• The Plant Pathology Journal
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• v.16 no.2
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• pp.118-124
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• 2000

The coat protein (CP) gene of kyuri green mottle mosaic virus zucchini strain (KGMMV-Z) isolated from zucchini (Cucurbita pepo) in Chonfu, Korea in 1999 was sequenced by the reverse transcription and polymerase chain reaction with degenerate and generate primers originated from tobamoviruses. The degenerate primers were very effective in amplification of KGMMV-Z CP region. The KGMMV-Z CP gene consisted of 486 nucleotides and had the same nucleotide length compared with those of cucurbit-infecting tobamoviruses. KGMMV-Z CP gene shared 43.8, 44.2, and 44.4% nucleotide sequence similarity with the CP gene of cucumber green mottle mosaic virus watermelon strain (CGMMZ-W), CGMMV-KW1, and CGMMV-SH, respectively, whereas three CGMMV strains among themselves showed 98.6-99.6% nucleotide similarity. The deduced amino acids of KGMMV-Z CP gene were 161 amino acid residues with the molecular weight of 17,181 daltons. The first 24 codons of KGMMV-Z CP gene corresponded to the sequences of the N-terminal amino acid of the viral capsid protein. The amino acid sequences of KGMMV-Z CP had 45.3% similarity compared with those of three CGMMV strains. However, the amino acid sequences of CGMMV strains were identical. These results showed that two cucurbit-infecting tobamovirus members, KGMMV-Z and CGMMV were genetically distantly related.

• The Plant Pathology Journal
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• v.31 no.4
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• pp.433-437
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• 2015

The infectious full-length cDNA clones of Cucumber green mottle mosaic virus (CGMMV) isolates KW and KOM, which were isolated from watermelon and oriental melon, respectively, were constructed under the control of the cauliflower mosaic virus 35S promoter. We successfully inoculated Nicotiana benthamiana with the cloned CGMMV isolates KW and KOM by Agrobacterium-mediated infiltration. Virulence and symptomatic characteristics of the cloned CGMMV isolates KW and KOM were tested on several indicator plants. No obvious differences between two cloned isolates in disease development were observed on the tested indicator plants. We also determined full genome sequences of the cloned CGMMV isolates KW and KOM. Sequence comparison revealed that only four amino acids (at positions 228, 699, 1212, and 1238 of the replicase protein region) differ between the cloned isolates KW and KOM. A previous study reported that the isolate KOM could not infect Chenopodium amaranticolor, but the cloned KOM induced chlorotic spots on the inoculated leaves. When compared with the previously reported sequence of the original KOM isolate, the cloned KOM contained one amino acid mutation (Ala to Thr) at position 228 of the replicase protein, suggesting that this mutation might be responsible for induction of chlorotic spots on the inoculated leaves of C. amaranticolor.

• The Plant Pathology Journal
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• v.21 no.3
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• pp.258-261
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• 2005

We conducted a survey on pepper virus diseases in 31 regions in Korea from November 2001 to December 2004. Using electron microscopy, test plant reaction, rapid immuno-filter paper assay (RIPA), reverse transcription-polymerase chain reaction (RT-PCR) and/or analysis of viral nucleotide sequences, we found a number of viruses from 1,056 samples that we collected. These included Cucumber mosaic virus (CMV), Pepper mottle virus (PepMoV), Pepper mild mottle virus (PMMoV), Broad bean wilt virus 2 (BBWV2), Tobacco mild green mosaic virus (TMGMV), and Tomato spotted wilt virus (TSWV). Of the samples analyzed, $343(32.5\%)$ were infected with CMV, $209(19.8\%)$ with PepMoV, $141(13.4\%)$ with PMMoV, $12(1.1\%)$ with BBWV2, $40(3.8\%)$ with TMGMV, $5(0.5\%)$ with TSWV, $153(14.5\%)$ with CMV and PepMoV, $54 (5.1\%)$ with CMV and PMMoV, $31(2.9\%)$ with PepMoV and PMMoV, $3(0.3\%)$ with CMV and BBWV2, $1(0.1\%)$ with CMV, PepMoV and BBWV2, $8(0.8\%)$ with CMV, PepMoV and PMMoV, and $30 (2.8\%)$ samples were infected with viruses which were not identified. CMV was the most predominant virus in all inspected fields and the number of the samples infected with PMMoV was relatively low as compared PepMoV infection level in pepper. TMGMV was only found in the southern part of Korea, while TSWV was isolated in Anyang and Yesan. However, we did not encounter in this survey the Alfalfa mosaic virus (AMV), Potato virus Y (PVY), Tobacco mosaic virus (TMV), and Pepper vein chlorosis virus (PVCV).

• Research in Plant Disease
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• v.13 no.1
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• pp.71-73
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• 2007

Field disease incidences of squash virus diseases in Jeonnam province were estimated to be 76.1% and of delayed planting on August-September (retarding culture) and on February-March (semi-forcing culture) on glass house were 55.0% and 0%, respectively, in 2000. Disease incidences of individual squash plant within a field were 100% and 3.6%, respectively, in wild culture and retarding culture. Total of 61 samples suspected to be infected with viruses were collected in 2000 and tested by RT-PCR using specific oligonulceotide primer sets designed for the detection of Cucumber green mottle mosaic virus (CGMMV), Kyuri green mottle mosaic virus (KGMMV), Zucchini yellow mosaic virus (ZYMV), Papaya ring spot virus (PRSV), Watermelon mosaic virus (WMV), and Cucumber mosaic virus (CMV). Each specific primer set for WMV, ZYMV, and PRSV amplified expected size of DNA fragments from 16, 10, and 2 samples in wild culture, respectively. Double or triple infection were observed in 7 samples tested. In contrast, each specific primer set for WMV, ZYMV, and PRSV confirmed virus infection from 7, 6, and 6 samples, respectively, in samples collected from semi-forcing culture. Double infection of WMV and PRSV was observed in only one sample. However, no DNA fragment was amplified from RT-PCR using CGMMV, KGMMV, and CMV specific oligonucleotide primer sets indicating no CGMMV, KGMMV, or CMV infection in squash fields in Jeonnam province in 2000.

• Research in Plant Disease
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• v.10 no.1
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• pp.53-57
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• 2004

Cucumber green mottle mosaic virus (CGMMV) was a major pathogen of watermelon and had affected seriously to watermelon production in Korea. Rapid and sensitive detection method of CGMMV associated with bottle gourd (Lagenafia siceraria) seeds was developed by using RT-PCR in this study. A pair of primeri Wmfl and Wmrl, specific for CGMMV was designed from coat protein gene sequences of CGMMV-W and used for amplifying 420 bp product in RT-PCR. To simplify the virus extraction procedure and reduce an inhibitor from the extract for the RT-PCR, some methods using ethanol precipitation, double filtration, polyethylene glycol precipitation and phenol/chloroform/isoamyl alcohol extraction procedure were compared and the phenol/chloroform/isoamyl alcohol extraction procedure was selected by its enhanced sensitivity. This detection method using the selected extraction step and the primers for RT-PCR could reliably detect an infected level of one CGMMV-infested seed in 1,000 seeds. This rapid and sensitive RT-PCR assay provides auseful tool for the specific detection of CGMMV in bottle gourd seed samples containing high levels of back-ground inhibitors.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2005.04a
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• pp.57.2-58
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• 2005

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2000.10a
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• pp.66.1-66
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• 2000

• Proceedings of the Korean Society of Plant Pathology Conference
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• 1999.10a
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• pp.85.2-85
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• 1999

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.149.2-150
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• 2003

Occurrences of virus diseases on paprika ( Capsicum annuum var. grossum) were surveyed in Joennam province from 1999 to 2003 and the collected samples showing virus-like symptoms were tested using ELISA. Virus diseases appeared 4.5％, 17.5％, and 4.9％ in 2000, 2002, and 2003, respectively. As the results of investigation of the seasonal incidence with the growing stages of plant, virus was not occurred at seedling stage and was slightly from the planting time to the first harvesting time, but was dramatically increased at the second harvesting time. Virus diseases were more severe on the vinyl house than on the green house. Pepper mild mottle virus (PMMoV) was severely occurred in 2000 but not after that year. Comparing the virus species, Pepper mottle virus (PepMoV) was 35.9％, Broad bean wilt virus (BBWV) was 14.1％, and Cucumber mosaic virus (CMV) was 10.9％ in 2002, and 76.0％, 11.1％, and 2.4％ in 2003, respectively.

• Korean Journal of Plant Tissue Culture
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• v.25 no.6
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• pp.519-524
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• 1998

A simple and reliable method to diagnose cucumber green mottle mosaic virus of watermelon in Korea (CGMMV-WK) was determined by RT-PCR, and coat protein gene for CGMMV-WK was cloned. Comparing to a method reported by Lee et al. (1996), the method developed here showed a better RT-PCR reaction. RT-PCR was possible by one step in the PCR reaction mixture that contains 20 pmol of primer, reverse transcriptase (30 unit), RNasin (5 unit) using the crude RNA solution. RT-PCR condition for specifically diagnosing CGMMV-WK was that cDNA was synthesized at 42$^{\circ}C$ for 45 min followed by pre-denaturation at 95$^{\circ}C$ for 2 min, and then PCR reaction was carried out with a programmed condition that consisted of 36 sequential cycles at 96$^{\circ}C$ for 30 sec, 6$0^{\circ}C$ for 30 sec, and 72$^{\circ}C$ for 1 min. A gene encoding the coat protein of CGMMV-WK was cloned and characterized. Nucleotide sequence of coat protein gene of CGMMV-WK shared 98.77% and 99.38% of sequence identity with those of CGMMV-W and CGMMV-SH, respecitvely, however, all of amino acid sequences were same.