Concerns about possible health hazards arising from human consumption of lettuce and other edible vegetable crops with high concentrations of nitrate have generated demands for a greater understanding of processes involved in its uptake and accumulation in order to devise more sustainable strategies for its control. This paper evaluates a proposed iso-osmotic mechanism for the regulation of nitrate accumulation in lettuce (Lactuca sativa L.) heads. This mechanism assumes that changes in the concentrations of nitrate and all other endogenous osmotica (including anions, cations and neutral solutes) are continually adjusted in tandem to minimise differences in osmotic potential of the shoot sap during growth, with these changes occurring independently of any variations in external water potential. The hypothesis was tested using data from six new experiments, each with a single unique treatment comprising a separate combination of light intensity, N source (nitrate with or without ammonium) and nitrate concentration carried out hydroponically in a glasshouse using a butterhead lettuce variety. Repeat measurements of plant weights and estimates of all of the main soluble constituents (nitrate, potassium, calcium, magnesium, organic anions, chloride, phosphate, sulphate and soluble carbohydrates) in the shoot sap were made at intervals from about 2 weeks after transplanting until commercial maturity, and the data used to calculate changes in average osmotic potential in the shoot. Results showed that nitrate concentrations in the sap increased when average light levels were reduced by between 30 and 49 % and (to a lesser extent) when nitrate was supplied at a supra-optimal concentration, and declined with partial replacement of nitrate by ammonium in the external nutrient supply. The associated changes in the proportions of other endogenous osmotica, in combination with the adjustment of shoot water content, maintained the total solute concentrations in shoot sap approximately constant and minimised differences in osmotic potential between treatments at each sampling date. There was, however, a gradual increase in osmotic potential (ie a decline in total solute concentration) over time largely caused by increases in shoot water content associated with the physiological and morphological development of the plants. Regression analysis using normalised data (to correct for these time trends) showed that the results were consistent with a 1:1 exchange between the concentrations of nitrate and the sum of all other endogenous osmotica throughout growth, providing evidence that an iso-osmotic mechanism (incorporating both concentration and volume regulation) was involved in controlling nitrate concentrations in the shoot
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