Manners J M, Penninckx I A, Vermaere K, Kazan K, Brown R L, Morgan A, Maclean D J, Curtis M D, Cammue B P, Broekaert W F
F.A. Janssens Laboratory of Genetics, Katholieke Universiteit Leuven, Heverlee-Leuven, Belgium.
Plant Mol Biol. 1998 Dec;38(6):1071-80. doi: 10.1023/a:1006070413843.
The plant defensin PDF1.2 has previously been shown to accumulate systemically via a salicylic acid-independent pathway in leaves of Arabidopsis upon challenge by fungal pathogens. To further investigate the signalling and transcriptional processes underlying plant defensin induction, a DNA fragment containing 1184 bp and 1232 bp upstream of the transcriptional and translational start sites, respectively, was cloned by inverse PCR. To test for promoter activity this DNA fragment was linked to the beta-glucuronidase (GUS)-encoding region of the UidA gene as a translational fusion and introduced into Arabidopsis ecotype C-24. Challenge of the transgenic plants with the fungal pathogens Alternaria brassicicola and Botrytis cinerea resulted in both local and systemic induction of the reporter gene. Wounding of the transgenic plants had no effect on GUS activity. Treatment of the transgenic plants with either jasmonates or the active oxygen generating compound paraquat strongly induced the reporter gene. In contrast, neither salicylate nor its functional analogues 2,6-dichloroisonicotinic acid and 1,2,3-benzothiodiazole-7-carbothioic acid S-methyl ester resulted in reporter gene induction. These results are consistent with the existence of a salicylic acid-independent signalling pathway, possibly involving jasmonates as regulators, that is triggered by pathogen challenge but not by wounding. The transgenic plants containing the PDF1.2-based promoter-reporter construct will provide useful tools for future genetic dissection of this novel systemic signalling pathway.
植物防御素PDF1.2先前已被证明,在受到真菌病原体攻击时,拟南芥叶片中可通过一条不依赖水杨酸的途径进行系统性积累。为了进一步研究植物防御素诱导背后的信号传导和转录过程,通过反向PCR克隆了一个DNA片段,该片段分别位于转录起始位点和翻译起始位点上游1184 bp和1232 bp处。为了检测启动子活性,将该DNA片段与UidA基因的β-葡萄糖醛酸酶(GUS)编码区连接作为翻译融合体,并导入拟南芥生态型C-24中。用真菌病原体芸苔链格孢和灰葡萄孢对转基因植物进行攻击,导致报告基因在局部和全身都被诱导。对转基因植物进行创伤处理对GUS活性没有影响。用茉莉酸酯或产生活性氧的化合物百草枯处理转基因植物,强烈诱导了报告基因。相反,水杨酸及其功能类似物2,6-二氯异烟酸和1,2,3-苯并噻二唑-7-硫代羧酸S-甲酯均未导致报告基因诱导。这些结果与存在一条不依赖水杨酸的信号传导途径一致,该途径可能涉及茉莉酸酯作为调节因子,由病原体攻击触发而非创伤触发。含有基于PDF1.2的启动子-报告基因构建体的转基因植物将为未来对这条新的系统性信号传导途径进行遗传剖析提供有用的工具。