Physiological Response of European Hornbeam Leaves to Nitrogen Dioxide Stress and Self-recovery

in Journal of the American Society for Horticultural Science

Plant leaves absorb atmospheric nitrogen dioxide (NO2) primarily via the stomata. Studies of changes in plant growth and physiology after exposure to NO2 are limited. Therefore, this study investigated the physiological response of Carpinus betulus (european hornbeam) chloroplasts after NO2 exposure using fumigation equipment that was able to control timing and record NO2 concentrations. The NO2 concentration was 6 µL·L−1. Seven treatment durations (0, 1, 6, 12, 24, 48, and 72 hours) were designed. After fumigation, plants recovered for 30 days under greenhouse conditions. The physiological response, stomatal behavior, thicknesses of palisade and spongy tissues, and chloroplast ultrastructure were measured. In the 48-hour and 72-hour NO2 treatment groups, the chloroplast contents and net photosynthesis rates of the leaves decreased, palisade and spongy tissues thickened, and chloroplast thylakoids swelled; however, the 1-hour NO2 treatment did not have a noticeable toxic effect on C. betulus leaves. After 30 days of recovery, the plants returned to their natural growth level by increasing the chloroplast content and enhancing net photosynthesis. Short durations and high concentrations of NO2 exposure had significantly negative impacts on the physiological response of C. betulus; however, this toxic effect of high NO2 concentrations on C. betulus can be recovered by restoration of unpolluted air. The results of this study may provide a scientific reference and an additional choice of plants species for the application of C. betulus in functional gardening design and ecological green space construction.

Contributor Notes

This work was supported by The National Natural Science Foundation of China (31770752), Jiangsu Province Agricultural Science and Technology Independent Innovation Funds (CX (16)1005-4), Postgraduate Research & Practice Innovation Program of Jiangsu Province, Jiangsu Province Engineering Technology Research Center Projects (BM2013478), Jiangsu Province Six Big Talent Peak Project (NY-029), and Fifth Stage Funded Research Projects of 333 in Jiangsu Province.

Corresponding author. E-mail: zhuzunling@njfu.edu.cn.

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Article Figures

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    A fume exhauster device with timing control and nitrogen dioxide (NO2) concentration recording capabilities for NO2 fumigation: 1 = NO2 gas cylinders; 2 = pressure-reducing valve; 3 = solenoid valve; 4 = microcomputer timer switch system; 5 = centrifugal fan; 6 = blower; 7 = air inlet pipe; 8 = NO2 gas sensor; 9 = computer terminal; 10 = glass partition; 11 = acrylic sheet; and 12 = fume gas chamber.

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    Diurnal change in photosynthesis [(A) net photosynthetic rate (Pn), (B) transpiration rate (Tr), (C) stomatal conductance (gS), and (D) intercellular CO2 concentration (Ci)] in leaves of european hornbeam grown under exposure to 6 µL·L−1 of NO2 for 0 [control (CK)], 1, 6, 12, 24, 48, and 72 h and after 30 d of self-recovery.

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    Photosynthesis irradiance (PAR) responses [(A) net photosynthetic rate (Pn), (B) transpiration rate (Tr), (C) stomatal conductance (gS), and (D) intercellular CO2 concentration (Ci)] in leaves of european hornbeam grown under exposure to 6 µL·L−1 of NO2 for 0 [control (CK)], 1, 6, 12, 24, 48, and 72 h and after 30 d of self-recovery; PPFD = photosynthetic photon flux density.

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    Scanning electron microscope (SEM) images of the stomata of european hornbeam leaves of the control group [CK (0 h)] exposed to 6 µL·L−1 of NO2 for 1, 24, and 72 h and after 30 d of self-recovery based on two scale bars (50 and 200 μm). (A) Microstructure of leaves without NO2 treatment. (B) Normal stomatal state of the leaves without NO2 treatment. (CE) Increased number of closed stomata on abaxial surfaces with the increased timing of NO2 treatment in comparison with the controls and self-recovered plants. (F) Stomatal state of the leaves after 30 d of recovery. UE = upper epidermis, PT = palisade tissue, ST = spongy tissue, LV = leaf vein, TR = trichomes.

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    Scanning electron microscope (SEM) images of palisade tissue and spongy tissue of the european hornbeam leaves of the control group [CK (0 h)] exposed to 6 µL·L−1 of NO2 for 1, 24, and 72 h and after 30 d of self-recovery based on two scale bars (50 and 100 μm). (A) Normal palisade tissue and spongy tissue in the leaves without NO2 treatment. (BD) Thicknesses of the palisade and spongy tissues increased after NO2 treatment compared with those of the controls and self-recovered plants. (E) Palisade tissue and spongy tissue of the leaves after 30 d of recovery.

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    Transmission electron microscope (TEM) micrographs of chloroplasts from european hornbeam leaves of the control group [CK (0 h)], 6 µL·L−1 NO2-treated (1, 24, and 72 h, respectively) and self-recovery plants. (A) CK plant with a discoidal chloroplast containing abundant well-compartmentalized grana. (B) Chloroplast with complete leaf structures in the 1-h treatment group. (C) Slightly changed chloroplasts with dilated thylakoid (yellow oval) in the 24-h treatment group. (D) Extensively damaged chloroplasts in the 72-h treatment group showing grossly dilated thylakoids (yellow oval). (E) Chloroplasts with complete leaf structures after 30 d of self-recovery. C = cytoplasm, M = mitochondria, P = plastoglobuli, Pl = primary leaf, S = starch grain, T = tonoplast, V = vacuole, CW = cell wall, G = grana stack.

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    Pairwise comparisons of the net photosynthetic rates among the control (0 h), 6 µL·L−1 NO2-treated (1, 6, 12, 24, 48, and 72 h), and self-recovery european hornbeam plants at different diurnal time points based on nonparametric Kruskal-Wallis one-factor analysis of variance. The yellow line indicates a significant difference between groups after Bonferroni correction: (A) 0800 hr; (B) 1000 hr; (C) 1200 hr; (D) 1400 hr; (E) 1600 hr; and (F) 1800 hr. 1 = control group; 2 = 1-h group; 3 = 6-h group; 4 = 12-h group; 5 = 24-h group; 6 = 48-h group; 7 = 72-h group; 8 = recovery group.

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    Pairwise comparisons of the transpiration rates among the control (0 h), 6 µL·L−1 NO2-treated (1, 6, 12, 24, 48, and 72 h), and self-recovery european hornbeam plants at different diurnal time points based on nonparametric Kruskal-Wallis one-factor analysis of variance. The yellow line indicates a significant difference between groups after Bonferroni correction: (A) 0800 hr; (B) 1000 hr; (C) 1200 hr; (D) 1400 hr; (E) 1600 hr; and (F) 1800 hr. 1 = control group; 2 = 1-h group; 3 = 6-h group; 4 = 12-h group; 5 = 24-h group; 6 = 48-h group; 7 = 72-h group; 8 = recovery group.

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    Pairwise comparisons of the gS among the control (0 h), 6 µL·L−1 NO2-treated (1, 6, 12, 24, 48, and 72 h) and self-recovery european hornbeam plants at different diurnal time points based on nonparametric Kruskal-Wallis one-factor analysis of variance. The yellow line indicates a significant difference between groups after Bonferroni correction: (A) 0800 hr; (B) 1000 hr; (C) 1200 hr; (D) 1400 hr; (E) 1600 hr; and (F) 1800 hr. 1 = control group; 2 = 1-h group; 3 = 6-h group; 4 = 12-h group; 5 = 24-h group; 6 = 48-h group; 7 = 72-h group; 8 = recovery group.

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    Pairwise comparisons of the intercellular CO2 concentrations between different diurnal time points in the same european hornbeam group. The yellow line indicates a significant difference between different diurnal time points after Bonferroni correction: (A) control group; (B) 1-h NO2-treated group; (C) 6-h NO2-treated group; (D) 12-h NO2-treated group; (E) 24-h NO2-treated group; (F) 48-h NO2-treated group; (G) 72-h NO2-treated group; and (H) recovery group. 1 = 0800 hr; 2 = 1000 hr; 3 = 1200 hr; 4 = 1400 hr; 5 = 1600 hr; 6 = 1800 hr.

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