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Simple steps to enable reproducibility: culture conditions affecting Chlamydomonas growth and elemental composition

Hui, Colleen and Schmollinger, Stefan and Strenkert, Daniela and Holbrook, Kristen and Montgomery, Hayden R. and Chen, Si and Nelson, Hosea M. and Weber, Peter K. and Merchant, Sabeeha S. (2022) Simple steps to enable reproducibility: culture conditions affecting Chlamydomonas growth and elemental composition. Plant Journal, 111 (4). pp. 995-1014. ISSN 0960-7412. doi:10.1111/tpj.15867.

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Even subtle modifications in growth conditions elicit acclimation responses affecting the molecular and elemental makeup of organisms, both in the laboratory and in natural habitats. We systematically explored the effect of temperature, pH, nutrient availability, culture density and access to CO₂ and O₂ in laboratory-grown algal cultures on growth rate, the ionome, and the ability to accumulate Fe. We found algal cells accumulate Fe in alkaline conditions, even more so when excess Fe is present, coinciding with a reduced growth rate. Using a combination of Fe-specific dyes, X-ray fluorescence microscopy and NanoSIMS, we show that the alkaline-accumulated Fe was intracellularly sequestered into acidocalcisomes, which are localized towards the periphery of the cells. At high photon flux densities, Zn and Ca specifically over-accumulate, while Zn alone accumulates at low temperatures. The impact of aeration was probed by reducing shaking speeds and changing vessel fill-levels; the former increased the Cu quota of cultures, the latter resulted in a reduction in P, Ca, and Mn at low fill-levels. Trace element quotas were also affected in stationary phase, where specifically Fe, Cu and Zn accumulate. Cu accumulation here depends inversely on the Fe concentration of the medium. Individual laboratory strains accumulate Ca, P and Cu to different levels. All together, we identified a set of specific changes to growth rate, elemental composition, and the capacity to store Fe in response to subtle differences in culturing conditions of Chlamydomonas, affecting experimental reproducibility. Accordingly, we recommend that these variables be recorded and reported as associated metadata.

Item Type:Article
Related URLs:
URLURL TypeDescription
Schmollinger, Stefan0000-0002-7487-8014
Strenkert, Daniela0000-0002-3420-1332
Nelson, Hosea M.0000-0002-4666-2793
Merchant, Sabeeha S.0000-0002-2594-509X
Additional Information:© 2022 Wiley. Issue Online: 16 August 2022; Version of Record online: 11 July 2022; Accepted manuscript online: 14 June 2022; Manuscript accepted: 03 June 2022; Manuscript revised: 01 June 2022; Manuscript received: 15 February 2022.
Subject Keywords:ICP; iron; elemental composition; ionome; stress; Chlamydomonas reinhardtii
Issue or Number:4
Record Number:CaltechAUTHORS:20220708-319697100
Persistent URL:
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:115430
Deposited By: Tony Diaz
Deposited On:08 Jul 2022 23:15
Last Modified:14 Sep 2022 19:36

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