The determination of activity of the enzyme Rubisco in cell extracts of the dinoflagellate alga Symbiodinium sp. by manganese chemiluminescence and its response to short-term thermal stress of the alga

Plant, Cell & Environment
Ross McC LilleyAnthony W D Larkum

Abstract

The dinoflagellate alga Symbiodinium sp., living in symbiosis with corals, clams and other invertebrates, is a primary producer in coral reefs and other marine ecosystems. The function of the carbon-fixing enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) in dinoflagellates is difficult to study because its activity is rapidly lost after extraction from the cell. We report procedures for the extraction of Rubisco from Symbiodinium cells and for stable storage. We describe a continuous assay for Rubisco activity in these crude cell extracts using the Mn(2+) chemiluminescence of Rubisco oxygenase. Chemiluminescence time courses exhibited initial transients resembling bacterial Form II Rubisco, followed by several minutes of linearly decreasing activity. The initial activity was determined from extrapolation of this linear section of the time course. The activity of fast-frozen cell extracts was stable at -80 degrees C and, after thawing and storage on ice, remained stable for up to 1 h before declining non-linearly. Crude cell extracts bound [(14)C] 2-carboxy-D-arabitinol 1,5-bisphosphate to a high molecular mass fraction separable by gel filtration chromatography. After pre-treatment of Symbiodinium cell cultures ...Continue Reading

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Citations

Jul 3, 2013·The New Phytologist·Patrick BradingDavid J Suggett
Jul 16, 2013·Plant Science : an International Journal of Experimental Plant Biology·Ling HaoZhaohu Li
Feb 9, 2017·Plant Science : an International Journal of Experimental Plant Biology·Yongjiang SunHeng Zhai
Aug 9, 2017·Physiologia Plantarum·Arnold J Bloom, Petra Kameritsch
Oct 17, 2019·Global Change Biology·David J Suggett, David J Smith
Apr 5, 2018·Frontiers in Microbiology·Jinjin DiaoWeiwen Zhang
May 20, 2020·Science Advances·P BuergerM J H van Oppen
Aug 26, 2021·Trends in Microbiology·Justin Maire, Madeleine J H van Oppen

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