• 한국식물병리학회지
• /
• 제13권2호
• /
• pp.73-78
• /
• 1997

A virus was purified from Korean cactus (Chamaeaereus silverstrii f. Variegata Hort) showing necrotic spots and its biophysicochemical properties were examined. The typical necrotic spots were observed on healthy looking cactus inoculated with the purified virus and the various symptoms from necrotic spots to systemic were observed on 17 species of test plats used, which suggests that the purified virus has the normal infectivity and wide host range. Ultraviolet absorption scanning analysis revealed that the viral preparation had the typical curve with shoulder at 290 nm and its ratio of absorbances at 260 nm to 280 nm and extinction coefficient was 1.156 and 2.86, respectively. The virus represented one distinct and homogeneous band and molecular weight of viral coat protein was determined to 17, 000 Dalton by SDS-PAGE. The virus revealed rod-shaped virion (325$\times$20 nm) by electron microscopy. Based on these results, it has been thought that the virus purified in this study is the pathogenic agents causing necrotic spot symptom on Korean cacuts and one of the member of tobamovirus family.

• The Plant Pathology Journal
• /
• 제35권5호
• /
• pp.498-507
• /
• 2019

Odontoglossum ringspot virus (ORSV) is a member of the genus Tobamovirus. It is one of the most prevalent viruses infecting orchids worldwide. Earlier studies reported the genetic variability of ORSV isolates from Korea and China. However, the evolutionary rate, timescale, and phylogeographical analyses of ORSV were unclear. Twenty-one coat protein (CP) gene sequences of ORSV were determined in this study, and used them together with 145 CP sequences obtained from GenBank to infer the genetic diversities, evolutionary rate, timescale and migration of ORSV populations. Evolutionary rate of ORSV populations was $1.25{\times}10^{-3}nucleotides/site/y$. The most recent common ancestors came from 30 year ago (95% confidence intervals, 26-40). Based on CP gene, ORSV migrated from mainland China and South Korea to Taiwan island, Germany, Australia, Singapore, and Indonesia, and it also circulated within east Asia. Our study is the first attempt to evaluate the evolutionary rates, timescales and migration dynamics of ORSV.

• 한국자원식물학회:학술대회논문집
• /
• 한국자원식물학회 2022년도 추계학술대회
• /
• pp.68-68
• /
• 2022

The pepper mild mottle virus (PMMoV), belonging to the tobamovirus genus, is currently one of the most destructive pathogens in pepper production. Tobamoviruses have been classified in terms of increased pathogenicity as pathotypes P₀, P₁, P_1,2, P_1,2,3 and P_1,2,3,4, based on their ability to infect systemically Capsicum L⁰ , L¹ , L² , L³ and L⁴ resistant plants, respectively. Two hundred eighty pepper germplasms and 5 reference accessions known as resistant L alleles, were analyzed to select the resistance cultivars against PMMoV- P_1,2,3 (CV130614-2) using bioassay and genetic markers. The susceptible accessions showed systemic symptom when inoculated with PMMoV- P_1,2,3. However, accessions including IT223737, were resistant as they developed necrotic local lesions only on inoculated leaves, whereas no symptoms were observed on the upper leaves. Moreover, RT-PCR results for detecting the presence of virus were also negative. Thus, those accessions will be used as a novel source to facilitate introduction the resistant gene into commercial cultivars of pepper.

• The Plant Pathology Journal
• /
• 제21권2호
• /
• pp.167-171
• /
• 2005

We isolated the Cucumber green mottle mosaic virus(CGMMV) particles from watermelon leaves and designated as CGMMV-HY1 as a watermelon isolate and attempted to characterize the pathogenic isolate responsible for such an epidemic in watermelon and also to monitor dominant viral isolates in greenhouse. The watermelon plants infected with CGMMV generally showed mottle mosaic, mosaic, growth stunting, necrosis and deformed fruit. The reactions of indicator plants to CGMMV-HY1 were the local lesions on Nicotiana tabacum cv. White Burley, Nicotiana tabacum cv. Samsun, and Chenopodium amaranticola, and the mosaic symptoms only on Cucumis sativus, but the CGMMV-HY1 did not infect Nicotiana sylvesytis, Datura stramonium, Chenopodium quinoa, and Petunia hybrida. Purified virus particles were rod-shaped and about 300 nm long. The coat protein (CP) of purified CGMMV-HY1 was single band with molecular weight of about 16.5 kDa which was confirmed by western blot analysis probed with monoclonal antibody of CGMMV-HY1. The genomic and subgenomic RNAs of 6.4 kb and 0.75 kb were revealed by the electrophoresis on 1.2% formaldehydedenatured agarose gel. Viral and complementary CGMMV-specific primer sets were designed for spanning the genome using previously reported CGMMV sequences. A 464bp of CP gene of CGMMV-HY1 was amplified by RT-PCR and cloned into PGEM-T easy vector. The nucleotide sequence of CP gene of CGMMV-HY1 shared 98%, 99%, and 100% identities with that of CGMMV strains W, KOM, and KW respectively. Based on these results, we identified CGMMV-HY1 as a CGMMV isolate of watermelon, a member of Tobamovirus.

• 한국자원식물학회지
• /
• 제16권1호
• /
• pp.82-88
• /
• 2003

고추냉이에 모자이크 병징을 나타내는 이병주로부터 고추냉이 모자이크 바이러스를 분리하였다. 고추냉이 모자이크 바이러스의 genomic RNA를 추출하여 전체 유전자 구조를 결정하였다. 유전자 전체길이는 6,298 염기를 가지고 있었으며, 4개 ORF로 구성 되어 있었다. ORF 1은 180KD 단백질, ORF 2는 130KD 단백질 , ORF 3은 30KD 단백질, ORF4는 18KD로 외피단백질로 구성되어 있었다. ORF 유전자간에는 ORF4와 ORF 3 유전자간 130개의 염기, ORF 2와 ORF 3 유전자갈 20개 염기 그리고 ORF 1 과 ORF2 유전자간에는 40개의 염기로 overlaps되어 있었다 3'NCR부분은 238개 염기, 외피단백질은 537개 염기, 30KD 이동단백질은 825개 염기, 130KD 단백질은 1,896개 염기와 180k단백질의 2,958개의 염기로 구성되어 있었다. TMV-WTF전체 염기 서열의 유전자 상동성에서는 비교 유전자에서 미보고된 일본의 TMV-WSF와 러시아의 TMV-crucifer와 각각 98.6%와 82.4%로 매우 높았다.

• The Plant Pathology Journal
• /
• 제22권4호
• /
• pp.353-359
• /
• 2006

Transgenic Nicotiana benthamiana plants harboring the coat protein(CP) gene of Zucchini green mottle mosaic virus(ZGMMV) were chosen as a model host for the environmental risk assessment of genetically modified plants with virus resistance. This study was focused on whether new virus type may arise during serial inoculation of one point CP mutant of ZGMMV on the transgenic plants. In vitro transcripts derived from the non-functional CP mutant were inoculated onto the virus-tolerant and -susceptible transgenic N. benthamiana plants. Any notable viral symptoms that could arise on the inoculated transgenic host plants were not detected, even though the inoculation experiment was repeated a total of ten times. This result suggests that potential risk associated with the CP-expressiing transgenic plants may not be significant. However, cautions must be taken as it does not guarantee environmental safety of these CP-mediated virus-resistant plants, considering the limited number of the transgenic plants tested in this study. Further study at a larger scale is needed to evaluate the environmental risk that might be associated with the CP-mediated virus resistant plant.

• 한국식물병리학회지
• /
• 제10권3호
• /
• pp.228-234
• /
• 1994

Genomic RNA was extracted from Cy strain of odontoglossum ringspot tobamovirus (ORSV-Cy) isolated from infected leaves of tobacco cv. Samsun. Size of the genomic RNA was about 6.6 kb in length. The genomic RNA was fractionated using Sephadex G-50 column chromatography into 2 fractions. They were polyadenylated at their 3'-end using E. coli poly(A) polymerase. Polyadenylated viral RNA was recovered by oligo (dT) primer adapter containing NotI restriction site and Moloney murine leukemia virus SuperScript reverse transcriptase (RNase H-). Second-strand cDNA was synthesized by using E. coli DNA ligase, E. coli DNA polymerase I and E. coli RNase H. Recombinant plasmids containing cDNAs for ORSV-Cy RNA ranged from about 800 bp to 3,000 bp. Among the selected 238 recombinants, pORCY-124 clone was the largest one covering 3'-terminal half of the viral RNA. This clone contained two restriction sites for EcoRI and XbaI and one site for AccI, AvaI, BglII, BstXI, HindIII, PstI, and TthIII 1. respectively. The clone contained partial viral replicase, a full-length movement protein and a complete coat protein genes followed by a 3' untranslated region of 414 nucleotides based on restriction mapping and nucleotide sequencing analyses. Clones pORCY-028, -068, -072, -187 and -224 were overlapped with the pORCY-124. Clones pORCY-014 and -095 covered 5' half upstream from the middle region of the viral RNA, which was estimated based on restriction mapping and partial sequence analysis. Constructed cDNA library covered more than 90% of the viral genome.

• 식물병연구
• /
• 제7권2호
• /
• pp.80-85
• /
• 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.

• 식물병연구
• /
• 제13권2호
• /
• pp.82-87
• /
• 2007

박과 작물에 발생하는 종자전염성 막대형 바이러스인 CGMMV, KGMMV, ZGMMV 3종의 단독 및 동시 VC/RT-PCR 유전자 진단법을 개발하였다. CGMMV 등 3종의 바이러스에 대한 동시진단용 조합선발을 위하여 12조합 중에서 동시 진단용 프라이머 조합 CGMMV-C724, KGMMV-K513, ZGMMV-Z407A를 선발하였다. CGMMV등 3종에 대한 triplex VC/RT-PCR 유전자 진단법은 박, 수박, 오이, 멜론, 쥬키니 호박, 애호박, 담배(N. benthamiana) 작물의 식물체 즙액과 프라이머 간에 간섭현상 없이 특이적으로 진단되었다.

• The Plant Pathology Journal
• /
• 제29권3호
• /
• pp.260-273
• /
• 2013

The incidence and distribution of Tobacco mosaic virus (TMV) and related tobamoviruses was determined using an enzyme-linked immunosorbent assay on 1,926 symptomatic horticultural crops and 107 asymptomatic weed samples collected from 78 highly infected fields in the major horticultural crop-producing areas in 17 provinces throughout Iran. The results were confirmed by host range studies and reverse transcription-polymerase chain reaction. The overall incidence of infection by these viruses in symptomatic plants was 11.3%. The coat protein (CP) gene sequences of a number of isolates were determined and disclosed to be a high identity (up to 100%) among the Iranian isolates. Phylogenetic analysis of all known TMV CP genes showed three clades on the basis of nucleotide sequences with all Iranian isolates distinctly clustered in clade II. Analysis using the complete CP amino acid sequence showed one clade with two subgroups, IA and IB, with Iranian isolates in both subgroups. The nucleotide diversity within each subgroup was very low, but higher between the two clades. No correlation was found between genetic distance and geographical origin or host species of isolation. Statistical analyses suggested a negative selection and demonstrated the occurrence of gene flow from the isolates in other clades to the Iranian population.