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Open Access Methodology

Poinsettia protoplasts - a simple, robust and efficient system for transient gene expression studies

Andrea Pitzschke* and Helene Persak

Author Affiliations

Dept. Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, Muthgasse 18, University of Natural Resources and Life Sciences, 1190, Vienna, Austria

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Plant Methods 2012, 8:14  doi:10.1186/1746-4811-8-14

Published: 4 May 2012

Abstract

Background

Transient gene expression systems are indispensable tools in molecular biology. Yet, their routine application is limited to few plant species often requiring substantial equipment and facilities. High chloroplast and chlorophyll content may further impede downstream applications of transformed cells from green plant tissue.

Results

Here, we describe a fast and simple technique for the high-yield isolation and efficient transformation (>70%) of mesophyll-derived protoplasts from red leaves of the perennial plant Poinsettia (Euphorbia pulccherrima). In this method no particular growth facilities or expensive equipments are needed. Poinsettia protoplasts display an astonishing robustness and can be employed in a variety of commonly-used downstream applications, such as subcellular localisation (multi-colour fluorescence) or promoter activity studies. Due to low abundance of chloroplasts or chromoplasts, problems encountered in other mesophyll-derived protoplast systems (particularly autofluorescence) are alleviated. Furthermore, the transgene expression is detectable within 90 minutes of transformation and lasts for several days.

Conclusions

The simplicity of the isolation and transformation procedure renders Poinsettia protoplasts an attractive system for transient gene expression experiments, including multi-colour fluorescence, subcellular localisation and promoter activity studies. In addition, they offer hitherto unknown possibilities for anthocyan research and industrial applications.

Keywords:
Protoplast; Transient expression; High transformation efficiency; Chlorophyll autofluorescence; Promoter activity studies