• The Plant Pathology Journal
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• 제20권2호
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• pp.135-141
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• 2004

A filamentous virus was isolated from taro (Colocasia esculenta Schott) showing mosaic and chlorotic feather-ing symptoms in Chuncheon, Gangwon province in 2002. Based on ELISA, its appearance in electron microscope, serological relationships, and RT-PCR using specific primer and nucleotide sequence analysis of the CP gene, the isolated virus was identified as Dasheen mosaic virus (DsMV) and designated as Korean isolated (DsMV-Kr). DsMV was not serologically related to Zantedeschia mosaic virus (ZaMV), which has been reported to infect an Araceae plants. Since the coat protein revealed electrophoretic heterogeneity, about 42 kDa, 39 kDa and 31 kDa by SDS-PAGE, an improved purification method was established for the production of antisera against DsMV-Kr. The purification method used in this study may be effectively applied to the purification of other filamentous viruses.

• The Plant Pathology Journal
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• 제21권4호
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• pp.361-368
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• 2005

A Korean isolate of Pepper mild mottle virus (PMMoV-Kr) was isolated from a diseased hot pepper plant and its biological and molecular properties were compared to that of PMMoV-J and PMMo V -So The genomic RNA of PMMoV-Kr consists of 6,356 nucleotides. The nucleotide and amino acid sequences identities of four viral proteins and two noncoding regions among PMMoV-Kr, PMMoV-S and PMMoV-J were $96.9\%\;to\;100.0\%\;and\;97.5\%\;to\;98.6\%$, respectively. Full-length cDNA amplicon of PMMoV-Kr was directly amplified by RT-PCR with a set of 5'-end primer anchoring T7 RNA promoter sequence and 3'-end virus-specific primer. Capped transcript RNAs from the full-length cDNA clone were highly infectious and caused characteristic symptoms of wild type PMMoV when mechanically inoculated to systemic host plants such as Nicotiana benthamiana and pepper plants.

• The Plant Pathology Journal
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• 제20권2호
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• pp.147-154
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• 2004

A simple and reliable procedure for RT-PCR detection of Apple stem pitting virus (ASPV), Cherry rasp leaf virus (CRLV), and Cherry necrotic rusty mottle virus (CNRMV) was developed. Two virus specific primer sets for each virus were found to specifically detect each virus among fourteen sets of designed oligonucleotide primers. Total RNAs extracted from healthy and from ASPV-,CRLV- and CNRMV-infected plant tissues were used to synthesize cDNA using oligo dT primer and then amplified by virus-specific primers for each virus. Each primer specifically amplified DNA fragments of 578 bp and 306 bp products for ASPV (prAS CP-C and prAS CP-N primers, respectively); 697 bp and 429 bp products for CRLV (prCR4 and prCR5-JQ3D3 primers, respectively); and 370 bp and 257 bp products for CNRMV (prCN4 and prCN6-NEG 1 primers, respec-tively) by RT-PCR. DNA sequencing of amplified DNA fragments confirmed the nature of each amplified DNA. Altogether, these results suggest that these virus specific primer sets can specifically amplify viral sequences in infected tissues and thus indicate that they can be used for specific detection of each virus.

• The Plant Pathology Journal
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• 제19권3호
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• pp.159-161
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• 2003

The coat protein (CP) gene of Apple mosaic virus (ApMV), a member of the genus Ilarvirus, was selected for the design of virus-specific primers for amplification and molecular detection of the virus in cultivated apple. A combined assay of reverse transcription and polymerase chain reaction (RT-PCR) was performed with a single pair of ApMV-specific primers and crude nucleic acid extracts from virus-infected apple for rapid detection of the virus. The PCR product was verified by restriction mapping analysis and by sequence determination. The lowest concentration of template viral RNA required for detection was 100 fg. This indicates that the RT-PCR for detection of the virus is a 10$^3$times more sensitive, reproducible and time-saving method than the enzyme-linked immunosorbent assay. The specificity of the primers was verified using other unrelated viral RNAs. No PCR product was observed when Cucumber mosaic virus (Cucumovirus) or a crude extract of healthy apple was used as a template in RT-PCR with the same primers. The PCR product (669 bp) of the CP gene of the virus was cloned into the plasmid vector and result-ant recombinant (pAPCP1) was selected for molecule of apple transformation to breed virus-resistant transgenic apple plants as the next step. This method can be useful for early stage screening of in vitro plantlet and genetic resources of resistant cultivar of apple plants.

• The Plant Pathology Journal
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• 제27권3호
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• pp.291-296
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• 2011

In order to detect quarantine plant viruses, we developed reverse transcription-polymerase chain reaction (RT-PCR) primer pairs for five single-stranded (ss) plant RNA viruses that are not currently reported in Korea but could be potential harmful plant viral pathogens. Three viruses such as Chilli veinal mottle virus (ChiVMV), Colombian datura virus (CDV), and Tobacco etch virus (TEV) belong to the genus Potyvirus while Chrysanthemum stem necrosis virus (CSNV) and Iris yellow spot virus (IYSV) are members of the genus Tospovirus. To design RT-PCR primers, we used reported gene sequences corresponding to the capsid protein and polyprotein for ChiVMV, CDV, and TEV while using nucleocapsid protein regions for CSNV and IYSV. At least two different primer pairs were designed for each virus. Fifteen out of 16 primer pairs were successfully applied in detection of individual quarantine virus with high specificity and efficiency. Taken together, this study provides a rapid and useful protocol for detection of five quarantine viruses.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.144.2-145
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• 2003

This study was performed to investigate the incidence of Hosta uirus X (HVX), a Potexvirus, from cultivated hosta ornamental plants in Korea and to ascertain seed transmission of the virus from infected parent plant to progeny ones for breeding program of hosta plants. Infection rate of HVX in cultivated hostas was 25.6 ％ (11 out of 43 collected samples contained HVX) based on Western blot and RT-PCR detection methods. Most of HVX-infected hostas showed visible systemic leaf symptoms (mosaic, mottle, curling, stunting or combinations). Variability of HVX was confirmed by sequences of coat protein gene of individual isolates from different hostas. HVX was seed-transmitted on Hosta 'Blue Cadet'. The virus was detected from seeds, and sprouts and seedlings from the virus-contaminated seed sources. Over 7.5 ％ of seeds were HVX-contaminated surveyed in this study, Our data suggest that HVX can be transmitted by seed source, and indexing of the virus should be done for breeding program of Hosta.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.148.1-148
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• 2003

Natural virus infection of cultivated Daphe odora plants showing chlorosis and stunting was observed and their causal agent was investigated. An isolate of isometic virus was purified from infected leaf tissues, and it could infect systemic severe mosaic on Chenopodium quinoa and C. amaranticolor. cDNA library was generated from partially purified viral RNAs and oligo dT primer-pSPORTl system, and recombinant clones were selected and their inserts were sequenced randomly. Nucleotide sequences of the virus were analyzed by BLAST, and it was closely related to members of subgroup B in the genus Nepovirus. The sequence analysis suggest that the virus was identified as an isolate of Cycas necrotic stunt virus (CNSV) because it was 89.7 ％ and 94.7 ％ identical to known CNSV for the CP and 3' noncoding region, respecitively. RT-PCR was performed to screen disease incidence of CNSV in Daphe plants, and five out of 10 plants (50 ％) were infected by CNSV This is the first sequence information of CNSV from Daphe plants.

• 한국동물위생학회지
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• 제34권4호
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• pp.297-301
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• 2011

Rabies virus is transmitted most commonly through a bite from an infected dog. Especially, stray dogs that are not excluded from contact with rabid wild animals can become rabies vectors. Therefore, serological survey of rabies virus from stray dogs in Seoul Metropolitan City was carried out in this study. To investigate prevalence of rabies antibodies in the stray dogs, serum samples were taken from 500 stray dogs between April and December 2010. Antibodies to rabies virus were detected by indirect ELISA. Of 500 tested sera, 147 (29.4%) were positive to rabies virus. Prevalence rates of rabies antibodies(PRRA) in northern and southern Han river region of Seoul were 26.4% and 33.2% respectively. PRRA in male and female dogs were 33.6% and 26.1% respectively. PRRA in less than 1 year, 1~2< years, 2~3< years, 3~5< years, 5~10< years and over 10 years old dogs were 12.7%, 21.6%, 26.4%, 36.4%, 32.5% and 46.4%, respectively. These results indicates that antibody seroprevalence to rabies is still not enough to prevent rabies and rabies vaccination is required to enhance the seroprevalence in the dogs. To improve the situation, measures to raise public awareness of rabies and its prevention is needed. Also, reducing stray dogs and keeping companion animals from contact with wild animals are indispensable for the prevention of rabies.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2002년도 총회 및 추계학술발표회
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• pp.123.1-123
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• 2002

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2002년도 총회 및 추계학술발표회
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• pp.111.2-112
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• 2002