Negative effects on the leaves of submerged macrophyte and associated biofilms growth at high nitrate induced-stress.

High nitrate (NO-N) concentration is a growing aquatic risk concern worldwide. However, adverse effects of high NO-N concentration on submerged macrophytes-epiphytic biofilms are unclear. In this study, the alterations in physiological changes, biofilms formation and chemical compositions were investigated on leaves of Vallisneria asiatica exposed to different NO-N concentrations. The findings showed that 10 mg LNO-N resulted in low photosynthetic efficiency by inhibiting chlorophyll content 26.2 % and decreased intrinsic efficiency of photosystem II significantly at 14th day post treatment. Malondialdehyde, several antioxidant enzyme activities (i.e., superoxide dismutase, peroxidase and catalase), and secondary metabolites (i.e., phenolic compounds and anthocyanin) were all significantly up-regulated with 10 mg LNO-N, implied oxidative stress were stimulated. However, no significant alterations in these indicators were observed with 5 mg LNO-N. Compared to control, 10 mg LNO-N concentration significantly stimulated microbes growth in biofilm and reduced the roughness of leaf-biofilms surface, but it had little effect on the biofilms distribution (from single clone to blocks) as revealed by scanning electron microscope and multifractal analysis. Results from X-ray photoelectron spectroscopy analysis showed that the percentage of P, Cl, K and the ratio of O1 (-O-) /O2 (C = O) were higher in leaves of control than treatments with 10 mg LNO-N, indicating that 10 mg LNO-N concentration exhibited significant inhibition of chemical activity and nutrient uptake of the leaf surfaces. Overall, these results demonstrated that high NO-N does stimulate the biofilm growth and can cause negative impacts on submerged macrophytes growth.