Abstract
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.
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References
Babil P, Kondo S, Iwata H, Kushikawa S (2013) Intra-specific ploidy variations in cultivated Chinese yam (Dioscorea polystachya Turcz.). Trop Agr Dev 57:101–107
Bertolini E, Moreno A, Capote N, Olmos A, de Luis A, Vidal E, Pérez-Panadés J, Cambra M (2008) Quantitative detection of citrus tristeza virus in plant tissues and single aphids by real-time RT-PCR. Eur J Plant Pathol 120:177–188
Chang P-G, McLaughlin WA, Tolin SA (2011) Tissue blot immunoassay and direct RT-PCR of cucumoviruses and potyviruses from the same NitroPure nitrocellulose membrane. J Virol Methods 171:345–351
Dovas CI, Katis NI (2003a) A spot nested RT-PCR method for the simultaneous detection of members of the Vitivirus and Foveavirus genera in grapevine. J Virol Methods 107:99–106
Dovas CI, Katis NI (2003b) A spot multiplex nested RT-PCR for the simultaneous and generic detection of viruses involved in the etiology of grapevine leafroll and rugose wood of grapevine. J Virol Methods 109:217–226
Fuji S, Nakamae H (1999a) Complete nucleotide sequence of the genomic RNA of a Japanese yam mosaic virus, a new potyvirus in Japan. Arch Virol 144:231–240
Fuji S, Nakamae H (1999b) Detection of Japanese yam mosaic virus by ELISA and RT-PCR. Ann Phytopathol Soc Jpn 65:207–210 (in Japanese with English summary)
Fuji S, Furuya H, Naito H, Natsuaki KT (2001) Detection of potyviruses in yam plants cultivated in Japan. Jpn J Phytopathol 67:261–263 (in Japanese with English summary)
Fukami M, Kunitomo E, Mochizuki T (2014) Detection of Chrysanthemum stem necrosis virus (CSNV) by print-capture RT-PCR. Ann Rep Kanto-Tosan Plant Prot Soc 61:78–81 (in Japanese)
Fukumoto F, Tochihara H (1978) Chinese yam necrotic mosaic virus. Ann Phytopathol Soc Jpn 44:1–5 (in Japanese with English summary)
Fukuta S, Iida T, Mizukami Y, Ishida A, Ueda J, Kanbe M, Ishimoto Y (2003) Detection of Japanese yam mosaic virus by RT-LAMP. Arch Virol 148:1713–1720
Kajihara H, Muramoto K, Fuji S, Tanaka S, Ito S (2009) Simultaneous detection of Japanese yam mosaic virus and Yam mild mosaic virus from yam using a tube capture reverse transcription-polymerase chain reaction assay. J Gen Plant Pathol 75:72–75
Kameya-Iwaki M, Yamaguchi K, Hara T, Ito S, Fuji S, Kajihara H, Tanaka S (1999) Detection of Japanese yam mosaic virus from Chinese yam (Dioscorea opposita Thunb. cv. Ichoimo) by immunocapture PCR and differentiation of virulent and attenuated isolates by PCR-RFLP analysis. Ann Phytopathol Soc Jpn 65:494–497 (in Japanese with English summary)
Kondo T, Fuji S, Yamashita S, Kang D-K, Chang M-U (2005) Broad bean wilt virus 2 in yams. J Gen Plant Pathol 71:441–443
La Notte P, Minafra A, Saldarelli P (1997) A spot PCR technique for the detection of phloem-limited grapevine viruses. J Virol Methods 66:103–108
Mumford RA, Seal SE (1997) Rapid single-tube immunocapture RT-PCR for the detection of two yam potyviruses. J Virol Methods 69:73–79
Odu BO, d’A Hughes J, Shoyinka SA, Dongo LN (1999) Isolation characterization and identification of potyvirus from Dioscorea alata L. (water yam) in Nigeria. Ann Appl Biol 134:65–71
Olmos A, Dasí MA, Candresse T, Cambra M (1996) Print-capture PCR: a simple and highly sensitive method for the detection of Plum pox virus (PPV) in plant tissues. Nucl Acids Res 24:2192–2193
Osman F, Rowhani A (2006) Application of a spotting sample preparation technique for the detection of pathogens in woody plants by RT-PCR and real-time PCR (TaqMan). J Virol Methods 133:130–136
Papayiannis LC, Hunter SC, Iacovides T, Browa JK (2010) Detection of Cucurbit yellow stunting disorder virus in cucurbit leaves using sap extracts and real-time TaqMan® reverse transcription (RT) polymerase chain reaction (PCR). J Phytopathol 158:487–495
Acknowledgements
The authors thank Masanobu Fukami for helpful comments on this work and Hirokazu Ueno and the Sasayama-Shinonome High School students for providing virus-infected and virus-free Tsukuneimo samples. HK was supported through funding from the Promotion of Pre-University Research Activities in Science Program by the Japan Science and Technology Agency.
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Mochizuki, T., Iwamoto, I., Atarashi, A. et al. Rapid and low-cost diagnosis of Japanese yam mosaic virus infection in Chinese yam (Dioscorea polystachya) leaves by a print-capture RT-PCR. J Gen Plant Pathol 83, 189–196 (2017). https://doi.org/10.1007/s10327-017-0719-6
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DOI: https://doi.org/10.1007/s10327-017-0719-6