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A novel wound-responsive cis-element, VWRE, of the vascular system-specific expression of a tobacco peroxidase gene, tpoxN1
Authors:Katsutomo Sasaki  Hiroyuki Ito  Ichiro Mitsuhara  Susumu Hiraga  Shigemi Seo  Hirokazu Matsui  Yuko Ohashi
Institution:(1) Division of Plant Sciences, Organization of National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan;(2) Program for Promotion of Basic Research Activities for Innovative Bioscience (PROBRAIN), Toranomon Minato-ku, Tokyo, Japan;(3) Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan;(4) National Institute of Crop Science (NICS), National Agriculture and Food Research Organization (NARO), Tsukuba, Ibaraki 305-8518, Japan
Abstract:The wound-induced expression of tpoxN1, encoding a tobacco peroxidase, is unique because of its vascular system-specific expression and insensitivity to known wound-signal compounds such as jasmonic acid, ethylene, and plant hormones Sasaki et al. (2002) Plant Cell Physiol 43:108–117]. To study the mechanism of expression, the 2-kbp tpoxN1 promoter region and successive 5′-deletion of the promoter were introduced as GUS fusion genes into tobacco plants. Analysis of GUS activity in transgenic plants indicated that a vascular system-specific and wound-responsive cis-element (VWRE) is present at the −239/−200 region of the promoter. Gel mobility shift assays suggested that a nuclear factor(s) prepared from wounded tobacco stems binds a 14-bp sequence (−229/−215) in the −239/−200 region in a sequence-specific manner. A mutation in this 14-bp region of the −239 promoter fragment resulted in a considerable decrease in wound-responsive GUS activity in transgenic plants. An 11-bp sequence, which completely overlaps with the 14-bp sequence, was found in the 5′ distal region (−420/−410) and is thought to contribute to the wound-induced expression together with the 14-bp. The −114-bp core promoter of the tpoxN1 gene was indispensable for wound-induced expression, indicating that the 14-bp region is a novel wound-responsive cis-element VWRE, which may work cooperatively with other factors in the promoter.
Keywords:Wounding  Tobacco peroxidase  Promoter analysis                  cis-element  Ethylene-independent  Jasmonic acid-independent
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