• 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.1
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• pp.21-40
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• 2006

We examined nrDNA ITS sequences from 16 taxa of Polygonum sect. Persicaria(Polygonaceae) in Korea to infer relationships among the taxa within the section. A neighbor-joining tree obtained from the analysis of the ITS sequences suggest that the ITS region was useful inferring the phylogenetic relationships among the taxa. The neighbor-joining tree indicates that P.amphibium is clearly separated from the other Korean taxa. The tree also reveals the presence of five major groups in the Korean taxa of the section; 1) P. lapathifolium var. lapathifolium, 2) P. persicaria and P. viscoferum, 3) P. orientale and P. viscosum, 4) P. japonicum and 5) a group including the remaining taxa. these relationships depicted on the ITS tree are largely congruent with those inferred from morphological and anatomical characters.

• Journal of Life Science
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• v.26 no.11
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• pp.1341-1344
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• 2016

Dendropanax morbifera is an endemic tree species of Korea, it is restricted to the southern parts of Korea. The internal transcribed spacer (ITS) region of nuclear ribosomal DNA (nrDNA) for Dendropanax morbifera grown at Bogil-do, Korea was determined. We investigated the sequence-based phylogenetic relationships of plants related and clarified its taxonomical position. The determined sequences consisted of 689 residues. ITS1 was 222 bp long while ITS2 was 233 bp long. The 5.8S rDNA was 160 bp long. The ITS region sequences of the Dendropanax species included in this study were obtained from GenBank. Oreopanax polycephalus was used as the outgroup. A pairwise alignment was calculated using the Clustal X program. A phylogenetic tree was constructed by the neighbor-joining method using the Tree view program. Sequence similarities among species including D. morbifera Bogil-do isolate showed the range 92.6 to 99.7% in sequence-based phylogenetic analysis using total 615 base pairs of ITS1, 5.8S rDNA and ITS2. D. morbifera Bogil-do isolate exhibited the highest degree of relatedness to D. chevalieri, sharing 99.7% ITS region similarity. D. morbifera Bogil-do isolate also showed to D. trifidus, sharing 99.4% ITS region similarity.

• Korean Journal of Plant Taxonomy
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• v.45 no.2
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• pp.158-168
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• 2015

A study of the genus Pilea in Korea including five taxa was carried out using molecular phylogenetic methods. The majority of members of the genus Pilea in Korea are annual herbs, and they live in moist habitats, flowering in summer and fruiting in autumn. The results of a phylogenetic analysis using nrDNA and cpDNA supported the recognition of P. japonica, P. peploides, and P. taquetii. Pilea taquetii from Mt. Sanbangsan in Jeju was nested within P. hamaoi and P. mongolica clade instead of the P. taquetii clade, with P. taquetii from Mt. Jirisan also separated from the P. taquetii clade. This indicates that the separation is not geographical isolation, but is instead related to taxonomic problems. Therefore, further study of the P. taquetii group is necessary.

• Korean Journal of Plant Resources
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• v.24 no.4
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• pp.358-369
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• 2011

The internal transcribed spacer (ITS) regions of nuclear ribosomal DNA from genus Chrysosplenium were sequenced to address phylogenetic relationship. ITS including 5.8S sequence varied in length from 647 bp to 653 bp. Among them, 219 sites were variable sites with parsimony-informative. The aligned sequences were analyzed by maximum parsimony (MP) and neighbor-joining (NJ) methods. In the strict consensus trees of parsimony analysis, the monophyly of Chrysosplenium was supported by 100% bootstrap value. The first clade, C. pseudofauriei was at the basal position of the genus, and others formed two clades with high bootstrap support. The second clade included Ser. Pilosa and Ser. Oppositifolia and third clade included Ser. Alternifolia and Ser. Flagellifera. The NJ trees showed essentially the same topology. Finally, DNA sequences of ITS regions were useful phylogenetic marker in this genus. Based on the ITS and ridge seed morphological results, C. sphaerospermum Maxim. and C. valdepilosum (Ohwi) S.H. Kang & J.W. Han were discussed their scientific names and taxonomic positions.

• Korean Journal of Plant Taxonomy
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• v.44 no.1
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• pp.22-32
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• 2014

The petal and sepal micromorphology of five species of Glechoma (Lamiaceae) was investigated to evaluate their taxonomic significance, and a molecular phylogeny using the sequences of internal transcribed spacers (ITS) regions of nuclear ribosomal DNA was carried out to resolve their phylogenetic relationships. Stomatal complexes were mostly found in the inner and outer part of the sepal from all investigated taxa, and the size length of the guard cell was variable among the taxa. Five types of trichomes (uni-cellular non-glandular trichome, multi-cellular non-glandular trichome, short-stalked capitate glandular trichome, long-stalked capitate glandular trichome, and peltate glandular trichome) were variable among the taxa as well as their distribution and density. In molecular phylogenetic studies, the genus Glechoma was composed of three geographically distinct major monophyletic groups (Europe-U.S.A., China-Korea, Japan). G. longituba in Korea and China formed well-supported monophyletic group. G. hederacea in Europe and U.S.A. formed a monophyletic and well-supported clade with G. sardoa, which are endemic species in Italy, with G. hirsuta falling as a sister to this clade. However, G. grandis did not form any phylogenetic relationships with the remaining taxa. The ITS analyses provided taxonomic boundaries of taxa in Glechoma although the petal and sepal micromorphological characters provided weak evidences of the systematic value. As further studies, incorporating more DNA regions to the matrix including other additional morphological analysis will be significant to provide clearer taxonomic structure in Glechoma.

• The Korean Journal of Mycology
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• v.48 no.3
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• pp.329-335
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• 2020

From 2008 to 2017, Korean azalea (Rhododendron yedoense f. poukhanense) showing angular, necrotic leaf spots were found in Jeju and Hongcheon, Korea. The lesions occurred frequently, detracting from the beauty of the glossy green leaves of the plant and causing premature defoliation. Therefore, to identify the fungus associated with the lesions, morphological characterization and molecular phylogenetic analysis of actin (Act), translation elongation factor 1-alpha (EF), internal transcribed spacer (ITS), 28S nrDNA (LSU), and RNA polymerase II encoding the second largest subunit (RPB2) of the two representative isolates were performed. The phylogenetic tree inferred from the neighbor-joining method showed the isolates clustering in the Sphaerulina azaleae group. Therefore, the fungus associated with the angular leaf spots on the Korean azalea was identified as Sphaerulina azaleae.

• Korean Journal of Plant Resources
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• v.23 no.4
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• pp.293-302
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• 2010

Taxa of Artemisia collected in Korea were constructed by molecular phylogenetic analysis based on the internal transcribed spacer(ITS) regions of nrDNA. The length of the ITS sequences aligned using the clustal X program was 636~643 bp, and the lengths of the ITS1 and ITS2 regions were 251~255 bp and 217~222 bp, respectively. The total number of variable sites was 95 for the entire sequence, and a parsimony- informative site represented an efficacious site in ITS1 rather than in ITS2. The maximum parsimony tree as calculated by the MEGA 4 program was clustered into five clades. The taxa(A. capillaris, A. japonica var. japonica, A. japonica var. hallaisanensis, A. japonica subsp. littoricora) degenerated ovary of clade 1 was supported as the subgenus Dracunculus by Ling's classification system. The results show that A. nakaii and A. fukudo were quite similar genetically(Boostrap 99%) and that the scientific name of Korean A. dubia should be reconsidered. A. sp. distributed in Ganghwa province was grouped with A. argyi(Boostrap 89%). These results suggest that the molecular techniques used in this study could be useful for the phylogenetic analysis of Korean Artemisia herbs having variations in their morphological characteristics.

• Korean Journal of Plant Resources
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• v.25 no.1
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• pp.96-104
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• 2012

Phylogenetic analyses were conducted to evaluate relationships of 7 taxa of Korean Carpesium including three outgroup (Inula britannica L., Inula germanica L., Rhanteriopsis lannginosa (DC.) Rauschert) by using ITS (internal transcribed spacer) sequences of nuclear ribosomal DNA. Phylogenetic studies used maximum parsimony, neighbor-joining and maximum likelihood methods analysis. The length of the ITS sequences was 731 bp, and the lengths of the ITS1, ITS2 and 5.8S regions were 284~297 bp, 264~266 bp and 164 bp, respectively. The total number of variable sites was 111 for the entire sequences, and a parsimony informative sites of 64 are valid. Base change appeared variously in ITS1 rather than in ITS2. As the result, Korean Carpesium were formed monophyletic group and C. abrotanoides situated as the most basal clade. The results show that C. macrocephalum is closely related with C. triste. C. rosulatum has the closest relationship with C. glossophyllum. C. cernuum is close to C. divaricatum. These results suggest that the ITS data used in this study could be useful for the phylogenetic analysis of Korean Carpesium.

• Korean Journal of Plant Taxonomy
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• v.45 no.3
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• pp.243-253
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• 2015

Series Chinensis, Genus Iris, endemic to the far regions of East Asia, consists of four species and related varieties. This series is divided into two major groups (I. rossii and I. minutiaurea complex). In this study, the ITS region and matK gene sequences within nuclear ribosomal DNA and plastid DNA were analyzed in order to investigate the phylogenetic relationships among the I. minutiaurea complex (I. minutiaurea, I. odaesanensis, and I. koreana) and the taxonomic identities of a putative hybrid in Mt. Palgong. In the internal transcribed spacer (ITS1, 5.8S, and ITS2) region, a total of 106 cloned genomic sequences from three taxa were obtained to study the intragenomic polymorphisms of the ITS regions. Three taxa revealed high levels of intragenomic polymorphisms, indicative of incomplete nrDNA concerted evolution. This incomplete ITS concerted evolution in the series Chinensis may be linked to the recent species divergence and frequent interspecies hybridization of the series Chinensis. In the matK gene, three taxa were fairly separated by eleven variable sites. In eight individuals collected on Mt. Palgong, putative hybrids between I. odaesanensis and I. minutiaurea were clustered in the I. minutiaurea clade in the NJ (neighbor-joining) tree based on the matK gene. However, in the ITS tree, some of them were clustered in the I. odaesanensis clade and others were clustered in the I. minutiaurea clade. Therefore, the individuals on Mt. Palgong were formed by the hybridization between two taxa (I. odaesanensis and I. minutiaurea) and not through the lineage of I. koreana.