Cultured guard cell protoplasts for standard methods of molecular, biochemical, and proteomic analysis.

 

Guard cell protoplasts: isolation, culture, and regeneration of plants.

by Tallman G. (2006)

Gary Tallman

in Plant Cell Culture Protocols –  Methods Mol. Biol. 318, 233–252. doi: 10.1385/1-59259-959-1:233 –

PubMed Abstract | CrossRef Full Text | Google Scholar – 

https://link.springer.com/protocol/10.1385%2F1-59259-959-1%3A233

Abstract

Guard cell protoplasts have been used extensively in short-term experiments designed to elucidate the signal transduction mechanisms that regulate stomatal movements. The utility of guard cell protoplasts for other types of longer-term signal transduction experiments is just now being realized. Because highly purified, primary isolates of guard cell protoplasts are synchronous initially, they are uniform in their responses to changes in culture conditions. Such isolates have demonstrated potential to reveal mechanisms that underlie hormonal signalling for plant cell survival, cell cycle re-entry, reprogramming of genes during dedifferentiation to an embryogenic state, and plant cell thermotolerance.

Plants have been regenerated from cultured guard cell protoplasts of two species: Nicotiana glauca (Graham), tree tobacco, and Beta vulgaris, sugar beet.

Plants genetically engineered for herbicide tolerance have been regenerated from cultured guard cell protoplasts of B. vulgaris. The method for isolating, culturing, and regenerating plants from guard cell protoplasts of N. glauca is described here.

A recently developed procedure for large-scale isolation of these cells from as many as nine leaves per experiment is described. Using this protocol, yields of 1.5–2 × 107 per isolate may be obtained. Such yields are sufficient for standard methods of molecular, biochemical, and proteomic analysis.

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Published by

Willem Van Cotthem

Honorary Professor of Botany, University of Ghent (Belgium). Scientific Consultant for Desertification and Sustainable Development.

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