• The Plant Pathology Journal
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• 제16권1호
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• pp.43-47
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• 2000

Citrus tristeza virus (CTV) is the causal agent of one of the most important diseases of citrus. Recently, CTV has been detected in Cheju Island by ELISA. The coat protein (CP) gene of CTV isolated form Cheju Island was cloned by RT-PCR and the nucleotide was analyzed in this study. Citrus leaves were collected from trees showing decline symptoms from various region of Cheju Island in the summer of 1998 and 1999. The CP gene open reading frame is composed of 670 nucleotides and encodes a polypeptide of 223 amono acids. Sequence analysis the CP gene revealed that two CTV strains present in Cheju Island. Viruses collected form Sogwipo area and Cheju City area in 1999 ahowed 91-93% nucleotide sequence homology with CTV T36 strain. Viruses collected form Cheju City area in 1999 and Sogwipo City in 1998 showed 94-98% nucleotide sequence homology with CTV SY568 strain. A efficient viral RNA extraction methods was developed by modifying procedure for animal virus RNA purification methods and PCR product was detected form one tenth of RNA purified from as small as 45 mg fresh or frozen tissue.

• The Plant Pathology Journal
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• 제25권2호
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• pp.157-164
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• 2009

The complete nucleotide sequences of both RNA of an isolated Barley mild mosaic virus (BaMMV) from Iksan, Korea, have been determined. RNA1 was 7273 nucleotides long and encodes for a polyprotein of 2261 amino acids, which contains the eight putative functional proteins. RNA2 was 3520 nucleotides long and encodes for a polyprotein of 894 amino acids, which contains two functional proteins. Results of multiple sequence alignment showed 92.9% similarity with Na1 isolate, followed by Sil, UK(F), Asl1, Remis M and UK(M) isolates, respectively. Comparison of the BaMMV-Iks polyproteins with the corresponding proteins of BaMMV-Na1 isolates showed 95% amino acid sequence identity. The phylogenetic analysis revealed that Iks isolate was closely related to Na1 strain and have a common origin.

• 한국식물병리학회지
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• 제14권1호
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• pp.68-73
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• 1998

Barley mild mosaic virus (BaMMV-Kor) was isolated from the southern part of Korea, and by mechanical inoculation onto barley cultivars, purification and production of antibody. BaMMV-Kor purified form infected plants were filamentous particle, with 13 nm in diameter and 250∼300 nm and 500∼650 nm in length. Antibody of BaMMV-Kor was made by injecting the purified virus to the muscle of a rabbit. In gel-diffusion test, antibody to BaMMV-Kor created spur with BaMMV-Kal and BaMMV-M, but did not make spur with BaMMV-Kor infected New Golden, Ishukushirazu, Joshushiro Hadaka and Misato Golden, but did not infect Kashimamugi, Chikurin Ibaraki 1 and Mokusekko 3. In Korean barley cultivars, BaMMV-Kor infected most of the covered barley cultivars, but did not infect Saeolbori. It also infected naked barley cultivars except Chalbori and Hinssalbori. And all the beer barley cultivars were infected by BaMMV-Kor. BaMMV-Kor had two RNAs, RNA 1 (7.5 Kb) and RNA 2 (3.5 Kb), and coat protein (33 KDa).

• 식물병연구
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• 제23권1호
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• pp.65-68
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• 2017

2단계의 nested PCR 방법을 이용하여 보리 및 벼 재배 토양에서 Barley yellow mosaic virus (BaYMV)를 검출하였다. BaYMV 분절 RNA1 외피단백질 영역의 특이 프라이머로 1차 PCR을 하고 내부서열로부터 작성된 프라이머로 2차 PCR을 실시하여 확보된 372 bp의 PCR 산물이 BaYMV 외피단백질 영역과 98%-100% 염기서열이 일치하여 BaYMV를 검출할 수 있음을 확인하였다. 이 결과는 토양으로부터 BaYMV 검출에 관한 최초의 보고이며 토양전염성 바이러스의 정확한 진단과 예찰에 적용될 수 있을 것으로 생각한다.

• 식물병연구
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• 제23권2호
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• pp.193-201
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• 2017

국내의 토마토에서 발생하는 주요 바이러스는 Tomato chlorosis virus (ToCV), Tomato spotted wilt virus (TSWV), Cucumber mosaic virus (CMV), Pepper mottle virus (PepMoV), Tomato mosaic virus (ToMV)이다. 이들 바이러스를 진단하기 위해 프라이머 세트와 반응액을 포함하는 역전사중합반응(RT-PCR)법의 조건을 조사하였다. 공시한 바이러스에 특이적인 염기서열로부터 모두 46개 프라이머 세트를 설계하고, 이를 이용해 주형을 넣지 않은 RT-PCR에서 비특이 반응을 조사하였다. 이들 가운데 16개 조합을 건전한 토마토 RNA에 적용한 결과 프라이머 세트와 RT-PCR 반응액 간의 친화성이 비특이 반응 감소에 영향을 주었다. cDNA 합성과 관련된 인자와 RT-PCR 반응액 사이의 조합을 근거로 ToCV 진단을 위한 두 종류의 반응액을 선발하였다. ToCV 진단 시 수립된 조건을 나머지 바이러스 진단에 적용했을 때, 특이성이 높은 프라이머 세트 C029 (ToCV), C072 (TSWV), C070 (CMV), C048 (PepMoV), C065 (ToMV)를 선발할 수 있었다. 이들 프라이머 세트는 공시한 바이러스를 특이적으로 진단하는 데 유용할 것으로 판단된다.

• BMB Reports
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• 제32권1호
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• pp.82-85
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• 1999

The movement protein of cucumber mosaic virus (CMV) is required for cell-to-cell movement of viral RNA. The movement of viral RNA occurs through the plant intercellular connection, the plasmodesmata. The viral movement protein was known to be multi-functional. In this work, a series of deletion mutants of CMV movement protein gene were created to identify the functional domains. The mutated movement proteins were produced as inclusion body in E. coli, and purified and renatured. A polyclonal antibody was raised against the CMV-Kor strain (Korean isolate) movement protein expressed in E. coli. The ability of the truncated proteins to bind to ssRNA was assayed by UV cross-linking and gel retardation analyses. The results indicate that the domain between amino acids 118 and 160 of CMV movement protein is essential for ssRNA binding.

• The Plant Pathology Journal
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• 제18권6호
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• pp.313-316
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• 2002

Systemic virus symptoms caused by a Potexvirus were observed on leaves of infected hosta (Hasta spp.) plants cultivated in Seoul, Korea. Symptoms on diseased hosta plants include mosaic, mottle, irregular blotchy patches, and chlorotic spots on or distortion of the leaves. No other viruses, such as Cucumber mosaic virus, Lily symptomless virus, or Potyvirus, were detected from the same plants by electron microscopy and by Western blot and RT-PCR analyses, indicating that they were singly infected by the potexvirus. The symptoms differed among cultivars and species of hosta, and affected the quality of plants for commercialization, as well as, plant growth and flowering of susceptible cultivars. Most of the cultivars and species investigated were susceptible to the virus, while some were not infected by the virus at all. Purified virus particles were of filamentous type with unaggregated forms 540 nm in length, which is a typical potexviral morphology. The virus consisted of a single-stranded RNA molecule of 6 kb long for genome and single component of coat protein (CP) about 27 kDa. The CP strongly reacted with the antiserum against Hosta vims X (HVX), suggesting that the virus is an isolate of HVX. This is the first report of the occurrence and identification of HVX from hosta plants in Korea.

• Journal of Plant Biotechnology
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• 제47권2호
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• pp.164-171
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• 2020

Transgenic Alstroemeria plants resistant to Alstroemeria mosaic virus (AlMV) were generated through RNA-mediated resistance. To this end, the friable embryogenic callus (FEC) of Alstroemeria was induced from the leaf axil tissue and transformed with a DNA fragment containing the coat protein gene and 3'-nontranslated region of AlMV through an improved particle bombardment system. The bar gene was used as a selection marker. More than 300 independent transgenic FEC lines were obtained. Among these, 155 lines resistant to phosphinothricin (PPT) were selected under low stringent conditions. After increasing the stringency of PPT selection, 44 transgenic lines remained, and 710 somatic embryos from these lines germinated and developed into shoots. These transgenic shoots were then transferred to the greenhouse and challenged with AlMV. In total, 25 of the 44 lines showed some degree of resistance. PCR analysis confirmed the presence of the viral sequence. Virus resistance was observed at various levels. Establishment of an efficient transformation system for Alstroemeria will allow inserting transgenes into this plant to confer resistance to viral and fungal pathogens. Accordingly, this is the first report on the production of a transgenic virus-resistant Alstroemeria and lays the foundation for alternative management of viral diseases in this plant.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.113.1-113
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• 2003

A potexvirus, Hosta virus X (HVX-Kr), causing mosaic and mottle symptoms was isolated from hosta plants (Hosta spp.), and its entire genome RNA sequence was determined. in Korea using cDNA library and RACE methods. The genome of HVX encodes five open reading frames coding for viral replicase, triple gene block (TGB), and viral coat protein (CP) from the 5'to 3' ends, which is a typical genome structure of potexviruses. The 3-terminal region of the virus includes the TGBI (26 kDa), TGB2 (13 kDa), TGB3 (8 kDa), and 23 kDa coat protein (CP) and the 3-nontranslated region (NTR). The CP gene of the type isolate of HVX (HVX-U) was amplified by RT-PCR and its nucleotide sequence was determined. The CPs of HVX-Kr and HVX-U had 100％ and 98.9％ identical amino acids and nucleotides, respectively. Most of the regions of the genome HVX had over 50％ nucleotide identical to other sequenced potexviruses. This is the first report of complete genome sequence information of HVX and molecular evidence supporting the virus as a distinct species of the genus Potexvirus.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.115.2-116
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• 2003

Complete genomic nucleotide sequence of Daphe virus S (DVS), a member of the genus Carlavirus, causing leaf distortion and chlorotic spot disease symptoms in daphne plants, has been determined in this study. The genome of DVS contained six open reading fames coding for long viral replicase, triple gene block, 36 kDa viral coat protein (CP) and 12 kDa from the 5' to 3' ends, which is a typical genome structure of carlaviruses. Two Korean isolates of DVS isolates were 98.1％ and 93.6％ amino acid identical in the CP and 12kDa, respectively. The CP gene of DVS shares 25.2-55.2％ and 42.9-56.1％ similarities with that of 19 other carlaviruses at the amino acid and nucleotide levels, respectively. The 3'-proximal 12 kDa gene of DVS shares 20.2-57.8％ amino acid identities with that of 18 other members of the genus. The 3' noncoding region of DVS consists of 73 nucleotides with long excluding poly A tract, and shares 69.1-77.1％ identities to the known carlaviruses. In the phylogenetic analyses of the two proteins, DVS was closely related to Helenium virus S and Chrysanthemum virus B. This is the first complete sequence information for the DVS, and further confirms the classification of DVS as a distinct species of the genus Carlavirus.