Generation of fluorescent flowers exhibiting strong fluorescence by combination of fluorescent protein from marine plankton and recent genetic tools in Torenia fournieri Lind.

Abstract

Florescent proteins have been popularly used for studying genes and proteins of interest in various experiments at a cellular level, such as the analysis of intracellular localization and protein–protein interaction. However, the strength of fluorescence was insufficient for macro level observations of tissues or of the whole plant, and the fluorescent flowers that have been generated so far needed high-sensitive imaging equipment for the observation. Here we generated fluorescent Torenia flowers by the combined use of a high-performance fluorescent protein and the latest protein expression technologies, leading to the production of fluorescent proteins that can be easily and clearly observed. A coding sequence of a yellowish green fluorescent protein from the marine plankton Chiridius poppei (CpYGFP) was fused to the optimized sequences of the heat shock protein terminator and the 5′-untranslated region of the alcohol dehydrogenase gene of Arabidopsis to gain massive accumulation of the fluorescent protein. Strong fluorescence of CpYGFP was apparent in every part of the transgenic plant under the simple combination of a blue LED for excitation and an orange colored transparent acrylic filter for emission, while faint autofluorescence remained in the wild-type plants. By evaluating the combination of excitation wavelengths (excitation and emission filters) we were able to eliminate this undesired fluorescence. The fluorescent flowers could be used for ornamental purposes as well as for the analysis of fluorescent transgenic plants spatiotemporally in a nondestructive manner.