Planta

, Volume 220, Issue 1, pp 87–96 | Cite as

Polyphenol oxidase overexpression in transgenic Populus enhances resistance to herbivory by forest tent caterpillar (Malacosoma disstria)

Original Article

Abstract

In order to functionally analyze the predicted defensive role of leaf polyphenol oxidase (PPO; EC 1.10.3.1) in Populus, transgenic hybrid aspen (Populus tremula × P. alba) plants overexpressing a hybrid poplar (Populus trichocarpa × P. deltoides) PtdPPO1 gene were constructed. Regenerated transgenic plants showed high PPO enzyme activity, PtdPPO1 mRNA levels and PPO protein accumulation. In leaf disk bioassays, forest tent caterpillar (Malacosoma disstria) larvae feeding on PPO-overexpressing transgenics experienced significantly higher mortality and reduced average weight gain compared to larvae feeding on control leaves. However, this effect was observed only when older egg masses were used and the resulting larvae showed reduced growth and vigor. In choice tests, no effect of PPO overexpression was detected. Although PPO in poplar leaves is latent and requires activation with detergents or trypsin for full enzymatic activity, in caterpillar frass the enzyme was extracted in the fully activated form. This activation correlated with partial proteolytic cleavage, suggesting that PPO latency and activation during digestion could be an adaptive and defense-related feature of poplar PPO.

Keywords

Forest tent caterpillar Herbivore defense Plant–insect interaction Populus Polyphenol oxidase Transgenic aspen 

Abbreviations

CaMV

Cauliflower mosaic virus

DOPA

Dihydroxyphenylalanine

FTC

Forest tent caterpillar

GUS

β-Glucuronidase

LPI

Leaf plastochron index

PPO

Polyphenol oxidase

SDS

Sodium dodecyl sulfate

Notes

Acknowledgements

The authors are grateful to Raju Datla (Plant Biotechnology Institute, Saskatoon, Canada) for providing the plant transformation vectors and host strains, Dave Ellis (CellFor Inc, Victoria, Canada) and Lise Jouanin (INRA, Versailles, France) for providing the hybrid aspen clone, Bob McCron (Canadian Forestry Service, Sault Ste Marie, Canada) for the forest tent caterpillar egg masses, and Brad Binges (Center for Forest Biology, University of Victoria) for help with plant maintenance. Funding was provided by the Natural Sciences and Engineering Research Council (NSERC) of Canada (C.P.C.) and by an FS Chia Fellowship from the University of Alberta (J.W.).

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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  1. 1.Centre for Forest Biology and Department of BiologyUniversity of VictoriaVictoriaCanada
  2. 2.Department of Forest Genetics and Plant PhysiologySwedish University of Agricultural SciencesUmeaSweden

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