Assessing the interactive effects of microplastics and acid rain on cadmium toxicity in rice seedlings: Insights from physiological and transcriptomic analyses.

In heavy metal-contaminated areas, the simultaneous occurrence of increasing microplastic pollution and persistent acid rain poses a serious threat to food security. However, the mechanisms of combined exposure to microplastics (MP) and acid rain (AR) on the toxicity of cadmium (Cd) in rice seedlings remain unclear. Our study investigated the combined effects of exposure to polyvinyl chloride microplastics and AR (pH 4.0) on the toxicity of Cd (0.3, 3, and 10 mg/L) in rice seedlings. The results showed that at low Cd concentrations, the combined exposure had no significant effect, but at high Cd concentrations, it alleviated the effects of Cd stress. The combined application of MP and AR alleviated the inhibitory effects of Cd on seedling growth and chlorophyll content. Under high Cd concentrations (10 mg/L), the simultaneous addition of MP and AR significantly reduced the production of reactive oxygen species (ROS), the content of malondialdehyde (MDA), and the activity of the superoxide dismutase (SOD). Compared with AR or MP alone, the combination of MP and AR reduced root cell damage and Cd accumulation in rice seedlings. Transcriptomic analysis confirmed that under high Cd concentrations, the combination of MP and AR altered the expression levels of genes related to Cd transport, uptake, MAPK kinase, GSTs, MTs, and transcription factors, producing a synergistic effect on oxidative stress and glutathione metabolism. These results indicate that co-exposure to MP and AR affected the toxicity of Cd in rice seedlings and alleviated Cd toxicity under high Cd concentrations to some extent. These findings provide a theoretical basis for evaluating the toxicological effects of microplastic and acid rain pollution on crop growth in areas contaminated with heavy metals, and are important for safe agricultural production and ecological security.

Physiological and biochemical responses of tea seedlings (Camellia sinensis) to simulated acid rain conditions.

Tea (Camellia sinensis), widely planted in the south of China, and often exposed to acid rain. However, research concerning the impacts of acid rain on physiology and biochemistry of tea plants is still scarce. In this study, we investigated the influence of simulated acid rain (SAR) on plant height, root length, photosynthetic pigment, Fv/Fm, proline, malondialdehyde, antioxidant enzyme activity, total nitrogen, caffeine, catechins, and free amino acids. Our results showed that SAR at pH 4.5 did not hinder plant development because growth characteristics, photosynthesis, and ascorbate peroxidase and catalase activities did not decrease at this pH compared to those at the other investigated pH values. However, at pH 3.5 and pH 2.5, the activities of antioxidase and concentrations of malondialdehyde and proline increased significantly in response to the decrease of photosynthetic pigments and Fv/Fm. In addition, the increase in acidity increased total nitrogen, certain amino acid content (theanine, cysteine), and decreased catechin and caffeine contents, resulting in an imbalance of the carbon and nitrogen metabolisms. Our results indicated that SAR at pH 3.5 and pH 2.5 could restrict photosynthesis and the antioxidant defense system, causing metabolic disorders and ultimately affecting plant development and growth, but SAR at pH 4.5 had no toxic effects on tea seedlings when no other stress factors are involved.

Hydrolytic enzymes mediated lipid-DNA catabolism and altered gene expression of antioxidants under combined application of lead and simulated acid rain in Fenugreek (Trigonella foenum graecum L.) seedlings.

Understanding ill effects of simultaneous existence of various abiotic stresses, commonly observed due to various anthropogenic activities and global climate change these days, over plants growth, metabolic activity and yield responses are important for continued agricultural productivity and food security. In the present study, seedlings of Fenugreek (Trigonella foenum graecum L.) were subjected to lead (Pb, 1200 ppm) and/or simulated acid rain (SAR, pH 3.5) for 30 days, and were then analysed. The results revealed reduced growth, and total lipid and DNA contents, while enhanced Pb accumulation, biological concentration factor, biological accumulation coefficient, translocation factor, lipase activity, and levels of free fatty acid, conjugated diene, lipid hydroperoxide, DNA oxidation and DNase activity under Pb and/or SAR exposure. Additionally, activities and gene expression levels of antioxidants (superoxide dismutase, catalase, guaiacol peroxidase and ascorbate peroxidase) were enhanced in response to applied treatments. The results also suggested that inhibitions/ accelerations determined under joint addition of Pb and SAR were comparatively more profound than those measured under their single application. Additionally, root was more sensitive to Pb treatment, compared to both leaf and shoot. Hence, under simultaneous presence of two or more number of abiotic stresses, the strategy opted by plants for survival is chiefly governed by the interaction between prevailing stressors, which is then conceived by plants as a new state of stress.

Effect of exogenous calcium on growth, nutrients uptake and plasma membrane H+-ATPase and Ca2+-ATPase activities in soybean (Glycine max) seedlings under simulated acid rain stress.

Calcium (Ca) is one of essential elements for plant growth and development, and also plays a role in regulating plant cell physiology and cellular response to the environment. Here, we studied whether calcium played a role in enhancing tolerance of plants to acid rain stress by hydroponics and simulating acid rain stress. Our results show that acid rain (pH 4.5/pH 3.0) caused decreases in dry weight biomass, chlorophyll content and uptake of nutrients elements (NO3-, P, K, Mg, Zn and Mo) and an increase in membrane permeability of root. However, all parameters in soybean treated with exogenous calcium (5 mM) and acid rain at pH 4.5 were closed to the control levels. In addition, exogenous calcium (5 mM) alleviated the inhibition induced by pH 3.0 acid rain on the activity of plasma membranes H+-ATPase and the expression of GmPHA1 at transcriptional level, being benefiting to maintaining uptake of nutrients (NO3-, P, K, Mg, and Zn), and then lower the decrease in dry weight biomass and chlorophyll content. After a 5-day recovery (without acid rain stress), all parameters in soybean treated with acid rain at pH 3.0 and exogenous calcium were still worse than those of the control, but obviously better than those treated with acid rain at pH 3.0. Higher activity of plasma membrane H+-ATPase in soybean treated with acid rain at pH 3.0 and exogenous calcium was good to uptake of nutrients and promoted the recovery of soybean growth, compared with soybean treated with acid rain at pH 3.0. In conclusion, exogenous calcium could alleviate the inhibition caused by acid rain on soybean growth by increasing the activity of plasma membrane H+-ATPase for providing driving force to nutrient absorption, and its regulating effect was limited by intensity of acid rain. Furthermore, the application of exogenous calcium can be one of ways to alleviate the damage caused by acid rain to plants.

Evidence for episodic acidification effects on migrating Atlantic salmon Salmo salar smolts.

Field studies were conducted to determine levels of gill aluminium as an index of acidification effects on migrating Atlantic salmon Salmo salar smolts in the north-eastern U.S.A. along mainstem river migration corridors in several major river basins. Smolts emigrating from the Connecticut River, where most (but not all) tributaries were well buffered, had low or undetectable levels of gill aluminium and high gill Na(+) /K(+) -ATPase (NKA) activity. In contrast, smolts emigrating from the upper Merrimack River basin where most tributaries are characterized by low pH and high inorganic aluminium had consistently elevated gill aluminium and lower gill NKA activity, which may explain the low adult return rates of S. salar stocked into the upper Merrimack catchment. In the Sheepscot, Narraguagus and Penobscot Rivers in Maine, river and year-specific effects on gill aluminium were detected that appeared to be driven by underlying geology and high spring discharge. The results indicate that episodic acidification is affecting S. salar smolts in poorly buffered streams in New England and may help explain variation in S. salar survival and abundance among rivers and among years, with implications for the conservation and recovery of S. salar in the north-eastern U.S.A. These results suggest that the physiological condition of outmigrating smolts may serve as a large-scale sentinel of landscape-level recovery of atmospheric pollution in this and other parts of the North Atlantic region.

Response of plasma membrane H+-ATPase in rice (Oryza sativa) seedlings to simulated acid rain.

Understanding the adaptation of plants to acid rain is important to find feasible approaches to alleviate such damage to plants. We studied effects of acid rain on plasma membrane H(+)-ATPase activity and transcription, intracellular H(+), membrane permeability, photosynthetic efficiency, and relative growth rate during stress and recovery periods. Simulated acid rain at pH 5.5 did not affect plasma membrane H(+)-ATPase activity, intracellular H(+), membrane permeability, photosynthetic efficiency, and relative growth rate. Plasma membrane H(+)-ATPase activity and transcription in leaves treated with acid rain at pH 3.5 was increased to maintain ion homeostasis by transporting excessive H(+) out of cells. Then intracellular H(+) was close to the control after a 5-day recovery, alleviating damage on membrane and sustaining photosynthetic efficiency and growth. Simulated acid rain at pH 2.5 inhibited plasma membrane H(+)-ATPase activity by decreasing the expression of H(+)-ATPase at transcription level, resulting in membrane damage and abnormal intracellular H(+), and reduction in photosynthetic efficiency and relative growth rate. After a 5-day recovery, all parameters in leaves treated with pH 2.5 acid rain show alleviated damage, implying that the increased plasma membrane H(+)-ATPase activity and its high expression were involved in repairing process in acid rain-stressed plants. Our study suggests that plasma membrane H(+)-ATPase can play a role in adaptation to acid rain for rice seedlings.

Combined effects of lead and acid rain on photosynthesis in soybean seedlings.

To explore how lead (Pb) and acid rain simultaneously affect plants, the combined effects of Pb and acid rain on the chlorophyll content, chlorophyll fluorescence reaction, Hill reaction rate, and Mg(2+)-ATPase activity in soybean seedlings were investigated. The results indicated that, when soybean seedlings were treated with Pb or acid rain alone, the chlorophyll content, Hill reaction rate, Mg(2+)-ATPase activity, and maximal photochemical efficiency (F(v)/F(m)) were decreased, while the initial fluorescence (F 0) and maximum quantum yield (Y) were increased, compared with those of the control. The combined treatment with Pb and acid rain decreased the chlorophyll content, Hill reaction rate, Mg(2+)-ATPase activity, F(v)/F(m), and Y and increased F 0 in soybean seedlings. Under the combined treatment with Pb and acid rain, the two factors showed additive effects on the chlorophyll content in soybean seedlings and exhibited antagonistic effects on the Hill reaction rate. Under the combined treatment with high-concentration Pb and acid rain, the two factors exhibited synergistic effects on the Mg(2+)-ATPase activity, F 0, F v/F m, as well as Y. In summary, the inhibition of the photosynthetic process is an important physiological basis for the simultaneous actions of Pb and acid rain in soybean seedlings.

Interactive effects of cadmium and acid rain on photosynthetic light reaction in soybean seedlings.

Interactive effects of cadmium (Cd(2+)) and acid rain on photosynthetic light reaction in soybean seedlings were investigated under hydroponic conditions. Single treatment with Cd(2+) or acid rain and the combined treatment decreased the content of chlorophyll, Hill reaction rate, the activity of Mg(2+)-ATPase, maximal photochemical efficiency and maximal quantum yield, increased initial fluorescence and damaged the chloroplast structure in soybean seedlings. In the combined treatment, the change in the photosynthetic parameters and the damage of chloroplast structure were stronger than those of any single pollution. Meanwhile, Cd(2+) and acid rain had the interactive effects on the test indices in soybean seedlings. The results indicated that the combined pollution of Cd(2+) and acid rain aggravated the toxic effect of the single pollution of Cd(2+) or acid rain on the photosynthetic parameters due to the serious damage to the chloroplast structure.

Physiological, molecular, and cellular mechanisms of impaired seawater tolerance following exposure of Atlantic salmon, Salmo salar, smolts to acid and aluminum.

We examined the physiological, molecular, and cellular mechanisms of impaired ion regulation in Atlantic salmon, Salmo salar, smolts following acute acid and aluminum (Al) exposure. Smolts were exposed to: control (pH 6.5, 3.4 micrpg l(-1) Al), acid and low Al (LAl: pH 5.4, 11 microg l(-1) Al), acid and moderate Al (MAl: pH 5.3, 42 microg l(-1) Al), and acid and high Al (HAl: pH 5.4, 56 microg l(-1) Al) for two and six days. At each time-point, smolts were sampled directly from freshwater treatment tanks and after a 24h seawater challenge. Exposure to acid/MAl and acid/HAl led to accumulation of gill Al, substantial alterations in gill morphology, reduced gill Na(+)/K(+)-ATPase (NKA) activity, and impaired ion regulation in both freshwater and seawater. Exposure to acid/MAl for six days also led to a decrease in gill mRNA expression of the apical Cl(-) channel (cystic fibrosis transmembrane conductance regulator I), increased apoptosis upon seawater exposure, an increase in the surface expression of mitochondria-rich cells (MRCs) within the filament epithelium of the gill, but reduced abundance of gill NKA-positive MRCs. By contrast, smolts exposed to acid and the lowest Al concentration exhibited minor gill Al accumulation, slight morphological modifications in the gill, and impaired seawater tolerance in the absence of a detectable effect on freshwater ion regulation. These impacts were accompanied by decreased cell proliferation, a slight increase in the surface expression of MRCs within the filament epithelium, but no impact on gill apoptosis or total MRC abundance was observed. However, MRCs in the gills of smolts exposed to acid/LAl exhibited morphological alterations including decreased size, staining intensity, and shape factor. We demonstrate that the seawater tolerance of Atlantic salmon smolts is extremely sensitive to acute exposure to acid and low levels of Al, and that the mechanisms underlying this depend on the time-course and severity of Al exposure. We propose that when smolts are exposed to acid and moderate to high Al concentrations, impaired seawater tolerance results from extensive gill Al accumulation, damage to the epithelium, reduced MRC and transport protein abundance, and a synergistic stimulation of apoptosis in the gill upon seawater exposure. When smolts are exposed to acid and low levels of Al, loss of seawater tolerance appears to be independent of these mechanisms and may result instead from a shift in the phenotype of MRCs present in the gill epithelium.

Effect of simulated acid rain on the litter decomposition of Quercus acutissima and Pinus massoniana in forest soil microcosms and the relationship with soil enzyme activities.

With the continuing increase in human activities, ecologists are increasingly interested in understanding the effects of acid rain on litter decomposition. Two dominant litters were chosen from Zijin Mountain in China: Quercus acutissima from a broad-leaved forest and Pinus massoniana from a coniferous forest. The litters were incubated in microcosms and treated with simulated acid rain (gradient pH levels). During a six-month incubation, changes in chemical composition (i.e., lignin, total carbohydrate, and nitrogen), litter mass losses, soil pH values, and activities of degradative enzymes were determined. Results showed that litter mass losses were depressed after exposure to acid rain and the effects of acid rain on the litter decomposition rates of needles were higher than on those of leaves. Results also revealed that simulated acid rain restrained the activities of cellulase, invertase, nitrate reductase, acid phosphatase, alkaline phosphatase, polyphenol oxidase, and urease, while it enhanced the activities of catalase in most cases during the six-month decomposition process. Catalase and polyphenol oxidase were primarily responsible for litter decomposition in the broad-leaved forest, while invertase, nitrate reductase, and urease were primarily responsible for litter decomposition in the coniferous forest. The results suggest acid rain-restrained litter decomposition may be due to the depressed enzymatic activities. According to the results of this study, soil carbon in subtropical forests would accumulate as a long-term consequence of continued acid rain. This may presumably alter the balance of ecosystem carbon flux, nutrient cycling, and humus formation, which may, in turn, have multiple effects on forest ecosystems.

Photosynthesis and oxidative stress in the restinga plant species Eugenia uniflora L. exposed to simulated acid rain and iron ore dust deposition: potential use in environmental risk assessment.

The Brazilian sandy coastal plain named restinga is frequently subjected to particulate and gaseous emissions from iron ore factories. These gases may come into contact with atmospheric moisture and produce acid rain. The effects of the acid rain on vegetation, combined with iron excess in the soil, can lead to the disappearance of sensitive species and decrease restinga biodiversity. The effects of iron ore dust deposition and simulated acid rain on photosynthesis and on antioxidant enzymes were investigated in Eugenia uniflora, a representative shrub species of the restinga. This study aimed to determine the possible utility of this species in environmental risk assessment. After the application of iron ore dust as iron solid particulate matter (SPM(Fe)) and simulated acid rain (pH 3.1), the 18-month old plants displayed brown spots and necrosis, typical symptoms of iron toxicity and injuries caused by acid rain, respectively. The acidity of the rain intensified leaf iron accumulation, which reached phytotoxic levels, mainly in plants exposed to iron ore dust. These plants showed the lowest values for net photosynthesis, stomatal conductance, transpiration, chlorophyll a content and electron transport rate through photosystem II (PSII). Catalase and superoxide dismutase activities were decreased by simulated acid rain. Peroxidase activity and membrane injury increased following exposure to acid rain and simultaneous SPM(Fe) application. Eugenia uniflora exhibited impaired photosynthetic and antioxidative metabolism in response to combined iron and acid rain stresses. This species could become a valuable tool in environmental risk assessment in restinga areas near iron ore pelletizing factories. Non-invasive evaluations of visual injuries, photosynthesis and chlorophyll a fluorescence, as well as invasive biochemical analysis could be used as markers.

Responses of antioxidant enzyme and photosynthesis in rape seedling to the combined stresses of acid rain and ultraviolet-B radiation.

Effects of the simulated acid rain (AR) and ultraviolet-B (UV-B, 280-320 nm) radiation with a single or two ways simultaneously (AR + UV-B) on the antioxidant enzyme and photosynthesis of the rape seedlings were investigated by the hydroponic culture. The results of static experiment indicated that the tolerance of rape seedling to single stress (AR or UV-B) is stronger than that to dual stresses (AR + UV-B). Furthermore, the dual stresses had additive effect on catalase activity, and a synergistic effect on MDA content, net photosynthesis rate, water use efficiency as well as intercellular CO2 concentration. Meanwhile, it has an independent effect on chlorophyll content, stomatal conductance, and transpiration rate as well as membrane permeability. During 64 h restoration course, the dynamic change in the curves of physiological and biochemical indices were not identical, and none of them show a simple linear variation. According to the static and dynamic experiments, it was found that a responsive sequence of catalase activity, membrane permeability, MDA content and photosynthetic characteristics to the above-mentioned stresses was as follows: AR + UV-B > UV-B > AR.

Alleviation of aluminum toxicity to Rhizobium leguminosarum bv. viciae by the hydroxamate siderophore vicibactin.

Acid rain solubilises aluminum which can exert toxic effects on soil bacteria. The root nodule bacterium Rhizobium leguminosarum biovar viciae synthesises the hydroxamate siderophore vicibactin in response to iron limitation. We report the effect of vicibactin on the toxicity of aluminum(III) to R. leguminosarum and kinetic studies on the reaction of vicibactin with Al(III) and Fe(III). Aluminum (added as the nitrate) completely inhibited bacterial growth at 25 microM final concentration, whereas the preformed Al-vicibactin complex had no effect. When aluminum and vicibactin solutions were added separately to growing cultures, growth was partly inhibited at 25 microM final concentration of each, but fully inhibited at 50 microM final concentration of each. Growth was not inhibited at 50 microM Al and 100 microM vicibactin, probably reflecting the slow reaction between Al and vicibactin; this results in some aluminum remaining uncomplexed long enough to exert toxic effects on growth, partly at 25 microM Al and vicibactin and fully at 50 microM Al and vicibactin. At 100 microM vicibactin and 50 microM Al, Al was complexed more effectively and there was no toxic effect. It was anticipated that vicibactin might enhance the toxicity of Al by transporting it into the cell, but the Al-vicibactin complex was not toxic. Several explanations are possible: the Al-vicibactin complex is not taken up by the cell; the complex is taken up but Al is not released from vicibactin; Al is released in the cell but is precipitated immediately. However, vicibactin reduces the toxicity of Al by complexing it outside the cell.

Biochemical responses of the mycorrhizae in Pinus massoniana to combined effects of Al, Ca and low pH.

Biochemical responses of Pinus massoniana, with and without the inoculation mycorrhizal fungus Pisolithus tinctorius at the root, to artificial acid rain (pH 2.0) and various Ca/Al ratios were investigated. Some enzymes associated with the nutritive metabolism, such as acid phosphatase, alkaline phosphatase, nitrate reductase, mannitol dehydrogenase and trehalase, in the roots, stems and leaves of plant were obviously inhibited by the artificial acid rain and Al. After treatment with pH 2.0 + Ca/Al (0/1 or 1/10) artificial acid rain, the protein content in the organs was decreased. However, the activities of superoxide dismutase (SOD) and peroxidase (POD) and glutathione (GSH) concentrations were induced. It demonstrated that acid rain and Al could induce oxygen radicals in plant. Compared with the treatments with lower pH or Al, respectively, the combination of lower pH and Al concentration was more toxic to P. massoniana. Al toxicity could be ameliorated by the addition of Ca and the amelioration was the most when the ratio was 1/1 among the various Ca/Al ratio. Infection with mycorrhizal fungus P. tinctorius at the root of P. massoniana increased the ability of the plant to resist the toxicity of artificial acid rain and Al stress.

Differential responses of certain lichen species to sulfur-containing solutions under acidic conditions as expressed by the production of stress-ethylene.

To determine whether fluctuations in the concentration of ethylene produced by lichens exposed to sulfur-containing solutions at a low pH correlate with the tolerance/sensitivity of these lichens to air pollution, we measured the amount of ethylene produced by thalli soaked in H2SO4 and NaHSO3. The exposure of Hypogymnia physodes, Cladina stellaris, and Bryoria fuscescens to H2SO4 at a pH ranging between 4.0 and 2.0 did not produce changes in the concentration of ethylene in comparison with samples wetted with H2O at pH 6.8. The exposure of two pendulous lichens, Usnea hirta and Alectoria sarmentosa, to 1.0 and 5.0 mM H2SO4 at pH 2.7 and 2.0, respectively, stimulated only a slight increase of ethylene production, whereas another pendulous lichen, Bryoria fremontii, exposed to H2SO4 at pH 4.0-2.0 decreased its production of ethylene. The soaking of H. physodes, U. hirta, C. stellaris, and A. sarmentosa thalli in NaHSO3 at pH 4.0 gradually increased the production of ethylene. The exposure of B. fremontii and B. fuscescens to low NaHSO3 concentrations depressed the production of ethylene in these lichens. The indifference of H. physodes to H2SO4 under strong acidic conditions correlated with its resistance to SO2 in the air. In accordance with a model by D.M. Reid (In "Effects of Atmospheric Pollutants on Forests, Wetlands and Agricultural Ecosystems" (T.C. Hutchinson and K.M. Meema, Eds.), Vol G 16, pp. 241-245. NATO ASI Series, Springer-Verlag, Berlin and Heidelberg, 1987) referring to higher plants, it is suggested that sulfur-containing solutions under acidic conditions increase the solubility of particles containing heavy metals entrapped among the mycobiont hyphae in lichens. This may lead to an increase of the production of endogenous ethylene in lichens as they are exposed to sulfur-containing chemicals, to acidic rain, or to heavy metal-polluted air.