• The Plant Pathology Journal
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• 제24권2호
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• pp.159-163
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• 2008

Accurate identification of pine wood nematode, Bursaphelenchus xylophilus is a prerequisite to diagnose the pine wilt disease. However, a fungivorous nematode, B. mucronatus is highly similar to B. xylophilus and it is difficult to differentiate these two species by morphological features. A molecular diagnosis method, ITSRFLP was applied for the identification of B. xylophilus and B. mucronatus from Korea. Genomic DNA was extracted from a single individual nematode and ITS DNA was amplified by PCR. The size of PCR product was approximately 900bp and the sequence data were obtained after cloning. Amplified ITS was digested by 5 different restriction enzymes (Rsa I, Hae III, Msp I, Hinf I, and Alu I) and provided a discriminatory profile for B. xylophilus and B. mucronatus. Besides, B. mucro- natus was determined to have 2 different genotypes, East Asian type and European type also clearly separated by Rsa I and Hae III digestion. European type of B. mucronatus is recently collected from Pinus koraiensis and has not been reported before. ITS sequnce data were analyzed by Restriction Mapper program and the result supported ITS-RFLP pattern. These data indicated that PCRRFLP method is an accurate and simple way for identification of Bursaphelenchus species.

• 한국수산과학회지
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• 제48권5호
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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.

• The Plant Pathology Journal
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• 제35권6호
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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.

• 한국식물병리학회지
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• 제14권6호
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• pp.603-605
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• 1998

Soil and root samples were collected form the rhizoshpere of 11 different medicinal plants to determine the incidence, density and identification of root-knot nematode species associated with medicinal herbs. About 55% of medicinal herbs examined was found to be infested with root-knot nematodes. As a result of infection casued by three root-knot nematodes, M. hapla recorded 43.3% in medicinal herba whereas M. incognita and M. arenaria showed 7.9% and 3.7%, repectively. Forsythia koreana, Hemerocalis fulva, Hibuscus mutabilis and Petasites japonicus were the most severely infested herbs whereas Acanthopanax sessilflorus was least infested. Population of the second stage younger plants. Meloidogyne hapla, M. incognita and M. arenaria were the species associated with the medicinal herbs. The most abundant nematode observed in medicinal herbs was M. hapla and followed by M. incognita and M. arenaria. M. arenaria was observed firstly on Ficus carica, one of medicinal plant.

• Fisheries and Aquatic Sciences
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• 제19권5호
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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.

• The Plant Pathology Journal
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• 제21권1호
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• pp.83-85
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• 2005

Ditylenchus acris was isolated from diseased strawberry plants. Frequently, D. acris and Aphelenchoides fragariae occur together in a strawberry plant. Both species appeared very similar in the shape, length, swimming behavior and causing symptoms, and difficult to distinguish them by a stereomicroscope. But they were easily distinguished under a compound microscope especially by their tail shape, median bulb, vulva position, and bursa.

• Parasites, Hosts and Diseases
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• 제53권6호
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• pp.713-717
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• 2015

A 23-year-old female residing in a village of Cao Bang Province, North Vietnam, visited the Hospital of Hanoi Medical University in July 2013. She felt dim eyes and a bulge-sticking pain in her left eye for some days before visiting the hospital. In the hospital, a clinical examination, an eye endoscopy, and an operation were carried out. A nematode specimen was collected from the eye of this patient. The body of this worm was thin and long and measured $22.0{\times}0.3mm$. It was morphologically suggested as an immature female worm of Angiostrongylus cantonensis. By a molecular method using 18S rRNA gene, this nematode was confirmed as A. cantonensis. This is the first molecular study for identification of A. cantonensis in Vietnam.

• 한국응용곤충학회지
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• 제45권1호
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• pp.79-85
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• 2006

국내 주요감자재배지의 식물기생선충 발생상을 밝히기 위해 2004년과 2005년에 제주, 예산, 김천, 고령, 홍천, 평창, 김제, 밀양, 남원, 강릉, 인제 등 11개 감자 주산지를 대상으로 선충속별 종류와 밀도를 조사하였다. 채집한 토양시료 50개 중 30개에서 뿌리혹선충 유충이 검출되었으며 밀도는 12-69마리/토양 100cc 정도로 비교적 낮았다. 식물기생선충류 중 Pratylenchus sp., Helicotylenchus sp, Ditylenchus sp., Tylenchus sp., and Tylenchorhynchus sp.가 지역별로 검출되었으나 대부분 밀도는 매우 낮게 나타났다. 채집한 토양을 토마토 뿌리에 접종 후 뿌리혹선충을 배양하여 PCR-RLFP에 의한 정밀동정 결과 제주, 밀양, 고령에서 채집한 뿌리혹선충 암컷에서 500 bp의 PCR 산물을 얻었으며 제한 효소 Dra I을 처리한 결과 290 및 230 bp의 밴드가 형성되어 당근뿌리혹선충(Meloidogyne hapla)으로 동정하였다.

• 한국응용곤충학회지
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• 제53권1호
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• pp.51-57
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• 2014

최근 5년 동안 식물검역 현장에서 검출된 뿌리썩이선충속 선충을 분류하여 Pratylenchus crenatus, P. fallax, P. kumamotoensis, P. panamaensis, P. penetrans로 동정하였다. 이 종들의 진단형질은 구순부의 주름 수, 두부의 형태, 구침의 길이, 수컷의 유무, 측대의 구조, 수정낭의 형태, 후부자궁낭의 길이, 미부의 형태 등을 포함한다. 이들의 사진과 측정치 및 형태적 특징을 기술하였다.

• 농약과학회지
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• 제12권1호
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• pp.97-101
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• 2008

소나무재선충에 대한 살선충 활성을 나타내는 방선균의 탐색 및 동정을 위하여 2000여개의 방선균배양액 라이브러리로부터 살선충 활성을 탐색하였다. 현재 소나무재선충에 대한 생물학적 방제제로 사용되고 있는 Streptomyces avermitilis 균주 배양액의 살선충 활성과 비교하여 선발한 결과, 강력한 살선충 활성을 나타내는 균주 AW050027을 균주를 최종 선발하였다. 선발군주에 대하여 형태학적, 배양학적, 생리학적 특성 분석 및 분자계통분류학적 분석을 한 결과 선발군주는 Micromonospora coriariae $NAR01^T$ 균주와 98.9%의 상동성을 가진 균주로 동정되었다.