• Korean Journal of Plant Taxonomy
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• v.41 no.2
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• pp.119-129
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• 2011

Phylogenetic studies were conducted to evaluate the taxonomic relationships among 27 taxa, including 2 outgroups of Korean Campanulaceae, using PCR-RFLP analysis and ITS sequences. In the PCR-RFLP analysis, 15 restriction endonucleases produced 244 restriction sites and size variations from the chloroplast DNA, and 59 restriction sites (24%) showed polymorphism. The length of the ITS regions ranged from 588 bp to 797 bp. The sequence divergence including the outgroups is 0-39.36%. Phylogenetic analyses based on PCR-RFLP and ITS data suggest that Campanulaceae is monophyletic; Codonopsis and Platycodon forms an independent clade; the Peracarpa and Asyneuma clade is a sister to the Adenophora-Hanabusaya clade; Campanula is monophyletic; and Wahlenbergia basally branches within the ITS tree, whereas they are placed between Campanula and the Codonopsis-Platycodon clade in the PCR-RFLP tree; Hanabusaya is placed within the Adenophora clade; and Adenophora is paraphyletic and shows discordance to the infrageneric classifications based on morphological data. The present results show two data sets, largely congruent at the generic level, but their phylogenetic positions, in particular the Wahlenbergia and Hanabusaya and the infrageneric classifications in Adenophora, show some incongruence.

• Journal of Microbiology and Biotechnology
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• v.14 no.2
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• pp.390-394
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• 2004

The genetically modified glyphosate-tolerant soybean contains the following introduced DNA sequences: the EPSPS (5-enol-pyruvylshikimate-3-phosphate synthase) gene from Agrobacterium sp. strain CP4, the 35S promoter from the cauliflower mosaic virus, and the NOS terminator from Agrobacterium tumefaciens. In the present study, detection of these introduced DNAs was performed by amplification using the polymerase chain reaction (PCR). A multiplex PCR method was also applied to prevent false positive results. When primers for 35S promoter, nos3', CTP(chloroplast transit peptide), and CP4 EPSPS (EPSPS from Agrobacterium sp. CP4) were used, positive results were obtained in PCR reactions using DNA from genetically modified glyphosate-tolerant soybeans. There were no false positive results when using DNA from non-genetically modified soybeans. The CP4 EPSPS gene was detected when less than 125 pg glyphosate-tolerant soybean DNA was amplified. Lectin Lel and psb A were amplified from both non-genetically modified and genetically modified glyphosate-tolerant soybean DNA. Multiplex PCR was performed using different primer sets for actin Sacl, 35S promoter and CP4 EPSPS. The actin gene was detectable in both non-genetically modified and glyphosate-tolerant soybeans as a constant endogenous gene. Target DNAs for the 35S promoter, and CP4 EPSPS were detected in samples containing 0.01-0.1% glyphosate-tolerant soybean, although there were variations depending on primers by multiplex PCR. Soybean seeds from five plants of non-genetically modified soybean were co-cultivated for six months with those of genetically modified soybean, and they were analyzed by PCR. As a result, they were not positive for 35S promoter, nos3' or CP4 EPSPS. Therefore, these results suggest there was no natural crossing of genes between glyphosate-tolerant and non-genetically modified soybean during co-cultivation, which indicates that gene transfer between these plants is unlikely to occur in nature.

• Weed & Turfgrass Science
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• v.4 no.1
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• pp.18-25
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• 2015

A universal DNA barcoding for agricultural noxious weeds is a powerful technique for species identification without morphological knowledge, by using short sections of DNA from a specific region of the genome. Two standard barcode markers, chloroplast rbcL and matK, and a supplementary nuclear ribosomal Internal Transcribed Spacer (ITS) region were used to examine the effectiveness of the markers for Pooideae barcoding using 163 individuals of 29 taxa across 16 genera of Korean Pooideae. The rbcL and ITS revealed a good level of amplification and sequencing success while matK did not. Barcode gaps were 78.6% for rbcL, 96.2% for matK, and 91.7% for ITS, respectively. Resolving powers were 89.3% for rbcL, 92.3% for matK, and 79.1% for ITS. The matK obtained the best both barcode gap and resolving power. However, it should be considered not to employ matK for Pooideae barcode because of low rate of PCR amplification and sequencing success. As a single DNA marker, rbcL and ITS were reasonable for Pooideae barcode. Barcode gap and resolving power were increased when ITS was incorporated into the rbcL. The barcode sequences were deposited to the National Center for Biotechnology Information (NCBI) database for public use.

• Journal of the Korean Society of Marine Environment & Safety
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• v.24 no.7
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• pp.967-972
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• 2018

The genus Chattonella belonging to the class raphidophyceae, is a harmful algal bloom species. Recently, its occurrence has been increasing and expanding along the Korean coast. Species identification of the genus Chattonella only by morphological observation is difficult due to the lack of rigid cell walls. In this study, the morphological characteristics and genetic affinity of Chattonella sp. isolated from Deungnyang Bay in 2017 were examined. We carried out single-cell isolation from field samples then sequenced three different areas using the single-cell PCR method: 1) parts of ribosomal operon, the large subunit (LSU) of the rDNA, 2) the chloroplast-encoded subunit psaA of Photosystem I, and 3) rbcL encoding the large subunit of the Rubisco gene. The cells were morphologically very similar to the general genus Chattonella ($74.0{\pm}10.1{\mu}m$ in length, $33.1{\pm}3.6{\mu}m$ in width). The three partial gene sequences were insufficient to justify distinction at the species rank. However, they clustered at 99-100 % sequence similarity with C. marina, C. marina var. antiqua and C. marina var. ovata.

• Plant Biotechnology Reports
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• v.4 no.3
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• pp.223-228
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• 2010

Schizonepeta tenuifolia (Korean name "Hyung- Gae") is an oriental medicinal plant that is widely used in Korea, China and Japan. S. tenuifolia (Hyung-Gae) has many pharmacological activities and is mostly used for many medicinal preparations. The dried aerial part (spikes and stems) of three oriental medicinal plants, S. tenuifolia (Hyung-Gae), Agastache rugosa (Kwhak-Hyang) and Elsholtzia ciliata (Hyang-Yoo) belonging to the same family, mint family Labiaceae, have such similar shape and smell that it is difficult to differentiate between them. The trnL-F regions of chloroplast DNA of the three medicinal plants were sequenced and used as targets in multiplex PCR reaction to identify S. tenuifolia. After alignment of trnL-F sequences of the authenticated plant samples, one single nucleotide polymorphism (SNP) specific to S. tenuifolia was found. Based on this SNP, a new primer was designed that specifically amplifies the trnL-F region of S. tenuifolia. The established multiplex-PCR was proven to be effective in the differentiation of commercial S. tenuifolia samples from A. rugosa and E. ciliata. This rapid and accurate molecular method is highly promising for use in the food industry.

• ALGAE
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• v.35 no.2
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• pp.145-155
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• 2020

Specimens of red algae corresponding to the peyssonnelioid genus Sonderophycus were collected at Kure Atoll, Hawai'i, at a depth range of 88-94 m depth during mesophotic surveys of the Papahānaumokuākea Marine National Monument, Hawaiian Islands, and were analyzed using morphological and molecular approaches. Analyses of mitochondrial cytochrome oxidase subunit 1 and chloroplast rbcL DNA sequences demonstrated that the Hawaiian specimens were identical to one another yet distinct from the three other species currently recognized within the genus (S. capensis [Montagne] M. J. Wynne, S. coriaceus [Womersley & Sinkora] M. J. Wynne, and S. fervens Dixon), as well as the likely congener, Peyssonnelia caulifera Okamura, and are proposed here as a new species: Sonderophycus copusii A. R. Sherwood. Sonderophycus copusii is morphologically distinct from other members of the genus by the following combination of characters: the presence of occasional secondary perithallial growth, emergence of rhizoids from the hypobasal cuticle at a strongly acute angle, a lack of horizontally directed filaments in the lower perithallus, and the lack of a stipe. This is the first record of the genus Sonderophycus in the Hawaiian Islands. Sonderophycus copusii was documented as a dominant member of the algal community at Kure Atoll, and thus may play a significant ecological role in the deep-water benthic community of Kure Atoll, along the lines of reports of deep water peyssonnelioid beds in the Mediterranean, Red Sea, and Caribbean. This study further highlights the unexplored diversity of the Peyssonneliales in Hawai'i, and emphasizes more generally the degree of as yet undiscovered biodiversity of algae at mesophotic depths.

• Korean Journal of Plant Taxonomy
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• v.37 no.4
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• pp.477-502
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• 2007

The Aconitum alboviolaceum Kom. complex includes four controversial species described from Korea; A. albouiolaceum, A. pseudolaeue, A. longecassidatum, and A. quelpaertense. The main objective of this study was to determine the taxonomic identities and systematic relationships among the species in the A. albouiolaceum complex based on morphology, numerical analyses and DNA sequence analysis. In the present study, variations in the principal morphological characters and chloroplast DNA noncoding region sequences (psbA-trnH IGS, trnL intron, and trnL-trnF IGS) were examined for 95 individuals from 20 populations. Also, neighbor-joining analysis was adopted to infer their relationships. Morphological variation appeared to be considerably high but not to be related to geographic distribution. These morphological results suggest that reevaluation of key morphological characters are needed for the proper taxonomic treatment of the complex. The length of psbA-trnH IGS region ranged from 241 to 250 bp, that of the trnL intron from 526 to 532 bp, and that of the trnL-trnF IGS region from 466 to 472 by in all taxa. Nine haplotypes were recognized from the analysis. Seven populations shared more than two haplotypes, while other thirteen populations shared only one haplotype. In the phylogenetic analysis, the nine haplotypes formed four groups, separated A. sibiricum, one of the sister groups of the complex. There also was no distinct grouping pattern supporting the species and populations observed. These results suggest that introgression or speciation might have been involved in the formation of taxa of A. alboviolaceum complex.

• Applied Biological Chemistry
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• v.37 no.5
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• pp.361-369
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• 1994

To study the light-induced expression mechanism and protein transport into the chloroplast, a rice genomic clone (GrbcS) for the small subunit of ribulose 1,5-bisphosphate carboxylase (rbcS) was isolated and its nucleotide sequence was determined. Nucleotide sequence analysis of GrbcS revealed that the gene consists of two exons interrupted by an intron, encoding a protein of 175 amino acids including a transit peptide of 47 amino acids. These structural features of GrbcS are consistent with those of other rbcS genes from monocot species. Genomic Southern blot analysis suggested that the rbcS genes are present as a relatively small multigene family in the rice genome. Comparison of the nucleotide and deduced amino acid sequences to other rice rbcSs shows close sequence similaritiy. Conserved DNA sequences present in other light-responsive genes are also found in the 5’ upstream region of GrbcS such as G-box, 3AF1-binding site and GATA site. The possible function of these putative regulatory elements are discussed.

• Journal of Life Science
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• v.20 no.11
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• pp.1697-1703
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• 2010

Asarum consists of low-growing herbs and is a genus in the Aristolochiaceae family with species found in the north temperate zones with most species in Asia. We evaluated the nine taxa with the trnL - trnT region of the chloroplast genome to estimate phenotypic relationships within genus Asarum in Korea. Alignment of the DNA sequences required the addition of numerous gaps. Sequence variation within the Asarum was mostly due to nucleotide inserts/deletions, although several indels and inserts were found. Another source of sequence divergence was length variation due to stretches of short repeats that occur at the trnL - trnT region in all the Asarum. A + T content for nine Korean species of genus Asarum ranged between 74.7% and 78.3%. These values were higher than those for the angiosperm alignments of the total trnL and trnT region (64.5~67.1%). Within genus Asarum, A. patens was strikingly different from the others in the three phylogenetic analyses (MP, ML, and NJ). However, some internal nodes were poorly supported. Within Korean Asarum, four species were unsolved portions. Possible reasons for the striking non-congruence between the previous morphological traits and the trnL - trnT based on phylogeny were discussed.

• Korean Journal of Plant Taxonomy
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• v.39 no.3
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• pp.135-142
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• 2009

Phylogeny of the family Ophioglossaceae and a phylogenetic position of Mankyua were estimated through analyses of chloroplast rbcL gene sequences and spore morphology. Sequence analysis of the rbcL gene clearly indicated that there are two major lineages in the family Ophioglossaceae: Botrychioid lineage and Ophioglossoid lineage. The Botrichioid lineage is composed of three distinct clades: Botrychium, Helminthostachys and Mankyua, where Helminthostachys and Mankyua were placed as sister groups to the Botrychium. Within the genus Botrychium, subgenera Septridium and Botrychium were monophyletic, while taxa of subgen. Botrypus branched as sister of the two, successively, thus making a non-monophyletic group. Ophioglossum formed the Ophioglossoied lineage, where the subgen. Ophioglossum is monophyletic, while subgen. Cheiroglossa and Ophoderma formed a sister relationship with subgen. Ophioglossum. In terms of external morphology and spores, Mankyua is most similar to Helminthostachys, however, patristic distance in the cladogram and trophophore characteristics of the two genera are distinct. Therefore, Mankyua is a well defined genus within the family in terms of morphology as well as molecular phylogeny which places it in basal position of the Botrychioid lineage on the gene tree.