Development of a PCR Diagnostic System for Iris yellow spot tospovirus in Quarantine

Abstract

Iris yellow spot virus (IYSV) is a plant pathogenic virus which has been reported to continuously occur in onion bulbs, allium field crops, seed crops, lisianthus, and irises. In South Korea, IYSV is a “controlled” virus that has not been reported, and inspection is performed when crops of the genus Iris are imported into South Korea. In this study, reverse-transcription polymerase chain reaction (RT-PCR) and nested PCR inspection methods, which can detect IYSV, from imported crops of the genus Iris at quarantine sites, were developed. In addition, a modified positive plasmid, which can be used as a positive control during inspection, was developed. This modified plasmid can facilitate a more accurate inspection by enabling the examination of a laboratory contamination in an inspection system. The inspection methods that were developed in this study are expected to contribute, through the prompt and accurate inspection of IYSV at quarantine sites to the plant quarantine in South Korea.

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 RNA was reverse-transcribed at 42°C for 30 min. PCR was performed at 95°C for 1 min followed by 35 cycles of 95°C for 45 s, 55°C for 1 min, and 72°C for 1 min with a final extension at 75°C for 5 min [16]. PCR products (4 ll) were subjected to 1.2 % agarose gel electrophoresis.…”

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

“…Third, if the positive control sample group is not secured, PCR test errors and amplification or non-amplification are difficult to accurately assess [13]. Moreover, since the positive control group sample is infected with the virus, it may be difficult to distribute domestically and import [14]. Therefore, in this study, both RT-PCR and nested PCR methods were developed to accurately diagnose the plant quarantine seed-transmitted TRSV, which may infect diverse imported seeds.…”

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

“…그러나 많은 연구자들에 의해 개발 된 PCR 방법은 조건이 서로 상이하여 시료에서 다양한 바이 러스들을 검사할 경우 활용이 불편하였다. 이로 인하여 최근 식물바이러스 진단 시 검사조건을 통일시킨 user-base의 검 사방법이 개발 및 적용 (Lee, 2013;Shin, 2009;Shin and Rho, 2014 (Fig. 1B).…”