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

Plant Methods
Andrea Pitzschke, Helene Persak

Abstract

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. 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. The simplicity of the isolation and transformation procedure renders Poinsettia protoplasts an attractive sys...Continue Reading

References

Aug 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·B TinlandB Hohn
Jun 1, 1990·Plant Molecular Biology·R JeffersonM Bevan
Feb 8, 2005·Biotechnology Advances·Michael R DaveyKenneth C Lowe
Feb 13, 2009·Proteomics·Ganesh Kumar AgrawalRandeep Rakwal
Jul 18, 2009·Current Opinion in Plant Biology·Andrea PitzschkeHeribert Hirt
Oct 13, 2009·Proceedings of the National Academy of Sciences of the United States of America·Andrea PitzschkeHeribert Hirt

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Citations

Feb 18, 2014·International Journal of Molecular Sciences·Helene Persak, Andrea Pitzschke
Jan 16, 2016·International Journal of Molecular Sciences·Andrea PitzschkeGeorg J Seifert
Nov 22, 2016·Frontiers in Plant Science·Igor FesenkoVadim M Govorun

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Methods Mentioned

BETA
transfection
transgenic
fluorescence
myristoylation
PCR
light microscopy
electrophoresis
UV

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