박과류 관련 종자전염 검역바이러스 3종의 PCR 진단시스템 개발 Development of Diagnostic PCR System for Three Seedtransmitted Quarantine Viruses Associated with Cucurbitaceae원문보기
박과는 스쿼시, 호박, 애호박, 조롱박, 오이 및 수박 등 100여 속이 넘는 속이 포함된 식물과이며, 종자의 수입량이 매우 많은 작물이다. 이들이 우리나라로 수입될 때, 규제바이러스인 Squash mosaic virus (SqMV), Cucumber green mottle mosaic virus (CGMMV) 및 Kyuri green mottle mosaic virus (KGMMV)에 대하여 검역을 수행한다. 본 연구에서는 SqMV, CGMMV 및 KGMMV를 신속·높은 감도로 검출할 수 있는 특이적 RT-PCR 및 nested PCR프라이머 조합과 유전자변형-양성대조구 플라스미드를 개발하였다. 또한 본 연구에서 개발한 진단시스템을 2010년부터 2014년 상반기까지 현장적용하여, SqMV 47건, CGMMV 67건 및 KGMMV 17건을 검출하였다.
박과는 스쿼시, 호박, 애호박, 조롱박, 오이 및 수박 등 100여 속이 넘는 속이 포함된 식물과이며, 종자의 수입량이 매우 많은 작물이다. 이들이 우리나라로 수입될 때, 규제바이러스인 Squash mosaic virus (SqMV), Cucumber green mottle mosaic virus (CGMMV) 및 Kyuri green mottle mosaic virus (KGMMV)에 대하여 검역을 수행한다. 본 연구에서는 SqMV, CGMMV 및 KGMMV를 신속·높은 감도로 검출할 수 있는 특이적 RT-PCR 및 nested PCR 프라이머 조합과 유전자변형-양성대조구 플라스미드를 개발하였다. 또한 본 연구에서 개발한 진단시스템을 2010년부터 2014년 상반기까지 현장적용하여, SqMV 47건, CGMMV 67건 및 KGMMV 17건을 검출하였다.
The Cucurbitaceae are a plant family that consist of over a hundred genera, the most important of which are squash, pumpkin, zucchini, some gourds, cucumber and watermelon. These are among the top imported seeds in Korea. At the time of their import, the Squash mosaic virus (SqMV), the Cucumber gree...
The Cucurbitaceae are a plant family that consist of over a hundred genera, the most important of which are squash, pumpkin, zucchini, some gourds, cucumber and watermelon. These are among the top imported seeds in Korea. At the time of their import, the Squash mosaic virus (SqMV), the Cucumber green mottle mosaic virus (CGMMV) and the Kyuri green mottle mosaic virus (KGMMV) are designated as regulated viruses for quarantine in Korea. This study was conducted to develop specific primer sets for easy and rapid detection of SqMV, CGMMV and KGMMV. RT-PCR with the nested PCR primer sets and modified positive control plasmids were capable of highly sensitive detection and verification of such viruses. In addition, developed diagnostic PCR systems applied to quarantine sites detected 47 cases of SqMV, 67 cases of CGMMV and 17 cases of KGMMV between 2010 and the first half of 2014.
The Cucurbitaceae are a plant family that consist of over a hundred genera, the most important of which are squash, pumpkin, zucchini, some gourds, cucumber and watermelon. These are among the top imported seeds in Korea. At the time of their import, the Squash mosaic virus (SqMV), the Cucumber green mottle mosaic virus (CGMMV) and the Kyuri green mottle mosaic virus (KGMMV) are designated as regulated viruses for quarantine in Korea. This study was conducted to develop specific primer sets for easy and rapid detection of SqMV, CGMMV and KGMMV. RT-PCR with the nested PCR primer sets and modified positive control plasmids were capable of highly sensitive detection and verification of such viruses. In addition, developed diagnostic PCR systems applied to quarantine sites detected 47 cases of SqMV, 67 cases of CGMMV and 17 cases of KGMMV between 2010 and the first half of 2014.
* AI 자동 식별 결과로 적합하지 않은 문장이 있을 수 있으니, 이용에 유의하시기 바랍니다.
대상 데이터
For the species-specific primers, SqMV (4 forward and 5 reverse primers), CGMMV (8 forward and 7 reverse primers) and KGMMV (6 forward and 4 reverse primers) were searched (Supplementary Table S1); and 10 sets, 17 sets and 15 sets that were capable of PCR amplification were combined, respectively (Supplementary Table S2). The designed primers were synthesized by Bioneer (Korea).
성능/효과
2010 and the first half of 2014. Total 47 positive cases of SqMV were detected (2 cases in 2010, 18 cases in 2011, 11 cases in 2012, 3 cases in 2013 and 13 cases in the first half of 2014); 67 cases of CGMMV (30 cases in 2010, 22 cases in 2011, 4 cases in 2012, 6 cases in 2013 and 5 cases in the first half of 2014); and 17 cases of KGMMV (13 cases in 2010 and 4 cases in 2011). All crops with contaminant viruses were discarded or returned.
참고문헌 (19)
Animal, Plant and Fisheries Quarantine and Inspection Agency. 2013. List of plant quarantine viruses in Korea in newly revised in 2013. Res. Plant Dis. 19: 67-75.
Caruso P, Bertolini E, Cambra M, López MM. 2003. A new and sensitive co-operational polymerase chain reaction for rapid detection of Ralstonia solanacearum in water. J. Microbiol. Methods 55: 257-272.
Cho JD, Kim JS, Lee SH, Chung BN. 2007. Triplex virion capture (VC)/RT-PCR for three seed transmissible Tobamoviruses of CGMMV, ZGMMV and KGMMV occurring on Cucurbitaceae. Res. Plant Dis. 13: 82-87.
Hall TA. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp. Ser. 41: 95-98.
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.
Lee S, Cha M, Kim SM, Heo NY, Shin YG, Lee SH. 2014. Development of nucleotide primers for diagnostic RT-PCR and nested PCR detection of three seed-transmitted viruses (CRLV, SpLV and WClMV) in quarantine. J. Agric. Life Sci. 48: 75-83.
Lee S, Kang EH, Chu YM, Shin YG, Ahn TY. 2013a. Development of PCR diagnosis system for plant quarantine seedborne Wheat streak mosaic virus. Korean J. Microbiol. 49:112-117.
Lee S, Kang EH, Heo NY, Kim SM, Kim YJ, Shin YG. 2013b. Detection of Carnation necrotic fleck vrus and Carnation ring-spot virus using RT-PCR. Res. Plant Dis. 19: 36-44.
Lee S, Kang EH, Shin YG, Lee SH. 2013c. Development of RT-PCR and nested PCR for detection of four quarantine plant viruses belonging to Nepovirus. Res. Plant Dis. 19: 220-225.
Nelson M, McClelland M. 1992. Use of DNA methyltransferase/endonuclease enzyme combinations for megabase mapping of chromosomes. Methods Enzymol. 216: 279-303.
Pan YB, Burner DM, Legendre BL. 2000. An assessment of the phylogenetic relationship among sugarcane and related taxa based on the nucleotide sequence of 5S rRNA intergenic spacers. Genetica. 108: 285-295.
Park SJ, Lee JH, Nam M, Park CY, Kim JS, Lee JH, et al. 2011. Virus disease incidences and transmission ecology of oriental melons in seongju area. Res. Plant Dis. 17: 342−350.
Priou S, Gutarra L, Aley P. 2006. An improved enrichment broth for the sensitive detection of Ralstonia solanacearum (biovars 1 and 2A) in soil using DAS-ELISA. Plant Pathol. J. 55: 36-45.
Rural Development Administration. 2012. Present condition and henceforward prospect of seed industry in the world. J. Korea Soc. Seed Sci. Ind. 9: 32-38.
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.
Yang DC, Lee JS, Kim DW, Lim YP, Min BH. 1998. Efficient diagnosis of Cucumber Green Mottle Mosaic Virus in watermelon using RT-PCR and cloning of coat protein gene. Korean J. Plant Tissue Culture 25: 519-524.
※ AI-Helper는 부적절한 답변을 할 수 있습니다.