اثر سطوح مختلف مولیبدن و نیتروژن روی برخی خصوصیات مورفوفیزیولوژیکی اسفناج

Abstract
Nitrate accumulation is a common problem in most leafy vegetables and it happens when the amount of nitrate absorption exceeds the amount consumed by the plant. This study was conducted as a factorial experiment in the form of a completely randomized design and in three replications. The first factor included adding nitrate (calcium nitrate) to Hoagland's standard solution at three levels of 0, 10 and 20%, and the second factor was adding different concentrations of molybdenum from sodium molybdate salt at four levels of 0, 0.5, 1.5 and 3 µM. The measured traits included traits of some growth characteristics, total phenol and flavonoids, ascorbic acid, spinach leaf nitrate. According to the obtained results, fresh and dry weight of leaf, dry weight of root and percentage of dry matter increased by increasing calcium nitrate concentration up to 20% and molybdenum application up to 3 µM in nutrient solution. The simultaneous application of molybdenum and nitrogen had no significant effect on total phenolic, total flavonoid and ascorbic acid traits, and the highest amount of these traits was observed in 3 µM molybdenum. The highest amount of leaf nitrate (3247.4 mg kg^-1 FW) was observed in the treatment of 20% additional nitrate along with molybdenum. Based on the results, the use of molybdenum in nutrient solution had a reducing effect on nitrate accumulation in spinach leaf, therefore adding 3 µM sodium molybdate to Hoagland nutrient solution in hydroponic system for spinach production is recommended to obtain the maximum amount of antioxidant compounds and the minimum amount of nitrate accumulation.
Introduction
Plants take nitrogen (N) either as nitrate (NO₃⁻) or ammonium (NH₄⁺) form for various growth and developmental processes; however, NO₃⁻ is more important for such processes. For most of the crop plants, the nitrate form is mobile, less toxic, and can be stored in vacuoles. However, Nitrate must be reduced to NH₄⁺ before it can be utilized for the synthesis of amino acids, proteins, and other nitrogenous compounds in plant cells. Nitrate reductase (NR) and nitrite reductase (NiR), the key nitrate assimilatory enzymes, are located in cytosol and chloroplasts and catalyze nitrate reduction to NO₂⁻ followed by NO₂⁻ to NH₄⁺, respectively, in the leaf tissues. However, NH₄⁺ is directly assimilated to produce different amino acids by the mutual actions of glutamine synthetase (GS) and glutamate synthase (GOGAT) enzymes in a cyclic manner within the plant cells. Nitrate accumulation is a common problem in most leafy vegetables and it happens when the amount of nitrate absorption exceeds the amount consumed by the plant. Molybdenum (Mo) is an essential microelement for higher plants and also a metal component of the Mo-cofactor, (Moco) biosynthesis. Moco binds to Mo-requiring enzymes and optimizes their activities for normal functioning of plant growth and developmental processes. Molybdenum plays a significant role in N metabolism, which includes nitrate reduction, assimilation, and fixation, by regulating the NR and GS enzymes activities and expressions. During symbiotic N fixation, Mo acts as a cofactor for nitrogenase enzymes to catalyze the redox reaction to convert elemental N into ammonium ions.
Materials and methods
This study was conducted as a factorial experiment in the form of a completely randomized design and in three replications. The first factor included adding nitrate (calcium nitrate) to Hoagland's standard solution at three levels of 0, 10 and 20%, and the second factor was adding different concentrations of molybdenum from sodium molybdate salt at four levels of 0, 0.5, 1.5 and 3 µM. This research was conducted in the research greenhouse of Campus of Agriculture and Natural Resources Razi University. Senator spinach seeds were obtained from Fardin Kasht Alborz Institute. Spinach seeds were disinfected with 1% sodium hypochlorite for 5 minutes and then planted in seedling trays containing cocopeat and perlite with a volume ratio of 1:1. The measured traits included traits of some growth characteristics, total phenol and flavonoids, ascorbic acid, spinach leaf nitrate.
Results and discussion
 According to the obtained results, fresh and dry weight of leaf, dry weight of root and percentage of dry matter increased by increasing calcium nitrate concentration up to 20% and molybdenum application up to 3 µM in nutrient solution. The simultaneous application of molybdenum and nitrogen had no significant effect on total phenolic, total flavonoid and ascorbic acid traits, and the highest amount of these traits was observed in 3 µM molybdenum. The highest amount of leaf nitrate (3247.4 mg kg^-1 FW) was observed in the treatment of 20% additional nitrate along with molybdenum. The most important point in this research was the use of molybdenum to control nitrate accumulation, and the use of molybdenum in concentrations of 1.5 and 3 µM was able to reduce the amount of nitrate accumulation in the aerial parts to some extent, thus neutralizing the toxicity of excess nitrate. The use of sodium molybdate can be one of the recommended ways to reduce the accumulation of nitrates and at the same time enrich molybdenum in spinach and increase the nutritional value of this plant.
Conclusion
Nitrate and ammonium are the major forms of N that plants use for different growth and developmental processes. The present study revealed that Mo fertilizer plays a key role in N metabolism through regulating the activities and expressions of N-assimilating enzymes. In general, the results of this research showed that the increase in molybdenum concentration in hydroponic culture affected the growth and biochemical characteristics of spinach. Based on the results, the use of molybdenum in nutrient solution had a reducing effect on nitrate accumulation in spinach leaf, therefore adding 3 µM sodium molybdate to Hoagland nutrient solution in hydroponic system for spinach production is recommended to obtain the maximum amount of antioxidant compounds and the minimum amount of nitrate accumulation.