• Ocean and Polar Research
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• v.44 no.2
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• pp.161-177
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• 2022

Microalgae can attach to the surface of ships and then spread to various areas by means of ship transport. The introduction of invasive species through ships is recognized as a marine problem worldwide. Identification of attached microalgae is necessary to investigate such movement between countries through ships. In the present study, through analytical methods we reviewed research data to identify the taxa of domestic attached microalgae and assess the ecological impacts of such microalgae. A total of 87 genera and 153 species (143 species of diatoms, 10 species of cyanobacteria, and 4 genera of dinoflagellates) were identified as native attached microalgae in Korea, and diatoms accounted for 93% of the total. Most of these attached microalgae were identified through research on natural substrates such as seaweeds and bedrock, and some were also identified through experiments using artificial adherent plates. To date, there is no information on microalgae attached to international ships and introduced into Korea. Molecular genetic analysis and systematic management through on-site sampling of international ships, microscopic analysis, and meta-barcoding are necessary to assess the inflow and spread path of hull-attached marine alien species and evaluate the risk they pose to the domestic ecosystem.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.55 no.4
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• pp.478-483
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• 2022

A single specimen (522.0 mm in total length) of the family Muraenidae, collected from the waters off south-western Jeju-do Island in November 2021, was identified as Echidna nebulosa on the basis of morphological and molecular methods (DNA barcoding). This species is characterized by several morphological traits as follows: 10 rounded molar, median intermaxillary, and vomerine teeth; 4 predorsal vertebrae, 50 preanal vertebrae and 123 total vertebrae; and a pale background color with two rows of 17-20 blackish brown starry blotches along the head and body. As this is the first record of E. nebulosa in Korea, we propose the new Korean names, "Dung-Geun-Ni-Gom-Chi-Sog" for the genus Echidna, and "Beol-Kkok-Gom-Chi" for E. nebulosa.

• ALGAE
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• v.30 no.3
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• pp.217-222
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• 2015

Pythium species (Pythiales, Oomycetes) are well known as the algal pathogen that causes red rot disease in Pyropia / Porphyra species (Bangiales, Rhodophyta). Accurate species identification of the pathogen is important to finding a scientific solution for the disease and to clarify the host-parasite relationship. In Korea, only Pythium porphyrae has been reported from Pyropia species, with identifications based on culture and genetic analysis of the nuclear internal transcribed spacer (ITS) region. Recent fungal DNA barcoding studies have shown the low taxonomic resolution of the ITS region and suggested the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene as an alternative molecular marker to identify Pythium species. In this study, we applied an analysis of both the ITS and cox1 regions to clarify the taxonomic relationships of Korean Pythium species. From the results, the two closely related Pythium species (P. chondricola and P. porphyrae) showed the same ITS sequence, while the cox1 marker successfully discriminated P. chondricola from P. porphyrae. This is the first report of the presence of P. chondricola from the infected blade of Pyropia yezoensis in Asia. This finding of the algal pathogen provides important information for identifying and determining the distribution of Pythium species. Further studies are also needed to confirm whether P. chondricola and P. porphyrae are coexisting as algal pathogens of Pyropia species in Korea.

• ALGAE
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• v.33 no.2
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• pp.157-166
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• 2018

Cryptic species complexes are increasingly recognized in phycological research, obscuring taxonomy and raising questions about factors influencing speciation. A recent exploration of kelp genetic diversity on Haida Gwaii, British Columbia revealed the existence of a new species, Saccharina druehlii, which is cryptic with Saccharina sessilis. This suggests that molecular investigations further north may be required to elucidate the taxonomy and evolutionary history of this lineage. Although, for several decades, S. sessilis was considered a single highly variable species, its taxonomy has been far from straightforward. In particular, Hedophyllum subsessile (Areschoug) Setchell is now recognized as a synonym of S. sessilis in North America, but as a growth form of Saccharina bongardiana in Far East Russia. To resolve this taxonomic confusion, we sequenced mitochondrial (CO1-5P) and nuclear (internal transcribed spacer) markers of S. sessilis populations from the Aleutian Islands, Alaska, USA. Interestingly, none of our sequences matched S. sessilis sensu stricto. Instead, CO1-5P sequences from populations in the central and eastern Aleutians matched exactly S. druehlii with increasing sequence divergence occurring westward. Samples from Attu, the western-most island, composed a genetic group that clearly represents Kjellman's concept of Hafgygia bongardiana f. subsessilis and is distinct enough from S. druehlii and S. sessilis to potentially constitute a distinct species. Therefore, Saccharina subsessilis comb. nov. is proposed for this entity. Our results suggest the existence of a species complex at the crown node of S. sessilis and thus further investigation of Saccharina in Alaskan waters should be conducted to reconstruct the evolutionary history of this fascinating lineage.

• Journal of Plant Biotechnology
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• v.47 no.1
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• pp.15-25
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• 2020

Allium L. is one of the largest genera of the Amaryllidaceae family, with more than 920 species including many economically important species used as vegetables, spices, medicines, or ornamental plants. Currently, DNA barcoding tools are being successfully used for the molecular taxonomy of Allium. A total of 46 Allium species were collected from their native areas, and DNA was extracted using the IBRC DNA extraction kit. We used specific primers to PCR amplify matK. DNA sequences were edited and aligned for homology, and a phylogenetic tree was constructed using the neighbor-joining method. The results show thymine (38.5%) was the most frequent and guanine (13.9%) the least frequent nucleotide. The matK regions of the populations were quite highly conserved, and the amount of C and CT was calculated at 0.162 and 0.26, respectively. Analysis of the nucleotide substitution showed C-T (26.22%) and A-G (8.08%) to have the highest and lowest percent, respectively. The natural selection process dN/dS was 1.16, and the naturality test results were -1.5 for Tajima's D and -1.19 for Fu's Fs. The NJ dendrogram generated three distinct clades: the first contained Allium austroiranicum and A. ampeloprasum; the second contained A. iranshahrii, A. bisotunense, and A. cf assadi; and the third contained A. rubellum and other species. In this study, we tested the utility of the matK region as a DNA barcode for discriminating Allium. species.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.54 no.5
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• pp.685-690
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• 2021

Harmful shell-boring species of the genus Polydora (Polychaeta: Spionidae) were frequently reported from commercially important mollusk species in Korea, Japan and China. The traditional approach based on the morphological characteristics showed limitations for species discrimination among shell-boring species. Therefore, DNA barcoding was adopted to identify Polydora species using molecular markers. Two Polydora species (P. haswelli and P. hoplura) in abalone shells were reported from our previous molecular phylogenetic study. In this study, we additionally reported the presence of shell-boring Polydora haswelli in commercially sold shellfish. The taxon-specific cox1 marker used in this study successfully allowed the isolation of P. haswelli from cockle Scapharca subcrenata, mussel Mytilus galloprovincialis, oyster Crassostrea gigas and scallop Argopecten irradians. Polydora hoplura was not found in these shellfish species. The genetic variations were found on the intraspecific level of P. haswelli and the same genotype was also detected in different shellfish species. This result can provide information on a new host and accurate parasitic Polydora species. Moreover, this report can be used as the biodiversity data of Polydora species on the invasion and transition of harmful Polydora species in mollusk aquaculture farms.

• Horticulture, Environment, and Biotechnology : HEB
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• v.59 no.6
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• pp.865-873
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• 2018

Allium ochotense and Allium microdictyon are commonly known as 'Mountain garlic' and are popular, economically important species in many countries such as Korea, China, and Mongolia. Their leaves are used as culinary side dishes and in traditional medicines. In Korea, these two species are at risk of extinction due to damage to their natural habitat and thus, conservation and breeding programs are needed. However, their identification relies mostly on morphological data, which is limited and until recently, led to classifying these two species under A. victorialis. In the present study, a simple and reliable method of molecular identification was developed to distinguish A. ochotense from A. microdictyon that targets four barcoding regions: the internal transcribed spacer (ITS), the maturase K gene (matK), the chloroplast psbA-trnH intergenic region, and the ribulose-bisphosphate carboxylase large subunit gene (rbcL). Single nucleotide polymorphisms (SNPs) were found in ITS and matK regions, and species-specific primers were designed based solely on the SNP at position 680 of the ITS region that could differentiate A. ochotense from A. microdictyon. Using these primers in amplification refractory mutation system (ARMS)-PCR, A. ochotense, and A. microdictyon could be simultaneously and efficiently distinguished. This study is the first to report a simple, rapid, and efficient method for discriminating A. ochotense and A. microdictyon, indicating the utility of species-specific markers in the development of conservation and breeding programs.

• The Korean Journal of Mycology
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• v.49 no.4
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• pp.455-467
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• 2021

Macrofungi are valuable resources as novel drug candidates, new biomaterials, and edible materials. Recently, genetic approaches pertaining to macrofungi have been continuously growing for their identification, molecular breeding, and genetic engineering. However, purification and amplification of fungal DNA is challenging because of the rigid cell wall and presence of PCR inhibitory metabolites. Here, we established a direct PCR method to provide a rapid and efficient method for PCR-grade macrofungal DNA preparation applicable to both conventional PCR and real-time PCR. We first optimized the procedure of lysis and PCR using the mycelia of Lentinula edodes, one of the most widely consumed macrofungal species. Lysates prepared by neutralizing with (NH₄)₂SO₄ after heating the mycelia in a mixture of TE buffer and KOH at 65℃ for 10 min showed successful amplification in both conventional and real-time PCR. Moreover, the addition of bovine serum albumin to the PCR mixture enhanced the amplification in conventional PCR. Using this method, we successfully amplified not only internal transcribed spacer fragments but also low-copy genes ranging in length from 500 to 3,000 bp. Next, we applied this method to 62 different species (54 genera) of macrofungi, including edible mushrooms, such as Pleurotus ostreatus, and medicinal mushrooms such as Cordyceps militaris. It was found that our method is widely applicable to both ascomycetes and basidiomycetes. We expect that our method will contribute to accelerating PCR-based approaches, such as molecular identification, DNA marker typing, gene cloning, and transformant screening, in macrofungal studies.

• Korean Journal of Environmental Biology
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• v.40 no.4
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• pp.464-476
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• 2022

In recent years, macroalgal bloom occurs frequently in coastal oceans worldwide. It might be attributed to accelerating climate change. "Green tide" events caused by proliferation of green macroalgae (Ulva spp.) not only damage the local economy, but also harm coastal environments. These nuisance events have become common across several coastal regions of continents. In Korea, green tide incidences are readily seen throughout the year along the coastlines of Jeju Island, particularly the northeastern coast, since the 2000s. Ulva species are notorious to be difficult for morphology-based species identification due to their high degrees of phenotypic plasticity. In this study, to investigate temporal variation in Ulva community structure on Jeju Island between 2015 and 2020, chloroplast barcode tufA gene was sequenced and phylogenetically analyzed for 152 specimens from 24 sites. We found that Ulva ohnoi and Ulva pertusa known to be originated from subtropical regions were the most predominant all year round, suggesting that these two species contributed the most to local green tides in this region. While U. pertusa was relatively stable in frequency during 2015 to 2020, U. ohnoi increased 16% in frequency in 2020 (36.84%), which might be associated with rising sea surface temperature from which U. ohnoi could benefit. Two species (Ulva flexuosa, Ulva procera) of origins of Europe should be continuously monitored. The findings of this study provide valuable information and molecular genetic data of genus Ulva occurring in southern coasts of Korea, which will help mitigate negative influences of green tide events on Korea coast.

• 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.