• Journal of Microbiology and Biotechnology
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• 제22권12호
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• pp.1776-1781
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• 2012

Multiple viruses such as Lily symptomless virus (LSV), Lily mottle virus (LMoV), and cucumber mosaic virus (CMV) are the most prevalent viruses infecting lilies in Korea. Leaf samples and bulbs showing characteristic symptoms of virus infection were collected from Gangwon, Chungnam, and Jeju provinces of Korea in 2008-2011. Coat protein (CP) genes of LSV and LMoV were amplified from collected samples by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into a pET21d(+) expression vector to generate recombinant CPs. The resulting carboxy-terminal His-tagged CPs were expressed in Escherichia coli strain BL21(DE3) by isopropyl-1-thio-${\beta}$-D-galactoside induction. The recombinant proteins were purified using Ni-NTA agarose beads, and the purified proteins were used as an immunogen to produce polyclonal antibodies in rabbits. The resulting polyclonal antisera recognized specifically LSV and LMoV from infected plant tissues in Western blotting assays. Indirect enzymelinked immunosorbent assay and immunocapture RT-PCR using these polyclonal antisera were developed for the sensitive, efficient, economic, and rapid detection of Lily viruses. These results suggest that large-scale bulb tests and economic detection of Lily viruses in epidemiological studies can be performed routinely using these polyclonal antisera.

• 한국동물위생학회지
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• 제44권1호
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• pp.15-19
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• 2021

Senecavirus A (SVA), previously known as Seneca Valley virus, can cause vesicular disease and neonatal losses in pigs that is clinically indistinguishable from foot-and-mouth disease virus (FMDV). After the first case report in Canada in 2007, it had been restrictively identified in North America including United States. But, since 2015, SVA emerged outside North America in Brazil, and also in several the Asian countries including China, Thailand, and Vietnam. Considering the SVA occurrence in neighboring countries, there has been a high risk that Korea can be introduced at any time. In particular, it is very important in terms of differential diagnosis in the suspected case of vesicular diseases in countries where FMD is occurring. So far, several different molecular detection methods for SVV have been published but not validated as the reference method, yet. In this study, seven different molecular methods for detecting SVA were evaluated. Among them, the method by Flowler et al, (2017) targeted to 3D gene region with the highest sensitivity and no cross reaction with other vesicular disease agents including FMDV, VSV and SVD, was selected and applied further to antigen surveillance of SVA. A total of 245 samples of 157 pigs from 61 farms submitted for animal disease diagnose nationwide during 2018 were tested all negative. In 2018, no sign of SVA occurrence have been confirmed in Korea, but the results of the surveillance for SVA needs to be continued and accumulated at a high risk of SVA in neighboring countries.

• 한국응용곤충학회지
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• 제16권1호
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• pp.65-67
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• 1977

우리나라 중부이남 지역에서 벼 오갈병에 의한 피가 현저히 늘어나고 있는 실정이다. 본시험에서는 매개충을 사용, 순수분리하고 이를 접종하여 증식한 후 Toyoda 등의 순화방법을 개선하여 순화하였다. 그 결과 순화된 바이러스 함량은 ml당 3.12mg 이었다. 순화된 바이러스를 Adjuvant와 함께 토끼에 10-14일 간격으로 5회 주사하여 항혈청을 제조한 결과 1/4,096의 높은 역가를 나타내는 항혈청이 제조되었다.

• Journal of Plant Biotechnology
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• 제47권2호
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• pp.150-156
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• 2020

Weeds play an important role in the survival of viruses and are potential inoculum sources of viral diseases for crop plants. In this study, specimens of Pseudostellaria heterophylla exhibiting symptoms of the cucumber mosaic virus (CMV) were collected in Bonghwa, Korea. The characteristics of the disease were described and leaf RNA was extracted and sequenced to identify the virus. Three CMV contigs were obtained and PCR was performed using specific primer pairs. RNA from positive samples exhibiting CMV leaf symptoms was amplified to determine the coat protein. A sequence comparison of the coat protein gene from the CMV BH isolate shared the highest nucleotide identity (99.2%) with the CMV ZM isolate. Phylogenetic analysis showed that CMV-BH belonged to subgroup IA and that the most closely-related isolate was CMV-ZM. All test plants used for the biological assay were successfully infected with CMV and exhibited CMV disease symptoms such as blistering, mosaic, and vein yellowing. To our knowledge, this is the first report of CMV infection in P. heterophylla from Korea.

• 한국식물병리학회지
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• 제12권2호
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• pp.187-190
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• 1996

백합으로부터 total RNA를 분리하여 LSV 외피단백질 유전자의 551 bp에 해당하는 특정 염기서열을 증폭할 수 있는 primer로 RT-PCR를 수행하였다. 그 결과 Lilium oriental hybrid cvs. Miani, Marco Polo, Casablanca, Le Reve 품종에서 551 bp의 DNA 절편이 증폭되었고 이 절편의 염기서열을 분석한 결과 LSV외피단백질 유전자의 일부임을 확인할 수 있었다. 그러므로 RT-PCR 방법으로, 실험에 사용하 s4품종 모두 LSV에 감염되어 있음을 알 수 있었다.

• 농업과학연구
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• 제42권2호
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• pp.105-109
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• 2015

Andean potato latent virus (APLV)는 Group IV (+) sense ssRNA viruses, Tymovirus속으로 분류되는 식물 병원성 바이러스로, 주로 감자에 감염되며 국내 관리급 검역바이러스로 지정되어 있다. 본 연구에서는 검역현장에서 APLV를 신속 정확하게 진단할 수 있는 2개의 RT-PCR 프라이머 조합[조합 2 (404 bp)와 23 (501 bp)]과 각각의 nested PCR 조합($404{\rightarrow}259bp$ 및 $501{\rightarrow}349bp$)을 선발하였다. 또한, 진단에 양성대조군으로 활용할 수 있는 유전자변형 양성대조구를 개발하여 실험실 오염에 대한 검증이 가능하도록 하였다. 본 연구에서 개발한 PCR 기반 진단시스템은 향후 검역 현장에서 APLV를 검출하여 우리나라 식물검역에 기여할 것이라고 기대된다.

• 한국작물학회지
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• 제46권1호
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• pp.53-56
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• 2001

Barley Yellow Dwarf Virus (BYDV), an aphid-borne luteovirus, is a major plant pathogenic disease causing a huge economic loss in the grain production of a wide range of Gramineae species throughout the world. It has been recently reported that BYDV also occurred frequently in wheat field of Korea. Here, we performed to develop the detection and classification methods of BYDV strains that were accomplished by reverse transcription-polymerase chain reaction (RT-PCR). Since there are high variations among BYDV strains, three pairs of primers were designed to detect BYDV strains such as PAV (Vic-PAV and CN-PAV) and MAV (primer A) simultaneously, specifically Vic-PAV(primer B), and MAV (primer C) based on the genomic RNA sequences of BYDV strains previously published. The validity of the primers was confirmed using several BYDV strains obtained from CIMMYT. Though three BYDV strains were able to be detected using primer A, PCR products were not distinguished between two PAV strains. It was possible to separate them with a restriction enzyme, EcoRI, whose restriction site was present in the amplified DNA fragment from Vic-PAV, but not from CN-PAV.

• The Plant Pathology Journal
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• 제17권6호
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• pp.392-392
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• 2001

• 식물병연구
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• 제24권1호
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• pp.81-85
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• 2018

2016년 충북 충주에서 모자이크 병징을 보이는 초석잠을 채집하였다. 담배와 명아주에 즙액접종을 실시한 결과 담배에서 접종엽에 괴사반점이 나타나고 명아주 2종에서 상엽에 퇴록반점을 동반한 모자이크 병징이 관찰되었다. 기주반응과 결과를 통해 여러 바이러스의 복합감염을 추정할 수 있었고, 추정되는 바이러스의 확인을 위해 속특이 및 종특이 프라이머를 이용하여 RT-PCR을 수행하여 3종의 바이러스(AMV, CMV, TMV)를 검정하였다. 본 연구는 국내 초석잠에서 3종의 바이러스 감염을 확인하였고 CMV의 감염에 대한 첫 보고이다.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2000년도 추계 학술발표회
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• pp.63.2-63
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• 2000