Poster (Scientific congresses and symposiums)
A forward genetic screen to identify hydrogenase-deficient mutants in the unicellular green alga Chlamydomonas reinhardtii
Emonds-Alt, Barbara; Godaux, Damien; Cardol, Pierre et al.
2014The 20th World Hydrogen Energy Conference
 

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Keywords :
Chlamydomonas; hydrogen; mutagenesis
Abstract :
[en] The ability of the unicellular green alga Chlamydomonas reinhardtii to evolve molecular hydrogen (H2) is due to the presence of oxygen-sensitive Fe-hydrogenases (HydA1/2), expressed in anoxic conditions that drive the photosynthetic electron flow to reduce protons into H2. In order to identify new players involved in H2 photoproduction in Chlamydomonas, an insertion mutant library was generated using cassettes conferring resistance to hygromycin or paromomycin. Hydrogenase activity is physiologically relevant during a transition from dark anoxia to light. In dark anoxic conditions, the cellular redox poise is high and the photosynthetic electron transport chain is fully reduced. Upon illumination, hydrogenase activity allows the reoxidation of photosynthetic intersystem electron carriers until oxic conditions and carbon fixation ability are restored. We thus designed an in vivo fluorescence imaging screen based on the different kinetics of photosynthesis induction between wild type and hydrogenase-deficient mutants [1]. At this stage, three putative hydrogenase mutants have been identified on 10,000 transformants. Molecular characterization of the insertion site of the resistance cassette by TAIL-PCR and genetic analyses of the linkage between the antibiotic resistance and the fluorescence phenotype showed that one mutant was untagged with the resistance while two tagged mutants were deficient for the HydG assembly factor.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Emonds-Alt, Barbara ;  Université de Liège - ULiège > Département des sciences de la vie > Génétique et physiologie des microalgues
Godaux, Damien ;  Université de Liège - ULiège > Département des sciences de la vie > Génétique et physiologie des microalgues
Cardol, Pierre  ;  Université de Liège - ULiège > Département des sciences de la vie > Génétique et physiologie des microalgues
Remacle, Claire  ;  Université de Liège - ULiège > Département des sciences de la vie > Génétique et physiologie des microalgues
Language :
English
Title :
A forward genetic screen to identify hydrogenase-deficient mutants in the unicellular green alga Chlamydomonas reinhardtii
Publication date :
15 June 2014
Number of pages :
A0
Event name :
The 20th World Hydrogen Energy Conference
Event organizer :
International Association for Hydrogen Energy
Event place :
Gwangju Metropolitan city, South Korea
Event date :
du 15 juin 2014 au 20 juin 2014
Audience :
International
Funders :
FRIA - Fonds pour la Formation à la Recherche dans l'Industrie et dans l'Agriculture [BE]
Available on ORBi :
since 29 August 2014

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