• Horticultural Science & Technology
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• v.32 no.2
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• pp.235-240
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• 2014

Cucumber mosaic virus (CMV) is one of the most important viral diseases in pepper (Capsicum annuum L.), and several genes for resistance were reported in Capsicum spp. In Korea, a single dominant gene that is resistant to $CMV_{Fny}$ and $CMV_{P0}$ has been used for breeding. Recently, a new strain ($CMV_{P1}$) was reported that could infect cultivars resistant to both $CMV_{Fny}$ and $CMV_{P0}$. Therefore, breeding of more robust CMV-resistant cultivars is required. In this study, we surveyed the inheritance of $CMV_{P1}$ resistance and analyzed the location of the resistance loci. After $CMV_{P1}$ inoculation of various germplasms and breeding lines, one accession (ICPN18-8) showed no visual symptoms at 15 dpi (days post inoculation) but was susceptible after 45 dpi, and one resistant line (I7339) showed resistance until at 45 dpi. The latter line was used for tests of resistance inheritance. A total of 189 $F_2$ plants were examined, with 42 individuals showing resistance at 15 dpi and a phenotype segregation ratio close to 1:3 (resistant:susceptible plants). In a lateral ELISA test at 45 dpi, 11 plants showed resistance, and the segregation ratio was changed to 1:15. These results indicate that resistance in C. annuum 'I7339' is controlled by two different recessive genes; we named these resistance genes 'cmr3E' and 'cmr3L,' respectively. To locate these two resistant loci in the pepper linkage map, various RAPD, SSR, and STS markers were screened; only nine markers were grouped into one linkage group (LG). Only one RAPD primer (OPAT16) was distantly linked with cmr3E (22.3 cM) and cmr3L (20.7 cM). To develop more accurate markers for marker-assisted breeding, enriching for molecular markers spanning two loci will be required.

• Horticultural Science & Technology
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• v.31 no.6
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• pp.764-771
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• 2013

Recently, Cucumber mosaic virus (CMV)-P1 infection in pepper cultivation is very serious problem, which causes low marketability and yield. In this research, 56 domestic pepper cultivars including 20 PR (Phytophthora resistance) cultivars, 31 foreign pepper lines collected from USA and 112 genetic pepper resources form RDA Genebank were used for CMV-P1 resistance evaluation. Resistance evaluation was performed at 24 days and 51 days after artificial inoculation of peppers by enzyme-linked immunosorbent assay (ELISA). Among the 36 domestic cultivars 'Muhanjilju' were resistant to CMV-P1. All 20 PR cultivars and 29 foreign peppers except 'NuMex Twilight' and 'Chainese Giant' were susceptible. Among 112 pepper lines from RDA Genebank, nine pepper lines were resistant, and 17 pepper lines were moderately resistant, and 86 pepper lines were susceptible. Almost all domestic peppers on the market were highly susceptible to CMV-P1, whereas 17.2% of foreign pepper cultivars and genetic pepper lines from RDA Genebank were resistant or moderately resistant. Resistant pepper lines selected in this study can be used as genetic sources for breeding CMV-P1 resistant pepper.

• Horticultural Science & Technology
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• v.34 no.6
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• pp.924-939
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• 2016

The great economic losses caused by Cucumber mosaic virus (CMV) infection of peppers has led to the development of genetically modified (GM) CMV-resistant peppers. We developed virus-resistant pepper plants using Agrobacterium tumefaciens -mediated transformation. The expressed recombinant protein was purified using nickel-nitrilotriacetic acid resin and immunoaffinity chromatography, and purity was assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Immunoblot analysis revealed the purified CMV coat protein (CMV-CP) had a molecular mass of 25 kDa. After in-gel digestion and desalting, the internal peptide fragments of CMV-CP were sequenced by matrix-assisted laser desorption/ionization-time of flight. Most GM pepper and Escherichia coli BL21 internal peptides had identical peptide sequences and contained 137 of 183 whole peptides in CMV-CP. A quantitative enzyme-linked immunosorbent assay was performed to detect CMV-resistant GM peppers. We also provide basic information about the expressed protein in GM peppers for further safety assessment. The contents of soluble protein and CMV-CP were measured in GM and control peppers cultivated in three different areas of Korea. Statistical significance in terms of cultivation areas, harvest times, generations, and plant tissue origin were determined based on a P value of 0.05. The highest amount of CMV-CP was detected at the seedling stage from plant grown in each region. T3 and T5 showed significantly different levels of CMV-CP from T4 in leaves in the whorl stage. No statistical differences were observed among GM peppers at different stages of maturity in any cultivation area. The results from this study contribute to the safety evaluation of newly designed CMV-resistant GM peppers and provide a standard against which to compare other virus-resistant GM peppers.

• The Plant Pathology Journal
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• v.22 no.3
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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.

• Research in Plant Disease
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• v.21 no.2
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• pp.99-102
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• 2015

Cucumber mosaic virus (CMV) is one of the most destructive viruses in chilli pepper. An isolate of CMV was obtained from the chilli pepper cv. Chungyang showing top necrosis symptom in 2013 and designated as CMV-GTN. CMV-GTN was compared with the well-characterized isolate, CMV-Ca-P1, by investigating their amino acid sequences of the coat protein (CP) and biological reactions in several host plants. The CP of CMV-Ca-P1 composed of 217 amino acids but that of CMV-GTN composed of 218 amino acids by including additional valine in the $57^{th}$ amino acid position. Amino acid sequence similarity of the CP gene among CMV-GTN and other CMV isolates recorded in the GeneBank database ranged from 96% to 99%. CMV-GTN was selected as a representative isolate to screen the resistance pepper cultivars to CMV because it was highly pathogenic to tomatoes and peppers upon biological assays. The virulence of CMV-GTN was tested on 135 pepper cultivars which has been bred in Korea and compared with that of CMV-Ca-P1. Only the cv. Premium was resistant and three cvs. Hot star, Kaiser, and Good choice were moderately resistant to CMV-GTN, whereas two cvs. Baerotta and Kaiser were resistant to CMV-Ca-P1.

• Korean Journal of Plant Tissue Culture
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• v.24 no.3
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• pp.153-160
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• 1997

Cucumber mosaic virus (CMV) leads to a cause of poor crop productivity and quality. To solve this problem, we attempted to develop a virus-resistance tobacco plants by using viral coat protein (CP) gene. Transgenic tobacco plants expressing CMV CP gene were analysed by the resistance upon CMV infection. The virus-resistance was measured in $\textrm{T}_{1}$, generation by the inhibition of plant growth and the expression of the mosaic symptoms infected with CMV. The transgenic lines were divided into four groups: highly resistant, resistant, moderate and susceptible based on their growth and symptom severity. Out of 39 transgenic lines, 16 lines showed significant virus-resistance. And of resistant lines, 2 lines were designated highly resistant based on the facts that they achieved similar plant height to that of non-infected tobacco plants and showed lower disease symptom than that of other lines. The steady state level of CP RNA and coat protein level were measured by northern blot and immunoblot analysis. The CP RNA was highly accumulated in most resistant and moderate lines but barely detected in susceptible lines. The coat protein was detected in most lines regardless of their resistance to CMV. from this result, virus-resistance appeared to correlate more with CP RNA level than the level of coat protein. However, in two highly resistant lines, CP RNA level was unexpectedly low. This unexpected phenomenon need to be further investigated.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.4 no.1
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• pp.103-108
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• 1968

Resistance to various diseases of Korean native varieties were investigated in field. The results obtained are summarized as follows: 1) The varieties Ubangtchio and Osib-eubthio were moderately resistant to black shank, but Suantchio, Usultchio and Gwangtchio were slightly resistant. Dixie Bright 101, $H_2$ and Bright Yellow 4 were more resistant to black shank than any of the Korean native varieties. 2) Mokgitchio, Ubangtchio and Osib-eubtchio were moderately resistant to black root rot, but Useultchio Suantchio and Hoetchio were susceptible than Others. 3) Muktchio, Hoetchio and Ubangtchio were slightly resistant to wild fire. 4) Osib-ebthio was highly resistant to brown spot, but Ubangtchio, Hyangtchio and Mokgitchio were moderately resistant and Useultchio was susceptible to brown spot, but it was resistant than Bright ellow 4 or Bernhart 1000-1. 5) Hyangtchio was slightly resistant to CMV-Y. According to these results, except Virus disease, Ubangtchio was resistant to all of the above mentioned diseases. Osib-eubtchio, Hoetchio, Hyangtchio and Suantchio showed proferable disease resistant and these might be well utilized as breeding materials.

• Food Science and Preservation
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• v.22 no.6
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• pp.901-907
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• 2015

Genetically modified (GM) pepper H15 containing the gene for cucumber mosaic virus (CMV) coat protein (CP) and its control line non-GM pepper P2377 were investigated for their allergic risk. Amino acid sequence of the inserted gene product CMV-CP was compared with those of known allergens. No known allergen had greater than 35% amino acid sequence homology over an 80 amino acid window or more than 8 consecutive identical amino acids. Protein patterns of GM and non-GM pepper extracts were evaluated by SDS-PAGE, which showed similar distribution of protein bands for both GM and non-GM pepper. Antigen-antibody reactions were compared between GM and its non-transgenic parental control. ELISA and immunoblot analysis of sera from allergic patients showed some IgE reactivity; however, no differences were observed between GM pepper H15 and P2377. We therefore conclude that CMV-CP is less likely to be an allergen; the protein composition and allergenicity of the GM pepper H15 is not different from that of P2377 and safe as a commercial host.

• Journal of Environmental Impact Assessment
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• v.22 no.2
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• pp.125-134
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• 2013

Plant virus coat (CP) gene-mediated protection is one of the best known approaches to protect against virus resistant transgenic plants. Transgenic N. benthamiana plants containing the CP gene of Zucchini green mottle mosaic virus (ZGMMV) were used for the environmental risk assessment of the living modified (LM) plants with plant virus resistance. The most optimal co-infection method of both ZGMMV and CMV (Cucumber mosaic virus) on Non-LM and CP-expressing LM tobacco plants was established and co-infection of CMV and ZGMMV was confirmed by polymerase chain reaction (PCR). To address the effects of LM tobacco plants on the mutation of the virus, in-vitro transcripts of CP and Replicase (Rep) derived from CMV and/or ZGMMV were inoculated onto Non-LM or LM tobacco plants. Mutation frequency of CP and Rep from CMV and ZGMMV was examined through six serial passages in Non-LM and LM tobacco plants. Little actual frequency of mutation was estimated, probably due to the limited number of transgenic plants tested in this study. However, it does not suggest environmental safety of these CP-mediated LM plants. Further study at a larger scale is needed to evaluate the environmental risk associated with the CP-expressing LM plants.

• Research in Plant Disease
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• v.18 no.4
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• pp.290-297
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• 2012

In this study, total number of 1941 Capsicum accessions conserved at RDA Genebank was evaluated for their response to Cucumber mosaic virus (CMV). These accessions were composed with 9 species originated from 89 countries, included 839 Capsicum annuum, 277 C. baccatum, 395 C. chinense, 343 C. frutescens, 49 C. pubescens, and other 38 wild pepper species (C. chacoense, C. galapagoense, etc.). Resistant to CMV was screened with the 240H02SP6 SNP marker related to the Cmr1 (Cucumber mosaic resistance 1). Eighty nine accessions of pepper germplasm were resistant to CMV based on the marker. One hundred sixty two accessions showed heterozygosity. One thousand two hundred seventy accessions were susceptible to CMV. Four hundred twenty accessions did not show distinction by 240H02SP6 marker. These 89 resistant pepper germplasm can be used in a pepper breeding program against CMV.