Development and Practical Use of RT-PCR for Seed-transmitted Prune dwarf virus in Quarantine

Abstract

Among imported plants, seeds are the items that have many latent pathogens and are difficult to inspect. Also, they are the import and export items whose market is expected to expand. The biggest problem with seeds is viruses. Prune dwarf virus (PDV) is the virus that is commonly inspected in Prunus cerasifera, P. persica, P. armeniaca, P. mandshurica, P. cerasus, P. avium or P. serotina seeds. In this study, two RT-PCR primer sets, which can promptly and specifically diagnose plant quarantine seed-transmitted PDV, were developed; and nested PCR primers, where products amplify 739 and 673 nucleotides (nt), and an nested PCR-product, 305 nt, can be obtained as these products are amplified again, were developed. Also, a modified-positive control plasmid was developed, where the restriction enzyme XhoI, which can identify the contamination of samples from the control, was inserted. The method developed in this study has detected PDV in 18 cases since 2007, and is expected to continuously contribute to the plant quarantine in Korea.

Development of PCR Diagnostic System for Detection of the Seed-Transmitted Tobacco Ringspot Virus in Quarantine

“…RNA was extracted from the affected samples, cDNA was synthesized, DNA was extracted from healthy plants, and RT-PCR and electrophoresis were performed as previously described [5,18].…”

“…To manufacture the modified-positive control plasmid for use in the TRSV RT-PCR and nested PCR diagnosis systems was performed as previously described [18,19]. To confirm the inserted gene, TRSV RT-PCR set 2 was amplified using the manufactured modified-positive control plasmid as the template DNA.…”

Development of PCR-base Diagnostic System for the Detection of Andean potato latent virus

“…및 종자전 염을 하고 Fribourg et al, 1977), 핵산은 3개의 단백질을 암호화하며 총 염기서열의 길이는 약 6.4 kb이다 (Koenig and Lesemann, 1981;Bernal et al, 2000). APLV는 파파리자, 비름과, 명아주과, 박과 및 가지과에서 감염 보고사례 (Gibbs and Harrison, 1969;Koenig and Bode, 1978 (Lee et al, 2013a;Lee et al, 2013c;Lee and Shin, 2014;Lee et al, 2014a;Lee et al, 2015)되어 바이러스 검역실적이 증가하였으나 (Shin, 2009;Lee et al, 2013b) 참 고 문 헌…”

Development of a Specific Diagnostic System for Detecting Turnip Yellow Mosaic Virus from Chinese Cabbage in Korea

“…TYMV is also potential quarantine virus in Korea for imported crops. Since user-based RT-PCR diagnosis system was introduced, various viruses have been detected at the quarantine sites [10][11][12][13][14][15][16][17]. However, inspection method for TYMV has not been clarified.…”

“…Total RNAs of TYMV and reference viruses [Andean potato latent tymovirus (APLV) and Poinsettia mosaic tymovirus (PnMV)] were purchased from Bi-One (Korea). Primer3 program was used to establish the same inspection conditions as in quarantine sites [14]. Accordingly, 19 TYMV-specific primer sets were designed in forward and reverse directions ( Table 1).…”