Nitrogen Metabolism in Acorus calamus L. Leaves Induced Changes in Response to Microcystin-LR at Environmentally Relevant Concentrations.

Acorus calamus L., a semiaquatic plant with a high capacity to remove nitrogen and phosphorus from polluted water, is a potential candidate plant for use in the restoration of eutrophic aquatic ecosystems. However, it is not clear how microcystins (MCs), commonly found in eutrophic water, influence plant growth since the effects of MCs are likely to be dose and species dependent. The present study aimed to investigate the regulation of nitrogen metabolism, a key metabolic process related to plant growth, in the leaves of A. calamus L. exposed to microcystin-leucine-arginine (MC-LR) (1.0-29.8 µg/L). Nitrate (NO3-) uptake, assimilation and transformation was stimulated in the leaves of A. calamus L. when exposed to 1.0 µg/L MC-LR through the elevation of nitrate reductase (NR), glutamine synthetase (GS), glutamate synthase (GOGAT), glutamic-pyruvic transaminase (GPT), and glutamic-oxaloacetic transaminase (GOT) activity. Conversely, MC-LR inhibited nitrogen metabolism by decreasing NO3- uptake and the activities of enzymes related to nitrogen metabolism following exposure to MC-LR (9.9-29.8 µg/L) for 30 days, while, ammonium nitrogen (NH4+) content and glutamate dehydrogenase (GDH) activity increased significantly (p < 0.05, LSD test), when compared with the control group. Chronic exposure to MC-LR (9.9-29.8 µg/L) negatively influenced nitrogen metabolism in A. calamus L. leaves, which suggested that it may not be a suitable candidate species for use in the restoration of eutrophic aquatic ecosystems containing MC-LR at concentrations ≥ 9.9 µg/L.

Phytotoxicity and oxidative stress of perfluorooctanesulfonate to two riparian plants: Acorus calamus and Phragmites communis.

Despite previous efforts and the rapid progress on elucidating the impact of perfluorooctanesulfonate (PFOS) on the environment, its effects on riparian plants, a key component of aquatic ecosystems, are still poorly understood. A 48-day hydroponic experiment was carried out on two typical riparian species (Acorus calamus and Phragmites communis) to examine the toxic effects of PFOS on these plants. The results showed that, at high concentration (more than 10 mg L-1), PFOS could prevent chlorophyll accumulation (reduced by 13.7-22.2% at 10 mg L-1 PFOS and 22.4-30.0% at 50 mg L-1 PFOS for 48 days) and soluble protein synthesis (reduced by 2.3-9.0% at 10 mg L-1 PFOS and 10.6-26.8% at 50 mg L-1 PFOS for 48 days). Contrastingly, less than 1 mg L-1 of PFOS could induce chlorophyll accumulation (increased by 18.6% in A. calamus roots, 11.3% in A. calamus leaves, and 13.6% in P. communis roots at 1 mg L-1 PFOS for 3 days) and soluble protein synthesis (increased by 6.1% in A. calamus roots, 18.4% in A. calamus leaves, 9.7% in P. communis roots, 23.4% in P. communis stems, and 24.0% in P. communis leaves, at 1 mg L-1 PFOS for 6 days). In addition, PFOS led to oxidative stress, as revealed by the elevated concentrations of malonaldehyde and hydrogen peroxide, and reduced the activities of antioxidant enzymes such as superoxide dismutase (reduced by 10.3% in P. communis stems at 50 mg L-1 PFOS for 48 days), catalase (reduced by 20.6-50.3% in test species at 50 mg L-1 PFOS for 48 days), and peroxidase (reduced by 24.9-37.7% in test species at 50 mg L-1 PFOS for 48 days). The biomarkers of both plants changed rapidly in the first half of the experiment (0-24 days) and stabilized in the second half of the experiment (24-48 days). The risk and related factors of PFOS on riparian plants were evaluated by using these biomarkers. Experiments showed that P. communis was more resistant to low concentration (<10 mg L-1) of PFOS than A. calamus.

Effects of antibiotics on nitrogen uptake of four wetland plant species grown under hydroponic culture.

To investigate the effects of antibiotics on nitrogen removal and uptake by wetland plants, four typical macrophyte species, Cyperus alternifolius L., Typha angustifolia L., Lythrum salicaria L., and Acorus calamus L., were grown in hydroponic cultivation systems and fed wastewater polluted with 10 µg L-1 Ofloxacin (OFL) and Tetracycline (TET). Biomass production, nitrogen mass concentration, chlorophyll content, root exudates, and nitrogen removal efficiency of hydroponic cultivation were investigated. The results indicated that in all hydroponic systems, NH4+-N was entirely removed from the hydroponic substrate within 1 day and plant nitrogen accumulation was the main role of the removed NO3-. OFL and TET stimulated the accumulation of biomass and nitrogen of A. calamus but significantly inhibited the NO3--N removal ability of L. salicaria (98.6 to 76.2%) and T. augustifolia (84.3 to 40.2%). This indicates that A. calamus may be a good choice for nitrogen uptake in wetlands contaminated with antibiotics. OFL and TET improved the concentrations of total organic carbon (TOC), total nitrogen (TN), organic acid, and soluble sugars in root exudates, especially for oxalic acid. Considering the significant correlation between TOC of root exudates and nitrogen removal efficiency, the TOC of root exudates may be an important index for choosing macrophytes to maintain nitrogen removal ability in wetlands contaminated with antibiotics.

Stomatal Conductance Measurement for Toxicity Assessment in Zero-Effluent Constructed Wetlands: Effects of Landfill Leachate on Hydrophytes.

In this research, we explore for the first time the use of leaf stomatal conductance (gs) for phytotoxicity assessment. Plants respond to stress by regulating transpiration. Transpiration can be correlated with stomatal conductance when the water vapor pressure gradient for transpiration is constant. Thus, our working hypothesis was that the gs measurement could be a useful indicator of the effect of toxic compounds on plants. This lab-scale study aimed to test the measurement of gs as a phytotoxicity indicator. Our model plants were two common hydrophytes used in zero-effluent constructed wetlands for treating landfill leachate. The toxic influence of two types of leachate from old landfills (L1, L2) on common reed (Phragmites australis (Cav.) Trin. ex Steud.) and sweet flag (Acorus calamus L.) was tested. The gs measurements correlated well with plant response to treatments with six solutions (0 to 100%) of landfill leachate. Sweet flag showed higher tolerance to leachate solutions compared to common reed. The estimated lowest effective concentration (LOEC) causing the toxic effect values for these leachates were 3.94% of L1 and 5.76% of L2 in the case of reed, and 8.51% of L1 and 10.44% of L2 in the case of sweet flag. Leachate L1 was more toxic than L2. The leaf stomatal conductance measurement can be conducted in vivo and in the field. The proposed approach provides a useful parameter for indicating plant responses to the presence of toxic factors in the environment.

Effects of perchlorate on growth of four wetland plants and its accumulation in plant tissues.

Perchlorate contamination in water is of concern because of uncertainties about toxicity and health effects, impact on ecosystems, and possible indirect exposure pathways to humans. Therefore, it is very important to investigate the ecotoxicology of perchlorate and to screen plant species for phytoremediation. Effects of perchlorate (20, 200, and 500 mg/L) on the growth of four wetland plants (Eichhornia crassipes, Acorus calamus L., Thalia dealbata, and Canna indica) as well as its accumulation in different plant tissues were investigated through water culture experiments. Twenty milligrams per liter of perchlorate had no significant effects on height, root length, aboveground part weight, root weight, and oxidizing power of roots of four plants, except A. calamus, and increasing concentrations of perchlorate showed that out of the four wetland plants, only A. calamus had a significant (p<0.05) dose-dependent decrease in these parameters. When treated with 500 mg/L perchlorate, these parameters and chlorophyll content in the leaf of plants showed significant decline contrasted to control groups, except the root length of E. crassipes and C. indica. The order of inhibition rates of perchlorate on root length, aboveground part weight and root weight, and oxidizing power of roots was: A. calamus > C. indica > T. dealbata > E. crassipes and on chlorophyll content in the leaf it was: A. calamus > T. dealbata > C. indica > E. crassipes. The higher the concentration of perchlorate used, the higher the amount of perchlorate accumulation in plants. Perchlorate accumulation in aboveground tissues was much higher than that in underground tissues and leaf was the main tissue for perchlorate accumulation. The order of perchlorate accumulation content and the bioconcentration factor in leaf of four plants was: E. crassipes > C. indica > T. dealbata > A. calamus. Therefore, E. crassipes might be an ideal plant with high tolerance ability and accumulation ability for constructing wetland to remediate high levels of perchlorate polluted water.