关键词: 华重楼, 华重楼, 组织分离法, 匍枝根霉, 软腐病

Abstract: In order to efficiently find various signal transduction components in ocimene signaling pathway, this study uses forward genetic strategy, namely the technology system combining EMS mutagenesis and luciferase-luciferin in vivo fluorescence imaging detection technology for screening mutant. To do this, PR1pro::Luciferase expression cassette and PDF1.2pro::Luciferase expression cassette driven by PR1 gene promoter and PDF1.2 gene promoter respectively were constructed and the recombinant plasmids were transformed into Arabidopsis. By antibiotic selection and PCR verification, T₃ transgenic homozygous plants from several lines were obtained. Meanwhile, a suitable high sensitivity CCD camera, camera obscura and software were purchased, through assembling and debugging, a self-made, affordable in vivo fluorescence detector was successfully generated. After treated by jasmonic acid, salicylic acid or ocimene alone followed by fluorescent substrate spraying, homozygous transgenic plants were submitted to fluorescence detector, high intensity of induced-fluorescence signal in transgenic plants was detected, indicating that in vivo fluorescence imaging detection system was successfully constructed. These results provided a powerful forward genetic methodology and made it possible to use high-throughput selection of plants carrying mutations in ocimene signaling pathway components, as well as shed light on the mechanism of defense responses induced by ocimene.