• Journal of Korean Society of Forest Science
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• v.108 no.2
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• pp.200-207
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• 2019

The aim of this study was to provide the basic information necessary to identify Korean fir (Abies holophylla), momi fir (A. firma), and Nikko fir (A. homolepis), and other fir trees planted in South Korea that are protected by law. Analysis of the morphological characteristics of the needles from each sample was investigated. The shape of the needle-leaf tip from A. holophylla was acute, whereas that from A. firma and A. homolepis was emarginate and that from the protected fir trees was obtuse. The number of stomata on the needles was not significantly different between A. holophylla and A. firma, and the number of stomata on the needles from A. homolepis and the protected fir trees were highly similar. In addition, the genetic differences among the Abies species were analyzed using the sequences of five chloroplast DNA regions-matK, atpF-atpH, rpoC2-rps2, rpoC1, and psbA-trnH.The atpF-atpH and psbA-trnH regions were useful for discriminating A. firma from the other species, but there were no differences among A. holophylla, A. homolepis, and the protected fir trees. The same chloroplast sequences were found in both A. holophylla and A. homolepis, which suggests that additional genetic studies might be necessary to identify the Abies species planted in both South Korea and Japan.

• Journal of Plant Biotechnology
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• v.1 no.1
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• pp.46-55
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• 1999

The expressed sequence tags(ESTs) from immature seed of rice, Oryza sativa cv Milyang 23, were partially sequenced and analyzed by homology. As of 1998, the partial sequences of about 6,600 cDNA clones were analyzed from normal and normalized immature seed cDNA libraries. About 2,200 ESTs were putatively identified by BLASTX deduced amino acid sequence homology analysis. About 20% of them were putatively identified as storage proteins. Also the clones were highly homologous to genes involved particularly in starch biosynthesis, glycolysis, signal transduction and defenses. Compared to 35% of redundancy in the ESTs of normal cDNA library, that from the substracted library was 15%. The Korea Rice Genome Network is maintained to provide the updated information of sequences, their homologies and sequence alignments of ESTs. For the stable expression of transgene in rice, diverse vectors were developed for overexpression, targeting and gene dosage effect with transit peptides (Tp) and matrix attachment region (MAR) sequence from chicken lysozyme locus. The rice calli were transformed via Agrobacterium tumefaciens LBA4404(pSB1) with the triparental mating technique and selected by herbicide resistance. The green fluorescent protein(GFP) gene in expression vector under the control of rbcS promoter-Tp was overexpressed upto 10 % of the total soluble protein. In addition, the Tp-sGFP fusion protein was properly processed during translocation into chloroplast. The expression of sGFP in the presence of MAR sequences was analyzed with Northern and immunoblot analysis. All the lines in which sGFP transgene with MAR sequence, showed position independent and copy number-dependent expression, while the lines without MAR showed the varied level of expression with the integration site. Thus the MAR sequence significantly reduced the variation in transgene expression between independent transformants.

• Journal of Plant Biotechnology
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• v.43 no.4
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• pp.417-421
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• 2016

The complete chloroplast genome sequence of Panax ginseng breeding line 'G07006', showing higher salt tolerance, was confirmed by de novo assembly using whole genome next-generation sequences. The complete chloroplast (CP) genome size is 156,356 bp, including two inverted repeats (IRs) of 52,060 bp, separated by the large single-copy (LSC 86,174 bp) and the small single-copy (SSC 18,122 bp) regions. One hundred fourteen genes were annotated, including 80 protein-coding genes, 30 tRNA genes, and 4 rRNA genes. Among them, 18 sites were duplicated in the inverted repeat regions. By comparative analyses of the previously identified CP genome sequences of nine cultivars of P. ginseng and that of G07006, five useful SNPs were defined in this study. Since three of the five SNPs were cultivar-specific to Chunpoong and Sunhyang, they could be easily used for distinguishing from other ginseng accessions. However, on arranging SNPs according to their gene location, the G07006 genotype was 'GTGGA', which was distinct from other accessions. This complete chloroplast DNA sequence could be conducive to discrimination of the line G07006 (salt-tolerant) and further enhancement of the genetic improvement program for this important medicinal plant.

• Korean Journal of Plant Taxonomy
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• v.44 no.3
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• pp.202-207
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• 2014

Lonicera insularis Nakai (Caprifoliaceae) is Korean endemic plant that lives along the shore of Ulleungdo and Dokdo. The aim of this study is to construct a phylogenetic relationship within six species (L.maackii, L.chrysantha, L.subsessilis, L. harai, L. morrowii) of genus Lonicera L. and Wigela subsessilis as outgroup and intraspecific variation of L. insularis using chloroplast DNA five regions sequences. Sequence analysis revealed that both L. insularis and L. morrowii showed complete homologies in the intergenic regions of trnL-trnF, trnS-trnG, psbM-trnD and matK coding region. However, sequence in the petN-psbM intergenic region showed a single nucleotide difference between both species, thus we designated them as CP01 and CP02. The plants having CP01 are prevalent in the Ulleungdo and Dokdo, while L. insularis and L. morrowii from Ulleungdo and of Dokdo, and Japan have CP02. This confirmed the existence of two cp DNA lineages with different geographical distributions. We can infer the allopatric speciation by geographical barrier. The result will provide the important basal data to study speciation and specie evolution of ocean islands such as Ulleungdo and Dokdo.

• Horticultural Science & Technology
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• v.31 no.2
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• pp.219-223
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• 2013

DNA markers can determine the genotype of many species. Single nucleotide polymorphism (SNP) detection is difficult without sequencing but it becomes easier with sdCAPS method. Here an experiment was performed for developing molecular markers using two SNPs, CSatpB-SNP and CSycf1-SNP, of chloroplast in cucumber plants. Properly designed primers with nucleotide sequences for restriction enzymes proved success of PCR and efficacy of digestion by the restriction enzymes. Then these markers were used to study the genotyping of cucumber breeding lines and cultivars obtained from various sources in respect of their chilling stress response. We confirmed that a U.S. cucumber line, 'NC76' known to possess a nuclear factor for the chilling tolerance showed the chloroplast genotypes related to chilling tolerance. However all Korean cucumber cultivars tested in this study showed the chloroplast genotypes related to chilling susceptibility. In conclusion, to develop chilling tolerant cucumber, both maternal and a nuclear factors related to chilling tolerance should be transferred from 'NC76' when 'NC76' is used as a female source and other elite lines as recurrent parents.

• Journal of Species Research
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• v.12 no.3
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• pp.258-265
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• 2023

We conducted phylogenetic analyses using multiplexed inter-simple sequence repeat genotyping by sequencing and compared chloroplast DNA sequences among Ardisia japonica, A. pusilla, and morphologically intermediate plants found on Jeju Island, Korea. Our network analysis demonstrated that the intermediate plants were genetically positioned between A. japonica and A. pusilla. Our comparison of the intergenic spacer between the psbA and trnH genes in chloroplast DNA indicated that four nucleotide substitutions separate A. japonica and A. pusilla, whereas the intermediate plants exhibited the A. japonica haplotype. Our results suggest that the intermediate plants on Jeju Island represent a natural hybrid of A. japonica, as the maternal species, and A. pusilla, and that they are attributable to Ardisia×walkeri. This record constitutes the first documented occurrence of the hybrid taxon in Korea.

• ALGAE
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• v.28 no.4
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• pp.307-330
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• 2013

The genus Cryptomonas is easily recognized by having two flagella, green brownish color, and a swaying behavior. They have relatively simple morphology, and limited diagnostic characters, which present a major difficulty in differentiating between species of the genus. To understand species delineation and phylogenetic relationships among Cryptomonas species, the nuclear-encoded internal transcribed spacer 2 (ITS2), partial large subunit (LSU) and small subunit ribosomal DNA (rDNA), and chloroplast-encoded psbA and LSU rDNA sequences were determined and used for phylogenetic analyses, using Bayesian and maximum likelihood methods. In addition, nuclear-encoded ITS2 sequences were predicted to secondary structures, and were used to determine nine species and four unidentified species from 47 strains. Sequences of helix I, II, and IIIb in ITS2 secondary structure were very useful for the identification of Cryptomonas species. However, the helix IV was the most variable region across species in alignment. The phylogenetic tree showed that fourteen species were monophyletic. However, some strains of C. obovata had chloroplasts with pyrenoid while others were without pyrenoid, which used as a key character in few species. Therefore, classification systems depending solely on morphological characters are inadequate, and require the use of molecular data.

• Weed & Turfgrass Science
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• v.3 no.3
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• pp.183-189
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• 2014

The genus Echinochloa (L.) P. Beauv. comprised of approximately 30-40 species in the tropical and warm temperate regions of the world, including numerous interspecific and intraspecific types which make the genus difficult to identify. As an attempt to identify the species within the genus easier, the taxonomy of the genus Echinochloa, Poaceae in Korea was reviewed on the basis of sequencing data derived from nuclear ribosomal DNA internal transcribed spacer (ITS) and external transcribe spacer and chloroplast DNA trnL intron, trnL-F intergenic spacer and matK regions using a total of 46 accessions representing all the species in Korea. The results of maximum parsimony found separate lineage comprised of E. colona and E. frumentaceae which are not Korean species, but no resolution within Korean Echinochloa species, supporting the suggestion of Yamaguchi group that E. crus-galli, E. oryzoides, and E. esculenta should be considered to belong to the same species. However, the relationship between these three species and the other species, i.e. E. oryzicola should be better understood with more detail studies.

• The Korea Journal of Herbology
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• v.28 no.5
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• pp.1-6
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• 2013

Objectives : This study was conducted to identify the original Sinomini Caulis et Rhizoma plant among Stephania tetrandra, Cocculus trilobus, and Aristolochiae fangchi to develop the genetic marker for Sinomini Caulis et Rhizoma. Methods : Sinomenium acutum was identified by the classification and identification committee of the National Center for Standardization of Herbal Medicines. The chloroplast ndhF gene was amplified. We performed sequences alignment analysis of Sinomenium acutum, Stephania tetrandra, C. trilobus, and A. fangchi using BioEdit program. The SFR markers designed were consisted of SF01, SR04, and SR05 primers. Results : Many variations of Sinomeni Caulis et Rhizoma are currently commercialized as herbal medicine. We compared the base sequences of the ndhF intergenic space of chloroplast DNA with Sinomenium acutum, Stephania tetrandra, C. trilobus, and A. fangchi. According to the results, it showed that the nucleotide variations were seen in 30 genes of four species. Phylogenetic analysis revealed that 4 species were classified into five groups based on an inter-group divergence in nucleotide sequence of 9%. We developed SFR marker nucleotides enough to authenticate respective species and confirmed its application on the band size at 419 base pair. These sequence differences at corresponding positions were available genetic markers to identity the Sinomeni Caulis et Rhizoma. Conclusions : Base on these results, the ndhF region was effective in distinguishing Sinomini Caulis et Rhizoma The SFR genetic marker was useful for identifying Sinomini Caulis et Rhizoma with other species.

• Korean Journal of Plant Taxonomy
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• v.50 no.1
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• pp.8-16
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• 2020

Complete chloroplast genome sequences provide detailed information about any structural changes of the genome, instances of phylogenetic reconstruction, and molecular markers for fine-scale analyses. Recent developments of next-generation sequencing (NGS) tools have led to the rapid accumulation of genomic data, especially data pertaining to chloroplasts. Short reads deposited in public databases such as the Sequence Read Archive of the NCBI are open resources, and the corresponding chloroplast genomes are yet to be completed. The V. dilatatum complex in Korea consists of four morphologically similar species: V. dilatatum, V. erosum, V. japonicum, and V. wrightii. Previous molecular phylogenetic analyses based on several DNA regions did not resolve the relationship at the species level. In order to examine the level of variation of the chloroplast genome in the V. dilatatum complex, raw reads of V. dilatatum deposited in the NCBI database were used to reconstruct the whole chloroplast genome, with these results compared to the genomes of V. erosum, V. japonicum, and three other species in Viburnum. These comparative genomics results found no significant structural changes in Viburnum. The degree of interspecific variation among the species in the V. dilatatum complex is very low, suggesting that the species of the complex may have been differentiated recently. The species of the V. dilatatum complex share large unique deletions, providing evidence of close relationships among the species. A phylogenetic analysis of the entire genome of the Viburnum showed that V. dilatatum is a sister to one of two accessions of V. erosum, making V. erosum paraphyletic. Given that the overall degree of variation among the species in the V. dilatatum complex is low, the chloroplast genome may not provide a phylogenetic signal pertaining to relationships among the species.