Molecular Mechanisms Underlying the Acclimation of Chlamydomonas reinhardtii Against Nitric Oxide Stress

The acclimation mechanism of Chlamydomonas reinhardtii to nitric oxide (NO) was studied by exposure to S-nitroso-N-acetylpenicillamine (SNAP), a NO donor. Treatment with 0.1 or 0.3 mM SNAP transiently inhibited photosynthesis within 1 h, followed by a recovery, while 1.0 mM SNAP treatment caused irreversible photosynthesis inhibition and mortality. The SNAP effects are avoided in the presence of the NO scavenger, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-l-oxyl-3-oxide (cPTIO). RNA-seq, qPCR, and biochemical analyses were conducted to decode the metabolic shifts under NO stress by exposure to 0.3 mM SNAP in the presence or absence of 0.4 mM cPTIO. These findings revealed that the acclimation to NO stress comprises a temporally orchestrated implementation of metabolic processes: (1). modulation of NADPH oxidase (respiratory burst oxidase-like 2, RBOL2) and ROS signaling pathways for downstream mechanism regulation, (2). trigger of NO scavenging elements to reduce NO level; (3). prevention of photo-oxidative risk through photosynthesis inhibition and antioxidant defense system induction; (4). acclimation to nitrogen and sulfur shortage; (5). attenuation of transcriptional and translational activity together with degradation of damaged proteins through protein trafficking machinery (ubiquitin, SNARE, and autophagy) and molecular chaperone system for dynamic regulation of protein homeostasis. In addition, the expression of the gene encoding NADPH oxidase, RBOL2, showed a transient increase while that of RBOL1 was slightly decreased after NO challenge. It reflects that NADPH oxidase, a regulator in ROS-mediated signaling pathway, may be involved in the responses of Chlamydomonas to NO stress. In conclusion, our findings provide insight into the molecular events underlying acclimation mechanisms in Chlamydomonas to NO stress.

Acclimation of Chlamydomonas reinhardtii to Nitric Oxide Stress Related to Respiratory Burst Oxidase-Like 2

The acclimation mechanism of Chlamydomonas reinhardtii to nitric oxide (NO) was studied by exposure to S-nitroso-N-acetylpenicillamine (SNAP), a NO donor. Treatment with 0.1 or 0.3 mM SNAP transiently inhibited photosynthesis within 1 h, followed by a recovery without growth impairment, while 1.0 mM SNAP treatment caused irreversible photosynthesis inhibition and mortality. The SNAP effects are avoided in the presence of the NO scavenger, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-l-oxyl-3-oxide (cPTIO). RNA-seq, qPCR, and biochemical analyses were conducted to decode the metabolic shifts under sub-lethal NO stress by exposure to 0.3 mM SNAP in the presence or absence of 0.4 mM cPTIO. These findings revealed that the acclimation to NO stress comprises a temporally orchestrated implementation of metabolic processes: 1. trigger of NO scavenging elements to reduce NO level; 2. prevention of photo-oxidative risk through photosynthesis inhibition and antioxidant defense system induction; 3. acclimation to nitrogen and sulfur shortage; 4. degradation of damaged proteins through protein trafficking machinery (ubiquitin, SNARE, and autophagy) and molecular chaperone system for dynamic regulation of protein homeostasis. NO increased NADPH oxidase activity and respiratory burst oxidase-like 2 (RBOL2) transcript abundance, which were not observed in the rbol2 insertion mutant. Changes in gene expression in the rbol2 mutant and increased mortality under NO stress demonstrate that NADPH oxidase (RBOL2) is involved in the modulation of some acclimation processes (NO scavenging, antioxidant defense system, autophagy, and heat shock proteins) for Chlamydomonas to cope with NO stress. Our findings provide insight into the molecular events underlying acclimation mechanisms in Chlamydomonas to sub-lethal NO stress.

Photosynthetic responses of Halimeda scabra (Chlorophyta, Bryopsidales) to interactive effects of temperature, pH, and nutrients and its carbon pathways

In this study, we evaluated the interactive effects of temperature, pH, and nutrients on photosynthetic performance in the calcareous tropical macroalga Halimeda scabra. A significant interaction among these factors on gross photosynthesis (Pgross) was found. The highest values of Pgross were reached at the highest temperature, pH, and nutrient enrichment tested and similarly in the control treatment (no added nutrients) at 33 °C at the lowest pH. The Q10 Pgross values confirmed the effect of temperature only under nutrient enrichment scenarios. Besides the above, bicarbonate (HCO3−) absorption was assessed by the content of carbon stable isotope (δ13C) in algae tissue and by its incorporation into photosynthetic products, as well as by carbonic anhydrase (CA) inhibitors (Acetazolamide, AZ and Ethoxyzolamide, EZ) assays. The labeling of δ13C revealed this species uses both, CO2 and HCO3− forms of Ci relying on a CO2 Concentration Mechanism (CCM). These results were validated by the EZ-AZ inhibition assays in which photosynthesis inhibition was observed, indicating the action of internal CA, whereas AZ inhibitor did not affect maximum photosynthesis (Pmax). The incorporation of 13C isotope into aspartate in light and dark treatments also confirmed photosynthetic and non-photosynthetic the HCO3−uptake.

Performance of Chlorella Vulgaris Exposed to Heavy Metal Mixtures: Linking Measured Endpoints and Mechanisms

Microalgae growth inhibition assays are candidates for referent ecotoxicology as a fundamental part of the strategy to reduce the use of fish and other animal models in aquatic toxicology. In the present work, the performance of Chlorella vulgaris exposed to heavy metals following standardized growth and photosynthesis inhibition assays was assessed in two different scenarios: (1) dilutions of single heavy metals and (2) an artificial mixture of heavy metals at similar levels as those found in natural rivers. Chemical speciation of heavy metals was estimated with Visual MINTEQ software; free heavy metal ion concentrations were used as input data, together with microalgae growth and photosynthesis inhibition, to compare different effects and explain possible toxicity mechanisms. The final goal was to assess the suitability of the ecotoxicological test based on the growth and photosynthesis inhibition of microalgae cultures, supported by mathematic models for regulatory and decision-making purposes. The C. vulgaris algae growth inhibition test was more sensitive for As, Zn, and Pb exposure whereas the photosynthesis inhibition test was more sensitive for Cu and Ni exposure. The effects on growth and photosynthesis were not related. C. vulgaris evidenced the formation of mucilaginous aggregations at lower copper concentrations. We found that the toxicity of a given heavy metal is not only determined by its chemical speciation; other chemical compounds (as nutrient loads) and biological interactions play an important role in the final toxicity. Predictive mixture effect models tend to overestimate the effects of metal mixtures in C. vulgaris for both growth and photosynthesis inhibition tests. Growth and photosynthesis inhibition tests give complementary information, and both are a fast, cheap, and sensitive alternative to animal testing. More research is needed to solve the challenge of complex pollutant mixtures as they are present in natural environments, where microalgae-based assays can be suitable monitoring tools for pollution management and regulatory purposes.

An Ascophyllum nodosum-Derived Biostimulant Protects Model and Crop Plants from Oxidative Stress

Abiotic stresses, which at the molecular level leads to oxidative damage, are major determinants of crop yield loss worldwide. Therefore, considerable efforts are directed towards developing strategies for their limitation and mitigation. Here the superoxide-inducing agent paraquat (PQ) was used to induce oxidative stress in the model species Arabidopsis thaliana and the crops tomato and pepper. Pre-treatment with the biostimulant SuperFifty (SF) effectively and universally suppressed PQ-induced leaf lesions, H2O2 build up, cell destruction and photosynthesis inhibition. To further investigate the stress responses and SF-induced protection at the molecular level, we investigated the metabolites by GC-MS metabolomics. PQ induced specific metabolic changes such as accumulation of free amino acids (AA) and stress metabolites. These changes were fully prevented by the SF pre-treatment. Moreover, the metabolic changes of the specific groups were tightly correlating with their phenotypic characteristics. Overall, this study presents physiological and metabolomics data which shows that SF protects against oxidative stress in all three plant species.

H2O2/ABA signal pathway participates in the regulation of stomata opening of cucumber leaves under salt stress by putrescine

The stomatal-aperture is imperative for plant physiological metabolism. The function of polyamines (PAs) in stomatal regulation under stress environment largely remains elucidate. Herein, we investigated the regulatory mechanism of exogenous putrescine (Put) on the stomatal opening of cucumber leaves under salt stress. The results revealed that Put relieved the salt-induced photosynthetic inhibition of cucumber leaves by regulating stomatal-apertures. Put application increased hydrogen peroxide (H2O2) and decreased abscisic acid (ABA) content in leaves under salt stress. The inhibitors of diamine oxidase (DAO), polyamine oxidase (PAO), nicotinamide adenine dinucleotide phosphate oxidase (NADPH) are AG, 1,8-DO and DPI, respectively and pre-treatment with these inhibitors up-regulated key gene NCED of ABA synthase and down-regulated key gene GSHS of reduced glutathione (GSH) synthase. The content of H2O2 and GSH were decreased and ABA content was increased and its influenced trend is AG>1,8-DO>DPI. Moreover, the Put induced down-regulation of ABA content under salt stress blocked by treatment with H2O2 scavenger (DMTU) and GSH scavenger (CNDB). Additionally, the application of DMTU also blocked the increase of GSH content. Collectively, these results suggest that Put can regulate GSH content by promoting H2O2 generation through polyamine metabolic pathway, which inhibits ABA accumulation to achieve stomatal regulation under salt stress.HighlightExogenous putrescine alleviates photosynthesis inhibition in salt-stressed cucumber seedlings by regulating stomatal-aperture.