• Energy Research

Microalgae represent a carbon-neutral source of bulk biomass, for extraction of high-value compounds and production of renewable fuels. Due to their high metabolic activity and reproduction rates, species of the genus Chlorella are highly productive when cultivated in photobioreactors. However, wild-type strains show biological limitations making algal bioproducts expensive compared to those extracted from other feedstocks. Such constraints include inhomogeneous light distribution due to high optical density of the culture, and photoinhibition of the surface-exposed cells. Thus, the domestication of algal strains for industry makes it increasingly important to select traits aimed at enhancing light-use efficiency while withstanding excess light stress. Carotenoids have a crucial role in protecting against photooxidative damage and, thus, represent a promising target for algal domestication. We applied chemical mutagenesis to Chlorella vulgaris and selected for enhanced tolerance to the carotenoid biosynthesis inhibitor norflurazon. The NFR (norflurazon-resistant) strains showed an increased carotenoid pool size and enhanced tolerance towards photooxidative stress. Growth under excess light revealed an improved carbon assimilation rate of NFR strains with respect to WT. We conclude that domestication of Chlorella vulgaris, by optimizing both carotenoid/chlorophyll ratio and resistance to photooxidative stress, boosted light-to-biomass conversion efficiency under high light conditions typical of photobioreactors. Comparison with strains previously reported for enhanced tolerance to singlet oxygen, reveals that ROS resistance in Chlorella is promoted by at least two independent mechanisms, only one of which is carotenoid-dependent.

AbstractBackgroundMicroalgae are coming to the spotlight due to their potential applications in a wide number of fields ranging from the biofuel to the pharmaceutical sector. However, several factors such as low productivity, expensive harvesting procedures and difficult metabolite extractability limit their full utilization at industrial scale. Similarly to the successful employment of enzymatic arsenals from lignocellulolytic fungi to convert lignocellulose into fermentable sugars for bioethanol production, specific algalytic formulations could be used to improve the extractability of lipids from microalgae to produce biodiesel. Currently, the research areas related to algivorous organisms, algal saprophytes and the enzymes responsible for the hydrolysis of algal cell wall are still little explored.ResultsHere, an algal trap method for capturing actively growing microorganisms was successfully used to isolate a filamentous fungus, that was identified by whole-genome sequencing, assembly and annotation as a novelPenicilliumsumatraenseisolate. The fungus, classified asP.sumatraenseAQ67100, was able to assimilate heat-killedChlorellavulgariscells by an enzymatic arsenal composed of proteases such as dipeptidyl- and amino-peptidases, β-1,3-glucanases and glycosidases including α- and β-glucosidases, β-glucuronidase, α-mannosidases and β-galactosidases. The treatment ofC.vulgariswith the filtrate fromP.sumatraenseAQ67100 increased the release of chlorophylls and lipids from the algal cells by 42.6 and 48.9%, respectively.ConclusionsThe improved lipid extractability fromC.vulgarisbiomass treated with the fungal filtrate highlighted the potential of algal saprophytes in the bioprocessing of microalgae, posing the basis for the sustainable transformation of algal metabolites into biofuel-related compounds.

Additional file 1: Figure S1. Design of the algal trap. Figure S2. Morphological characteristics of the unknown fungal isolate. Figure S3. Extraction of genomic DNA (gDNA) from the unknown fungal isolate. Figure S4. Growth of P. sumatraense AQ67100 in different algal-supplemented media. Figure S5. Time-course analysis of GH activities from P. sumatraense AQ67100 cultures as grown in algal-supplemented media. Figure S6. Analysis of GH activities from P. sumatraense AQ67100 cultures upon growth in different cell wall polysaccharide-supplemented media. Figure S7. Evaluation of protease activity in C.v.-filtrate. Figure S8. Protein bands putatively ascribable to the main algal-degrading enzymes. Figure S9. Growth of P. sumatraense AQ67100 in a chitin-supplemented medium.

In light of ongoing climate changes in wine-growing regions, the selection of drought-tolerant rootstocks is becoming a crucial factor for developing a sustainable viticulture. In this study, M4, a new rootstock genotype that shows tolerance to drought, was compared from a genomic and transcriptomic point of view with the less drought-tolerant genotype 101.14. The root and leaf transcriptome of both 101.14 and the M4 rootstock genotype was analysed, following exposure to progressive drought conditions. Multifactorial analyses indicated that stress treatment represents the main factor driving differential gene expression in roots, whereas in leaves the genotype is the prominent factor. Upon stress, M4 roots and leaves showed a higher induction of resveratrol and flavonoid biosynthetic genes, respectively. The higher expression of VvSTS genes in M4, confirmed by the accumulation of higher levels of resveratrol in M4 roots compared with 101.14, was coupled to an up-regulation of several VvWRKY transcription factors. Interestingly, VvSTS promoter analyses performed on both the resequenced genomes highlighted a significantly higher number of W-BOX elements in the tolerant genotype. It is proposed that the elevated synthesis of resveratrol in M4 roots upon water stress could enhance the plant's ability to cope with the oxidative stress usually associated with water deficit.

AbstractIn this work, we investigated the molecular basis of autotrophic vs. mixotrophic growth of Chlorella sorokiniana, one of the most productive microalgae species with high potential to produce biofuels, food and high value compounds. To increase biomass accumulation, photosynthetic microalgae are commonly cultivated in mixotrophic conditions, adding reduced carbon sources to the growth media. In the case of C. sorokiniana, the presence of acetate enhanced biomass, proteins, lipids and starch productivity when compared to autotrophic conditions. Despite decreased chlorophyll content, photosynthetic properties were essentially unaffected while differential gene expression profile revealed transcriptional regulation of several genes mainly involved in control of carbon flux. Interestingly, acetate assimilation caused upregulation of phosphoenolpyruvate carboxylase enzyme, enabling potential recovery of carbon atoms lost by acetate oxidation. The obtained results allowed to associate the increased productivity observed in mixotrophy in C. sorokiniana with a different gene regulation leading to a fine regulation of cell metabolism.

Abstract Microalgae represent a potential sustainable source of molecules and materials and the improvement of the knowledge of their metabolic regulation is essential to maximize their potential. Nutrients deprivation stimulates the accumulation of reserve lipids, triacylglycerols but also inhibits photosynthesis with a negative impact on biomass production and therefore overall productivity. In this work, the seawater microalga Nannochloropsis gaditana was cultivated long term in a semi-continuous system where the concentrations of nitrogen or phosphorus were limiting but still sufficient to sustain growth indefinitely, to highlight the response to a long-term nutrient limitation and distinguishing it from the one to acute short-term stress. N. gaditana cells can acclimate to chronic nutrients limitation maintaining photosynthetic activity while also accumulating lipids. Both nitrogen and phosphorus limitation induced an increase of triacylglycerols content, although not by the induction of the synthesis of fatty acids but rather by modulating the fluxes of reduced carbon molecules toward lipid biosynthesis. Photosynthetic activity was maintained under P limitation while this was strongly affected by nitrogen depletion, where proteins of photosynthetic apparatus were largely reduced in content but still maintained their functionality and were able to achieve half of the biomass productivity with 30% of the nitrogen supply.