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Plant and Cell Physiology, 2002, Vol. 43, No. 3 266-274
© 2002 Oxford University Press

Induced Expression of a Temperature-Sensitive Leucine-Rich Repeat Receptor-like Protein Kinase Gene by Hypersensitive Cell Death and Wounding in Tobacco Plant Carrying the N Resistance Gene

Naoko Ito1,3, Reona Takabatake1, Shigemi Seo1, Susumu Hiraga1,4, Ichiro Mitsuhara1,2 and Yuko Ohashi1,2,5

1 Core Research of Evolutionary Science and Technology, 2-3 Surugadai, Kannda, Chiyoda-Ku, Tokyo, 101-0062 Japan 2 Molecular Genetics Department, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, 305-8602 Japan

A gene encoding a receptor-like protein kinase was isolated as the gene induced in the early period of N gene-dependent hypersensitive cell death in tobacco leaves. The kinase domain expressed as a glutathione S-transferase fusion protein was capable of autophosphorylation, indicating that this gene encodes an active protein kinase. A high level of the transcript accumulated before necrotic lesion formation in tobacco mosaic virus (TMV)-inoculated tobacco leaves carrying the N gene but it was low in a tobacco cultivar lacking the N gene. A small but reproducible increase in the transcript was found 1–2 h after a temperature shift from 30°C to 20°C even in healthy leaves, suggesting the gene expression is temperature sensitive. The gene was named WRK for wound-induced receptor-like protein kinase, because the transcript increased to a maximum within 15–30 min of wounding. In suspension cultured tobacco cells, an increase in the transcript was found 15 min after transfer to a new medium, but it was suppressed under high osmotic pressures. The wound-induced WRK accumulation was enhanced by cycloheximide treatment, but not by known defense signal compounds (salicylic acid, jasmonic acid, 1-aminocyclopropan-1-carboxylic acid and abscisic acid) and some plant hormones. Thus, WRK is a wound-inducible and temperature-sensitive protein kinase gene induced before hypersensitive cell death probably through unknown signaling pathways.

3 Present address: Niigata Univ. of Health and Welfare, Niigata, 950-3198 Japan.

4 Present address: Graduate School of Agriculture, Hokkaido University, Sapporo, 060-8589 Japan.

5 Corresponding author: E-mail, yohashi@affrc.go.jp; Fax, +81-298-38-7469.


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