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http://dx.doi.org/10.7744/cnujas.2015.42.2.099

Development of a diagnostic system to detect potato virus T using RT-PCR and nested PCR  

Lee, Si Won (Environmental Infrastructure Research Department, National Institute of Environmental Research)
Shin, Yong-Gil (Incheon International Airport Regional Office, Animal and Plant Quarantine Agency)
Lee, Jin-Young (Incheon International Airport Regional Office, Animal and Plant Quarantine Agency)
Kim, Young-Suk (Yeongsan River Environment Research Center, National Institute of Environmental Research)
Yang, Mi Hee (Environmental Infrastructure Research Department, National Institute of Environmental Research)
Choi, In-Cheol (Environmental Infrastructure Research Department, National Institute of Environmental Research)
Publication Information
Korean Journal of Agricultural Science / v.42, no.2, 2015 , pp. 99-103 More about this Journal
Abstract
Potato virus T (PVT) is a plant pathogen in the family Betaflexiviridae, group IV single-stranded positive sense RNA viruses. The major host of PVT is potato, and it has been reported in Ullucus tuberosus, Oxalis tuberosa and Tropaeolum tuberosum. This study aimed at developing reverse transcription (RT)-polymerase chain reaction (PCR) and nested PCR techniques for specific detection of PVT. Finally, Two RT-PCR primer sets were developed and verified. The RT-PCR products were amplified to 734 (PVT RT-PCR primer set 6) and 828 bp (PVT RT-PCR primer set 29) long to detect PVT. The nested PCR primer sets [PVT-N70/C20 ($734{\rightarrow}315bp$) and PVT-N75/C30 ($828{\rightarrow}529bp$)] were developed which are high sensitivity and verification for detection of PVT. Furthermore, a modified-positive control plasmid is use to verify contamination of laboratory in PVT detection. This study supported the diagnose PVT in potato or PVT related hosts.
Keywords
Betaflexiviridae; Nested PCR; Potato virus T; PVT; RT-PCR;
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Times Cited By KSCI : 4  (Citation Analysis)
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1 Caruso P, Bertolini E, Cambra M, Lopez, MM. 2003. A new and sensitive co-operational polymerase chain reaction for rapid detection of Ralstonia solanacearum in water. Journal of Microbiological Methods 55:257-272.   DOI   ScienceOn
2 Hall TA. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41:95-98.
3 Kim D, Hyun J, Hwang H, Lee S. 2000. RT-PCR detection of Citrus tristeza virus from early satsuma mandarin and yuzu Cheju Island. Plant Pathology Journal 16:48-51.
4 Kim YJ, Park S, Yie SW, Kim KH. 2005. RT-PCR detection of dsRNA Mycoviruses infecting Pleurotus ostreatus and Agaricus blazei Murrill. Plant Pathology Journal 21:343-348.   DOI   ScienceOn
5 Lizarraga C, Querci M, Santa Cruz M, Bartolini I, Salazar LF. 2000. Other natural hosts of Potato virus T. Plant Disease 84:736-738.   DOI   ScienceOn
6 Lee S. 2013. A study of molecular biological detection methods for seed-transmitted viruses in quarantine. Ph. D. thesis. Dankook University, Cheonan, Chungcheongnam-do, Korea.
7 Lee S, Cha M, Kim SM, Heo NY, Shin YG, Lee SH. 2014. Development of nucleotide primers for dignostic RT-PCR and nested PCR detection of three seed-transmitted viruses (CRLV, SpLV and WClMV) in quarantine. Journal of Agriculture & Life Science 48:75-83.
8 Lee S, Kang EH, Chu YM, Shin YG, Ahn TY. 2013. Development of PCR diagnosis system for plant quarantine seedborne Wheat streak mosaic virus. Korean Journal of Microbiology 49:112-117.   DOI   ScienceOn
9 Lee S, Shin YG. 2014. Development and practical use of RT-PCR for seed-transmitted Prune dwarf virus in quarantine. Plant Pathology Journal 30:178-182.   DOI   ScienceOn
10 Nelson M, McClelland M. 1992. Use of DNA methyltransferase/endonuclease enzyme combinations for megabase mapping of chromosomes. Methods in Enzymology 216:279-303.   DOI
11 Shin YG. 2009. An advanced quarantine system for the inspection of seed-borne viruses in Korea. Ph.D. Thesis, Kyungpook National University, Daegu, Gyeongsangbuk-do, Korea.
12 Shin YG, Rho JY. 2014. Development of a PCR diagnostic system for Iris yellow spot tospovirus in quarantine. Plant Pathology Journal 30:440-444.   DOI   ScienceOn
13 Stein A, Loebensten G, Koenic R. 1979. Detection of Cucumber mosaic virus and Bean yellow mosaic virus in gladiolus by enzyme linked immuno sobent assay (ELISA). Plant Disease Report 63:185-188.