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A method was described to determine root growth respiration and root maintenance respiration rate of plants, grown in culture solutions of high and low oxygen concentrations, during linear growth. Root growth respiration of aerobically grown plants was three to four times lower in the flood-intolerant Senecio jacobaea L. than in the flood-tolerant species. Senecio aquaticus Hill. Root growth respiration of Senecio aquaticus Hill decreased by a factor two to three upon transplantation to a culture solution of low oxygen tension. The difference in root growth respiration between aerobically and anaerobically grown Senecio aquaticus Hill was ascribed to the presence of a highly active non-phosphorylating oxidase.

During the night, shoot nitrate concentration in spinach (Spinacia oleracea L. cv. Vroeg Reuzenblad) increased due to increased uptake of nitrate by the roots. When the plants were subjected to a one night “low light’period at 35 μmol m−2 s−1, the shoot nitrate concentration did not increase and was reduced by 25% compared to control plants in the dark. The major contribution to this decrease was located in the leaf blades, where the nitrate concentration was decreased by 60%, while the petiole nitrate concentration decreased by only 9%. Nitrate accumulated in the leaf blade vacuoles during a dark night, but this was not the case during the “low light’period. This decrease in vacuolar nitrate concentration, compared to control plants in the dark, was not caused by increased amounts of leaf blade nitrate reductase (NR; EC 1.6.6.1). During a “low light’night period, the cytoplasmic soluble carbohydrate concentration was increased compared to the control plants in the dark. Calculations showed in situ NR activity to be higher than in the control plants in the dark. This increase in NR activity, however, was not large enough to account for the total difference found in the shoot nitrate concentration. Net uptake of nitrate by the roots was increased during the initial hours of the dark night, while vacuolar nitrate concentration in the leaf blades increased at the same time. During the “low light’night period, however, net uptake of nitrate by the roots did not increase, and vacuolar nitrate concentration did not change. We conclude that nitrate uptake by the roots and vacuolar nitrate concentration in the leaf blades are tightly coupled. The decreased shoot nitrate concentration is mainly caused by a reduction in net uptake of nitrate by the roots. During the “low light’night period, carbohydrates and malic acid partly replaced vacuolar nitrate. A “low light’period one night prior to harvest provides a valuable tool to reduce shoot nitrate concentrations in spinach grown in greenhouses in the winter months.

Root growth respiration of Senecio aquaticus Hill (flood-tolerant) and Senecio jacobaea L. (flood-sensitive) was calculated, assuming different P: O ratios. The growth respiration values were calculated on the basis of the chemical composition of root and shoot dry matter, in combination with published data on the energy costs of biosynthetic and transport processes. The comparison between calculated and experimental values suggests a relatively low efficiency of ATP utilization in the roots of the flood-tolerant species. Root growth respiration of S. congestus (R.Br.)DC., which is also flood-tolerant, and Plantago lanceolata L. were also determined. The data showed that not all the flood-tolerant species investigated had high root growth respiration values. An “overflow model’ is proposed to explain observed differences in root growth respiration between species.

Under poor light conditions, as normally used during winter production of greenhouse vegetables, the nitrate concentration in the shoot of spinach (Spinacia oleracea L. cv. Vroeg Reuzenblad) showed a diurnal rhythm. This rhythm was mainly caused by a decrease during the day, followed by an increase during the night in the leaf blade nitrate concentration. Nitrate was mainly located in the vacuoles of the leaf blades. A strong correlation was found between net uptake of nitrate by the roots and the nitrate concentration in the leaf blade vacuoles. The nitrate concentration in the leaf blades increased during the initial hours of the night. This increase was caused by a marked increase in the net uptake rate of nitrate by the roots during the first hours of the dark period. During the second part of the night both net uptake rate of nitrate by the roots and the vacuolar nitrate concentration in the leaf blades remained constant. We conclude that nitrate is taken up for osmotic purposes when light conditions are poor because of a lack of organic solutes. During the night, nitrate influx into the vacuole is needed for replacement of organic solutes, which are metabolized during the night, and possibly also for leaf elongation growth. During the day, vacuolar nitrate may be exchanged for newly synthesized organic solutes and be metabolized in the cytoplasm. A strong diurnal rhythm in nitrate reductase (NR; EC 1.6.6.1.) activity was absent, due to the poor light conditions, and in vitro NR activity was not correlated with nitrate flux from the roots. In vivo NR activity also lacked a strong diurnal rhythm, but it was calculated that in situ nitrate reduction was much lower during the night, so that the major nitrate assimilation took place during the day.

Experiments were designed to provide information about the physiological basis of flood-tolerance in Senecio species. The oxygen concentration in roots of S. jacobaea L., S. viscosus L. and S. vulgaris L. became almost zero after transplantation to a solution of low oxygen concentration, and it was concluded that the flood-sensitivity of these Senecio species could be due to insufficient oxygen transport from the shoots to the roots. The oxygen concentration in the roots of the flood-tolerant S. congestus (R.Br.) DC., growing in a solution of low oxygen tension, was almost sufficient to maintain oxygen utilization at the rate observed in roots of plants, grown in an air-saturated solution. Oxygen utilization by roots of the flood-tolerant S. aquaticus Hill, growing in a solution of low oxygen tension, was inhibited 50%. However, the oxygen concentration in the roots of this species remained high enough to maintain cytochrome oxidase activity and oxidative phosphorylation at the rate observed in roots from an air-saturated environment. The activity of a second (“alternative”) oxidase must have been drastically reduced. Alternative NADH-oxidizing enzymes, like nitrate reductase which was induced by anaerobiosis in roots of S. aquaticus, might replace the regulatory function of the alternative oxidase. — Thus, in S. aquaticus root porosity and root length contributed to the maintenance of an oxygen concentration which was sufficient for uninhibited cytochrome oxidase activity and oxidative phosphorylation rate in roots growing in a solution of low oxygen tension.