Red fluorescent protein DsRed from Discosoma sp. as a reporter protein in higher plants

Abstract

GFP from Aequorea victoria is a standard genetic marker widely used to visualize cellular events in a noninvasive manner. For simultaneous imaging of different processes, in vivo mutants of GFP with shifted wavelength spectra (e.g., blue fluorescent protein) are conventionally used. The recently reported red fluorescent protein from Discosoma sp., DsRed, represents a new marker that can be used together with GFP variants for multicolor imaging. DsRed is an interesting marker protein for use in plants because of its red-shifted wavelength spectrum that will avoid damaging cells and tissues by excitation light. In this report, we show that DsRed is an excellent marker in higher plants in spite of the interfering red autofluorescence of chlorophyll, which can be eliminated by using the appropriate filter sets. Transient expression of DsRed1-C1 and a soluble-modified, red-shifted GFP variant has been carried out both individually and jointly in the epidermal cells of three different Nicotiana species and Chenopodium quinoa, which gives rise to dual labeling in plants. For this purpose, a human codon-optimized variant of DsRed has been adopted for expression in plants. Moreover, the DsRed reporter gene was expressed by using a labeled plant viral vector derived from an infectious full-length clone of potato virus X.