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Temperature is a crucial factor affecting microalgae CO 2 capture and utilization. However, an in-depth understanding of how microalgae respond to temperature stress is still unclear. In particular, the regulation mechanism under opposite temperature (heat and cold) stress had not yet been reported. In this study, the physicochemical properties and transcription level of related genes of microalgae Auxenochlorella protothecoides UTEX 2341 under heat and cold stress were investigated. Heat stress (Hs) caused a drastic increase of reactive oxygen species (ROS) in UTEX 2341. As key elements responded to Hs, superoxide dismutase (SOD) enzyme increased by 150%, 70%, and 30% in activity, and nitric oxide (NO) grew by 409.6%, 212.5%, and 990.4% in content compared with the control at 48 h, 96 h, 168 h. Under cold stress (Cs), ROS increased in the early stage and decreased in the later stage. As key factors responded to Cs, proline (Pro) increased respectively by 285%, 383%, and 81% in content, and heat shock transcriptional factor HSFA1d increased respectively by 161%, 71%, and 204% in transcript level compared with the control at 48 h, 96 h, 168 h. Furthermore, the transcript level of antioxidant enzymes or antioxidant coding genes was consistent with the changing trend of enzymes activity or antioxidant content. Notably, both glutathione (GSH) and heat shock protein 97 (hsp 97) were up-regulated in response to Hs and Cs. In conclusion, GSH and hsp 97 were the core elements of UTEX 2341 in response to both Hs and Cs. SOD and NO were the key elements that responded to Hs, while proline and HSFA1d were the key elements that responded to Cs. This study provided a basis for the understanding of the response mechanism of microalgae under temperature stress and the improvement of the microalgae tolerance to temperature stress., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Microalgae are the largest CO 2 fixer and O 2 producer on the earth and occupy an increasingly important position in human life and production. Various environmental factors have a significant impact on the growth and metabolism of microalgae. As global warming intensifies, heat stress has become a crucial factor affecting the microalgae industry. However, till now, it has not been clear how microalgae sensed the temperature stress, transmitted stress signals and adjusted in intracellular metabolic pathways. In this study, the growth of microalgae Auxenochlorella protothecoides UTEX2341 was inhibited at 32 °C, but the single cell dry weight increased. The cell component analyses showed that both the carbohydrate and total protein content decreased significantly, while the lipid content increased by 158%. Meanwhile, the intracellular Ca 2+ concentration increased continuously, with a maximum increase of 1.65 times. According to the transcriptome analyses, the up-regulation of Ca 2+ influx channel protein mid1-complementing activity 1 (MCA1) gene and the down-regulation of efflux channel protein cation exchanger 1(CAX) and autoinhibited Ca 2+ -ATPase 1 (ACA1) genes in cytoplasmic membrane jointly facilitated the increase of Ca 2+ in the cytoplasm. Coexpression network analysis indicated that the fluctuation of Ca 2+ in the cytoplasm could activate the expression of transcription factors MYB3 and AP2-4 through calmodulin (CAM) and calcium-dependent protein kinase (CDPK), and then regulate glycerol-3-phosphate acyltransferases (GPAT) at the beginning of TAG synthesis and diacylglycerol acyltransferase (DGAT)/phospholipid: diacylglycerol acyltransferase (PDAT) in the last step of TAG synthesis. Furthermore, the addition of Ca 2+ specific chelator BAPTA-AM inhibited the expression of GPAT, which was consistent with the decrease in microalgae lipid content. The results proved that Ca 2+ participated in the regulation of microalgae TAG synthesis under heat stress, which provided a new view for the understanding of the microalgae lipid accumulation mechanism., (Copyright © 2022 Elsevier Inc. All rights reserved.)

The ultra-high Cd polluted environment is a special habitat in nature. Analysis of the biological adaptation and resistance mechanism of Auxenochlorella protothecoides UTEX234 to ultra-high Cd stress would offer some inspiring understanding on Cd detoxification mechanism and help discovering highly active bioremediation agents. In this study, integrated analyses of the transcriptome, multi-physiological and biochemical data and fatty acid profilings of UTEX2341 were performed for the first time. It was found that exogenous Ca ions could alleviate Cd stress. Manganese-dependent superoxide dismutase and peroxidase also participated in intracellular detoxification. And non-enzymatic antioxidants rather than one specific enzymatic antioxidant were suggested to be used as "core antioxidants", which witnessed better performance in Cd detoxification. In addition, Cd stress improved sixteen alkane value and biofuel yield and quality., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Algae lipid production combined with heavy metal removal is a cost-effective and environment-friendly method for algae biofuel production and hazardous waste treatment. Chlorella minutissima UTEX 2341 had strong resistance to cadmium, copper, manganese and zinc ions under heterotrophic culture condition and could efficiently remove them through intracellular accumulation and extracellular immobilization. Meanwhile, lipid accumulation was not inhibited by heavy metals. Instead, the algae lipid content significantly increased by 21.07% and 93.90%, respectively with the addition of cadmium and copper. Furthermore, the heavy metal residue in lipid was within μg range and satisfied the commercial standard. This artificial wastewater-algae biofuel-heavy-metal integrated utilization technology offered a new alternative solution to the problems of energy shortage and environmental pollution. [ABSTRACT FROM AUTHOR]

Abstract: High production cost is a major obstacle to the extensive use of microalgae biodiesel. To cut the cost and achieve higher biomass productivity, Chlorella minutissima UTEX2341 was cultured under photoheterotrophic conditions. With the carbon, nitrogen and phosphorus concentration of 26.37, 2.61 and 0.03gL−1 d−1 respectively, a maximum biomass productivity of 1.78gL−1 d−1 was obtained, which was 59 times more than that cultured under autotrophic condition. The lipid productivity reached 0.29gL−1 d−1, which was 11.9 times higher than the highest value reported by . The conversion rate of microalgae lipids to FAME was found to be elevated from 45.65% to 62.97% and the FAME productivity increased from 1.16 to 180.68mgL−1 d−1 after the optimization. 94% of the fatty acid of C. minutissima UTEX2341 was found to be composed of palmitic, oleic, linoleic and γ linoleic and the unsaturated fatty acids were the main parts (79.42%). [Copyright &y& Elsevier]

• Cd2+ inhibited the most of basic metabolism pathways except lipid metabolism. • Seven transcription factors with glutathionylated sites linked to TAG metabolism. • Transcription factors MYB, NF-YB and ABI5 regulated DGAT expression. • Triacylglycerol maintained lipid homeostasis by balancing fatty acids saturation. • Microalgae activated TAG synthesis by GSH then resisted the Cd stress together. Under ultra-high cadmium (Cd) stress, large amounts of glutathione are produced in Auxenochlorella protothecoides UTEX 2341, and the lipid content increases significantly. Glutathione is the best reductant that can effectively remove Cd, but the relationship between lipid accumulation and the cellular response to Cd stress has not been ascertained. Integrating analyses of the transcriptomes and lipidomes, the mechanism of lipid accumulation to Cd tolerance were studied from the perspectives of metabolism, transcriptional regulation and protein glutathionylation. Under Cd stress, basic metabolic pathways, such as purine metabolism, translation and pre-mRNA splicing process, were inhibited, while the lipid accumulation pathway was significantly activated. Further analysis revealed that the transcription factors (TFs) and genes related to lipid accumulation were also activated. Analysis of the TF interaction sites showed that ABI5, MYB_rel and NF-YB could further regulate the expression of diacylglycerol acyltransferase through glutathionylation/deglutathionylation, which led to increase of the triacylglycerol (TAG) content. Lipidomes analysis showed that TAG could help maintain lipid homeostasis by adjusting its saturation/unsaturation levels. This study for the first time indicated that glutathione could activate TAG synthesis in microalga A. protothecoides , leading to TAG accumulation and glutathione accumulation under Cd stress. Therefore, the accumulation of TAG and glutathione can confer resistance to high Cd stress. This study provided insights into a new operation mode of TAG accumulation under heavy metal stress. [Display omitted] [ABSTRACT FROM AUTHOR]

Highlights: [•] Environmental factors were proved to be helpful to lipid accumulation. [•] NaCl, Fe3+ and nitrogen starvation together made lipid content increase to 2.5 times. [•] Both high and low temperature could promote lipid accumulation. [•] UTEX 2341 was proved to be a potential species for high-quality biodiesel production. [ABSTRACT FROM AUTHOR]