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Mutagenesis and phenotypic selection as a strategy toward domestication of Chlamydomonas reinhardtii strains for improved performance in photobioreactors

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Abstract

Microalgae have a valuable potential for biofuels production. As a matter of fact, algae can produce different molecules with high energy content, including molecular hydrogen (H2) by the activity of a chloroplastic hydrogenase fueled by reducing power derived from water and light energy. The efficiency of this reaction, however, is limited and depends from an intricate relationships between oxygenic photosynthesis and mitochondrial respiration. The way toward obtaining algal strains with high productivity in photobioreactors requires engineering of their metabolism at multiple levels in a process comparable to domestication of crops that were derived from their wild ancestors through accumulation of genetic traits providing improved productivity under conditions of intensive cultivation as well as improved nutritional/industrial properties. This holds true for the production of any biofuels from algae: there is the need to isolate multiple traits to be combined and produce organisms with increased performances. Among the different limitations in H2 productivity, we identified three with a major relevance, namely: (i) the light distribution through the mass culture; (ii) the strong sensitivity of the hydrogenase to even very low oxygen concentrations; and (iii) the presence of alternative pathways, such as the cyclic electron transport, competing for reducing equivalents with hydrogenase and H2 production. In order to identify potentially favorable mutations, we generated a collection of random mutants in Chlamydomonas reinhardtii which were selected through phenotype analysis for: (i) a reduced photosynthetic antenna size, and thus a lower culture optical density; (ii) an altered photosystem II activity as a tool to manipulate the oxygen concentration within the culture; and (iii) State 1–State 2 transition mutants, for a reduced cyclic electron flow and maximized electrons flow toward the hydrogenase. Such a broad approach has been possible thanks to the high throughput application of absorption/fluorescence optical spectroscopy methods. Strong and weak points of this approach are discussed.

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Abbreviations

Chl:

Chlorophyll

Car:

Carotenoid

LHC:

Light-harvesting complex

H2 :

Hydrogen

Chl a/b :

Chlorophyll a to b ratio

Fo/Fv/Fm :

Basal/variable/maximum fluorescence

St:

State transition

PSI (II):

Photosystem I(II)

WT:

Wild type

DCMU:

3-(3,4-Dichlorophenyl)-1,1-dimethylurea

NaN3 :

Sodium azide

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Acknowledgments

This study was supported by the Italian Ministry of Agriculture, Project IDROBIO.

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  1. Claudia Catalanotti

    Present address: Department of Plant Biology, Carnegie Institution for Science, 260 Panama Street, Stanford, CA, 94305, USA

Authors and Affiliations

  1. Dipartimento di Biotecnologie, Università di Verona, Strada Le Grazie 15, 37134, Verona, Italy

    Giulia Bonente, Cinzia Formighieri, Manuela Mantelli & Roberto Bassi

  2. Italian National Agency for New Technologies, Energy and Sustainable Development (ENEA), Casaccia Research Center, Via Anguillarese 301, 00123, Rome, Italy

    Claudia Catalanotti & Giovanni Giuliano

  3. Dipartimento di Biologia, Università di Padova, Via U. Bassi 58/B, 34134, Padova, Italy

    Tomas Morosinotto

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  1. Giulia Bonente
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Correspondence to Roberto Bassi.

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Bonente, G., Formighieri, C., Mantelli, M. et al. Mutagenesis and phenotypic selection as a strategy toward domestication of Chlamydomonas reinhardtii strains for improved performance in photobioreactors. Photosynth Res 108, 107–120 (2011). https://doi.org/10.1007/s11120-011-9660-2

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