• Proceedings of the Plant Resources Society of Korea Conference
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• 2021.04a
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• pp.22-22
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• 2021

큰졸방제비꽃(Viola kusanoana)의 엽록체 DNA 염기서열을 밝히고자 차세대염기서열분석법(NGS)을 이용하여 분석하였다. 재료는 경상북도 울릉군 나리분지에 자생하는 개체의 잎을 사용하였다. 염기서열 분석결과, 총 길이는 158,644 bp 였고, GC함량은 36.3%로 분석되었다. 구간별로는 LSC (Large single copy)지역이 86,999 bp (GC content: 33.9%)였고 SSC (Small single copy)지역은 17,439 bp (GC content: 29.9%)으로 분석되었으며 IR (Invertied repeats)지역은 27,103 bp (GC content: 42.2%)로 확인되었다. 유전자는 protein coding gene 77개, tRNA gene 30개, rRNA 4개 등 총 111개로 이는 선행 연구된 제비꽃속 8개 분류군과 유전자의 순서와 방향이 모두 일치하였다. 이를 통해 제비꽃속의 엽록체 게놈의 유전자는 상당히 보존되어 있음을 확인하였다.

• Journal of Life Science
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• v.21 no.10
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• pp.1452-1459
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• 2011

Citrus is a common term and genus (Citrus) of flowering plants in the rue family, Rutaceae. Citrus is believed to have originated in the part of Southeast Asia bordered by Northeastern India, Myanmar (Burma) and the Yunnan province of China. The taxonomy and systematics of the genus are complex and the precise number of natural species is unclear, as many of the named species are clonally propagated hybrids, and there is genetic evidence that even some wild, true-breeding species are of a hybrid origin. One of the most popular sequences for phylogenetic inference at the generic and infrageneric levels in plants is the chloroplast trnL-trnF region. We evaluated the seven taxa with the trnL-trnF region to estimate phenotypic relationships within the genera Citrus, Poncirus, and Fortunella in Korea. Alignment of the DNA sequences required the addition of numerous gaps. Sequence variation within Citrus was mostly due to insertion/deletion. Within the genus Citrus, C. lomonia and C. sinensis were relatives and sistered with C. aurantium in the four phylogenetic analyses (MP, ML, ME, and NJ). However, some external nodes were poorly supported.

• Journal of Microbiology and Biotechnology
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• v.16 no.2
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• pp.256-263
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• 2006

In the model legume Medicago truncatula, two mutants, sickle and sunn, exhibit morphologically and genetically distinct hypernodulation phenotypes. However, efforts to isolate the single recessive and single semidominant genes for sickle and sunn, respectively, by map-based cloning have so far been unsuccessful, partly due to the absence of clones that enable walks from linked marker positions. To help resolve these difficulties, a new bacterial artificial chromosome (BAC) library was constructed using BamHI-digested genomic fragments. A total of 23,808 clones were collected from ligation mixtures prepared with double-size-selected high-molecular-weight DNA. The average insert size was 116 kb based on an analysis of 88 randomly selected clones using NotI digestion and pulsed-field gel electrophoresis. About 18.5% of the library clones lacked inserts. The frequency of the BAC clones carrying chloroplast or mitochondrial DNA was 0.98% and 0.03%, respectively. The library represented approximately 4.9 haploid M. truncatula genomes. Hybridization of the BAC clone filters with a $C_{0}t-l$ DNA probe revealed that approximately 37% of the clones likely carried repetitive sequence-enriched DNA. An ordered array of pooled BAC DNA was screened by polymerase chain reactions using 13 sequence-characterized molecular markers that belonged to the eight linkage groups. Except for two markers, one to five positive BAC clones were obtained per marker. Accordingly, the sickle- and sunn-linked BAC clones identified herein will be useful for the isolation of these biotechnologically important genes. The new library will also provide clones that fill the gaps between preexisting BAC contigs, facilitating the physical mapping and genome sequencing of M. truncatula.

• Korean Journal of Plant Taxonomy
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• v.40 no.3
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• pp.157-162
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• 2010

Sequences of the trnL/F intergenic spacer of chloroplast DNA were used to investigate the intraspecific evoution and phylogeography of Sedum takesimense (Crassulaceae). The trnL/F intergeneric spacer sequences from 32 individuals of S. takesimense were either 291 bp (17 samples "without indel" in the following) or 297 bp (15samples "with indel 1") in length due to an indel of 6 bp. Two main cpDNA haplotypes were detected within S. takesimense. The haplotype with indel was found on Ulleung Island and without indel on Ulleung Island and Dok Island. This confirmed the existence of two cpDNA lineages with different geographical distributions. The cpDNA sequence analysis also suggested a putative long-distance dispersal event between Ulleung Island and Dok Island.

• Korean Journal of Plant Taxonomy
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• v.49 no.2
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• pp.127-139
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• 2019

The purported four hybrid origins of Asplenium in Korea were tested based on morphological, cytological and DNA sequence data. Asplenium castaneo-viride, A. ${\times}$ uiryeongse, A. ${\times}$ montanus, and A. ${\times}$ kitazawae share several morphological characteristics with the Asian walking fern A. ruprechtii and related taxa as parents and show a sympatric distribution with the putative parents, raising the possibility of hybrid origins: A. castaneo-viride (A. ruprechtii and A. incisum), A. ${\times}$ uiryeongse (A. ruprechtii and A. pekinense), A. ${\times}$ montanus (A. ruprechtii, A. trichomanes, and A. incisum), and A. ${\times}$ kitazawae (A. ruprechtii and A. sarelii). We investigated flow cytometry and chloroplast DNA sequence data (rbcL, rps4-trnS, and rps4-trnS intergenic spacer) to clarify the hybridization and origin of each hybrid. In the flow cytometry analyses, A. ruprechtii shows diploid (2x) only, whereas A. castaneo-viride (3x, 4x), A. ${\times}$ uiryeongse (3x), A. ${\times}$ montanus (3x, 4x), and A. ${\times}$ kitazawae (2x, 4x) exhibit polyploidy, suggesting hybrid events along speciation. The rbcL and rps4-trnS and rps4-trnS intergenic spacer data suggest that A. ruprechtii is one the maternal ancestors of all four hybrids. In addition, the rps4-trnS and rps4-trnS intergenic spacer data indicate that A. incisum is also the maternal ancestor of A. ${\times}$ kitazawae and A. ${\times}$ montanus, proposing multiple hybridization events for these two hybrids. In A. ${\times}$ montanus, morphological features such as the leaf forms and sympatric distributions of the species also support the multimaternal hypothesis, but the morphological features of A. ${\times}$ kitazawae must be examined with consideration of hybrid events. To clarify the complex hybrid evolutionary lineages of the four Asplenium hybrids, further research with taxon sampling and molecular markers should be conducted.

• Journal of Applied Biological Chemistry
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• v.44 no.4
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• pp.167-172
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• 2001

A cDNA fragment encoding the chloroplastic fructose-1, 6-bisphosphatase (FBPase) was cloned via PCR from the cDNA library of pea leaves. The cloned cDNA, about 1.05 kbp without signal sequence, was introduced into a pET-28a vector for expression in E. coli strain BL21(DE3)pLysS. The recombinant FBPase was purified through $Ni^+-NTA$ affinity chromatography and characterized. Molecular mass of the monomer was about 42,000. Enzymatic activity of the purified enzyme as the native pea chloroplastic FBPase was the highest at alkaline pH (pH 9.0). The recombinant enzyme was activated by a reducing agent DTT and was insensitive to AMP. The activation energy (Ea) and Arrehenius frequency factor were 42.67 kcal/mol and $2.65{\times}10^{14}/s$, respectively, slightly higher than those of the native enzyme. $K_M$ and $V_{max}$ were $99.98{\mu}M$ and $52.9{\mu}M/min$, respectively, which were comparable with the native enzyme.

• ALGAE
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• v.29 no.4
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• pp.267-277
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• 2014

Green tides occur every year in the Yellow Sea (YS), and numerous investigations are proceeding on various aspects of the phenomenon. We have identified bloom-forming species collected from diverse locations in the YS using morphological traits and the chloroplast gene for the large subunit of ribulose-1,5-bisphosphate carboxylase (rbcL). Morphological and rbcL sequence data analyses characterized the blooming species on both sides of the YS as belonging to the Ulva linza-procera-prolifera (LPP) complex clade or U. prolifera of earlier reports. However, U. procera within the LPP complex must be regarded as synonym of U. linza. Moreover, U. prolifera in free-floating samples collected from the Qingdao coast in 2009 was clearly in a distinct clade from that of the blooming species. Therefore, U. linza is the main green tide alga in the YS and has the procera-morphology. The green drift mats in the southeastern part of the YS (southwest sea of Korea) consisted predominantly of U. linza and rarely of U. compressa or U. prolifera.

• ALGAE
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• v.34 no.4
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• pp.267-275
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• 2019

Three new species of the genus Micractinium were collected from five localities on the South Shetland Islands in maritime Antarctica, and their morphological and molecular characteristics were investigated. The vegetative cells are spherical to ellipsoidal and a single chloroplast is parietal with a pyrenoid. Because of their simple morphology, no conspicuous morphological characters of new species were recognized under light microscopy. However, molecular phylogenetic relationships were inferred from the concatenated small subunit rDNA, and internal transcribed spacer (ITS) sequence data indicated that the Antarctic microalgal strains are strongly allied to the well-supported genus Micractinium, including M. pusillum, the type species of the genus, and three other species in the genus. The secondary structure of ITS2 and compensatory base changes were used to identify and describe six Antarctic Micractinium strains. Based on their morphological and molecular characteristics, we characterized three new species of Micractinium: M. simplicissimum sp. nov., M. singularis sp. nov., and M. variabile sp. nov.

• Journal of Applied Biological Chemistry
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• v.44 no.1
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• pp.6-11
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• 2001

An ${\omega}-3$ fatty acid desaturase gene which is involved in de novo synthesis of -Iinolenate was isolated from cDNA library of Perilla frutescens. A cDNA library was constructed with mRNA extracted from perilla seeds of 12 DAF. The cDNA clone consisting of 1317-bp open reading frame encoding 438 amino acids with a relative MW of 50kDa, was isolated and showed 65-83% similarities to other known genes. This cDNA is deduced to encode a plastidal ${\omega}-3$ fatty acid desaturase based on the fact that it has higher homology to plastidal ones than to microsomal ones and its N-terminal sequence shares several characteristics of transit peptides of chloroplast proteins. Southern blot analysis of genomic DNA indicated that more than one gene or alleles for ${\omega}-3$ fatty acid desaturase are present in the genome of perilla. Northern blot analysis showed that the ${\omega}-3$ fatty acid desaturase gene is mainly revealed in early developing seeds and has different expression patterns depending on tissue types compared to the microsomal ones.

• Korean Journal of Plant Taxonomy
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• v.50 no.3
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• pp.253-261
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• 2020

Carex splendentissima U. Kang & J. M. Chung, endemic to the Korean peninsula, is characterized by staminate terminal spikes and glabrous elliptic perigynia. Based on its broad leaves, androgynous spikes, and tri-stigmatic features, the species has been placed in Carex sect. Siderostictae Franch. ex Ohwi, an East Asian section and a basal group in the genus. To clarify the monophyly and phylogenetic position of the species, a molecular study using the internal transcribed spacer region of nuclear ribosomal DNA and chloroplast DNA (trnL-F) data was conducted. The DNA sequence data of ten taxa in sect. Siderostictae and closely related taxa (two taxa in sect. Surculosae) with outgroups were analyzed based on maximum parsimony and maximum likelihood (ML) criteria. In the analyses, C. splendentissima was monophyletic and placed within the Siderostictae clade (sect. Siderostictae + two species of sect. Surculosae), forming a clade with C. ciliatomarginata and C. pachygyna (endemic to Japan). The clade (C. splendentissima + C. ciliatomarginata + C. pachygyna) shows evidence of diploidy. Furthermore, C. splendentissima is a sister to C. ciliatomarginata in the ML tree, and the two taxa have staminate terminal spikes. This study also updates the distribution of C. splendentissima and provides keys to the four Korean taxa in sect. Siderostictae. To conserve the endemic species C. splendentissima, further research on its genetic and ecological features should be conducted at the population level.