Uptake of a Fluorescent Dye as a Swift and Simple Indicator of Organelle Intactness: Import-competent Chloroplasts from Soil-grown Arabidopsis

• DOI: 10.1177/002215540405200514
• Publication Date: 2011-04-01T07:28:21Z
• Authors: Alexander Schulz; Jürgen Knoetzel; Henrik V. Scheller; Alexandra Mant
• Keywords: Organelle

We developed a rapid and reliable technique for specifically staining intact chloroplasts using the fluorescent dye carboxyfluorescein diacetate. Intact, import-competent were isolated simply rapidly from soil-grown Arabidopsis thaliana plants, with yields of 20 ± 5 μg chlorophyll per g FW, greater than previously reported Arabidopsis. Traditional chloroplast isolation buffers sometimes contain low concentrations (<10 mM) sodium ascorbate as general-purpose anti-oxidant, but we found that only in presence high (50–100 initial grinding buffer import-competent.