RT-PCR Detection of Three Non-reported Fruit Tree Viruses Useful for Quarantine Purpose in Korea

Park, Mi Ri; Kim, Kook Hyung

doi:2004.20.2.147

Plant Pathol J > Volume 20(2); 2004 > Article

The Plant Pathology Journal 2004;20(2):147-154.
DOI: https://doi.org/10.5423/PPJ.2004.20.2.147 Published online June 30, 2004.

RT-PCR Detection of Three Non-reported Fruit Tree Viruses Useful for Quarantine Purpose in Korea

Mi Ri Park, Kook Hyung Kim

Abstract

A simple and reliable procedure for RT-PCR detection of Apple stem pitting virus (ASPV), Cherry rasp leaf virus (CRLV), and Cherry necrotic rusty mottle virus (CNRMV) was developed. Two virus specific primer sets for each virus were found to specifically detect each virus among fourteen sets of designed oligonucleotide primers. Total RNAs extracted from healthy and from ASPV-, CRLV- and CNRMV-infected plant tissues were used to synthesize cDNA using oligo dT primer and then amplified by virus-specific primers for each virus. Each primer specifically amplified DNA fragments of 578 bp and 306 bp products for ASPV (prAS CP-C and prAS CP-N primers, respectively); 697 bp and 429 bp products for CRLV (prCR4 and prCR5-JQ3D3 primers, respectively); and 370 bp and 257 bp products for CNRMV (prCN4 and prCN6-NEG 1 primers, respectively) by RT-PCR. DNA sequencing of amplified DNA fragments confirmed the nature of each amplified DNA. Altogether, these results suggest that these virus specific primer sets can specifically amplify viral sequences in infected tissues and thus indicate that they can be used for specific detection of each virus.

Key Words: Apple stem pitting virus, ASPV, Cherrynecrotic rusty motle virus, CNRMV, Cherry rasp Leafvirus, CRLV, quarantine