Truncated chlorophyll antenna size of the photosystems?a practical method to improve microalgal productivity and hydrogen production in mass culture

Polle, Juergen; Saradadevi, K.; Jin, EonSeon; Masuda, Tatsuru; Melis, Anastasios

doi:10.1016/s0360-3199(02)00116-7

Cited by 180 publications

(88 citation statements)

References 29 publications

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“…High CO 2 atmosphere was achieved by incubating the plates in BBL GasPak CO 2 pouches (BectonDickinson, Franklin Lakes, NJ). In addition to these 14 treatments, the maximum chlorophyll fluorescence of the dark-grown TAP stock cultures was measured using video imaging (Polle et al, 2002).…”

Section: Screeningmentioning

confidence: 99%

Functional Genomics of Eukaryotic Photosynthesis Using Insertional Mutagenesis of Chlamydomonas reinhardtii

et al. 2005

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The unicellular green alga Chlamydomonas reinhardtii is a widely used model organism for studies of oxygenic photosynthesis in eukaryotes. Here we describe the development of a resource for functional genomics of photosynthesis using insertional mutagenesis of the Chlamydomonas nuclear genome. Chlamydomonas cells were transformed with either of two plasmids conferring zeocin resistance, and insertional mutants were selected in the dark on acetate-containing medium to recover lightsensitive and nonphotosynthetic mutants. The population of insertional mutants was subjected to a battery of primary and secondary phenotypic screens to identify photosynthesis-related mutants that were pigment deficient, light sensitive, nonphotosynthetic, or hypersensitive to reactive oxygen species. Approximately 9% of the insertional mutants exhibited 1 or more of these phenotypes. Molecular analysis showed that each mutant line contains an average of 1.4 insertions, and genetic analysis indicated that approximately 50% of the mutations are tagged by the transforming DNA. Flanking DNA was isolated from the mutants, and sequence data for the insertion sites in 50 mutants are presented and discussed.As with other model organisms, the availability of genome sequence data is revolutionizing and revitalizing research into the biology of the unicellular green alga Chlamydomonas reinhardtii Ledford et al., 2005). Over the past four decades, many fundamental insights into the structure, function, assembly, and regulation of the photosynthetic apparatus have come from studies of Chlamydomonas, which offer several advantages for the genetic dissection of eukaryotic photosynthesis (for review, see Davies and Grossman, 1998; Hippler et al., 1998;Grossman, 2000;Rochaix, 2001). First and foremost, photosynthesis is fully dispensable in Chlamydomonas, as cells can grow heterotrophically in the dark using acetate as a sole carbon source. Cells grown in the dark, however, still synthesize and assemble a fully functional photosynthetic apparatus. This allows the isolation and analysis of mutants that are unable to perform photosynthesis, and lightsensitive mutants can be maintained in complete darkness. Because Chlamydomonas is predominantly maintained in a haploid form, it is not necessary to generate homozygous nuclear mutants, and mutants affecting photosynthesis can be screened immediately following mutagenesis. Chlamydomonas has an easily controlled and rapid sexual cycle (approximately 2 weeks) with the possibility of tetrad analysis, which facilitates genetic analysis. Its rapid cell-doubling time (approximately 10 h) and microbial lifestyle mean that it is easy to grow homogeneous cultures on any scale, simplifying physiological and biochemical characterization in comparison to multicellular land plants (Ledford et al., 2005). By way of example, the application of inhibitors and generators of various types of reactive oxygen species results in uniform uptake of the chemical by each cell. In land plants, multicellularity leads to differential ...

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Section: Screeningmentioning

confidence: 99%

Functional Genomics of Eukaryotic Photosynthesis Using Insertional Mutagenesis of Chlamydomonas reinhardtii

et al. 2005

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“…Work is ongoing to improve light conversion efficiency through the truncation of light-harvesting antenna complexes, an approach already proven using PNS bacteria (see below, section 2.1.2) (Akkerman et al 2002;Polle et al 2002). Other approaches are to study and develop O 2 -tolerant hydrogenases (Ghirardi et al 2005;Tosatto et al 2008) and to express clostridial hydrogenase in non-heterocystous cyanobacteria, the aim being to engineer the rapid and ATPindependent (hydrogenase-mediated) H 2 production by direct photolysis in a fast-growing host organism, possibly overcoming O 2 -inhibition through strong expression .…”

Section: 11: Photoautotrophic Microorganismsmentioning

confidence: 99%

Integrating dark and light bio-hydrogen production strategies: towards the hydrogen economy

Redwood

Paterson-Beedle

Macaskie

2008

Rev Environ Sci Biotechnol

140

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Biological methods of hydrogen production are preferable to chemical methods because of the possibility to use sunlight, CO 2 and organic wastes as substrates for environmentally benign conversions, under moderate conditions. By combining different microorganisms with different capabilities, the individual strengths of each may be exploited and their weaknesses overcome. Mechanisms of bio-hydrogen production are described and strategies for their integration are discussed. Dual systems can be divided broadly into wholly light-driven systems (with microalgae/cyanobacteria as the 1 st stage) and partially light-driven systems (with a dark, fermentative initial reaction). Review and evaluation of published data suggests that the latter type of system holds greater promise for industrial application. This is because the calculated land area required for a wholly light-driven dual system would be too large for either centralised (macro-) or decentralised (micro-)energy generation. The potential contribution to the hydrogen economy of partially light-driven dual systems is overviewed alongside that of other bio-fuels such as bio-methane and bio-ethanol.Redwood et al. -Dual systems for Bio-H 2 -Reviews in Environ. Sci. Bio/Technol. 8:149 http://dx

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“…It has been shown that high-density cultures of microalgae with a truncated Chl antenna size are photosynthetically more productive under bright sunlight due to the elimination of overabsorption and wasteful dissipation of excess energy Ueda, 1997, 1999;Melis et al, 1999;Polle et al, 2002Polle et al, , 2003Melis, 2009). Identification of genes that confer a permanently truncated light-harvesting antenna size phenotype in plants and algae is thus of interest, as they could be applied in efforts to improve solar-to-product conversion efficiencies (Mitra and Melis, 2008;Melis, 2009;Ort et al, 2011).…”

mentioning

confidence: 99%

Truncated Photosystem Chlorophyll Antenna Size in the Green Microalga Chlamydomonas reinhardtii upon Deletion of the TLA3-CpSRP43 Gene

et al. 2012

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The truncated light-harvesting antenna size3 (tla3) DNA insertional transformant of Chlamydomonas reinhardtii is a chlorophylldeficient mutant with a lighter green phenotype, a lower chlorophyll (Chl) per cell content, and higher Chl a/b ratio than corresponding wild-type strains. Functional analyses revealed a higher intensity for the saturation of photosynthesis and greater light-saturated photosynthetic activity in the tla3 mutant than in the wild type and a Chl antenna size of the photosystems that was only about 40% of that in the wild type. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western-blot analyses showed that the tla3 strain was deficient in the Chl a/b light-harvesting complex. Molecular and genetic analyses revealed a single plasmid insertion in chromosome 4 of the tla3 nuclear genome, causing deletion of predicted gene g5047 and plasmid insertion within the fourth intron of downstream-predicted gene g5046. Complementation studies defined that gene g5047 alone was necessary and sufficient to rescue the tla3 mutation. Gene g5047 encodes a C. reinhardtii homolog of the chloroplast-localized SRP43 signal recognition particle, whose occurrence and function in green microalgae has not hitherto been investigated. Biochemical analysis showed that the nucleus-encoded and chloroplast-localized CrCpSRP43 protein specifically operates in the assembly of the peripheral components of the Chl a/b light-harvesting antenna. This work demonstrates that cpsrp43 deletion in green microalgae can be employed to generate tla mutants with a substantially diminished Chl antenna size. The latter exhibit improved solar energy conversion efficiency and photosynthetic productivity under mass culture and bright sunlight conditions.

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Truncated chlorophyll antenna size of the photosystems?a practical method to improve microalgal productivity and hydrogen production in mass culture

Cited by 180 publications

References 29 publications

Functional Genomics of Eukaryotic Photosynthesis Using Insertional Mutagenesis of Chlamydomonas reinhardtii

Functional Genomics of Eukaryotic Photosynthesis Using Insertional Mutagenesis of Chlamydomonas reinhardtii

Integrating dark and light bio-hydrogen production strategies: towards the hydrogen economy

Truncated Photosystem Chlorophyll Antenna Size in the Green Microalga Chlamydomonas reinhardtii upon Deletion of the TLA3-CpSRP43 Gene

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