Key Laboratory of Marine Chemistry Theory and Technology, Ocean University of China, Qingdao, 266100, China.
College of Chemistry and Chemical Engineering, Ocean University of China, Qingdao, 266100, China.
J Phycol. 2021 Jun;57(3):955-966. doi: 10.1111/jpy.13141. Epub 2021 Mar 24.
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 in vitro 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 in vitro analysis method showed it was optimal for the NRA assay when the extraction time was 6 min, enzymatic reaction time 30 min, volume of phenazine methosulfate (PMS) solution 50 μL, NADH concentration 0.36 mM, and KNO concentration 10 mM. The maximal NRA (NRA ) 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 5 d at 30 and 25°C and 7 d at 20°C and 9 d 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 positive correlation 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.
为了更好地了解温度对浒苔生长和硝酸还原酶活性(NRA)的影响及其相互关系,在实验室条件下研究了 5 种不同温度(10、15、20、25 和 30°C)的影响。本研究建立了一种优化的浒苔 NRA 体外分析方法。在不同处理下,评估了 NRA、硝酸盐浓度、pH 值、细胞内硝酸盐和亚硝酸盐浓度以及 POC/PON。体外分析方法的结果表明,当提取时间为 6 min、酶促反应时间为 30 min、吩嗪甲硫酸酯(PMS)溶液体积为 50 μL、NADH 浓度为 0.36 mM 和 KNO3 浓度为 10 mM 时,最适合 NRA 测定。在 10、15 和 20°C(低温)组中,第 2 天 NRA 达到最大值,在 25 和 30°C(高温)组中第 1 天 NRA 达到最大值。高温下藻类生长更早结束,30 和 25°C 下 5 天后结束,20°C 下 7 天后结束,15°C 下 9 天后结束,10°C 下藻类在培养期间一直在生长。在 10-20°C 范围内培养的浒苔生长周期较长,当温度超过 15°C 时,NRA 随温度升高而降低,藻生长与 NRA 呈正相关。本研究支持 NRA 是温度变化对浒苔吸收和代谢氮营养能力影响的合适替代指标。