Cell and tissue specific monitoring of jasmonates in plant interactions with pathogens and symbionts

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Jasmonic acid (JA) and its derivatives collectively known as jasmonates belong to a family of lipid-derived signaling molecules that regulates many aspects of plant life, including defense against herbivores and pathogens, but also symbiotic interactions with mycorrhizal fungi. Jasmonates alter gene expression positively or negatively in a regulatory network. Most functions of jasmonates have been identified through analysis of mutants that are JA-insensitive or affected in the biosynthesis of JA. To broaden our knowledge on the mechanism of JA perception in response to pathogens and symbionts, the development but also the utilization of a detection system for JA at cell and tissue specific level are the main goals of this project. The approaches used for detection of JA in transgenic plants are complementary and will employ bimolecular fluorescent complementation (BiFC) using the in vivo interaction of COI1 with JAZ proteins as well as a synthetic JA-specific promoter fused to nuclear encoded GFP to visualize early and late JA-responsive cells, respectively. After isolation of nuclei from these cells by fluorescence-activated cell sorting (FACS), transcript profiling will be carried 2 out. Time-dependent and cell-specific patterns of gene expression as a read-out of cell-specific functions will lead to identification of new JA-responsive genes. Additionally, a screen for small noncoding RNAs regulating transcript levels of JA-responsive genes will be performed. At the end, the identification of new regulatory elements in JA-signaling is expected.