Abstract

To better understand the effect of temperature on the growth and nitrate reductase activity (NRA) of Ulva prolifera and their relationships, the effects of five different temperatures (10, 15, 20, 25, and 30°C) were investigated in a laboratory setup. In this study, an optimization invitro analysis method for Ulva prolifera NRA was developed. Under different treatments, the NRA, nitrate concentration, pH, the intracellular nitrate and nitrite concentrations, and the POC/PON were evaluated. The results of the invitro analysis method showed it was optimal for the NRA assay when the extraction time was 6min, enzymatic reaction time 30min, volume of phenazine methosulfate (PMS) solution 50μL, NADH concentration 0.36mM, and KNO3 concentration 10mM. The maximal NRA (NRAmax ) appeared on the 2nd day in the 10, 15, and 20°C (low-temperature) groups and on the 1st day in the 25 and 30°C (high-temperature) groups. The algal growth ended earlier at a high temperature, ending after 5d at 30 and 25°C and 7d at 20°C and 9d at 15°C, and the alga at 10°C had been growing during the incubation period. Ulva prolifera cultivated in a range of 10-20°C had a long growth cycle and the NRA decreased with increasing temperature when exceeded 15°C, a positivecorrelation between algal growth and NRA was observed. This study supports NRA is a suitable proxy of the effects of temperature changes on the ability of Ulva prolifera to uptake and metabolize nitrogen nutrients.

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