. cuspidata var. nana were strongly supported as a monophyletic group in neighbor-joining analysis. Taxus cuspidata var. latifolia showed 100% sequence identity to related taxa. Korean endemic T. caespitosa is also distinguishable from related taxa by prostrate stems and spiral arrangement of leaf. The examinations of external morphology and DNA barcoding study suggest that the taxonomic position of T. cuspidata var. latifolia should be maintained as a variety of T. cuspidata.