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

茉莉酸（Jasmonic acid， JA）及其衍生物统称为茉莉酸盐（jasmonates），属于一类脂质衍生的信号分子，调节植物生命的许多方面，包括防御食草动物和病原体，以及与菌根真菌的共生相互作用。茉莉酸盐在调节网络中积极或消极地改变基因表达。大多数茉莉酸盐的功能是通过分析对茉莉酸不敏感或受茉莉酸生物合成影响的突变体来确定的。为了扩大我们对病原和共生体的JA感知机制的认识，开发和利用细胞和组织特异性水平的JA检测系统是本项目的主要目标。用于检测转基因植物中JA的方法是互补的，将采用双分子荧光互补（BiFC），利用COI1与JAZ蛋白的体内相互作用，以及合成的与核编码GFP融合的JA特异性启动子，分别观察JA反应的早期和晚期细胞。通过荧光活化细胞分选（FACS）从这些细胞中分离细胞核后，将进行转录物分析2。时间依赖性和细胞特异性基因表达模式作为细胞特异性功能的读出将导致鉴定新的ja反应基因。此外，还将筛选调节ja应答基因转录水平的小非编码rna。最后，我们期待在ja信号传导中发现新的调控元件。

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.