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Fusion genetic analysis of jasmonate‐signalling mutants in Arabidopsis
Author(s) -
Jensen Anders B.,
Raventos Dora,
Mundy John
Publication year - 2002
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.0960-7412.2001.01241.x
Subject(s) - mutant , reporter gene , biology , microbiology and biotechnology , staurosporine , jasmonate , methyl jasmonate , phosphatidate , arabidopsis , fusion protein , gene expression , biochemistry , gene , kinase , protein kinase a , protein kinase c , recombinant dna , diacylglycerol kinase
Summary Jasmonates induce plant‐defence responses and act to regulate defence‐related genes including positive feedback of the lipoxygenase 2 ( LOX2 ) gene involved in jasmonate synthesis. To identify jasmonate‐signalling mutants, we used a fusion genetic strategy in which the firefly luciferase ( FLUC ) and Escherichia coli β‐glucuronidase (GUS) reporters were expressed under control of the jasmonate‐responsive LOX2 promoter. Spatial and temporal patterns of reporter expression were determined initially, and revealed that JA‐responsive expression from the LOX2 promoter required de novo protein synthesis. Reporter activity was also induced by the protein kinase inhibitor staurosporine and antagonized by the protein phosphatase inhibitor okadaic acid. FLUC bio‐imaging, RNA gel‐blot analysis and progeny analyses identified three recessive mutants that underexpress the FLUC reporter, designated jue1 , 2 and 3 , as well as two recessive mutants, designated joe1 and 2 , that overexpress the reporter. Genetic analysis indicated that reporter overexpression in the joe mutants requires COI . joe1 responded to MeJA with increased anthocyanin accumulation, while joe2 responded with decreased root growth inhibition. In addition, reporter induction and endogenous LOX2 expression by staurosporine was absent in joe2 .