Copper and zinc differentially affect root glutathione accumulation and phytochelatin synthase gene expression of Rhizophora mucronata seedlings: Implications for mechanisms underlying trace metal tolerance.

Mangroves are susceptible to contamination due to their proximity to shores and human activities. Exposure to excessive trace metals can disturb their physiological functions and may eventually lead to death. Rhizophora mucronata is a common species growing in the mangrove forests of Thailand. Previous studies have shown that seedlings of R. mucronata are tolerant of trace metal and that they accumulate a large metal content in their root tissue. However, knowledge of their tolerance mechanisms is still lacking. To elicit the role of metal detoxification and sequestration by phytochelatins (PC) in the roots of R. mucronata seedlings, the impacts of Cu and Zn exposure were assessed on 1) physiological characteristics 2) the concentration of glutathione (GSH), a precursor of PC and 3) the level of the transcripts encoding phytochelatin synthase (PCS), the key enzyme for PC biosynthesis. Seedlings of R. mucronata were exposed to Cu and Zn in a hydroponic experiment (200 mg Cu or Zn/L in 1/4× Hoagland solution containing 8‰ NaCl, single addition). We found that both trace metals were largely accumulated in the roots. Only Cu-treated seedlings showed a decrease in the photosynthetic efficiency, in line with observed toxicity symptoms (i.e. bent stems and slight wilting of leaves). Metal accumulation, however, did not induce oxidative stress in the roots as indicated by similar level of total reactive species and lipid peroxidation across treatments. The GSH content in the roots exposed to Cu was significantly reduced while no change was observed in Zn-exposed roots. Coordinated semi-quantitative PCR and RT-qPCR revealed pcs down-regulation in Cu-treated roots, whereas Zn-treated roots showed a down-regulation on day 1 and a subsequent recovery on day 5. Failure of detoxification and sequestration of excess Cu due to GSH limitation and down-regulation of pcs may lead to the phytotoxic effects observed in Cu-treated plants. Our results suggest that both GSH and PC play an important role in trace metal tolerance in R. mucronata seedlings.

Tolerance Mechanisms to Copper and Zinc Excess in Rhizophora mucronata Lam. Seedlings Involve Cell Wall Sequestration and Limited Translocation.

Rhizophora mucronata is a common mangrove growing in habitats subjected to heavy metal (HM) contamination. Understanding their physiological responses to copper (Cu) and zinc (Zn) excess and underlying tolerance mechanisms is crucial to assess impacts of metal pollution on mangrove community. Seedlings were treated with Cu or Zn (0, 50 or 100 mg per plant) by means of a single addition. At day 3 and 7, Cu and Zn accumulation, photosynthetic efficiency, superoxide dismutase and peroxidase activity, non-protein thiols, reactive oxygen species and lipid peroxidation in roots and leaves were measured. R. mucronata restricted Cu and Zn translocation, thus accumulated HM mainly in roots while kept the leaves unaffected. However, high root HM did not induce oxidative stress nor anti-oxidative defense as HM were largely deposited in cell wall. We concluded that HM tolerance strategies of R. mucronata seedlings are exclusion and restriction of translocation to the vital photosynthetic tissue.

The role of O2 as an electron acceptor alternative to CO2 in photosynthesis of the common marine angiosperm Zostera marina L.

This study investigates the role of O2 as an electron acceptor alternative to CO2 in photosynthesis of the common marine angiosperm Zostera marina L. Electron transport rates (ETRs) and non-photochemical quenching (NPQ) of Z. marina were measured under saturating irradiance in synthetic seawater containing 2.2 mM DIC and no DIC with different O2 levels (air-equilibrated levels, 3 % of air equilibrium and restored air-equilibrated levels). Lowering O2 did not affect ETR when DIC was provided, while it caused a decrease in ETR and an increase in NPQ in DIC-free media, indicating that O2 acted as an alternative electron acceptor under low DIC. The ETR and NPQ as a function of irradiance were subsequently assessed in synthetic seawater containing (1) 2.2 mM DIC, air-equilibrated O2; (2) saturating CO2, no O2; and (3) no DIC, air-equilibrated O2. These treatments were combined with glycolaldehyde pre-incubation. Glycolaldehyde caused a marked decrease in ETR in DIC-free medium, indicating significant electron flow supported by photorespiration. Combining glycolaldehyde with O2 depletion completely suppressed ETR suggesting the operation of the Mehler reaction, a possibility supported by the photosynthesis-dependent superoxide production. However, no notable effect of suppressing the Mehler reaction on NPQ was observed. It is concluded that during DIC-limiting conditions, such as those frequently occurring in the habitats of Z. marina, captured light energy exceeds what is utilised for the assimilation of available carbon, and photorespiration is a major alternative electron acceptor, while the contribution of the Mehler reaction is minor.

Unraveling the physiological responses of morphologically distinct corals to low oxygen.

Background: Low oxygen in marine environments, intensified by climate change and local pollution, poses a substantial threat to global marine ecosystems, especially impacting vulnerable coral reefs and causing metabolic crises and bleaching-induced mortality. Yet, our understanding of the potential impacts in tropical regions is incomplete. Furthermore, uncertainty surrounds the physiological responses of corals to hypoxia and anoxia conditions. Methods: We initially monitored in situ dissolved oxygen (DO) levels at Kham Island in the lower Gulf of Thailand. Subsequently, we conducted a 72-hour experimental exposure of corals with different morphologies-Pocillopora acuta, Porites lutea, and Turbinaria mesenterina-to low oxygen conditions, while following a 12/12-hour dark/light cycle. Three distinct DO conditions were employed: ambient (DO 6.0 ± 0.5 mg L-1), hypoxia (DO 2.0 ± 0.5 mg L-1), and anoxia (DO < 0.5 mg L-1). We measured and compared photosynthetic efficiency, Symbiodiniaceae density, chlorophyll concentration, respiratory rates, primary production, and calcification across the various treatments. Results: Persistent hypoxia was observed at the study site. Subsequent experiments revealed that low oxygen levels led to a notable decrease in the maximum quantum yield over time in all the species tested, accompanied by declining rates of respiration and calcification. Our findings reveal the sensitivity of corals to both hypoxia and anoxia, particularly affecting processes crucial to energy balance and structural integrity. Notably, P. lutea and T. mesenterina exhibited no mortality over the 72-hour period under hypoxia and anoxia conditions, while P. acuta, exposed to anoxia, experienced mortality with tissue loss within 24 hours. This study underscores species-specific variations in susceptibility associated with different morphologies under low oxygen conditions. The results demonstrate the substantial impact of deoxygenation on coral growth and health, with the compounded challenges of climate change and coastal pollution exacerbating oxygen availability, leading to increasingly significant implications for coral ecosystems.

Comparative effects of heat stress on photosynthesis and oxidative stress in Halophila ovalis and Thalassia hemprichii under different light conditions.

This study investigated the physiological responses of two tropical seagrass species, Halophila ovalis and Thalassia hemprichii, to heat stress under varying light conditions in a controlled 5-day experiment. The experimental design included four treatments: control, saturating light, heat stress under sub-saturating light, and heat stress under saturating light (combined stress). We assessed various parameters, including chlorophyll fluorescence, levels of reactive oxygen species (ROS), antioxidant enzyme activities, and growth rates. In H. ovalis, heat stress resulted in a significant reduction in the maximum quantum yield of photosystem II (Fv/Fm) regardless of the light condition. However, the effects of heat stress on the effective quantum yield of photosystem II (ɸPSII) were more pronounced under saturating light conditions. In T. hemprichii, saturating irradiance exacerbated the heat stress effects on Fv/Fm and ɸPSII, although the overall photoinhibition was less severe than in H. ovalis. Heat stress led to ROS accumulation in H. ovalis and reduced the activity of superoxide dismutase (SOD) and ascorbate peroxidase in the sub-saturating light condition. Conversely, T. hemprichii exhibited elevated SOD activity under saturating light. Heat stress suppressed the growth of both seagrass species, regardless of the light environment. The Biomarker Response Index indicated that H. ovalis displayed severe effects in the heat stress treatment under both light conditions, while T. hemprichii exhibited moderate effects in sub-saturating light and major effects in saturating light conditions. However, the Effect Addition Index revealed an antagonistic interaction between heat stress and high light in both seagrass species. This study underscores the intricate responses of seagrasses, emphasizing the importance of considering both local and global stressors when assessing their vulnerability.

Comparative study on anatomical traits and gas exchange responses due to belowground hypoxic stress and thermal stress in three tropical seagrasses.

BACKGROUND: The ability to maintain sufficient oxygen levels in the belowground tissues and the rhizosphere is crucial for the growth and survival of seagrasses in habitats with highly reduced sediment. Such ability varies depending on plant anatomical features and environmental conditions. METHODS: In the present study, we compared anatomical structures of roots, rhizomes and leaves of the tropical intertidal seagrasses, Cymodocea rotundata, Thalassia hemprichii and Halophila ovalis, followed by an investigation of their gas exchange both in the belowground and aboveground tissues and photosynthetic electron transport rates (ETR) in response to experimental manipulations of O2 level (normoxia and root hypoxia) and temperature (30 °C and 40 °C). RESULTS: We found that C. rotundata and T. hemprichii displayed mostly comparable anatomical structures, whereas H. ovalis displayed various distinctive features, including leaf porosity, number and size of lacunae in roots and rhizomes and structure of radial O2 loss (ROL) barrier. H. ovalis also showed unique responses to root hypoxia and heat stress. Root hypoxia increased O2 release from belowground tissues and overall photosynthetic activity of H. ovalis but did not affect the other two seagrasses. More pronounced warming effects were detected in H. ovalis, measured as lower O2 release in the belowground tissues and overall photosynthetic capacity (O2 release and dissolved inorganic carbon uptake in the light and ETR). High temperature inhibited photosynthesis of C. rotundata and T. hemprichii but did not affect their O2 release in belowground tissues. Our data show that seagrasses inhabiting the same area respond differently to root hypoxia and temperature, possibly due to their differences in anatomical and physiological attributes. Halophila ovalis is highly dependent on photosynthesis and appears to be the most sensitive species with the highest tendency of O2 loss in hypoxic sediment. At the same time, its root oxidation capacity may be compromised under warming scenarios.

Integrated biomarker responses of rice associated with grain yield in copper-contaminated soil.

Copper (Cu) contamination in soil is an environmental issue that affects rice growth and development. This study investigated changes in photosynthetic capacities in combination with integrated biomarker responses at different growth stages of rice (Oryza sativa L. var. Hom Bai Toey) exposed to various concentrations of Cu. A randomized complete block design with four replications was used. Exposure to high copper concentrations of 200 Cu mg kg-1 of soil and more resulted in a marked decline in the photosynthetic efficiency of Photosystem II (Phi2) but increased yield of non-photochemical quenching (PhiNPQ) and yield of non-regulatory energy dissipation (PhiNO) at tillering and flowering stages. In addition, these concentrations induced a delay in the flowering of rice, as a consequence of stress experienced in early growth stage. Significant lipid peroxidation and leaf area reduction were observed with 400 Cu mg kg-1 treatment at flowering stage. Rice grain yield decreased significantly at copper concentrations of 200 and 400 mg kg-1. Overall, excess copper inhibited photosynthetic capacity, growth, and development of rice in the early growth stage, and synergistic effects of yield components contributed to final grain yield reduction at harvesting stage. In addition, calculated integrated biomarker response (IBR) values reflect well the severity of Cu toxicity with a decreasing order from tillering stage to harvesting stage.

Calcification in Three Common Calcified Algae from Phuket, Thailand: Potential Relevance on Seawater Carbonate Chemistry and Link to Photosynthetic Process.

Calcifying macroalgae contribute significantly to the structure and function of tropical marine ecosystems. Their calcification and photosynthetic processes are not well understood despite their critical role in marine carbon cycles and high vulnerability to environmental changes. This study aims to provide a better understanding of the macroalgal calcification process, focusing on its relevance concerning seawater carbonate chemistry and its relationship to photosynthesis in three dominant calcified macroalgae in Thailand, Padina boryana, Halimeda macroloba and Halimeda opuntia. Morphological and microstructural attributes of the three macroalgae were analyzed and subsequently linked to their calcification rates and responses to inhibition of photosynthesis. In the first experiment, seawater pH, total alkalinity and total dissolved inorganic carbon were measured after incubation of the macroalgae in the light and after equilibration of the seawater with air. Estimations of carbon uptake into photosynthesis and calcification and carbon release into air were obtained thereafter. Our results provide evidence that calcification of the three calcified macroalgae is a potential source of CO2, where calcification by H. opuntia and H. macroloba leads to a greater release of CO2 per biomass weight than P. boryana. Nevertheless, this capacity is expected to vary on a diurnal basis, as the second experiment indicates that calcification is highly coupled to photosynthetic activity. Lower pH as a result of inhibited photosynthesis under darkness imposes more negative effects on H. opuntia and H. macroloba than on P. boryana, implying that they are more sensitive to acidification. These effects were worsened when photosynthesis was inhibited by 3-(3,4-dichlorophenyl)-1,1-dimethylurea, highlighting the significance of photosynthetic electron transport-dependent processes. Our findings suggest that estimations of the amount of carbon stored in the vegetated marine ecosystems should account for macroalgal calcification as a potential carbon source while considering diurnal variations in photosynthesis and seawater pH in a natural setting.

Temperature and concentration of ZnO particles affect life history traits and oxidative stress in Daphnia magna.

Temperature affects physiological processes in organisms and the toxicity of chemicals. The widespread industrial use of ZnO causes contamination in aquatic ecosystems. This study aimed to investigate the chronic toxicity of ZnO at different temperatures using Daphnia magna as a model organism. The chronic toxicity of five different concentrations of ZnO was assessed at 23 °C and 28 °C. The results showed that higher concentrations of ZnO inhibited growth, production of first clutch eggs and juvenile accumulation at both 23 °C and 28 °C. Growth rate, numbers of first clutch eggs and juvenile accumulation were lower at 28 °C than at 23 °C. We also observed the levels of malondialdehyde (MDA) and superoxide dismutase (SOD) activity. At higher concentrations of ZnO, oxidative stress was induced leading to increase MDA level and decrease SOD activity at 28 °C. These findings indicated that high temperature and high concentration of ZnO inhibited the activity of enzymatic proteins. Nonetheless, among all treatments, the accumulation of zinc in D. magna was not significantly different. Our results suggested that both ZnO and higher temperature induced oxidative stress in D. magna. As a result, MDA concentration increased, SOD activity changed and the growth and reproduction of D. magna was adversely affected.

Excess copper promotes photoinhibition and modulates the expression of antioxidant-related genes in Zostera muelleri.

Copper (Cu) is an essential micronutrient for plants and as such is vital to many metabolic processes. Nevertheless, when present at elevated concentrations, Cu can exert toxic effects on plants by disrupting protein functions and promoting oxidative stress. Due to their proximity to the urbanised estuaries, seagrasses are vulnerable to chemical contamination via industrial runoff, waste discharges and leachates. Zostera muelleri is a common seagrass species that forms habitats in the intertidal areas along the temperate coast of Australia. Previous studies have shown the detrimental effects of Cu exposure on photosynthetic efficiency of Z. muelleri. The present study focuses on the impacts of sublethal Cu exposure on the physiological and molecular responses. By means of a single addition, plants were exposed to 250 and 500 µg Cu L-1 (corresponding to 3.9 and 7.8 µM, respectively) as well as uncontaminated artificial seawater (control) for 7 days. Chlorophyll fluorescence parameters, measured as the effective quantum yield (ϕPSII), the maximum quantum yield (Fv/Fm) and non-photochemical quenching (NPQ) were assessed daily, while Cu accumulation in leaf tissue, total reactive oxygen species (ROS) and the expression of genes involved in antioxidant activities and trace metal binding were determined after 1, 3 and 7 days of exposure. Z. muelleri accumulated Cu in the leaf tissue in a concentration-dependent manner and the bioaccumulation was saturated by day 3. Cu exposure resulted in an acute suppression of ϕPSII and Fv/Fm. These two parameters also showed a concentration- and time-dependent decline. NPQ increased sharply during the first few days before subsequently decreasing towards the end of the experiment. Cu accumulation induced oxidative stress in Z. muelleri as an elevated level of ROS was detected on day 7. Lower Cu concentration promoted an up-regulation of genes encoding Cu/Zn-superoxide dismutase (Cu/Zn-sod), ascorbate peroxidase (apx), catalase (cat) and glutathione peroxidase (gpx), whereas no significant change was detected with higher Cu concentration. Exposure to Cu at any concentration failed to induce regulation in the expression level of genes encoding metallothionein type 2 (mt2), metallothionein type 3 (mt3) and cytochrome c oxidase copper chaperone (cox17). It is concluded that chlorophyll fluorescence parameters provide timely probe of the status of photosynthetic machinery under Cu stress. In addition, when exposed to a moderate level of Cu, Z. muelleri mitigates any induced oxidative stress by up-regulating transcripts coding for antioxidant enzymes.