American Potato Journal

, Volume 70, Issue 3, pp 185–199 | Cite as

Immunodetection of Polyphenol oxidase in glandular trichomes ofS. berthaultii, S. tuberosum and their hybrids

  • Stanley P. Kowalski
  • Robert L. Plaisted
  • John C. Steffens


Type A glandular trichomes confer insect resistance to the wild potato,S. berthaultii Hawkes by oxidatively polymerizing their contents upon breakage, resulting in insect entrapment. Polyphenol oxidase (PPO) is responsible for the oxidative polymerization of Type A glandular trichome exudate.S. tuberosum L. trichomes possess low amounts of PPO and lack the ability to either polymerize their contents or entrap insects. Polyclonal antibodies raised against purifiedS. berthaultii glandular trichome PPO were used to develop an enzyme-linked immunosorbent assay (ELISA) to quantify PPO content ofS. berthaultii, S tuberosum, and hybrid type A glandular trichomes. The PPO ELISA and immunoblots were used to quantify PPO expression as a function of leaf development and genetic background. PPO accumulates to its maximal levels in trichomes prior to completion of leaflet expansion. In advanced breeding generations, trichome PPO is expressed at levels varying from that found inS. tuberosum to that found inS. berthaultii. By serving as an initial screen for trichome PPO, this ELISA may be useful to assist rapid introgression of this insect resistance trait into adaptedS. tuberosum backgrounds.

Additional Key Words

Solanum berthaultii Solanum tuberosum polyphenol oxidase insect resistance host plant resistance potato glandular trichomes 


El tipo A de tricomas glandulares confiere resistencia a los insectos, en la papa silvestreSolanum berthaultii, porque al romperse, su contenido sufre una polimerización oxidativa que lo convierte en una trampa para los insectos. La oxidasa Polyphenol (PPO) es responsable de la polimerización oxidativa del exudado de los tricomas glandulares tipo A. Los tricomas deS. tuberosum poseen bajas cantidades de PPO y carecen de la capacidad de polimerizar su contenido o de atrapar a los insectos. Los anticuerpos policlonales logrados contra la PPO purificada de los tricomas glandulares deS. berthaultii fueron utilizados para desarrollar una prueba serológica por medio de conjugados enzimáticos (ELISA) para cuantificar el contenido de PPO de los tricomas glandulares tipo A enS. berthaultii, S. tuberosum e híbridos. Las pruebas ELISA e inmunológica en membranas de nitrocelulosa para PPO fueron utilizadas para cuantificar la PPO como una función del desarrollo foliar y base genética. La PPO se acumula en los tricomas a sus máximos niveles antes de completarse la total expansión de los foliolos. En generaciones avanzadas de mejoramiento, la PPO de los tricomas se presenta a niveles que varían desde aquellos encontrados enS. tuberosum hasta los deS. berthaultii. Sirviendo como una evaluatión y selectión inicial para PPO en los tricomas, esta ELISA puede ser útil para ayudar a una rápida introgresión de este atributo de resistencia a los insectos en las bases adaptadas deS. tuberosum.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Literature Cited

  1. 1.
    Ausubel, F., R. Brent, R.E. Kingston, D.D. Moore, J.A. Smith, J.G. Seidman and K. Struhl. 1987. Current Protocols in Molecular Biology. Greene Publishing Associates, Brooklyn, NY.Google Scholar
  2. 2.
    Ave, D.A., N.T. Eannetta and W.M. Tingey. 1986. A modified enzymic browning assay for potato glandular trichomes. Am Potato J 63:553–558.CrossRefGoogle Scholar
  3. 3.
    Dubernet, M. and P. Ribereau-Gayon. 1974, Isoelectric point changes inVitus vinifera catechol oxidase. Phytochem 13:1085–1087.CrossRefGoogle Scholar
  4. 4.
    Esterbauer, H., E. Schwarzl and M. Hayn. 1977. TNB: A rapid assay for catechol oxidase and laccase using 2-nitro-5-thiobenzoic acid. Anal Biochem 77:486–494.PubMedCrossRefGoogle Scholar
  5. 5.
    Gibson, R.W. 1974. Aphid-trapping glandular hairs on hybrids ofSolanum tuberosum andS. berthaultii. Potato Res 17:152–154.CrossRefGoogle Scholar
  6. 6.
    Goffreda J.C., M.A. Mutschier, D.A. Ave, W.M. Tingey and J.C. Steffens. 1989. Aphid deterrence by glucose esters in glandular trichome exudate of the wild tomato,Lycopersicon pennellii. J Chem Ecol 15:2135–2147.CrossRefGoogle Scholar
  7. 7.
    Gregory, P., W.M. Tingey, D.A. Ave and P.Y. Bouthyette. 1986. Potato glandular trichomes: a physiochemical defense mechanism against insects.In: Natural Resistance of Plants to Pests, Roles of Allelochemicals. M.B. Green, P.A. Hedin. (Eds.). American Chemical Society, Washington, D.C. pp. 160–167.Google Scholar
  8. 8.
    Hutcheson, S.W. and B.B. Buchanan. 1980. Polyphenol oxidation byVicia faba chloroplast membranes. Plant Physiol 66:1150–1154.PubMedGoogle Scholar
  9. 9.
    King, R.R., Y. Pelletier, R.P. Singh and L.A. Calhoun. 1986. 3,4-Di-0-isobutyry-6-0-caprylsucrose: the major component of a novel sucrose ester complex from the type B glandular trichomes ofSolanum berthaultii Hawkes (PI 473340). J Chem Soc Chem Commun 14:1078–1079.CrossRefGoogle Scholar
  10. 10.
    King, R.R., R.P. Singh and A. Boucher. 1987. Variation in sucrose esters from the type B glandular trichomes of certain wild potato species. Am Potato J 64:529–534.CrossRefGoogle Scholar
  11. 11.
    Kowalski, S.P., J.B. Bamberg, W.M. Tingey and J.C. Steffens. 1990. Inheritance of polyphenol oxidase in type A glandular trichomes ofSolanum berthaultii. J Heredity 81:475–478.Google Scholar
  12. 11a.
    Kowalski, S.P., N.T. Eannetta, A.T. Hirzel and J.C. Steffens. 1992. Purification and characterization of polyphenol oxidase from glandular trichomes ofSolanum berthaultii. Plant Physiol 100:677–684.PubMedCrossRefGoogle Scholar
  13. 12.
    Lanker, T., T.G. King, S.W. Arnold and W.H. Flurkey. 1987. Active, inactive and in vitro synthesized forms of polyphenoloxidase during leaf development. Physiol Plantarum 69:323–329.CrossRefGoogle Scholar
  14. 13.
    Lieberei, R., B. Bole and J. Voight. 1981. Serological studies on phenolase from spinach leaves. Phytochem 20:2109–2116.CrossRefGoogle Scholar
  15. 14.
    Mayer, A.M. and E. Harel. 1979. Review: polyphenol oxidases in plants. Phytochem 26:1–20.Google Scholar
  16. 15.
    Mehlenbacher, S.A., R.L. Plaisted and W.M. Tingey. 1983. Inheritance of glandular trichomes withSolanum berthaultii. Am Potato J 60:699–708.CrossRefGoogle Scholar
  17. 16.
    Mehlenbacher, S.A., R.L. Plaisted and W.M. Tingey. 1984. Heritability of trichome density and droplet size in interspecific potato hybrids and relationship to aphid resistance. Crop Sci 224:320–322.Google Scholar
  18. 17.
    Neal, J.J., W.M. Tingey and J.C. Steffens. 1990. Sucrose esters of carboxylic acids in glandular trichomes ofSolanum berthaultii deter settling and feeding by the green peach aphid. J Chem Ecol 16:487–497.CrossRefGoogle Scholar
  19. 18.
    Plaisted, R.L. 1980. PotatoIn: Hybridization of crop plants. (Fehr, W.R. and H.H. Hadley, Eds.). Madison, Wisconsin: American Society of Agronomy, Crop Science Society of America; 483–494.Google Scholar
  20. 19.
    Plaisted, R.L., W.M. Tingey and J.C. Steffens. 1991. Colorado potato beetle resistance derived fromSolanum berthaultii. Am Potato J 68:629 (Abstract).Google Scholar
  21. 20.
    Robertson, E.F., H.K. Dannelly, P.J. Malloy and H.C. Reeves. 1987. Rapid isoelectric focusing in a vertical polyacrylamide minigel system. Anal Biochem 167:290–294.PubMedCrossRefGoogle Scholar
  22. 21.
    Ryan, J.D., P. Gregory and W.M. Tingey. 1983. Glandular trichomes: enzymic browning assays for improved selection of resistance to the green peach aphid. Am Potato J 60:861–868.CrossRefGoogle Scholar
  23. 22.
    Tingey, W.M. and R.W. Gibson. 1978. Feeding and mobility of the potato leafhopper impaired by glandular trichomes ofSolanum berthaultii andSolanum polyadenium J Econ Entomol 71:856–858.Google Scholar
  24. 23.
    Tingey, W.M., R.L. Plaisted, J.E. Laubengayer, and S.A. Mehlenbacher. 1982. Green peach aphid resistance by glandular trichomes inSolanum tuberosum XS. berthaultii hybrids. Am Potato J 59:241–251.CrossRefGoogle Scholar
  25. 24.
    Tingey, W.M. 1991. Potato glandular trichomes: defensive activity against insect attack.In: Naturally Occurring Pest Bioregulators. P.A. Hedin Ed. American Chemical Society, Washington, DC. pp 126–135.Google Scholar

Copyright information

© Springer 1993

Authors and Affiliations

  • Stanley P. Kowalski
    • 1
  • Robert L. Plaisted
    • 1
  • John C. Steffens
    • 1
  1. 1.Department of Plant BreedingCornell UniversityIthaca

Personalised recommendations