Ascorbic acid regulates nitrogen, energy, and gas exchange metabolisms of alfalfa in response to high-nitrate stress

The effects of exogenous ascorbic acid (AsA) on the growth parameters, nitrogen metabolism, energy status, and photosynthetic gas exchange in alfalfa under high-nitrate stress were studied. The seedlings treated with the control, 200 mmol L-1 nitrates (HN) or 200 mmol L-1 nitrate + 0.1 mmol L-1 AsA (HN+ AsA), were sampled on days 0 and 10 after treatments. AsA was sprayed on the leaves, while HN was conducted by watering. Both of them were performed once every other day for three times in total. The results revealed that in the HN treatment, the growth parameters were the lowest; total phosphorus (TP), nitrogen-related enzyme activities, soluble protein (SP), adenosine triphosphate (ATP), and energy charge (EC) were reduced; and photosynthetic rate (Photo), conductance to H2O (Cond), transpiration rate (Trmmol), instantaneous water use efficiency (WUE), and apparent CO2 use efficiency (CUE) were also inhibited; and total nitrogen (TN), nitrate-nitrogen (NO3--N), ammonium-nitrogen (NH4+-N), adenosine diphosphate (ADP), adenosine monophosphate (AMP), and intercellular CO2 concentration (Ci) were increased compared with the control. However, these parameters changed conversely in the HN+ AsA treatment. In addition, there was a good curve regression equation relationship between TN and NO3--N, TN and NH4+-N, NO3--N and NH4+-N, respectively. It indicates that AsA improves the growth parameters, nitrogen-related enzyme activities, energy metabolism, and photosynthesis, whereas it inhibits the toxicity of excess NO3--N and NH4+-N accumulations, thereby promoting the growth of alfalfa under high-nitrate stress. These metabolisms are closely related to each other during the regulatory process in alfalfa. Hence, AsA has potential to be applied to improve the growth of alfalfa under high-nitrate stress.