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THE PLANT CELL, Vol 7, Issue 3 249-257, Copyright © 1995 by American Society of Plant Biologists
Use of a Gene Expression System Based on Potato Virus X to Rapidly Identify and Characterize a Tomato Pto Homolog That Controls Fenthion Sensitivity
CMT. Rommens, J. M. Salmeron, D. C. Baulcombe and B. J. Staskawicz
Department of Plant Biology, 111 Koshland Hall, University of California, Berkeley, California 94720
A novel transient gene expression system was used to study both the tomato
disease resistance gene Pto and a Pto homolog designated Fen. The gene
expression system was based on potato virus X (PVX). Tomato plants that
were both susceptible to strains of Pseudomonas syringae pv tomato carrying
the corresponding avirulence gene avrPto and insensitive to the insecticide
fenthion were infected with in vitro-generated transcripts of PVX
derivatives containing either Pto or Fen. Expression of the Pto gene from
the virus genome failed to elicit P. s. tomato resistance, indicating that
the PVX system is not suitable for the study of Pto. However, expression of
the Fen gene resulted in sensitivity to fenthion. The utility of the PVX
gene expression system was further demonstrated through structure/function
studies of the Fen gene. A correlation was shown between Fen protein kinase
activity and the ability of this protein to confer fenthion sensitivity to
tomato. Furthermore, it was demonstrated that mutation of a putative
N-terminal myristoylation site, proposed to be involved in membrane
targeting, rendered the Fen protein inactive. Analysis of a Pto-Fen
chimeric gene allowed the fenthion sensitivity domain to be localized to
the C-terminal part of the Fen protein. Interestingly, expression of the
Fen kinase from the PVX genome in Nicotiana spp resulted in a
fenthion-independent necrotic response. Our results support the involvement
of the Fen gene in a signal transduction pathway(s).
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