• 식물병연구
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• 제7권2호
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• pp.80-85
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• 2001

이 연구는 고랭지 나리에서 발생하는 바이러스의 병징, 종류 및 계통별 방병률을 조사 분석하고 효과적인 검정방법을 개발하고자 수행하였다. 고랭지 나리에서 발생하는 바이러스의 병징은 모자이크, 축엽, 퇴록반점, 줄무의, 라인패턴을 나타내었으며, 증상별 분포는 모자이크가 43.8%, 축엽이 29.2%, 퇴록반점이 10.9%였다. 바이러스 종류별로는 Lily symptomless virus(LSV), Cucumber mosaic virus(CMV), Lily mottle virus(LMoV) 등 6가지 바이러스가 전자현미경으로 검정되었다. 지역별로는 강릉(왕산)이 대관령보다 바이러스 이병률이 높았으며, 계통별 바이러스 이병률은 오리엔탈 계통(카사블랑카, 마르코폴로)이 아시아틱 계통(솔레미오, 플라토)보다 2~4배 높았다. 바이러스 진단방법으로는 기존의 PT-PCR보다 개선된 one-step RT-PCR 검정이 시간을 줄이면서 민감도가 뛰어나 가장 효과적이었다.

• The Plant Pathology Journal
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• 제25권3호
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• pp.225-230
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• 2009

Lily mottle virus (LMoV) causes flower quality reduction in Lilium spp. The coat protein gene was RT-PCR-amplified from total RNA extracted from infected lily leaves and the amplified fragment was cloned into the pRSET expression vector tagged with a His-MBP. The plasmid of recombinant coat protein was used to transform an Escherichia coli strain pLysS and was expressed. The coat protein was purified by affinity chromatography using a Ni-NTA resin. The identity of the purified protein was confirmed by SDS-PAGE. The in vitro-expressed protein was used for immunization of mice. The polyclonal and monoclonal antibodies reacted specifically for the detection of LMoV in lily extracts in Western blot. Moreover the monoclonal antibodies reacted with lily extracts in DAS-ELISA with no unspecific or heterologous reactions against other non-serologically related viruses, but the polyclonal antibodies revealed a weak reaction against both infected lily and healthy control.

• The Plant Pathology Journal
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• 제36권2호
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• pp.170-178
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• 2020

The Lily mottle virus (LMoV) impedes the growth and quality of lily crops in Lanzhou, China. Recently Arabis mosaic virus (ArMV) has been detected in LMoV-infected plants in this region, causing plant stunting as well as severe foliar symptoms, and likely posing a threat to lily production. Consequently, there is a need to develop simple, sensitive, and reliable detection methods for these two viruses to prevent them from spreading. Reverse transcription (RT) loop-mediated isothermal amplification (LAMP) assays have been developed to detect LMoV and ArMV using two primer pairs that match six conserved sequences of LMoV and ArMV coat proteins, respectively. RT-LAMP assay results were visually assessed in reaction tubes using green fluorescence and gel electrophoresis. Our assays successfully detected both LMoV and ArMV in lily plants without the occurrence of viral cross-reactivity from other lily viruses. Optimal conditions for LAMP reactions were 65℃ and 60℃ for 60 min for LMoV and ArMV, respectively. Detection sensitivity for both RT-LAMP assays was a hundredfold greater than that of our comparative RT-polymerase chain reaction assays. We have also found this relatively rapid, target specific and sensitive method can also be used for samples collected in the field and may be especially useful in regions with limited or no laboratory facilities.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2002년도 총회 및 추계학술발표회
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• pp.118.2-119
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• 2002

• 원예과학기술지
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• 제32권4호
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• pp.544-549
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• 2014

Lily symptomless virus (LSV), Lily mottle virus (LMoV), and Cucumber mosaic virus (CMV) are the most prevalent viruses infecting lilies in Korea. Leaf and bulb samples showing characteristic symptoms of virus infection were collected in 2012, and 80 field samples were analyzed by reverse transcription polymerase chain reaction (RT-PCR). The infection frequencies were 79% for LMoV, 5% for LSV, and 3% for CMV. The LMoV coat protein gene was amplified and cloned into the pET21d(+) expression vector to develop serological diagnostic tools to detect LMoV. The resulting carboxy-terminal His-tagged coat proteins were expressed in Escherichia coli strain BL21 (DE3) by induction with IPTG. The recombinant proteins were purified using Ni-NTA agarose beads and used as an antigen to produce polyclonal antibodies in laying hens. The resulting egg yolk immunoglobulin (IgY) specifically recognized LMoV from infected plant tissues in immunoblotting assays and had comparable sensitivity to that of a mammalian antibody. In addition, method of immunocapture RT-PCR using this IgY was developed for sensitive, efficient, and rapid detection of LMoV. Based on these results, large-scale bulb tests and detection of LMoV in epidemiological studies can be performed routinely using this IgY. This is the first report of production of a polyclonal IgY against a plant virus and its use for diagnosis.

• The Plant Pathology Journal
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• 제29권3호
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• pp.338-343
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• 2013

A detection system based on a multiplex reverse transcription (RT) polymerase chain reaction (PCR) was developed to simultaneously identify multiple viruses in the lily plant. The most common viruses infecting lily plants are the cucumber mosaic virus (CMV), lily mottle virus (LMoV), lily symptomless virus (LSV). Leaf samples were collected at lily-cultivation facilities located in the Kangwon province of Korea and used to evaluate the detection system. Simplex and multiplex RT-PCR were performed using virus-specific primers to detect single- or mixed viral infections in lily plants. Our results demonstrate the selective detection of 3 different viruses (CMV, LMoV and LSV) by using specific primers as well as the potential of simultaneously detecting 2 or 3 different viruses in lily plants with mixed infections. Three sets of primers for each target virus, and one set of internal control primers were used to evaluate the detection system for efficiency, reliability, and reproducibility.

• 식물병연구
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• 제9권4호
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• pp.237-241
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• 2003

신나팔나리(Lilium x fomolongi) 뇌산 품종 실생묘에 Cucumber mosaic virus(CMV), Lily mottle virus(LMoV) 및 Lily symptomless virus(LSV)를 인공적으로 접종하여 생육에 미치는 영향을 조사하였다. 또한 재배포장에서 자연감염된 바이러스 병징과 감염율을 조사하였다. 재배포장에서 가장 많이 관찰되는 병징은 LMoV 감염에 의한 잎 모틀 증상이었다. 바이러스 감염율은 6년 이상 재 사용된 구근으로 재배하는 포장의 경우 CMV, LSV, LMoV 바이러스 가운데 두 종류 이상의 바이러스에 의한 복합 감염율은 80% 가량이었다. 신나팔나리 (Lilium x fomolongi) 품종 뇌산에 CMV-Li1과 LMoV-Li 복합감염시 건전주에 비해 초장 14%, 생체중 38%, 꽃 길이가 15% 감소하였다. CMV-Li1과 LMoV-Li 단독감염시에도 생체중은 건전주에 비해 각각 21.8%, 28.4%의 유의성 있는 감소를 보였다.

• 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.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.107.2-108
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• 2003

Viral disease symptoms were investigated in the field grown Longiflorum hybrid cultivars, and the damages caused by infection with Lily mottle virus (LMoV) and Cucumber mosaic virus (CMV) were assessed by comparing growth of plants produced from seeds of Longiflorum hybrid cultivar both infected by artificial inoculation and free from infection with theses viruses. Dominant symptom caused by spotaneous infection with LMoV and CMV in the field was mottle combined with chlorotic stripe on leaves. LMoV developed brownish necrotic lesion on floral leaves. The incidence of viral disease by mixed infection with LMoV, CMV or Lily symptomless virus (LSV) in the filed grown Longiflorum hybrid cultivar, cultivated for more than 6 years, was 80 to 84 percent. In comparison with virus-free plants, plants doubly infected with CMV and LMoV by artificial inoculation decreased stem length by 14 percent and fresh weight by 38 percent. In conclusion, flower quality and the stem length of Longiflorum hybrid cultivar were affected by LMoV and CMV infection.

• 미생물학회지
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• 제45권3호
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• pp.251-256
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• 2009

2008~2009년에 강원, 충남, 제주 지역의 백합 재배 농가에서 바이러스 감염 증상을 보이는 백합의 잎과 구근을 채취하였으며 RT-PCR 방법으로 Lily mottle virus (LMoV), Lily symptomless virus (LSV), Cucumber mosaic virus (CMV) 등 3 종류의 바이러스를 검출 하였다. LSV 12주, LMoV 20주, CMV 1주가 검출 되었으며 LSV에 감염된 12개의 식물체 중 7개에서는 LMoV도 검출되어 복합 감염된 것을 확인하였다. LMoV와 LSV에 의한 복합 감염은 엽맥투명화, 잎말림, 반점, 모자이크, 황화 줄무늬 등 단독 감염보다 심각한 병징을 나타내었으며 좋지 않은 환경에서 저장된 구근에서도 복합 감염이 관찰되었다. 채집된 식물체는 LMoV 감염이 가장 많았으며 Lily virus X(LVX)에 의한 감염은 검출되지 않았다. 7개 분리주의(LMoV 4주, LSV 2주, CMV 1주) 외피 단백질 유전자를 증폭한 후 염기서열을 분석하여 국내에서 발표된 백합 바이러스 염기서열(LSV:AJ516059, CMV: AJ296154)과 비교 하였으며 LMoV는 국내에서 처음으로 염기서열을 결정하여 기존에 보고된 염기서열(AJ564636)과 비교하였다. 분리주는 기존에 보고된 바이러스와 95~99%의 뉴클레오티드 염기서열 유사성을 보였으며, 이들 분리주의 분자 생물학적 특성을 밝히고 신속하고 정확한 바이러스 진단을 위해서는 전체 염기 서열 분석이 필요한 것으로 판단되었다.