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

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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.

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Fig. 1a–d
Fig. 2a–c
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Fig. 4
Fig. 5a,b
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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

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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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  1. Jiehua Wang

    Present address: Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, Umea, Sweden

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  1. Centre for Forest Biology and Department of Biology, University of Victoria, PO Box 3020, Stn CSC, Victoria, BC, V8W 3N5, Canada

    Jiehua Wang & C. Peter Constabel

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  1. Jiehua Wang
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Correspondence to C. Peter Constabel.

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Wang, J., Constabel, C.P. Polyphenol oxidase overexpression in transgenic Populus enhances resistance to herbivory by forest tent caterpillar (Malacosoma disstria). Planta 220, 87–96 (2004). https://doi.org/10.1007/s00425-004-1327-1

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