Percoll-purified and photosynthetically active chloroplasts from Arabidopsis thaliana leaves

Plant Physiol Biochem. 2008 Nov;46(11):951-5. doi: 10.1016/j.plaphy.2008.06.009. Epub 2008 Jun 21.

Abstract

The availability of the complete genome sequence of Arabidopsis thaliana and of large collections of insertion mutants paved the way for systematic studies of gene functions in this organism, thus requiring adapting biochemical and physiological tools to this model plant. For physiological analysis of photosynthesis, methods combining high level of chloroplast purity and preservation of the photosynthetic activity were missing. Here, we describe a rapid method (less than 1h) to obtain Percoll-purified and photosynthetically active chloroplasts from Arabidopsis leaves retaining almost 90% of the Vmax of photosynthesis measured in the starting leaves from plants grown under a light intensity of 150mumolphotonm(-2)s(-1) and 80% of their initial photosynthetic rate after 3h of storage.

MeSH terms

  • Arabidopsis / chemistry*
  • Blotting, Western
  • Centrifugation, Isopycnic
  • Chloroplasts / physiology*
  • Electrophoresis, Polyacrylamide Gel
  • Oxygen / metabolism
  • Photosynthesis / physiology*
  • Plant Leaves / chemistry
  • Povidone
  • Silicon Dioxide

Substances

  • Percoll
  • Silicon Dioxide
  • Povidone
  • Oxygen