• Current Research on Agriculture and Life Sciences
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• 제9권
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• pp.115-125
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• 1991

우리나라와 일본(日本)에서 야생 또는 재배되고 있는 약용식물(藥用植物)의 바이러스병을 조사한 결과 33종의 식물이 자연상태에서 오이 모자이크 바이러스(CMV)에 감염되어 있음을 알았다. 이들 중에서 개맨드라미(Celosia argenteia)와 쇠비름(Portulaca oleracea)의 모자이크병(가칭(假稱)), 쥐방울덩굴(Aristolochia debilis)과 번행초(Tetragonia expansa)의 괴저(壞疽)모자이크병(가칭(假稱)), basella(Basella rubra)윤문병(輪紋病)(가칭(假稱)), 석결명(Cassia torosa)과 시호(Bupleurum falcatum), 당귀(Angelica acutiloba), 구릿대(A. keiskei), 회향(Foeniculum vulgare), peucedanum(Peucedanum japanicum)의 반문병(斑紋病)(가칭(假稱))등 11종의 바이러스병명(病名)을 새로히 명명(命名)하였다.

• 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.

• 한국식물병리학회지
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• 제14권6호
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• pp.630-635
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• 1998

Tobacco mosaic tobamovirus (TMV), cucumber mosaic cucumovirus (CMV), cucumber green mottle mosaic tobamovirus (CGMMV) and zucchini yellow mosaic potyvirus (ZYMV) from individual fruits and seeds of hot pepper and cucumber were detected by the reverse transcription-polymerase chain reaction (RT-PCR). The dilution end-points for RT-PCR in curde sap from TMV. and CMV - infected hot pepper leaves and CMV - and CGMMV-infected cucumber leaves were 10-5. However, the amount of PCR product obtained from preparation of ZYMV-infected cucumber leaf was 10-fold lower than those of CMV or CGMMV-infected cucumber leaves. In hot pepper, both TMV and CMV were detected in all parts of the fruit wall tissue, but the yields of PCR products in the fruit stalk and its surrounding tissues were higher than those of the end parts of the fruit. On the other hand, in cucumber fruit infected with CMV, CGMMV or ZYMV, the fruit wall tissue and seed located in both stalk and end parts showed higher yields of PCR products than those of intermediate parts. Of five viruses that were analysed, only TMV in hot pepper seed, and CGMMV and CMV in cucumber seed were detected in testa parts.

• The Plant Pathology Journal
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• 제20권3호
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• pp.212-219
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• 2004

Three isolates of Cucumber mosaic virus (CMV) from lily plants showing mosaic and distortion symptoms were detected by reverse-transcriptase polymerase chain reaction (RT-PCR) using primers specific to Cucumovirus genus namely, LK-CMV, LK4-CMV, and LKS-CMV. Restriction enzymes patterns of the RT-PCR products revealed that the lily isolates belonged to subgroup IA of CMV. In terms of biological properties, the lily isolates have highly similar but distinct pathogenicity as reported in other lily strains and ordinary strains of CMV. To characterize the molecular properties, cDNAs containing coat protein (CP) gene and 3' non-coding region (NCR) of RNA3 for the isolates were cloned and their nucleotide sequences were determined. The CP similarity (218 amino acids) was highly homologous (>97％) with that of subgroup I CMV strains. However, an additional 20-nulcleotide long segment was only present in 3' NCR of lily isolates, which form an additional stem-loop RNA structure. By using chimeric construct exchange cDNA containing 3'NCR of LK-CMV into the full-length cDNA clone of RNA3 of Fny-CMV, this additional segment may prove to be significant in the identification and fitness of the virus in lily plants. The pathology of zucchini squash infected by F1F2L3-CMV, a pseudorecombinant virus was showed to change drastically the severe mosaic and stunting symptom into a mild chlorotic spot on systemic leave, compared with Fny-CMV. To delimit the sequence of RNA3 affected the pathology, various RNA3 chimeras were constructed between two strains of CMV. The symptom determinants of F1F2L3-CMV were mapped to the positions amino acid 234, 239, and 250 in 3a movement protein (MP). RNA3 chimeras changed the sequences encoding three amino acids were resulted in alteration of systemic symptom.

• The Plant Pathology Journal
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• 제21권2호
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• pp.142-148
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• 2005

Whereas most of isolates of Cucumber mosaic virus(CMV) can induce green mosaic systemic symptoms on zucchini squash, foliar symptoms of a pepper isolate of CMV (Pf-CMV)-infected zucchini squash revealed systemic chlorotic spots. To assess this biological property, infectious full-length cDNA clones of Pf-CMV were constructed using long-template RT-PCR. The complete nucleotide sequences of RNA2 and RNA3 of Pf-CMV were determined from the infectious fulllength cDNA clones, respectively. RNA 2 and RNA3 of Pf-CMV contain 3,070 nucleotides and 2,213 nucleotides, respectively. Overall sequence homology of two RNAs revealed high similarity (90%) between CMV strains, and 60% similarity to those of Tomato aspermy virus and Peanut stunt virus strains. By sequence analysis with known representative strains of CMV, Pf- CMV belongs to a typical member of CMV subgroup IA. The virus has high evolutionary relationship with Fny-CMV, but the pathology of Pf-CMV in zucchini squash was quite different from that of Fny-CMV. The pesudorecombinant virus, F1P2P3, induced chlorotic spot leaf symptom and timing of systemic symptom in squash plants, similar to the plants infected by Pf-CMV. No systemic symptoms were observed when Pf-CMVinoculated cotyledons were removed at 5 days postinoculation (dpi) while Fny-CMV showed systemic symptom at 2 dpi. These results suggest that the pepper isolate of CMV possesses unique pathological properties distinguishable to other isolates of CMVs in zucchini squash.

• The Plant Pathology Journal
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• 제18권2호
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• pp.85-92
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• 2002

A new isolate of Cucumber mosaic virus (CMV), identified as Li-CMV was isolated from a diseased Korean native lily (Lithium tsingtauense Gilg). Biological and serological properties of Li-CMV were characterized, and reverse transcription-polymerase chain reaction (RT-PCR) analysis, restriction enzyme profiling of RT-PCR products, and nucleotide sequence analysis of RNA3 of the virus were performed in this study. Remarkable differences in symptoms between Li-CMV and ordinary CMV strains were found in tobacco plants and Datura stramonium. Li-CMV-infected tobacco plants (cv. Xanthi-nc and cv. Samsun) induced chlorotic ringspots on uninoculated upper leaves, and the symptom expression was delayed or faint whereas, ordinary CMV strains induced green mosaic symptoms on the plant. Systemic infections were observed on Nicotiana benthamiana with severe mosaic symptom. Restriction mapping analysis of RT-PCR products using MspI showed that Li-CMV belonged to CMV subgroup I. A full-length CDNA copy of RNA3 for the virus was amplified by RT-PCR, cloned, and its complete nucleotide sequence was determined. The RNA3 of Li-CMV was 2, 232 nucleotides long, and consisted of two open reading frames of 843 and 657 bases encoding 3a protein (movement protein) and coat protein, respectively. Results of this study indicate that Li-CMV is a novel strain and belongs to subgroup I of CMV in the genus Cucumovirus.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.148.2-149
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• 2003

Cucumber fruit mottle mosaic tobamovirus (CFMMV) causes severe mosaic symptoms with yellow mottling on leaves and fruits, and occasionally severe wilting of cucumber plants. No genetic source of resistance against this virus has been identified. The genes coding for the coat protein or the putative 54-kDa replicase were cloned into binary vectors under control of the SVBV promoter. Agrobacterium-mediated transformation was peformed on cotyledon explants of a parthenocarpic cucumber cultivar with superior competence for transformation. R1 seedlings were evaluated for resistance to CFMMV infection by lack of symptom expression, back inoculation on an alternative host and ELISA. From a total of 14 replicase-containing R1 lines, 8 exhibited immunity, while only 3 resistant lines were found among a total of 9 CP-containing lines. Line 144 homozygous for the 54-kDa replicase was selected for further resistance analysis. Line 144 was immune to CFMMV infection by mechanical and graft inoculation, or by root infection following planting in CFMMV-contaminated soil. Additionally, line 144 showed delay of symptom appearance following infection by other cucurbit-infecting tobamoviruses. Infection of line 144 plants with various potyviruses and cucumber mosaic cucumovirus did not break the resistance to CFMMV. The mechanism of resistance of line 144 appears to be RNA-mediated, however the means is apparently different from the gene silencing phenomenon. Homozygote line 144 cucumber as rootstock demonstrated for the first time protection of a non-transformed scion from soil inoculation with a soil borne pathogen, CFMMV.

• The Plant Pathology Journal
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• 제22권3호
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• pp.265-270
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• 2006

Cultivated hot pepper crops showing severe mosaic symptom were found in Korea in 2004 and their causal agent was identified as Cucumber mosaic virus (CMV). These pepper crops was resistant to the virus in the filled, and they belonged to pathotype 0 (P0) resistant pepper. Resistance screening of selected pepper plants showed that a pepper isolate of CMV was the P0 resistance-breaking virus. This P0 resistance-breaking isolate of CMV, named as Ca-P1, was isolated from leaves of the virus-infected Capsicum annuum cv. Manidda that showed systemic severe mosaic symptom. Ca-P1-CMV could induce systemic mosaic symptoms on P0-susceptible (P0-S) and P0-resistant (P0-R) cultivars whereas an ordinary strain (Fny-CMV) could not infect P0-R. This result suggests that Ca-P1-CMV can overcome P0 resistant pepper cultivars. To analyze its genome sequence, the complete nucleotide sequence of RNA3 of Ca-P1-CMV was determined from the infectious full-length cDNA clone of the virus. RNA3 of Ca-P1-CMV consisted of 2,219 nucleotides. Overall sequence homology of RNA3-encoded two viral proteins (movement protein and coat protein) revealed high similarity (75.2-97.2%) with the known CMV strains. By sequence analysis with known representative strains of CMV, Ca-P1-CMV belongs to a typical member of CMV subgroup IB. The resistance and resistance-breaking mechanisms of pepper and counterpart CMV, respectively, remain to be investigated, which will enrich the genetic resources and accelerate CMV-resistant pepper breeding programs.