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Plant and Cell Physiology, 1979, Vol. 20, No. 1 145-155
© 1979

Article

Firefly assay of adenine nucleotides from algae: Comparison of extraction methods

C.-M. Larsson and T. Olssonl1

Institute of Botany, University of Stockholm S-106 91 Stockholm, Sweden

Pools of ATP, ADP and AMP in algae were compared after extraction with Trisbuffer, ethanol, chloroform, sulfuric acid, trichloroacetic acid and perchloric acid. Experimental organisms were Anacystis nidulans, Euglena gracilis, Chlorella kessleri and Scenedesmus obtusiusculus. Adenine nucleotides were determined by firefly luciferase assay.

Extraction of illuminated algae in situ with the different methods resulted in great variations in the proportions between ATP, ADP and AMP. The sum of adenine nucleotides varied less. Extraction with trichloroacetic acid or perchloric acid generally yielded the highest energy charge and ATP/ADP ratio. Experiments with illuminated Anacystis and Scenedesmus showed that extraction with perchloric acid in situ gave higher yields of ATP in the extracts than if samples were withdrawn for extraction (time for sample withdrawal {small tilde}1 sec). For Anacystis, it was demonstrated that the extraction methods giving the highest proportions of ATP were suitable for studies on fluctuations of the adenine nucleotide pools at transitions from dark to light.

1 Permanent address: National Defence Research Institute, Department 4, S-172 04 Sundbyberg, Sweden.

(Received September 4, 1978; )
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