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PLANT PHYSIOLOGY , Vol 101, Issue 1 201-208, Copyright © 1993 by American Society of Plant Biologists
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MOLECULAR BIOLOGY AND GENE REGULATION |
Abolition of an Inducible Highly Anionic Peroxidase Activity in Transgenic Tomato
B. A. Sherf, A. M. Bajar and P. E. Kolattukudy
The Ohio State Biotechnology Center, 1060 Carmack Road, Columbus, Ohio 43210
Locally induced expression of a highly anionic peroxidase has previously
been correlated temporally and spatially with suberization of tissues
responding to pathogen assault, wounding, or exogenously applied abscisic
acid or fungal elicitors. DNA sequences corresponding to the 5[prime]
regions of two tomato (Lycopersicon esculentum) genes encoding homologous
anionic peroxidases were fused, inserted into a pTi-based plasmid designed
to express a composite antisense transcript, and introduced into tomato via
Agrobacterium-mediated transformation. RNA gel-blot analyses showed high
expression of the antisense transcript in most transgenic plants and no
detectable induction of native anionic peroxidase transcripts in wounded or
abscisic acid or pathogen-treated tissues. Plants and fruits expressing the
antisense transcript appeared normal in all respects. Electrophoretic
analysis of anionic proteins from selected transgenic plants showed no
detectable anionic peroxidase protein or activity. Depolymerization of
polymeric material from the wound periderm of transgenic tomato fruits and
analysis of the aliphatic products by gas-liquid chromatography/mass
spectrometry showed that the content and composition of C16/C18
[omega]-hydroxy and dicarboxylic acids, characteristic of suberin, were not
affected by the absence of the anionic peroxidase. Autofluorescence
generated from cell wall phenolics at the wound lesion was also not
affected by the absence of the highly anionic peroxidase.
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