The promoter of the plant defensin gene PDF1.2 from Arabidopsis is systemically activated by fungal pathogens and responds to methyl jasmonate but not to salicylic acid.
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.
Manners
J M
JM
F.A. Janssens Laboratory of Genetics, Katholieke Universiteit Leuven, Heverlee-Leuven, Belgium.
Penninckx
I A
IA
Vermaere
K
K
Kazan
K
K
Brown
R L
RL
Morgan
A
A
Maclean
D J
DJ
Curtis
M D
MD
Cammue
B P
BP
Broekaert
W F
WF
D000085:Acetates / Q000494:pharmacology*; D000528:Alternaria / Q000472:pathogenicity*; D000595:Amino Acid Sequence; D000935:Antifungal Agents; D017360:Arabidopsis / Q000235:genetics* / Q000382:microbiology*; D001483:Base Sequence; D020171:Botrytis / Q000472:pathogenicity*; D003517:Cyclopentanes / Q000494:pharmacology*; D017931:DNA Primers; D023082:Defensins; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D017930:Genes, Reporter; D005966:Glucuronidase / Q000096:biosynthesis / Q000235:genetics; D007539:Isonicotinic Acids / Q000494:pharmacology; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense; D054883:Oxylipins; D010937:Plant Growth Regulators / Q000494:pharmacology*; D018515:Plant Leaves; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000187:drug effects; D014176:Protein Biosynthesis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D020156:Salicylic Acid / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects; D014158:Transcription, Genetic / Q000187:drug effects*; D015533:Transcriptional Activation
D000085:Acetates; D000935:Antifungal Agents; D003517:Cyclopentanes; D017931:DNA Primers; D023082:Defensins; D007539:Isonicotinic Acids; D016376:Oligonucleotides, Antisense; D054883:Oxylipins; D010937:Plant Growth Regulators; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; C077261:2,6-dichloroisonicotinic acid; C072239:methyl jasmonate; D005966:Glucuronidase; D020156:Salicylic Acid