Effects of low temperature and highlight stress on lipid accumulation and cell structure of Tropidoneis maxima.

Tropidoneis maxima is a marine diatom with a rapid growth rate that produces high levels of lipids. To explore whether the lipid content could be further enhanced, cultures were first incubated under optimal conditions and then stressed under low temperature (10°C), a high light intensity level (80 µmol/m2 ·s), and the two factors together (interaction treatment). The results indicated that high light intensity and the temperature-light interaction exhibited greater impacts on lipid synthesis of T. maxima than low temperature. The two stress treatments increased lipid content by 17.16% and 16.6% compared to the control. In particular, higher biomass concentration was obtained with high light intensity (1.082 g L-1 ) and low temperature (1.026 g L-1 ). Moreover, high light intensity (9.06%) and interaction (10.3%) treatments yielded lower starch content compared to low temperature (14.27%) at the end of the stress culture. After 3 days of stress culture, the high light intensity treatment resulted in a 97.01% increase in cell wall thickness and an 18.46% decrease in cell diameter. The results suggest that high light intensity stress on T. maxima would open a new approach to cost-effective biolipid production.

Carbon and Nitrogen Allocation between the Sink and Source Leaf Tissue in Response to the Excess Excitation Energy Conditions.

Plants are inevitably exposed to extreme climatic conditions that lead to a disturbed balance between the amount of absorbed energy and their ability to process it. Variegated leaves with photosynthetically active green leaf tissue (GL) and photosynthetically inactive white leaf tissue (WL) are an excellent model system to study source-sink interactions within the same leaf under the same microenvironmental conditions. We demonstrated that under excess excitation energy (EEE) conditions (high irradiance and lower temperature), regulated metabolic reprogramming in both leaf tissues allowed an increased consumption of reducing equivalents, as evidenced by preserved maximum efficiency of photosystem II (ФPSII) at the end of the experiment. GL of the EEE-treated plants employed two strategies: (i) the accumulation of flavonoid glycosides, especially cyanidin glycosides, as an alternative electron sink, and (ii) cell wall stiffening by cellulose, pectin, and lignin accumulation. On the other hand, WL increased the amount of free amino acids, mainly arginine, asparagine, branched-chain and aromatic amino acids, as well as kaempferol and quercetin glycosides. Thus, WL acts as an important energy escape valve that is required in order to maintain the successful performance of the GL sectors under EEE conditions. Finally, this role could be an adaptive value of variegation, as no consistent conclusions about its ecological benefits have been proposed so far.

Enhanced astaxanthin production of Haematococcus pluvialis strains induced salt and high light resistance with gamma irradiation.

This study was conducted to increase the productivity of biomass that contains high astaxanthin content by developing a mutant Haematococcus pluvialis strain with strong environmental tolerance. H. pluvialis has a low cell-growth rate and is vulnerable to stressors such as salinity or light intensity, which may hinder large-scale commercial cultivation. A mutant M5 strain selected through 5000-Gy gamma irradiation showed improved biomass and astaxanthin production under high-salinity and high-light intensity conditions. With enhanced SOD activity and overexpressed astaxanthin biosynthesis genes (lyc, crtR-b, bkt2), M5 demonstrated an increase in biomass and astaxanthin productivity by 86.70 % and 66.15 %, respectively compared to those of untreated cells. Also, the omega-3 content of M5 increased by 149.44 % under 40 mM CaCl2 compared to the untreated cells. Finally, even when subjected to high-intensity light irradiation for the whole life cycle, the biomass and astaxanthin concentration increased by 84.99 % and 241 %, respectively, compared to the wild-type cells.

Effect of Rapid Polymerization on Water Sorption and Solubility of Bulk-fill Composites.

Objectives: The aim of the study was to examine the impact of rapid high-intensity polymerization on water sorption and solubility of a new generation of bulk-fill composite materials. Material and methods: Five materials were tested: a conventional composite Filtek Z250 (3M, St. Paul, USA) and four bulk-fill composites, Filtek One Bulk Fill (3M), Tetric PowerFill (Ivoclar Vivadent, Schaan, Liechtenstein), Tetric PowerFlow (Ivoclar Vivadent), SDR Plus (Dentsply, Konstanz, Germany). Composite specimens with a 9-mm diameter and a 2-mm height (n = 6) were polymerized with a high-intensity curing unit (Bluephase® PowerCure, Ivoclar Vivadent). The control group was polymerized for a total of 40 s on both sides (1193 mW / cm2) and 3s group for 3 s on one side (3053 mW/cm2). Water sorption and solubility were measured by ISO 4049 method up to 30 days of immersion. The results were statistically analyzed using one-way ANOVA with Tukey post-hoc correction. Different polymerization protocols for the same material were compared by t-test (p <0.05). Results: The 3s polymerization protocol increased the solubility of all materials. Filtek One Bulk Fill showed the highest water sorption and solubility values, and Tetric PowerFlow proved to be a stable material with low values of tested parameters. During immersion, a saturation of specimens was achieved in low-viscosity bulk-fill composites within two weeks, while in other materials it was not achieved within 30 days, or more, following the immersion. Conclusions: Rapid polymerization caused an increase in solubility, which could affect the biocompatibility of the investigated materials not intended for the 3s curing. SDR Plus and Filtek One Bulk Fill should not be polymerized with rapid high-intensity curing due to increased solubility that exceeds ISO 4049:2009 limits.

HPR1 Is Required for High Light Intensity Induced Photorespiration in Arabidopsis thaliana.

High light intensity as one of the stresses could lead to generation of large amounts of reactive oxygen species (ROS) in plants, resulting in severe plant growth retardation. The photorespiration metabolism plays an important role in producing and removing a variety of ROS, maintaining the dynamic balance of the redox reaction, and preventing photoinhibition. Arabidopsis hydroxypyruvate reductase 1 (HPR1) is a primary metabolic enzyme in the photorespiration cycle. However, the role of HPR1 in plants response to high light is not clear. Here, we found that the expression of HPR1 could be induced by high light intensity. The growth and photosynthetic capacity of hpr1 mutants are seriously affected under high light intensity. The absence of HPR1 suppresses the rates of photorepair of Photosystem II (PSII), aggravates the production of ROS, and accelerates photorespiration rates. Moreover, the activity of ROS scavenging enzymes in the hpr1 mutants is significantly higher. These results indicate that HPR1 is involved in plant response to high light intensity and is essential for maintaining the dynamic balance of ROS and photorespiration.

How Light Reactions of Photosynthesis in C4 Plants Are Optimized and Protected under High Light Conditions.

Most C4 plants that naturally occur in tropical or subtropical climates, in high light environments, had to evolve a series of adaptations of photosynthesis that allowed them to grow under these conditions. In this review, we summarize mechanisms that ensure the balancing of energy distribution, counteract photoinhibition, and allow the dissipation of excess light energy. They secure effective electron transport in light reactions of photosynthesis, which will lead to the production of NADPH and ATP. Furthermore, a higher content of the cyclic electron transport components and an increase in ATP production are observed, which is necessary for the metabolism of C4 for effective assimilation of CO2. Most of the data are provided by studies of the genus Flaveria, where species belonging to different metabolic subtypes and intermediate forms between C3 and C4 are present. All described mechanisms that function in mesophyll and bundle sheath chloroplasts, into which photosynthetic reactions are divided, may differ in metabolic subtypes as a result of the different organization of thylakoid membranes, as well as the different demand for ATP and NADPH. This indicates that C4 plants have plasticity in the utilization of pathways in which efficient use and dissipation of excitation energy are realized.

Design and Analysis of Native Photorespiration Gene Motifs of Promoter Untranslated Region Combinations Under Short Term Abiotic Stress Conditions.

Quantitative traits are rarely controlled by a single gene, thereby making multi-gene transformation an indispensable component of modern synthetic biology approaches. However, the shortage of unique gene regulatory elements (GREs) for the robust simultaneous expression of multiple nuclear transgenes is a major bottleneck that impedes the engineering of complex pathways in plants. In this study, we compared the transcriptional efficacies of a comprehensive list of well-documented promoter and untranslated region (UTR) sequences side by side. The strength of GREs was examined by a dual-luciferase assay in conjunction with transient expression in tobacco. In addition, we created suites of new GREs with higher transcriptional efficacies by combining the best performing promoter-UTR sequences. We also tested the impact of elevated temperature and high irradiance on the effectiveness of these GREs. While constitutive promoters ensure robust expression of transgenes, they lack spatiotemporal regulations exhibited by native promoters. Here, we present a proof-of-principle study on the characterization of synthetic promoters based on cis-regulatory elements of three key photorespiratory genes. This conserved biochemical process normally increases under elevated temperature, low CO2, and high irradiance stress conditions and results in ∼25% loss in fixed CO2. To select stress-responsive cis-regulatory elements involved in photorespiration, we analyzed promoters of two chloroplast transporters (AtPLGG1 and AtBASS6) and a key plastidial enzyme, AtPGLP using PlantPAN3.0 and AthaMap. Our results suggest that these motifs play a critical role for PLGG1, BASS6, and PGLP in mediating response to elevated temperature and high-intensity light stress. These findings will not only enable the advancement of metabolic and genetic engineering of photorespiration but will also be instrumental in related synthetic biology approaches.

Molybdenum disulfide nanoparticles concurrently stimulated biomass and ß-carotene accumulation in Dunaliella salina.

Molybdenum disulfide nanoparticles (MoS2 NPs) hold tremendous properties in wide domain of applications. In this study, the impact of MoS2 NPs was investigated on algal physiological and metabolic properties and a two-stage strategy was acquired to enhance the commercial potential of Dunaliella salina. With 50 µg/L of MoS2 NPs exposure, cellular growth and biomass production were promoted by 1.47- and 1.33-fold than that in control, respectively. MoS2 NPs treated cells were subject to high light intensity for 7 days after 30 days of normal light cultivation, which showed that high light intensity gradually increased ß-carotene content by 1.48-fold. Furthermore, analyses of primary metabolites showed that combinatorial approach significantly altered the biochemical composition of D. salina. Together, these findings demonstrated that MoS2 NPs at an optimum concentration combined with high light intensity could be a promising approach to concurrently enhance biomass and ß-carotene production in microalgae.

Crosstalk between sterol and neutral lipid metabolism in the alga Haematococcus pluvialis exposed to light stress.

The presence, biosynthesis and functional role of sterols in the green microalga Haematococcus pluvialis remain poorly understood. In this work we studied the effect of high-light (HL) stress on sterol synthesis in H. pluvialis UTEX 2505 cells. HL stress induced the synthesis of sterols in parallel with that of triacylglycerides (TAG), giving rise to the synthesis of cholesterol over that of phytosterols. Blockage of the carotenogenic 1-deoxy-D-xylulose 5-phosphate (MEP) pathway is shown to be involved in HL-induced sterol synthesis. In addition, high irradiance exposure induced MEP- and fatty acid (FA)-biosynthetic transcripts. The pharmacological inhibition of these pathways suggests a possible feedback regulation of sterol and FA homeostasis. Finally, both lipid classes proved crucial to the adequate photosynthetic performance of H. pluvialis grown under HL intensity stress. Our findings reveal new insights into H. pluvialis lipid metabolism that contribute to the development of value-added bioproducts from microalgae.

Autophagy-Related 2 Regulates Chlorophyll Degradation under Abiotic Stress Conditions in Arabidopsis.

Chloroplasts are extraordinary organelles for photosynthesis and nutrient storage in plants. During leaf senescence or under stress conditions, damaged chloroplasts are degraded and provide nutrients for developing organs. Autophagy is a high-throughput degradation pathway for intracellular material turnover in eukaryotes. Along with chloroplast degradation, chlorophyll, an important component of the photosynthetic machine, is also degraded. However, the chlorophyll degradation pathways under high light intensity and high temperature stress are not well known. Here, we identified and characterized a novel Arabidopsis mutant, sl2 (seedling lethal 2), showing defective chloroplast development and accelerated chlorophyll degradation. Map-based cloning combined with high-throughput sequencing analysis revealed that a 118.6 kb deletion region was associated with the phenotype of the mutant. Complementary experiments confirmed that the loss of function of ATG2 was responsible for accelerating chlorophyll degradation in sl2 mutants. Furthermore, we analyzed chlorophyll degradation under abiotic stress conditions and found that both chloroplast vesiculation and autophagy take part in chlorophyll degradation under high light intensity and high temperature stress. These results enhanced our understanding of chlorophyll degradation under high light intensity and high temperature stress.

Dynamic regulation of anthocyanin biosynthesis at different light intensities by the BT2-TCP46-MYB1 module in apple.

Teosinte branched1/cycloidea/proliferating (TCP) transcription factors play a broad role in plant growth and development, but their involvement in the regulation of anthocyanin biosynthesis is currently unclear. In this study, anthocyanin biosynthesis induced by different light intensities in apple (Malus domestica) was found to be largely dependent on the functions of the MdMYB1 and MdTCP46 transcription factors. The expression of MdTCP46 was responsive to high light intensity, and under these conditions it promoted anthocyanin biosynthesis by direct interactions with MdMYB1 that enhanced the binding of the latter to its target genes. MdTCP46 also interacted with a bric-a-brac/tramtrack/broad (BTB) protein, MdBT2, that is responsive to high light intensity, which ubiquitinated MdTCP46 and mediated its degradation via the 26S proteasome pathway. Our results demonstrate that the dynamic regulatory module MdBT2-MdTCP46-MdMYB1 plays a key role in modulating anthocyanin biosynthesis at different light intensities in apple, and provides new insights into the post-transcriptional regulation of TCP proteins.

Effect of High Light Intensity Bleaching Protocol versus Descending Light Intensities Bleaching Protocol on Post Bleaching Teeth Sensitivity: A Randomized Clinical Trial.

AIM: The study aimed to compare teeth sensitivity and shade after bleaching protocol with descending different light intensities versus bleaching protocol with the same high light intensity. MATERIAL AND METHODS: Sample size was twenty-four patients. Each group consisted of twelve patients. Group, I patients received bleaching protocol of descending different light intensities. Group II patients received bleaching protocol with the same high light intensity; both groups used the same home bleaching gel kit for seven days according to manufacturer instructions and protocol. Baseline records were digital photographs, teeth sensitivity and teeth shade for 12 anterior teeth. Teeth sensitivity was assessed using five points verbal rating scale and Standardized 100 mm Visual analogue scale after 1 day, after 2 days and after 1 week. Teeth shades for twelve anterior teeth were recorded by VITA Easy Shade V (VITA Zahnfabrik H. Rauter GmbH & Co. KG, Germany) after 1 week by VITA Easy Shade V. Mann-Whitney test (non-parametric test, 2 independent samples) was used to compare teeth sensitivity between both bleaching protocols at each period. A paired t-test (parametric test, 2 related samples) was performed to compare the colour change in shade guide units (SGU) and ∆E values within high light intensity bleaching protocol. While Wilcoxon Signed-Rank test (non-parametric test, 2 related samples) was used to compare colour change light intensities bleaching protocol. Comparison of bleaching effectiveness (∆SGU and ∆Ediff) between both bleaching protocols was performed by the Mann-Whitney test. RESULTS: Descending light intensities protocol showed a lower teeth sensitivity than high light intensity protocol after 1 and 2 days. There was no teeth sensitivity reported at 1-week post-bleaching. Regarding the teeth shade, descending light intensities protocol had a little higher effect on colour change in shade guide units (SGU) than high light intensity protocol effect. Both bleaching protocols showed there was no significant difference in ∆SGU recorded after bleaching between high and descending light intensities protocols. CONCLUSION: Descending different light intensities protocol showed a lower teeth sensitivity than high same light intensity protocol. Descending light intensities protocol had a little higher effect on colour change in shade guide units (SGU) than high light intensity protocol effect.

A Holistic Approach to Model the Kinetics of Photocatalytic Reactions.

Understanding and modeling kinetics is an essential part of the optimization and implementation of chemical reactions. In the case of photocatalytic reactions this is mostly done one-dimensionally, i.e., only considering the effect of one parameter at the same time. However, as discussed in this study, many of the relevant reaction parameters have mutual interdependencies that call for a holistic multi-dimensional approach to accurately model and understand their influence. Such an approach is described herein, and all the relevant equations given so that researchers can readily implement it to analyze and model their reactions.

Effects of temperature and its combination with high light intensity on lipid production of Monoraphidium dybowskii Y2 from semi-arid desert areas.

Temperature and light intensity are important environmental factors influencing microalgae for biodiesel production. The aim of present work was to study the effects of temperature (15 °C, 25 °C, and 35 °C) and its combination with high light intensity (HL, 400 µmol photon m-2 s-1) on lipid production of Monoraphidium dybowskii Y2 which was isolated from desert. The results demonstrated that algal growth was only inhibited at 15 °C. Promoted lipid content and decreased Fv/Fm were observed in 15 °C and 35 °C. Cellular carbohydrate, protein conversion and membrane lipid (MGDG, DGDG and SQDG) remodeling contributes for lipid accumulation. Stress combined temperatures with HL are benefit for lipid production, especially desired neutral lipid productivity all exceed 40 mg L-1 d-1. Fatty acids compositions of C16:0 and C18:1 were further promoted under 15 °C or 35 °C combined with HL. Thus, M. dybowskii Y2 will well-adapted to outdoors cultivation for biodiesel production.

Enhancing Carbohydrate Productivity of Chlorella sp. AE10 in Semi-continuous Cultivation and Unraveling the Mechanism by Flow Cytometry.

Accumulated carbohydrate in microalgae is promising feedstock for bioethanol fermentation. Selection of suitable cultivation conditions in semi-continuous cultivation is critical to achieve a high carbohydrate productivity. In the current study, the effects of macro-nutrient (nitrogen, phosphorus, and sulfur) limitations and light intensity were evaluated for the carbohydrate accumulations of Chlorella sp. AE10 under 10% CO2 conditions. It was shown that nitrogen limitation and high light intensity were effective for improving carbohydrate productivity. The average carbohydrate and biomass productivity in semi-continuous cultivation with 1/4 N medium and 1000 µmol photons m-2 s-1 was 0.673 and 0.93 g L-1 day-1, respectively. Sulfur and phosphorus limitations could improve the carbohydrate content but they could not enhance the carbohydrate productivity. The cell cycle progression and chlorophyll a were investigated using flow cytometry (FCM). The results showed that macro-nutrient limitation and high light intensity indeed influenced cell cycle progression and led to the formation of polyploid cells along with the carbohydrate accumulation in a certain range. FCM was rapid and accurate method to investigate the operation conditions why 1/4 N, 2 days as a cycle, and high light intensity were optimal ones. In addition, the remaining high level of photosynthesis activity was also important for achieving a high carbohydrate productivity. Dynamic tracking of carbohydrate accumulation is helpful for establishment of a semi-continuous cultivation for enhancing carbohydrate productivity in microalgae.

Concentration of phenolic compounds is increased in lettuce grown under high light intensity and elevated CO2.

The present study was focused on lettuce, a widely consumed leafy vegetable for the large number of healthy phenolic compounds. Two differently-pigmented lettuce cultivars, i.e. an acyanic-green leaf cv. and an anthocyanic-red one, were grown under high light intensity or elevated CO2 or both in order to evaluate how environmental conditions may affect the production of secondary phenolic metabolites and, thus, lettuce quality. Mild light stress imposed for a short time under ambient or elevated CO2 concentration increased phenolics compounds as well as antioxidant capacity in both lettuce cvs, indicating how the cultivation practice could enhance the health-promoting benefits of lettuce. The phenolic profile depended on pigmentation and the anthocyanic-red cv. always maintained a higher phenolic amount as well as antioxidant capacity than the acyanic-green one. In particular, quercetin, quercetin-3-O-glucuronide, kaempferol, quercitrin and rutin accumulated under high light or high CO2 in the anthocyanic-red cv., whereas cyanidin derivatives were responsive to mild light stress, both at ambient and elevated CO2. In both cvs total free and conjugated phenolic acids maintained higher values under all altered environmental conditions, whereas luteolin reached significant amounts when both stresses were administered together, indicating, in this last case, that the enzymatic regulation of the flavonoid synthesis could be differently affected, the synthesis of flavones being favored.

High light induced changes in organization, protein profile and function of photosynthetic machinery in Chlamydomonas reinhardtii.

The green alga Chlamydomonas (C.) reinhardtii is used as a model organism to understand the efficiency of photosynthesis along with the organization and protein profile of photosynthetic apparatus under various intensities of high light exposure for 1h. Chlorophyll (Chl) a fluorescence induction, OJIPSMT transient was decreased with increase in light intensity indicating the reduction in photochemical efficiency. Further, circular dichroism studies of isolated thylakoids from high light exposed cells showed considerable change in the pigment-pigment interactions and pigment-proteins interactions. Furthermore, the organization of supercomplexes from thylakoids is studied, in which, one of the hetero-trimer of light harvesting complex (LHC) II is affected significantly in comparison to other complexes of LHC's monomers. Also, other supercomplexes, photosystem (PS)II reaction center dimer and PSI complexes are reduced. Additionally, immunoblot analysis of thylakoid proteins revealed that PSII core proteins D1 and D2 were significantly decreased during high light treatment. Similarly, the PSI core proteins PsaC, PsaD and PsaG were drastically changed. Further, the LHC antenna proteins of PSI and PSII were differentially affected. From our results it is clear that LHCs are damaged significantly, consequently the excitation energy is not efficiently transferred to the reaction center. Thus, the photochemical energy transfer from PSII to PSI is reduced. The inference of the study deciphers the structural and functional changes driven by light may therefore provide plants/alga to regulate the light harvesting capacity in excess light conditions.

Carbon allocation from source to sink leaf tissue in relation to flavonoid biosynthesis in variegated Pelargonium zonale under UV-B radiation and high PAR intensity.

We studied the specific effects of high photosynthetically active radiation (PAR, 400-700 nm) and ecologically relevant UV-B radiation (0.90 W m(-2)) on antioxidative and phenolic metabolism by exploiting the green-white leaf variegation of Pelargonium zonale plants. This is a suitable model system for examining "source-sink" interactions within the same leaf. High PAR intensity (1350 µmol m(-2) s(-1)) and UV-B radiation induced different responses in green and white leaf sectors. High PAR intensity had a greater influence on green tissue, triggering the accumulation of phenylpropanoids and flavonoids with strong antioxidative function. Induced phenolics, together with ascorbate, ascorbate peroxidase (APX, EC 1.11.1.11) and catalase (CAT, EC 1.11.1.6) provided efficient defense against potential oxidative pressure. UV-B-induced up-regulation of non-phenolic H2O2 scavengers in green leaf sectors was greater than high PAR-induced changes, indicating a UV-B role in antioxidative defense under light excess; on the contrary, minimal effects were observed in white tissue. However, UV-B radiation had greater influence on phenolics in white leaf sections compared to green ones, inducing accumulation of phenolic glycosides whose function was UV-B screening rather than antioxidative. By stimulation of starch and sucrose breakdown and carbon allocation in the form of soluble sugars from "source" (green) tissue to "sink" (white) tissue, UV-B radiation compensated the absence of photosynthetic activity and phenylpropanoid and flavonoid biosynthesis in white sectors.

The effects of taurine on vigabatrin, high light intensity and mydriasis induced retinal toxicity in the pigmented rat.

The overall purpose of this study was to establish a model that may be used for examining the effect of Vigabatrin-induced retinal toxicity in pigmented rats, and subsequently examine the possible effects of taurine on the retinal toxicity. In the first part of the study, pigmented Long Evans rats were subjected to combinations of induced mydriasis, low/high light intensities (40/2000 lx) and oral administration of near-MTD (Maximum Tolerated Dose) doses (200 mg/kg/day) of Vigabatrin for up to 6 weeks. The combination of mydriasis and high light intensity applied to Long Evans rats resulted in retinal damage that was increased by the administration of Vigabatrin. In the second part of the study Long Evans rats were subjected to combinations of induced mydriasis and high/low light intensity (40/2000 lx) while being orally administered low (30 mg/kg/day) or high (200 mg/kg/day) doses of Vigabatrin for up to 6 weeks. In addition, selected groups of animals were administered taurine via the drinking water (20 mg/ml), resulting in systemic taurine concentrations of approximately threefold the endogenous concentration. The combined results of the studies demonstrate that retinal damage can be induced in pigmented animals when combining mydriasis and high light intensity. Retinal damage was functionally evaluated by electroretinography (ERG), then confirmed by histopathology. While depending on mydriasis and high light intensity, administration of Vigabatrin increased the retinal toxicity and resulted in the formation of rosette-like structures in the retina in a dose-related manner. Administration of taurine did not alleviate the Vigabatrin-induced retinal toxicity, as demonstrated either functionally by ERG or morphologically, although systemic concentrations of 3-fold the endogenous levels were reached, and it was thus not possible to demonstrate a protective effect of taurine in these pigmented animals.