Fluorescent Proteins in Poplar: A Useful Tool to Study Promoter Function and Protein Localization

K. Nowak; N. Luniak; S. Meyer; J. Schulze; R. R. Mendel; R. Hänsch

doi:10.1055/s-2004-815730

Plant Biology, Table of Contents

Plant Biol (Stuttg) 2004; 6(1): 65-73
DOI: 10.1055/s-2004-815730

Original Paper

Georg Thieme Verlag Stuttgart · New York

Fluorescent Proteins in Poplar: A Useful Tool to Study Promoter Function and Protein Localization

K. Nowak¹ , N. Luniak¹ , S. Meyer¹ , J. Schulze¹ , R. R. Mendel¹ , R. Hänsch¹

¹Institut für Pflanzenbiologie, Technische Universität Braunschweig, Braunschweig, Germany

Abstract

The jellyfish (Aequorea victoria) green fluorescent protein (GFP) and its variants (CFP [cyan] and YFP [yellow]) were successfully used as a vital marker system for the transformation of hybrid poplar (Populus tremula × P. alba). Our results show that, in this woody plant, fluorescent proteins can be expressed: (i) transiently in protoplasts after PEG-mediated transformation, as well as in leaf cells after particle bombardment, and (ii) stably in callus cells and plants after Agrobacterium-mediated transformation. For these studies, we constructed vectors permitting easy recloning of any promoter fragments of interest. Confocal laser scanning microscopy was used both for visualization and differentiation between the different colours of the GFP variants and autofluorescence of chlorophyll and lignified xylem vessels. Peroxisomes were chosen as target organelles for GFP translocation via the peroxisomal targeting sequence PTS1 because this allowed us to concentrate the fluorochrome in the small volume of a few peroxisomes, giving a bright fluorescence over background noise.

Key words

GFP - PTS1 - reporter gene expression - transgenic poplar - Agrobacterium - protoplasts - particle bombardment - CLSM

Full Text

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R. R. Mendel

Institut für Pflanzenbiologie
Technische Universität Braunschweig

Humboldtstraße 1

38106 Braunschweig

Germany

Email: r.mendel@tu-bs.de

Section Editor: H. Rennenberg