, Volume 220, Issue 1, pp 105–117 | Cite as

Antisense downregulation of polyphenol oxidase results in enhanced disease susceptibility

  • Piyada ThipyapongEmail author
  • Michelle D. Hunt
  • John C. Steffens
Original Article


Polyphenol oxidases (PPOs; EC or EC catalyze the oxidation of phenolics to quinones, highly reactive intermediates whose secondary reactions are responsible for much of the oxidative browning that accompanies plant senescence, wounding, and responses to pathogens. To assess the impact of PPO expression on resistance to Pseudomonas syringae pv. tomato we introduced a chimeric antisense potato PPO cDNA into tomato (Lycopersicon esculentum L.). Oxidation of caffeic acid, the dominant o-diphenolic aglycone of tomato foliage, was decreased ca. 40-fold by antisense expression of PPO. All members of the PPO gene family were downregulated: neither immunoreactive PPO nor PPO-specific mRNA were detectable in the transgenic plants. In addition, the antisense PPO construct suppressed inducible increases in PPO activity. Downregulation of PPO in antisense plants did not affect growth, development, or reproduction of greenhouse-grown plants. However, antisense PPO expression dramatically increased susceptibility to P. syringae expressing the avirulence gene avrPto in both Pto and pto backgrounds. In a compatible (pto) interaction, plants constitutively expressing an antisense PPO construct exhibited a 55-fold increase in bacterial growth, three times larger lesion area, and ten times more lesions cm−2 than nontransformed plants. In an incompatible (Pto) interaction, antisense PPO plants exhibited 100-fold increases in bacterial growth and ten times more lesions cm−2 than nontransformed plants. Although it is not clear whether hypersusceptibility of antisense plants is due to low constitutive PPO levels or failure to induce PPO upon infection, these findings suggest a critical role for PPO-catalyzed phenolic oxidation in limiting disease development. As a preliminary effort to understand the role of induced PPO in limiting disease development, we also examined the response of PPO promoter::β-glucuronidase constructs when plants are challenged with P. syringae in both Pto and pto backgrounds. While PPO B inducibility was the same in both compatible and incompatible interactions, PPO D, E and F were induced to higher levels and with different expression patterns in incompatible interactions.


Antisense Disease resistance Lycopersicon Polyphenol oxidase Pseudomonas 









Polyphenol oxidase


Reactive oxygen species


Salicylic acid



We are grateful to Dr. S.M. Newman for providing the transgenic tomato carrying the PPO A, B, D, E and F promoter::GUS constructs. We also thank Drs. S.D. Tanksley and A. Frary for providing P. syringae culture and the inoculation procedure, Dr. E.D. Earle for the use of microscope facilities, and C. Dougherty for the transmission electron microscope work. This work was supported by grants from the United States Department of Agriculture National Research Initiative (grant no. 91-37301-6571) and the Binational Agricultural Research and Development Fund (grant no. US 1870-90) to J.C.S.


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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Piyada Thipyapong
    • 1
    • 2
    Email author
  • Michelle D. Hunt
    • 1
    • 3
  • John C. Steffens
    • 1
    • 4
  1. 1.Department of Plant BreedingCornell UniversityIthacaUSA
  2. 2.Suranaree University of TechnologyNakhon RatchasimaThailand
  3. 3.CropsolutionResearch Triangle ParkUSA
  4. 4.Syngenta BiotechnologyResearch Triangle ParkUSA

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