• Title/Summary/Keyword: Cytochrome c oxidase subunit 1

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Identification of Heterodera glycines (Tylenchida; Heteroderidae) Using qPCR

  • Ko, Hyoung-Rai;Kang, Heonil;Park, Eun-Hyoung;Kim, Eun-Hwa;Lee, Jae-Kook
    • The Plant Pathology Journal
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    • v.35 no.6
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    • pp.654-661
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    • 2019
  • The soybean cyst nematode, Heterodera glycines, is a major plant-parasitic nematode that has caused important economic losses to Korea's soybean production. Four species of cyst nematodes, H. schachtii, H. glycines, H. trifolii, and H. sojae, all belong to schachtii group are coexist in field soil in Korea. The rapid identification of the nematode is crucial for preventing crop damage and in decision making for controlling this nematode. This study aimed to develop a species-specific primer set for quantitative PCR (qPCR) assay of H. glycines. The specific primer set (HGF1 and HGR1) for H. glycines was designed based on the cytochrome c oxidase subunit I (COI) sequence of mitochondrial DNA. After optimization, it is possible to identify the H. glycines using a qPCR assay with DNA extracted from a single cyst and single second-stage juvenile (J2). The specificity was confirmed by the absence of SYBR fluorescent signals of three other Heterodera species. A serial dilution of DNA extracted from a single cyst was obtained for the sensitivity test. The result showed that the standard curve of the test had a highly significant linearity between DNA concentration and Ct value (R2 = 0.996, slope = -3.49) and that the detection limit concentration of DNA of the primer set was 10 pg of DNA per reaction. Our findings suggested that H. glycines could be distinguished from H. sojae and other Heterodera species when a qPCR assay is used with a specific primer set.

New Record of Uraspis uraspis and Redescription of Uraspis helvola (Pisces: Carangidae) from Korea (한국산 전갱이과 어류 1미기록종, Uraspis uraspis 및 Uraspis helvola의 재기재)

  • Yeo, Sooeun;Kim, Jin-Koo
    • Korean Journal of Ichthyology
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    • v.28 no.1
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    • pp.57-64
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    • 2016
  • The fish species Uraspis helvola and U. uraspis (family Carangidae, Perciformes) were collected from Korea, and described and compared based on morphological and molecular characters. Uraspis helvola and U. uraspis were clearly distinguished by differences in the beginning point of the straight lateral lines scales (dorsal fin soft rays 12th~13th in U. helvola vs. 15th~16th in U. uraspis), and in the naked area on the breast extends to the pectoral fin base (naked area half way in U. helvola vs. naked area widely connected in U. uraspis). Molecular analysis using 530 base-pairs of mitochondrial DNA cytochrome c oxidase subunit I gene strongly supported the morphological identification. We described it as the new Korean record, and proposed the new Korean name "min-jeon-gaeng-i-sok" for the genus Uraspis, and "heuk-gi-min-jeon-gaeng-i" for the species U. uraspis.

Genetic identification of anisakid nematodes isolated from largehead hairtail (Trichiurus japonicus) in Korea

  • Kim, Jeong-Ho;Nam, Woo-Hwa;Jeon, Chan-Hyeok
    • Fisheries and Aquatic Sciences
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    • v.19 no.5
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    • pp.26.1-26.8
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    • 2016
  • Background: The nematode species belonging to genus Anisakis occur at their third larval stage in numerous marine teleost fish species worldwide and known to cause accidental human infection through the ingestion of raw or undercooked fish or squids. They may also draw the attention of consumers because of the visual impact of both alive and dead worms. Therefore, the information on their geographical distribution and clear species identification is important for epidemiological survey and further prevention of human infection. Results: For identification of anisakid nematodes species isolated from largehead hairtail (Trichiurus japonicus), polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of internal transcribed spacers of ribosomal DNA were conducted. Mitochondrial cytochrome c oxidase subunit 2 gene was also sequenced, and phylogenetic analysis was conducted. From the largehead hairtail (n = 9), 1259 nematodes were isolated in total. Most of the nematodes were found encapsulated throughout the viscera (56.2 %, 708/1259) or moving freely in the body cavity (41.5 %, 523/1259), and only 0.3 % (4/1259) was found in the muscles. By PCR-RFLP, three different nematode species were identified. Anisakis pegreffii was the most dominantly found (98.7 %, 1243/1259) from the largehead hairtail, occupying 98.7 % (699/708) of the nematodes in the mesenteries and 98.1 % (513/523) in the body cavity. Hybrid genotype (Anisakis simplex ${\times}$ A. pegreffii) occupied 0.5 %, and Hysterothylacium sp. occupied 0.2 % of the nematodes isolated in this study. Conclusions: The largehead hairtail may not significantly contribute accidental human infection of anisakid nematode third stage larvae because most of the nematodes were found from the viscera or body cavity, which are not consumed raw. But, a high prevalence of anisakid nematode larvae in the largehead hairtail is still in concern because they may raise food safety problems to consumers. Immediate evisceration or freezing of fish after catch will be necessary before consumption.

Distribution of Eggs and Larvae in Coastal Waters of Korea (우리나라 연안해역 난 및 자치어의 분포특성)

  • Baek, Jeong-Ik;Ji, Hwan-Sung;Yu, Hyo-Jae;Hwang, Kang-Seok;Kim, Doo-Nam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.54 no.4
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    • pp.467-479
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    • 2021
  • The distribution and abundance of fish eggs and larvae were investigated from February to December 2020 along the coastal waters of Korea. The eggs and larvae were identified using the mitochondrial DNA cytochrome c oxidase subunit I (mtDNA COI) and 16s rRNA gene. During the study period, eggs of overall 45 taxa belonging to 26 families were collected and larvae of overall 39 taxa belonging to 23 families were collected. In Yeongil Bay, eggs of Engraulis japonicus, which accounted for 83.9% of the total population, was the most dominant species, followed by Sardinops sagax (4.0%), Repomucenus valenciennei (3.8%) and E. japonicus larvae, which accounted for 34.9% of the total population. These were followed by Sebastiscus marmoratus (31.0%). In Gomso Bay, E. japonicus eggs accounted for 61.7% of the total population, followed by Sillago japonica (14.0%), Johnius grypotus (8.8%) and Pholis fangi larvae, which accounted for 53.5% of the total population, followed by Ammodytes personatus (34.1%). In Jinhae Bay, E. japonicus eggs accounted for 86.0% of the total population, followed by Leiognathus nuchalis (4.1%), Konosirus punctatus (3.7%) and E. japonicus larvae, which accounted for 48.7% of the total population, followed by Parablennius yatabei (21.6%).

Discovery of novel haplotypes from wild populations of Kappaphycus (Gigartinales, Rhodophyta) in the Philippines

  • Roleda, Michael Y.;Aguinaldo, Zae-Zae A.;Crisostomo, Bea A.;Hinaloc, Lourie Ann R.;Projimo, Vicenta Z.;Dumilag, Richard V.;Lluisma, Arturo O.
    • ALGAE
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    • v.36 no.1
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    • pp.1-12
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    • 2021
  • As the global demand for the carrageenophyte Kappaphycus is steadily increasing, its overall productivity, carrageenan quality, and disease resistance are gradually declining. In the face of this dilemma, wild Kappaphycus populations are viewed as sources of new cultivars that could potentially enhance production; therefore, assessment of their diversity is crucial. This study highlights the morphological and genetic diversity of wild Kappaphycus species obtained from two sites in the Philippines. Nucleotide alignments of available 5' region of the mitochondrial cytochrome c oxidase subunit I (COI-5P) and cox2-3 spacer sequences of Kappaphycus confirmed the presence of K. alvarezii in Guiuan, Eastern Samar and K. striatus in Bolinao, Pangasinan. Based on the concatenated sequences of the COI-5P and the cox2-3 spacer, nine novel haplotypes were observed along with other published haplotypes. However, there was no relationship between haplotype and morphology. These newly recognized haplotypes indicate a reservoir of unutilized wild genotypes in the Philippines, which could be taken advantage of in developing new cultivars with superior traits. DNA barcodes generated from this study effectively expand the existing databank of Kappaphycus sequences and can provide insights in elucidating the genetic diversity of Kappaphycus species in the country.

First Record of the Eared Blacksmelt, Lipolagus ochotensis (Bathylagidae, Osmeriformes) Larvae from the Southern Coastal Waters of Jejudo Island, Korea (제주도 남부해역에서 채집된 Bathylagidae (바다빙어목) Lipolagus ochotensis 자어의 한국 첫기록)

  • Moon Joo Yoon;Hwan-Sung Ji
    • Korean Journal of Ichthyology
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    • v.35 no.1
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    • pp.57-63
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    • 2023
  • Four individuals of Lipolagus ochotensis larvae (13.4~21.3 mm SL), belong to the family Bathylagidae, were collected by a Bongo net from the southern waters off Jejudo Island, Korea in February to March 2018. L. ochotensis is characterized by a elongated and compressed body, the eye stalks, series of melanophores on posterior of body, dorsal fin origin above the middle of the body. A molecular analysis based on 625 base pairs sequences in the mitochondrial DNA cytochrome c oxidase subunit I region shows that specimens were closely matched to adult L. ochotensis (genetic distance=0.024). We report the first record of family Bathylagidae, genus Lipolagus, L. ochotensis in Korean waters, and suggest their new Korean names, "Sim-hae-bing-eo-gwa", "Geom-eun-bing-eo-sok", and "Geom-eun-bbyam-bing-eo", respectively.

Review of the Scientific Name for Two Species of Genus Pampus (Stromateidae) in Korea (한국산 병어속(병어과) 어류 2종의 학명 검토)

  • Yeon-Myeong Lee;Jin-Koo Kim
    • Korean Journal of Ichthyology
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    • v.35 no.4
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    • pp.244-252
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    • 2023
  • This study compared the morphological and molecular characteristics of the two Pampus spp. in Korea, Japan and China to clarify the long-confused taxonomic status of the two Korean pomfrets named as "Byeong-eo" and "Deok-dae". Among them "Byeong-eo" was identified as Pampus punctatissimus in having the following morphological traits: patch of wavy ridges extending beyond the anterior tip of the pectoral fin, the groove on the lower ridge of gill cover absent, and the number of vertebrae 34. On the other hand, "Deok-dae" was identified as Pampus argenteus in having the following morphological traits: patch of wavy ridges not reaching to the anterior tip of the pectoral fin, the groove on the lower ridge of gill cover reaching to the lower jaw, and the number of vertebrae 38~42. In addition, "Byeong-eo" almost corresponded to P. punctatissimus from Japan in the mitochondrial DNA cytochrome c oxidase subunit I sequences (genetic distance=0.1%), while "Deokdae" showed genetic distance of 0.3% when compared to P. argenteus from China. Our comprehensive results suggest the validify of P. punctatissimus for "Byeong-eo", and P. argenteus for "Deok-dae".

Population Characteristics of the Venomous Giant Jellyfish, Nemopilema nomurai, found in the Yellow and Northern East China Seas (황해 중앙부와 동중국해 북부 해역에서의 대형 독성 노무라입깃해파리의 개체군 특성 연구)

  • Soo-Jung Chang;Jang-Seu Ki
    • Journal of Environmental Science International
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    • v.33 no.1
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    • pp.87-95
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    • 2024
  • The giant jellyfish, Nemopilema nomurai, is an endemic species found in Northeast Asian waters and their population structures, such as size and genetics, and their environmental characteristics were investigated. N. nomurai was obtained from the Yellow and Northern East China Seas during the summers of 2006, 2007, and 2009. In the northern Yellow Sea, small-sized jellyfish were found to be dominant and towards the southern seas, the size of the jellyfish increased. In the northern East China Sea, only one mode of jellyfish was found in May, and the number of modes increased up-to five in July. However, at the center of the Yellow Sea, one or two modes were found in July, 2007. Thus, different jellyfish populations were present in the northern East China Sea and the Yellow Sea. However, based on first appearance and a cohort analysis using the bell diameter, the jellyfish population in the northern Yellow Sea might be recognized as a distinct group that differed from those found in the northern East China Sea. Furthermore, mitochondrial DNA sequences (cytochrome c oxidase subunit I) of N. nomurai were, determined and compared with genetic structures obtained from jellyfish in the Yellow Sea. The genetic diversity of N. nomurai was highest in the regions around the northern East China Sea and at the center of the Yellow Sea and was the lowest around the northern Yellow Sea. Thus, N. nomurai populations in the Yellow Sea and northern East China Sea might be different concerning their seeding places.

Discordance between Morphological and Molecular Variations of the Genus Macroramphosus (Macroramphosidae) from Korea (한국산 대주둥치속(대주둥치과) 어류의 형태와 분자 변이의 불일치)

  • Sohn, Min-Soo;Kim, Jin-Koo
    • Korean Journal of Ichthyology
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    • v.32 no.4
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    • pp.199-209
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    • 2020
  • In order to clarify the taxonomic status of the Korean Macroramphosus species, which were previously confused, we investigated morphological and molecular variations of Macroramphosus (18 individuals) from Korea, and Macroramphosus (35 individuals) from Japan and Taiwan, and compared with those of M. scolopax from type locality (Mediterranean Sea). Although the Korean and Japanese specimens of Macroramphosus were clearly divided into two types in the first dorsal spine length (22.8~32.1% in A-type vs. 15.6~21.4% in B-type), distance between the first dorsal fin and second dorsal fin (6.4~9.7% vs. 8.6~13.3%), and body depth (20.0~28.0% vs. 17.3~22.6%), no genetic differences among all individuals of longspine snipefish between them were found at the specific level [d=0.0~3.3% in control region (CR); 0.0~1.3% in cytochrome b (cytb); 0.0~0.5% in cytochrome c oxidase subunit I (COI)]. Whereas, they were well distinguished in genetics (9.9~11.5% in CR; 3.8~4.6% in cytb; 1.2~3.6% in COI) from those of M. scolopax in Mediterranean Sea. It needs the scientific name of the longspine snipefish (M. scolopax) in Korea be changed as M. japonicus (and/or M. sagifue). However, our results could not find evidence of consistency between morphological and mitochondrial DNA variations which suggests that their differentiation event may occur fairly recently. Further studies using more sensitive markers such as microsatellite are needed to clarify the degree of gene flow between the two types.

Development of Species-Specific PCR Primers for the Rapid and Simultaneous Identification of the Six Species of Genus Takifugu

  • Dong, Chun Mae;Park, Yeon Jung;Noh, Jae Koo;Noh, Eun Soo;An, Cheul Min;Kang, Jung-Ha;Park, Jung Youn;Kim, Eun-Mi
    • Development and Reproduction
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    • v.23 no.4
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    • pp.367-375
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    • 2019
  • Pufferfish (Takifugu spp.) are economically important edible marine fish. Mistakes in pufferfish classification can lead to poisoning; therefore, accurate species identification is critical. In this study, we used the mtDNA cytochrome c oxidase subunit I gene (COI) to design specific primers for six Takifugu species among the 21 domestic or imported pufferfish species legally sold for consumption in Korea. We rapidly and simultaneously identified these pufferfish species using a highly efficient, multiplex polymerase chain reaction (PCR) system with the six species-specific primers. The results showed that species-specific multiplex PCR (multiplex species-specific polymerase chain reaction; MSS-PCR) either specifically amplified PCR products of a unique size or failed. MSS-PCR yielded amplification fragment lengths of 897 bp for Takifugu pardalis, 822 bp for T. porphyreus, 667 bp for T. niphobles, 454 bp for T. poecilonotus, 366 bp for T. rubripes, and 230 bp for T. xanthpterus using the species-specific primers and a control primer (ca. 1,200 bp). We visualized the results using agarose gel electrophoresis to obtain accurate contrasts of the six Takifugu species. MSS-PCR analysis is easily performed and provides identification results within 6 h. This technique is a powerful tool for the discrimination of Takifugu species and will help prevent falsified labeling, protect consumer rights, and reduce the risk of pufferfish poisoning..