• Korean Journal of Fisheries and Aquatic Sciences
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• v.48 no.5
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• pp.731-738
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• 2015

Nematode infection in the epithelial tissue of cultured rockfish Sebastes schlegeli was first reported in 2012. Since then, nematode infections have caused serious economic losses in rockfish aquaculture on the west coast of Korea. Taxonomic and life cycle information for this parasite are currently unknown. In this study, 18S rRNA and cytochrome c oxidase subunit I (COI) genes were used for molecular identification and polymerase chain reaction (PCR) to detect the invisible stages of this parasite. Nucleotide sequences of the 18S rRNA of the rockfish nematode showed 98% identity with that of Philometra morii. Therefore, this rockfish nematode was classified to the Philometridae family. However, we could not identify it to genus level using 18S rRNA. Its COI nucleotide sequences shared 85% and 82% identities with those of Bursaphelenchus sinensis and Philometra overstreeti, respectively. In addition, two gene-specific primer sets were designed based on the 18S rRNA gene to detect the intermediate host and nematode larvae. These primers were specific to this rockfish nematode without cross-reacting to other pathogens. The detection limit of the PCR assay using these primers was 1,000 copies of nematoda plasmid DNA. Therefore, the PCR assay described here is suitable for the detection of nematode DNA within rockfish. In addition, this PCR assay could be used to detect nematode larvae and the intermediate host.

• Proceedings of the National Institute of Ecology of the Republic of Korea
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• v.3 no.4
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• pp.221-226
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• 2022

A filarial nematode was found in a blood sample of an Anas falcata individual collected in South Korea in 2018. Phylogenetic analysis based on partial cytochrome C oxidase subunit I (COI) sequences placed the nematode as a novel genus of the family Onchocercidae and as closely related to Mansonella species, Chandlerella quiscali, and filarial nematodes recently reported in avian species. However, different phylogenetic relationship was observed in the NADH dehydrogenase subunit 5 and 12S rRNA-based phylogenetic trees, which might indicate the filarial nematode found in this study was not defined to belong to the known specific genera of the family Onchocercidae. The screening of 105 additional avian blood samples retrieved only one 12S rRNA-targeting polymerase chain reaction (PCR)-positive sample, which indicates that filarial nematode infection is rare in wild birds or that it occurs below the detection limit of PCR in blood samples. Nevertheless, considering the recent findings about ancient interactions between birds and human pathogenic filarial nematodes and their pathogenic potential in several avian species, additional exploration of novel filarial nematodes in wild birds remains necessary.

• Korean journal of applied entomology
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• v.29 no.1
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• pp.14-19
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• 1990

Nematodes and insects associated with dead trees were surveyed on the 238 dead trees belonging to 19 tree species in Pusan, Gyeongnam, Gyeongbuk, Chunnam, and Chonbuk provinces from April to September of 1989. Pine wood nematode, Bursaphelenchus xylophilus was found only in Pusan but B. mucronatus was collected at Chinju and Chinhae. The 13 nematode species in 9 genera were identified. Out of them, Diplogasteroides dimidius, Rhabdontolaimus adephagus, R. janae, Mikoletzkya diluta, M. ruminis, M. langcaudaa, Parasitorhabditis hylurgi, Panagrolaimus concolor, Panagrodontus dentatus, Prothalonema intermedium, and marcrolaimus canadensis were recorded for the first time in Korea. Insects collected from dead trees were 5 orders, 9 families, 25 genera, and 27 species. of them, the Coleoptera were the most collected insects by the 3 families, 19 genera and 22 species. The Scolytidae were 12 species in 10 genera. Hypothenemus eruditus was firstly collected from Campylotropis macrocarpa, Lespedeza maximowizi, Forsythia ovata, Meliosma oldhami, Securinega suffruticosa, Broussonetia kazinoki, and Cornus walteri. The maximum number of pine wood nematode was separated from the abdomen of Monchamus alternatus, the pine woodnematode vector. The maximum number of nematodes per an adult of M. alternatus was 127,535, minimum 2,616, and average 42,817.

• 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.

• Korean journal of applied entomology
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• v.25 no.3 s.68
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• pp.159-167
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• 1986

White tip nematode damage to rice was surveyed at 111 $My{\check{o}}n$ from 36 Gun in Kyongsangnam-Do, $Ky{\check{o}}ngsangbuk-Do\;and\;Ch{\check{o}}llanam-Do$. Rate of damaged stem by the nematode was highest at $Ch'angny{\check{o}}ng-Gun\;in\;Ky{\check{o}}ngsangnam-Do$ as 28.3%, $Y{\check{o}}ngch'on-Gun\;in\;Ky{\check{o}}ngsangbuk-Do$ as 69.3% and $Pos{\check{o}}ng-Gun\;in\;Ch{\check{o}}llanam-Do$ as 40.6%. In pot experiment, no white tip symptoms were appeared in Tongil hybrids except $W{\check{o}}np'ung$ by 3.4% but a great number of the nematode was detected in seeds while symptom and a fair amount of the nematode were observed in Japonica cultivars. The number of nematode in Shingwang cultivar of Tongil hybrids and Dongin cultivar of Japonica was highest by 1997, by 1023 individuals per 10g of seed respectively. Detection rate of the nematode was different depended upon peeling of the husks. Less than 50% was detected from unhulling seed compared with hulled seed except 8 cultivars such as $Shins{\check{o}}nchal$, Hangangchal, $S{\check{o}}gwang,\;Y{\check{o}}ngpung$, Milyang 23, $Such{\check{o}}ng,\;Taech'{\check{o}}ng\;and\;S{\check{o}}nam$. More effective detection was oftained with temperature level at $25^{\circ}C$ and increased with immersing time regardless of peeling the husks.

• The Plant Pathology Journal
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• v.33 no.2
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• pp.184-192
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• 2017

Tylenchulus semipenetrans is an important and widespread plant-parasitic nematode of citrus worldwide and can cause citrus slow decline disease leading to significant reduction in tree growth and yield. Rapid and accurate detection of T. semipenetrans in soil is important for the disease forecasting and management. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed to detect T. semipenetrans using DNA extracted from soil. A set of five primers was designed from the internal transcribed spacer region (ITS1) of rDNA, and was highly specific to T. semipenetrans. The LAMP reaction was performed at $63^{\circ}C$ for 60 min. The LAMP product was visualized directly in one reaction tube by adding SYBR Green I. The detection limit of the LAMP assay was $10^{-2}J2/0.5g$ of soil, which was 10 times more sensitive than conventional PCR ($10^{-1}J2/0.5g$ of soil). Examination of 24 field soil samples revealed that the LAMP assay was applicable to a range of soils infested naturally with T. semipenetrans, and the total assay time was less than 2.5 h. These results indicated that the developed LAMP assay is a simple, rapid, sensitive, specific and accurate technique for detection of T. semipenetrans in field soil, and contributes to the effective management of citrus slow decline disease.

• Proceedings of the Korean Society of Applied Entomolohy Conference
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• 2001.05a
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• pp.130-130
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• 2001

• Journal of the Korean Society of Surveying, Geodesy, Photogrammetry and Cartography
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• v.40 no.4
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• pp.251-260
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• 2022

Although the number of trees affected by pine wilt disease is decreasing, the affected area is expanding across the country. Recently, with the development of deep learning technology, it is being rapidly applied to the detection study of pine wilt nematodes and dead trees. The purpose of this study is to efficiently acquire deep learning training data and acquire accurate true values to further improve the detection ability of U-Net models through learning. To achieve this purpose, by using a filtering method applying a step-by-step deep learning algorithm the ambiguous analysis basis of the deep learning model is minimized, enabling efficient analysis and judgment. As a result of the analysis the U-Net model using the true values analyzed by period in the detection and performance improvement of dead pine trees of wilt nematode using the U-Net algorithm had a recall rate of -0.5%p than the U-Net model using the previously provided true values, precision was 7.6%p and F-1 score was 4.1%p. In the future, it is judged that there is a possibility to increase the precision of wilt detection by applying various filtering techniques, and it is judged that the drone surveillance method using drone orthographic images and artificial intelligence can be used in the pine wilt nematode disaster prevention project.

• Korean journal of applied entomology
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• v.52 no.4
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• pp.327-333
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• 2013

Plant parasitic nematodes were isolated from 274 soil samples collected from soybean fields in Korea. Nematode importance rating in soybean is proposed based on this study and by reviewing other reports. Soybean cyst nematode, Heterodera glycines is the most important nematode species and rated as $1^{st}$ because it detected from 38%(range 25-51%) of soil samples with high density except Jeju province. Root-knot nematode, Meloidogyne spp. is rated $2^{nd}$ and is more widely distributed in southern provinces, Gyeongnam, Jeonnam, and Jeju province (detection rate ranged 16-44%). Pratylenchus is rated $3^{rd}$ and is more frequently detected from northern provinces such as Gyeonggi and Gangwon (detection rate ranged 10-13%). Helicotylenchus is rated $4^{th}$ and is particularly important in Jeju province (detection rate is 62% and numbers averaged 571 nematodes/$300cm^3$ soil), which is the main production area for sprouting soybean in Korea. Tylenchorhynchus, Paratylenchus, Criconema, Criconemoide, Mesocriconema, Ogma, Xiphinema, Paratrichodorus, and Trichodorus occur in low frequency and density, thus they are rated to $5^{th}$. We propose to delete following nematode genus from the list of soybean parasitic nematode in Korea because their parasitism on soybean is unconfirmed or negligible; Tylenchus, Aphelenchus, Aphelenchoides, Ditylenchus, Hirschmanniella immamuri, Basiria graminophila, Psilenchus, and Pseudhalenchus. Therefore, we revised 30 species in 16 genera as soybean parasitic nematodes in Korea. Importance rating is $1^{st}$ Heterodera glycines, $2^{nd}$ Meloidogyne spp., $3^{rd}$ Pratylenchus spp. and $4^{th}$ Helicotylenchus spp. especially in Jeju province.

• Korean journal of applied entomology
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• v.57 no.1
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• pp.7-13
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• 2018

Currently, there is no available tool that rapidly diagnoses pine wood nematode (PWN)-infected pine trees in the field. In this study, we synthesized and purified PWN Galectin, which might be an antigen specific to PWN, using the Baculovirus expression system. We used PWN Galectin as an antigen for generating 1,464 fusion hybridoma cell lines secreting monoclonal antibodies (Mabs). Among them, we selected 62 fusion hybridoma cell lines showing high reactivity to PWN Galectin. We further selected 12 fusion hybridoma cell lines showing high reactivity to the standard PWN-infected pine tree phosphate buffered saline (PBS) extract. Additionally, two fusion hybridoma cell lines showing no or extremely low reactivity were used as controls. The selected fusion hybridoma cell lines were subjected to limiting dilutions for selecting and establishing Mab-secreting cell lines showing higher reactivity to the standard PWN-infected pine tree extract than to the standard normal pine tree PBS extract. Moreover, the selected fusion hybridoma cell lines were further selected based on their higher reactivity to PWN protein extracts than to three non-pathogenic nematode protein extracts. The Mab-secreting cell lines established in this study could be used to develop rapid diagnostic tools that can be used in the field or in laboratories for detecting PWN-infected pine trees or PWN.