To reveal the mechanism of submerged plants decline in progressively eutrophicated freshwaters, physiological responses of Vallisneria natans to epiphytic algae were studied in simulation lab by measuring plant physiological indexes of chlorophyll content, malondialdehyde (MDA) content, and superoxide dismutase (SOD) activity based on a 2 × 4 factorial design with two epiphytic conditions (with epiphytic algae and without) and four levels of N and P concentrations in water (N-P[mg.L(-1)]: 0.5, 0.05; 2.5, 0.25; 4.5, 0.45; 12.5, 1.25). Compared with control (non-presence of epiphytic algae), chlorophyll contents of V. natans were significantly decreased (p < 0.01) for the presence of epiphytic algae under any concentrations of N and P in water bodies. While the presence of epiphytic algae induced peroxidation of membrane lipids, MDA contents of V. natans had significantly increased (p < 0.05) by comparing with control. SOD activity significantly enhanced (p < 0.05) with the presence of epiphytic algae in the treatments of T2 and T3 in the whole culture process by comparing with control, sometimes reaching an extremely significant level (p < 0.01). However, in the treatments of T1 and T4, SOD activity had no obvious change with the presence of epiphytic algae (p < 0.05) by comparing with control. At the end of the experiment, the effects of epiphytic algae on chlorophyll content and SOD activity in the leaves of V. natans were increased at first and then decreased with the concentrations of N and P in water, and MDA content became higher with the increase of N and P. concentrations. Repeated measurement data testing showed that the effects of epiphytic algae on the chlorophyll content and MDA content and SOD activity were significant, respectively (p < 0.001), the effects of epiphytic algae were combining with effects of concentrations of N and P (p < 0.001), respectively, and their interaction (p < 0.001). Our observations confirmed that this prediction: the growth of epiphytic algae directly produced adverse effects on physiology of V. natans and epiphytic algal biomass were positively correlated with nutrient available in the water column.
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