Involvement of zeaxanthin and of the Cbr protein in the repair of photosystem II from photoinhibition in the green alga Dunaliella salina

AbstractA light-sensitive and chlorophyll (Chl)-deficient mutant of the green alga Dunaliella salina (dcd1) showed an amplified response to irradiance stress compared to the wild-type. The mutant was yellow–green under low light (100 μmol photons m−2 s−1) and yellow under high irradiance (2000 μmol photons m−2 s−1). The mutant had lower levels of Chl, lower levels of light harvesting complex II, and a smaller Chl antenna size. The mutant contained proportionately greater amounts of photodamaged photosystem (PS) II reaction centers in its thylakoid membranes, suggesting a greater susceptibility to photoinhibition. This phenotype was more pronounced under high than low irradiance. The Cbr protein, known to accumulate when D. salina is exposed to irradiance stress, was pronouncedly expressed in the mutant even under low irradiance. This positively correlated with a higher zeaxanthin content in the mutant. Cbr protein accumulation, xanthophyll cycle de-epoxidation state, and fraction of photodamaged PSII reaction centers in the thylakoid membrane showed a linear dependence on the chloroplast ‘photoinhibition index’, suggesting a cause-and-effect relationship between photoinhibition, Cbr protein accumulation and xanthophyll cycle de-epoxidation state. These results raised the possibility of zeaxanthin and Cbr involvement in the PSII repair process through photoprotection of the partially disassembled, and presumably vulnerable, PSII core complexes from potentially irreversible photooxidative bleaching

Comparison of Marine Microalgae Culture Systems for Fuels Production and Carbon Sequestration

The dual problems of global fossil fuels supplies and global warming focus attention on the need to develop technologies that can provide large amounts of renewable fuels without contributing to global warming. The capture of power plant flue gas CO2 using microalgae cultures is one potential technology that could meet this objective. The central R&D issues are the design and operation of low-cost algal mass culture systems and the development of algal strains and cultivation techniques that can achieve very high biomass productivities. The major objective of this project was to develop mass culture techniques that could result in greatly increased biomass productivities, well above the about 50 metric tons per hectare per year (mt/ha/y) currently achievable. In this project, two marine microalgae species, the diatom Cyclotella sp.. and the green alga Tetraselmis sp., were cultivated on seawater in both open ponds and closed photo bioreactors, under a variety of different cultivation conditions. Simultaneous operation of the closed photo bioreactors and open ponds demonstrated similar productivities, under the same operating conditions. Thus the very expensive closed systems do not provide any major or inherent advantages in microalgae production over open ponds. Mutants of Cyclotella sp. were developed that exhibited reduced pigment content, which theoretically would result in greatly increased productivities when grown under full sunlight. However, in open ponds, these mutant strains exhibited similar productivities as the parental strains. The mutant strains all grew relatively slowly, suggesting that additional mutations masked whatever inherent potential for increased productivities may have resulted from the reduced pigment content. Research is still required to develop improved low pigment strains. When open pond cultures were exposed to intermittent sunlight, by partially covering the ponds with slats, solar conversion efficiencies increased dramatically, by over 50%. Although such techniques are not directly applicable to practical processes, the experiments demonstrated the inherent potential of algal mass cultures to achieve very high productivities. Nitrogen limited pond cultures demonstrated that it is possible to produce biomass with a potentially high content of carbohydrates or oils (although these were not directly measured in these experiments), without reducing achievable productivities. This suggested that microalgae biomass suitable for conversion to biofuels (ethanol or biodiesel) could be produced without compromising productivity. Experiments combining both light modulation and nitrogen limitation indicated possibly synergistic effects. The goal of developing practical and economic processes for the sustainable production of renewable fuels with microalgae pond cultures using power plant flue gases as sources of CO2 was advanced by these studies, but requires more work. Most important is the research, development and demonstration in outdoor pond cultures of algal strains with low pigment content. Such strains are the most likely approach to achieve, in combination with the other mass culture techniques investigated in this study, the very high productivities, above 100 mt/ha/y (45 t/acre/y), that are the goal in this field. The projected economics for such a process suggests that, as for higher plant biofuel production, microalgae biofuels production should be developed as a multiproduct process providing additional higher value co-products

Phytoene Accumulation in the Novel Microalga Chlorococcum sp. Using the Pigment Synthesis Inhibitor Fluridone

Carotenoids are lipophilic pigments found in plants and algae, as well as some bacteria, archaea, and fungi that serve two functions—(1) as light harvesting molecules—primary carotenoids, and (2) as antioxidants, acting against reactive oxygen species–secondary carotenoids. Because of their strong antioxidant properties, they are also valuable for the development of anti-aging and photo-protective cosmetic applications. Of particular interest is the carotenoid phytoene, for its colorless and UV absorption characteristics. In this study, we targeted a reduction of phytoene desaturase (PDS) activity with the pigment-inhibiting herbicide 1-methyl-3-phenyl-5-[3- (trifluoromethyl)phenyl]pyridin-4-one (fluridone), which leads to the over-accumulation of phytoene in the recently characterized microalgal strain Chlorococcum sp. (UTEX B 3056). After post-incubation with fluridone, phytoene levels were measured at ~33 ug/mg cell tissue, as opposed to non-detectable levels in control cultures. Hence, the novel microalga Chlorococcum sp. is a viable candidate for the production of the high-value carotenoid phytoene and subsequent applications in cosmeceuticals, as well as more obvious nutraceutical and pharmaceutical applications

Draft Nuclear Genome, Complete Chloroplast Genome, and Complete Mitochondrial Genome for the Biofuel/ Bioproduct Feedstock Species Scenedesmus obliquus Strain DOE0152z

The green alga Scenedesmus obliquus is an emerging platform species for the industrial production of biofuels. Here, we report the draft assembly and annotation for the nuclear, plastid, and mitochondrial genomes of S. obliquus strain DOE0152z

The \u3ci\u3eChlorella variabilis\u3c/i\u3e NC64A Genome Reveals Adaptation to Photosymbiosis, Coevolution with Viruses, and Cryptic Sex

Chlorella variabilis NC64A, a unicellular photosynthetic green alga (Trebouxiophyceae), is an intracellular photobiont of Paramecium bursaria and a model system for studying virus/algal interactions. We sequenced its 46-Mb nuclear genome, revealing an expansion of protein families that could have participated in adaptation to symbiosis. NC64A exhibits variations in GC content across its genome that correlate with global expression level, average intron size, and codon usage bias. Although Chlorella species have been assumed to be asexual and nonmotile, the NC64A genome encodes all the known meiosis-specific proteins and a subset of proteins found in flagella. We hypothesize that Chlorella might have retained a flagella-derived structure that could be involved in sexual reproduction. Furthermore, a survey of phytohormone pathways in chlorophyte algae identified algal orthologs of Arabidopsis thaliana genes involved in hormone biosynthesis and signaling, suggesting that these functions were established prior to the evolution of land plants. We show that the ability of Chlorella to produce chitinous cell walls likely resulted from the capture of metabolic genes by horizontal gene transfer from algal viruses, prokaryotes, or fungi. Analysis of the NC64A genome substantially advances our understanding of the green lineage evolution, including the genomic interplay with viruses and symbiosis between eukaryotes

Draft Nuclear Genome Sequence of the Halophilic and Beta-Carotene- Accumulating Green Alga Dunaliella salina Strain CCAP19/18

The halotolerant alga Dunaliella salina is a model for stress tolerance and is used commercially for production of beta-carotene (pro-vitamin A). The presented draft genome of the genuine strain CCAP19/18 will allow investigations into metabolic processes involved in regulation of stress responses, including carotenogenesis and adaptations to life in high-salinity environments

Review of the cultivation program within the National Alliance for Advanced Biofuels and Bioproducts

The cultivation efforts within the National Alliance for Advanced Biofuels and Bioproducts (NAABB)were developed to provide four major goals for the consortium, which included biomass production for downstream experimentation, development of new assessment tools for cultivation, development of new cultivation reactor technologies, and development of methods for robust cultivation. The NAABB consortium test beds produced over 1500 kg of biomass for downstream processing. The biomass production included a number of model production strains, but also took into production some of the more promising strains found through the prospecting efforts of the consortium. Cultivation efforts at large scale are intensive and costly, therefore the consortium developed tools and models to assess the productivity of strains under various environmental conditions, at lab scale, and validated these against scaled outdoor production systems. Two new pond-based bioreactor designs were tested for their ability to minimize energy consumption while maintaining, and even exceeding, the productivity of algae cultivation compared to traditional systems. Also, molecular markers were developed for quality control and to facilitate detection of bacterial communities associated with cultivated algal species, including the Chlorella spp. pathogen, Vampirovibrio chlorellavorus,which was identified in at least two test site locations in Arizona and New Mexico. Finally, the consortium worked on understanding methods to utilize compromised municipal waste water streams for cultivation. This review provides an overview of the cultivation methods and tools developed by the NAABB consortium to produce algae biomass, in robust low energy systems, for biofuel production

Review of the algal biology program within the National Alliance for Advanced Biofuels and Bioproducts

In 2010,when the National Alliance for Advanced Biofuels and Bioproducts (NAABB) consortiumbegan, littlewas known about themolecular basis of algal biomass or oil production. Very fewalgal genome sequenceswere available and efforts to identify the best-producing wild species through bioprospecting approaches had largely stalled after the U.S. Department of Energy\u27s Aquatic Species Program. This lack of knowledge included how reduced carbon was partitioned into storage products like triglycerides or starch and the role played bymetabolite remodeling in the accumulation of energy-dense storage products. Furthermore, genetic transformation and metabolic engineering approaches to improve algal biomass and oil yields were in their infancy. Genome sequencing and transcriptional profiling were becoming less expensive, however; and the tools to annotate gene expression profiles under various growth and engineered conditions were just starting to be developed for algae. It was in this context that an integrated algal biology program was introduced in the NAABB to address the greatest constraints limiting algal biomass yield. This review describes the NAABB algal biology program, including hypotheses, research objectives, and strategies to move algal biology research into the twenty-first century and to realize the greatest potential of algae biomass systems to produce biofuels