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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
Article

Abstract

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 

Compendio

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.

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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

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