Summary
A tissue culture method was developed for the eradication of three important potato viruses, PVX, PVY, and PVS, from the Russet Norkotah variety and two of its strains (TXNS 112 and TXNS 278). The method combined the use of liquid medium, thermotherapy, and chemotherapy. Initially, virus-free plants were inoculated with PVX, PVY, and PVS and, after 10 d, tested quantitatively for virus titer by ELISA to determine the initial virus concentration. The apical tip and the roots were removed from the inoculated plants, and only stem sections from inoculated plants were cultured in liquid medium containing MS inorganic salts, vitamins, and ribavirin (40 µM or 80 µM). After 5 d, half of the plants were subjected to thermotherapy and half were kept at room temperature. At 6 wk, the uppermost node (5–7 mm) was removed and recultured, and plants were tested then and again after 6 wk using ELISA to identify the virus-free plants.
Ribavirin alone eradicated the viruses from some plants; however, more virus-free plants were obtained when the treatments included heat. Additionally, thein vitro culture technique using liquid medium promoted rapid lateral shoot elongation and resulted in significantly faster plant production. Also this approach, which required less skilled labor, produced more plants than the meristem culture method for virus eradication. A detailed procedure for elimination of multiple viruses is described. This procedure resulted in production of more than 10% virus-free plants.
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References
Banttari, E. E.; Ellis, P. J.; Khurana, S. M. Management of diseases caused by viruses and viruslike pathogens. In: Crow, R. C., ed. Potato health management. St. Paul, MN: APS Press; 1993: 1127–1133.
Bijman, J. Potatoes: Improving disease resistance and quality. Biotech. Dev. Monitor. 12:3–5; 1992.
Brown, C. R.; Kwiatkowski, S.; Martin, M. W., et al. Eradication of PVS from potato clones through excision of meristems fromin vitro, heattreated shoot tips. Am. Potato J. 65:633–638; 1988.
Cassells, A. C.; Long, R. D. The elimination of potato viruses X, Y, S and M in meristem and explant cultures of potato in the presence of Virazole. Potato Res. 25:165–173; 1982.
Chen, W. Q.; Johnson, B.; Sherwood, J. L. A two step process for the regeneration ofArachis spp. by shoot tip culture of greenhouse-grown plants. Peanut Sci. 17:25–27; 1990.
Clark, M. F.; Adams, A. N. Characteristics of the microplate method of enzyme-linked immunosorbent assay for the detection of plant viruses. J. Gen. Virol. 34:475–483; 1977.
Close, R. Some effects of other viruses and of temperature on the multiplication of potato virus X. Ann. Appl. Biol. 53:151–164; 1964.
Conrad, P. L. Potato virus S-free plants obtained using antiviral compounds and nodal segment culture of potato. Am. Potato J. 68:507–513; 1991.
Dawson, W. O.; White, J. L.; Grantham, G. L. Effect of heat treatment upon cowpea chlorotic mottle virus ribonucleic acid replication. Phytopathology 68:1042–1048; 1978.
Dodds, J. H.; Silva-Rodriguez, D.; Bryan, J. E. Transport, receipt and propagation ofin vitro potato plantlets. International Potato Center (CIP), Lima, Peru: 1986:14.
Griffiths, H. M.; Slack, S. A. Potato virus elimination by heat and ribavirin. Phytopathology Abstract 78:1616; 1988.
Griffiths, H. M.; Slack, S. A.; Dodds, J. H. Effect of chemical and heat therapy on virus concentrations inin vitro potato plantlets. Can. J. Bot. 68:1515–1521; 1990.
Kassanis, B. Heat inactivation of leaf-roll virus in potato tubers. Ann. Appl. Biol. 37:339–341; 1950.
Klein, R. E.; Livingston, C. H. Eradication of potato virus X from potato by ribavirin treatment of cultured potato shoot tips. Am. Potato J. 59:359–365; 1982.
Lozoya-Saldana, H.; Dawson, W. O. The use of constant and alternating temperature regimes and tissue culture to obtain PVS-free potato plants. Am. Potato J. 59:221–230; 1982.
Mellor, F. C.; Stace-Smith, R. Virus strain differences in eradication of potato viruses X and S. Phytopathology 60:1587–1590; 1970.
Miller, J. C., Jr. Potato, Irish. The McGraw Hill Encyclopedia of Science and Technology. Vol. 14. New York: McGraw-Hill; 1992: 222–223.
Miller, J. C.; Smallwood, D. G.; Miller J. P., et al. Norgold Russet and Norgold Russet Strain M: Additional evidence for genetic dissimilarity. Am. Potato J. 72:273–286; 1995.
Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15:473–497; 1962.
Oshima, N.; Livingston, C. The effects of antiviral chemicals on potato virus -X-I. Am. Potato J. 38:294–299; 1961.
Sanchez, G. E.; Slack, S. A.; Dodds, J. H. Response of selectedSolanum species to virus eradication therapy. Am. Potato J. 68:299–315; 1991.
Slack, S. A. Pathogen free plants by meristem-tip culture. Plant Dis. 64:15–17; 1980.
Streeter, D. G.; Witkowski, J. T.; Khare, G. P., et al. Mechanism of action of 1-β-D-ribofuranosyl-1,2,4-triazole-3-carboxamide (Virazole), a new broad-spectrum antiviral agent. Proc. Nat; Acad. Sci. 70:1174–1178; 1973.
Wambugu, F. M.; Secor, G. A.; Gudmestad, N. C. Eradication of potato virus Y and S from potato by chemotherapy of cultured axillary bud tips. Am. Potato J. 62:667–672; 1985.
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Zapata, C., Miller, J.C. & Smith, R.H. Anin vitro procedure to eradicate potato viruses X, Y, and S from Russet Norkotah and two of its strains. In Vitro Cell Dev Biol - Plant 31, 153–159 (1995). https://doi.org/10.1007/BF02632012
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DOI: https://doi.org/10.1007/BF02632012