Rapid and low-cost diagnosis of Japanese yam mosaic virus infection in Chinese yam (Dioscorea polystachya) leaves by a print-capture RT-PCR

• Published in: Journal of general plant pathology : JGPP Vol. 83; no. 4; pp. 189 - 196
• Main Authors: Mochizuki, Tomofumi, Iwamoto, Itsuki, Atarashi, Aya, Ohki, Satoshi T., Kikukawa, Hiroyuki
• Format: Journal Article
• Language: English
• Published: Tokyo Springer Japan 01-07-2017; Springer Nature B.V
• Subjects: Agriculture; Biomedical and Life Sciences; Cellulose esters; Cellulose nitrate; Diagnosis; Diagnostic systems; Dioscorea japonica; Dioscorea polystachya; Leaves; Life Sciences; Membranes; Microbiology; Pathogens; Plant diseases; Plant Pathology; Plant virus diseases; Polymerase chain reaction; Ribonucleic acid; RNA; Technique; Vegetables; Viruses; RNA preparation; Print-capture RT-PCR; Diagnosis; Nitrocellulose membrane; Chinese yam
• ISSN: 1345-2630; 1610-739X
• DOI: 10.1007/s10327-017-0719-6

A rapid diagnostic method for
Japanese yam mosaic virus
(JYMV) infection in Chinese yam (
Dioscorea polystachya
) by a print-capture RT-PCR was developed. The cut surface of a rolled leaf of a JYMV-infected Chinese yam was stamped onto a nitrocellulose membrane. The clipped membrane was boiled in eluting solution to elute the RNA for use as the template in RT-nested PCR. Although JYMV RNA was detectable in most infected plants in the first PCR of the print-capture RT-PCR, it was detected from all samples in the nested PCR. The tissue-printed membranes could be preserved for at least 3 months at 4 °C for JYMV detection.