Abstract
This report describes a simple, rapid and inexpensive procedure for sampling large numbers of dormant tubers for analysis of potato leafroll luteovirus (PLRV) infection. The procedure uses a common electric drill to simultaneously remove and macerate tuber-eye samples for detection of PLRV by the enzyme-linked immunosorbant assay (ELISA) and the polymerase chain reaction (PCR). By using these sampling and analysis approaches, 19 of 20 different PLRV isolates were detected in dormant tubers from plants with primary infections. Results from the dormant tuber analysis, were verified by planting the tubers and testing leaf tissue by ELISA and PCR. Similar sampling and testing done on healthy dormant tubers and sprouts from the tubers consistently gave negative results as expected.
Similar content being viewed by others
Literature Cited
Alves, A.M.C. and A.R. Figueira 1991. Efeito da desfolha precoce sobre a producao e incidencia de virus em tres cultivares de batata. Ciencia e Pratica, 15(2): 127–136.
Camargo F.W.P., and A.R. Mazzei. 1996. Bataticultura no Mercosul. Producao e Mercado no Brasil e na Argentina. Informacoes Economicas, SP, 26(3):53–67.
Casper, R. 1977. Detection of potato leafroll virus in potato andPhysalis floridana by enzyme linked immunosorbent assay (ELISA). Phytopathology. Z. 90:364–368.
Clark, M.F. and A.N. Adams. 1977. Characteristics of the microplate method of enzyme linked immunosorbent assay for the detection of plant viruses. Journal of General Virology 34:475–483.
Costa, A.S. 1965. Molestias de virus da batata. Tudo sobre batata. Boletim do Campo 190: 68–93
Ehlers, U.; H.J. Vetten, and H.L. Paul. 1983. Detection of potato leafroll virus in primarily infected tubers by enzyme-linked immunosorbent assay. Phytopathology. Z 107:37–46.
Ehlers, U. and H.J. Vetten. 1984. Detection of PLRV, PVA and PVY in potato tubers by ELISA. Potato Research 27:101–102.
Figueira, A.R. 1995. Viroses da batata e suas implicacoes na producao de batata-semente do Estado de Minas Gerais: historico do problema e solucoes. SummaPhytopathologica 21:268–269.
Gugerli, P. 1979. Le test immuno-enzymatique (ELISA) et son application pour la diagnostic rapide des viroses de la pomme de terre. Revue suisse d’Agriculture 11:253–260.
Kojima, M. and H. Lapierre. 1988. Potato leafroll virus (PLRV). In: Smith I M., Dunez J., Phillips D.H., Lelliot, R.A. and Archer S.A. (eds.) European Handbook of Plant Diseases. pp. 23–24. Blackwell Scientific Publications, Oxford.
Slack, S.A. 1995. Potato viruses with some implications for production and processing in the United States: a history of problems and solutions. Summa Phytopathologica 21(3–4): 273–275.
Souza-Dias, J.A.C. de; A.S. Costa, and A.M. Nardin. 1993. Potato leafroll virus in solanaceous weeds in Brazil explains severe outbreaks of the disease in absence of known potato donor sources. Summa Phytopathologica 19(2):80–85.
Souza-Dias, J.A.C. de, P. Russo, L. Miller, and S.A. Slack. 1998. Comparison of nucleotide sequences from three potato leafroll virus (PLRV) isolates collected in Brazil. AJPR (in press).
Sutula, C.L., J.M. Gilllet, S.M. Morrissey, and D.C. Ramsdell. 1986. Interpreting ELISA data and establishing the positive-negative threshold. Plant Disease 70:722–726.
Tamada, T. and B.D. Harrison. 1980. Factors affecting the detection of potato leafroll virus in potato foliage by enzyme-linked immunosorbent assay. Ann Appl Biol 95:209–219.
Wang, AM., M.V. Doyle, and D.F. Mark. 1989. Quantitation of mRNA by the polymerase chain reaction. Proc Natl Acad Sci USA. 86:9717–9721.
Author information
Authors and Affiliations
Corresponding author
Additional information
The research was funded in part by a grant from FAPESP, No. 96/03698-1 and CNIq/RHAE, No. 260014/96.6.
Rights and permissions
About this article
Cite this article
de Souza-Dias, J.A.C., Russo, P., Betti, J.A. et al. Simplified extraction method for ELISA and PCR detection of potato leafroll luteovirus primary infection in dormant potato tubers. Am. J. Pot Res 76, 209–213 (1999). https://doi.org/10.1007/BF02854223
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02854223