Welcome to Chinese Agricultural Science Bulletin,

Chinese Agricultural Science Bulletin ›› 2015, Vol. 31 ›› Issue (14): 102-107.doi: 10.11924/j.issn.1000-6850.casb15020082

Previous Articles     Next Articles

Establishment of a Living Fluorescence Detection System for High-throughput Screening Mutants of Different Components in the Ocimene Signaling Pathway

Xu Jian1,2, Li Lang2,3, Liu Qiwen1, Liu Chunlin2,3, Ruan Ying1,2   

  1. (1College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128;2Hunan Provincial Key Laboratory of Plant Epigenetics and Molecular Biology, Hunan Agricultural University, Changsha 410128; 3Agricultural College, Hunan Agricultural University, Changsha 410128)
  • Received:2015-02-13 Revised:2015-04-09 Accepted:2015-03-25 Online:2015-06-02 Published:2015-06-02

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.