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Xanthophyll cycle conversion and the antioxidant system in the peel of apple fruit (Malus ×domestica Borkh. `Liberty') were monitored in the field over a diurnal course at about 3 months after full bloom. Compared with leaves, sun-exposed peel of apple fruit had much lower photosystem II operating efficiency at any given photon flux density (PFD) and a larger xanthophyll cycle pool size on a chlorophyll basis. Zeaxanthin (Z) level increased with rising PFD in the morning, reached the highest level during midday, and then decreased with falling PFD for the rest of the day. At noon, Z accounted for >90% of the xanthophyll cycle pool in the fruit peel compared with only 53% in leaves. Efficiency of excitation transfer to PSII reaction centers (F _v′/F _m′) was negatively related to the level of Z in fruit peel and leaves throughout the day. In fruit peel, the antioxidant enzymes in the ascorbate-glutathione cycle, ascorbate peroxidase (APX), monodehydroascorbate reductase (MDAR), dehydroascorbate reductase (DHAR) and glutathione reductase (GR) showed a diurnal pattern similar to that of incident PFD. In contrast, the activities of APX and GR in leaves did not change significantly during the day although activities of both MDAR and DHAR were higher in the afternoon than in the morning. In both fruit peel and leaves, superoxide dismutase activity did not change significantly during the day; catalase activity showed a diurnal pattern opposite to that of PFD with much lower activity in fruit peel than in leaves. The total ascorbate pool was much smaller in fruit peel than in leaves on an area basis, but the ratio of reduced ascorbate to oxidized ascorbate reached a maximum in the early afternoon in both fruit peel and leaves. The total glutathione pool, reduced glutathione and the ratio of reduced glutathione to oxidized glutathione in both fruit peel and leaves also peaked in the early afternoon. We conclude that the antioxidant system as well as the xanthophyll cycle responds to changing PFD over the course of a day to protect fruit peel from photooxidative damage.

One-year-old `Concord' grapevines (Vitis labruscana Bailey) were fertigated with 0, 5, 10, 15, or 20 mm N in a modified Hoagland's solution for 8 weeks during summer. Half of the vines fertigated at each N concentration were sprayed with 3% foliar urea twice in late September while the rest served as controls. Four vines from each treatment combination were destructively sampled during dormancy to determine the levels and forms of N and carbohydrates. Nitrogen fertigation during the summer did not significantly alter vine N concentration whereas foliar urea application in the fall significantly increased vine N concentration. In response to foliar urea application, concentrations of both free amino acid-N and protein-N increased, but the ratio of protein-N to free amino acid-N decreased. Arginine was the most abundant amino acid in free amino acids and proteins, and its concentration was linearly correlated with vine N concentration. Concentrations of total nonstructural carbohydrates (TNC) decreased slightly in response to N supply from fertigation. Foliar urea application in the fall significantly decreased TNC concentration at each N fertigation level. Starch, glucose, and fructose decreased in response to foliar urea applications, but sucrose concentration remained unaffected. Approximately 60% of the carbon decrease in TNC caused by foliar urea application was recovered in proteins and free amino acids. We conclude that free amino acids account for a larger proportion of the N in vines sprayed with foliar urea compared with the unsprayed vines, but proteins remain as the main form of N storage. In response to foliar urea application, part of the carbon from TNC is incorporated into proteins and free amino acids, leading to a decrease in the carbon stored in TNC and an increase in the carbon stored in proteins and free amino acids.

The objective of this study was to quantify how photoprotective mechanisms in the leaves of `Concord' grapevines (Vitis labruscana Bailey) respond to a range of iron (Fe) supply. Own-rooted, 1-year-old container-grown vines were fertigated twice weekly for 11 weeks with a complete nutrient solution containing 1, 10, 20, 50, or 100 μm Fe from ferric ethylenediamine di (o-hydroxyphenylacetic) acid (Fe-EDDHA). Leaf total Fe content did not increase in response to Fe supply; however, “active” Fe (extracted with 2,2′-dipyridyl) and chlorophyll (Chl) increased on a leaf area basis as applied Fe increased. At the lowest active Fe level, leaf absorptance and the efficiency of excitation transfer (F_v′/F_m′) was lower, and nonphotochemical quenching (NPQ) was significantly greater. Photosystem II (PSII) quantum efficiency decreased curvilinearly, and the proportion of PSII reaction centers in the open state (qP) decreased linearly as active Fe content decreased. On a Chl basis, the xanthophyll cycle pool size [violaxanthin (V) + antheraxanthin (A) + zeaxanthin (Z)], lutein, and β-carotene increased curvilinearly as active Fe decreased, and neoxanthin (Neo) increased at the lowest Fe level. On a leaf area basis, as active Fe decreased, V+A+Z and β-carotene decreased curvilinearly, and lutein and Neo decreased linearly. At noon, conversion of V to A and Z increased as active Fe decreased. On a Chl basis, activities of antioxidant enzymes superoxide dismutase (SOD), monodehydroascorbate reductase (MDAR), and dehydroascorbate reductase (DHAR) increased curvilinearly, and glutathione reductase (GR) activity increased linearly as active Fe levels declined. Ascorbate peroxidase (APX) and catalase (CAT), on a Chl basis, were relatively constant. On a leaf area basis, a decrease in active Fe increased SOD and MDAR activity, whereas APX, CAT, DHAR and GR activity decreased. Antioxidant metabolites ascorbate (AsA), dehydroascorbate (DAsA), reduced glutathione (GSH) and oxidized glutathione (GSSG) also increased in response to Fe limitation when expressed on a Chl basis, whereas on a leaf area basis AsA and DAsA decreased and GSH increased curvilinearly. The GSH:GSSG ratio increased as active Fe declined, whereas the AsA:DAsA ratio did not change. In conclusion, both photoprotective mechanisms, xanthophyll cycle-dependent thermal dissipation and the ascorbate-glutathione antioxidant system, are enhanced in response to Fe deficiency to cope with excess absorbed light. In a low soil pH tolerant species such as V. labruscana, the foliar antioxidant system was upregulated in response to excess absorbed light from Fe deficiency-induced chlorosis, and there was no evidence of an increase in oxidative stress from high rates of applied Fe-EDDHA.

One-year-old `Concord' grapevines (Vitis labruscana Bailey) were fertigated with 0, 5, 10, 15, or 20 mm nitrogen by using a modified Hoagland's solution for 8 weeks during active vine growth in summer. Half of the vines at each N concentration were sprayed with 3% foliar urea twice in late September while the rest served as controls. After natural leaf fall, all the vines were overwintered in a cold room (2 to 4 °C). Four vines from each treatment were destructively sampled before budbreak for reserve N and carbohydrate analysis. The remaining vines were supplied with either no N or sufficient N (10 mm N) from 2 weeks before bloom to 1 month after bloom. All the vines were destructively harvested at 1 month after bloom. Total amount of N in dormant vines increased with increasing N fertigation concentration. Total nonstructural carbohydrates (TNC) increased with increasing N fertigation concentration from 0 to 10 mm, and then leveled off with further rises in N supply. Foliar urea application increased total N but decreased TNC of dormant vines at each given N fertigation level. When no N was provided during the regrowth period, vine total leaf area, fruit yield, and total dry weight increased with increasing N supply from fertigation the previous year. Vines sprayed with foliar urea the previous fall produced a larger total leaf area, a higher yield, and a higher total vine dry weight at each given N fertigation concentration. Providing vines with sufficient N during the regrowth period significantly increased total leaf area, fruit yield, and vine total dry weight across the previous N fertigation concentrations, but vines sprayed with foliar urea still had a larger leaf area, a higher yield, and a higher total vine dry weight at each given N fertigation concentration. Therefore, we conclude that both vegetative growth and fruiting of young `Concord' vines are largely determined by reserve nitrogen, not by reserve carbohydrates, and that current-season N supply plays a very important role in sustaining vine growth and development, especially fruit growth.

To determine the cause of a characteristic zonal chlorosis of `Honeycrisp' apple (Malus ×domestica Borkh.) leaves, we compared CO₂ assimilation, carbohydrate metabolism, the xanthophyll cycle and the antioxidant system between chlorotic leaves and normal leaves. Chlorotic leaves accumulated higher levels of nonstructural carbohydrates, particularly starch, sorbitol, sucrose, and fructose at both dusk and predawn, and no difference was found in total nonstructural carbohydrates between predawn and dusk. This indicates that carbon export was inhibited in chlorotic leaves. CO₂ assimilation and the key enzymes in the Calvin cycle, ribulose 1,5-bisphosphate carboxylase/oxygenase, NADP-glyceraldehyde-3-phosphate dehydrogenase, phosphoribulokinase, stromal fructose-1,6-bisphosphatase, and the key enzymes in starch and sorbitol synthesis, ADP-glucose pyrophosphorylase, cytosolic fructose-1,6-bisphosphatase, and aldose 6-phosphate reductase were significantly lower in chlorotic leaves than in normal leaves. However, sucrose phosphate synthase activity was higher in chlorotic leaves. In response to a reduced demand for photosynthetic electron transport, thermal dissipation of excitation energy (measured as nonphotochemical quenching of chlorophyll fluorescence) was enhanced in chlorotic leaves under full sun, lowering the efficiency of excitation energy transfer to PSII reaction centers. This was accompanied by a corresponding increase in both xanthophyll cycle pool size (on a chlorophyll basis) and conversion of violaxanthin to antheraxanthin and zeaxanthin. The antioxidant system, including superoxide dismutase and ascorbate peroxidase and the ascorbate pool and glutathione pool, was up-regulated in chlorotic leaves in response to the increased generation of reactive oxygen species via photoreduction of oxygen. These findings support the hypothesis that phloem loading and/or transport is partially or completely blocked in chlorotic leaves, and that excessive accumulation of nonstructural carbohydrates may cause feedback suppression of CO₂ assimilation via direct interference with chloroplast function and/or indirect repression of photosynthetic enzymes.

One-year-old grapevines (Vitis labrusca L. `Concord') were supplied twice weekly for 5 weeks with 0, 5, 10, 15, or 20 mm nitrogen (N) in a modified Hoagland's solution to generate a wide range of leaf N status. Both light-saturated CO₂ assimilation at ambient CO₂ and at saturating CO₂ increased curvilinearly as leaf N increased. Although stomatal conductance showed a similar response to leaf N as CO₂ assimilation, calculated intercellular CO₂ concentrations decreased. On a leaf area basis, activities of key enzymes in the Calvin cycle, ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), NADP-glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoribulokinase (PRK), and key enzymes in sucrose and starch synthesis, fructose-1,6-bisphosphatase (FBPase), sucrose phosphate synthase (SPS), and ADP-glucose pyrophosphorylase (AGPase), increased linearly with increasing leaf N content. When expressed on a leaf N basis, activities of the Calvin cycle enzymes increased with increasing leaf N, whereas activities of FBPase, SPS, and AGPase did not show significant change. As leaf N increased, concentrations of glucose-6-phosphate (G6P), fructose-6-phosphate (F6P), and 3-phosphoglycerate (PGA) increased curvilinearly. The ratio of G6P/F6P remained unchanged over the leaf N range except for a significant drop at the lowest leaf N. Concentrations of glucose, fructose, and sucrose at dusk increased linearly with increasing leaf N, and there was no difference between predawn and dusk measurements. As leaf N increased, starch concentration increased linearly at dusk, but decreased linearly at predawn. The calculated carbon export from starch degradation during the night increased with increasing leaf N. These results showed that 1) grapes leaves accumulated less soluble carbohydrates under N-limitation; 2) the elevated starch level in low N leaves at predawn was the result of the reduced carbon export from starch degradation during the night; and 3) the reduced capacity of CO₂ assimilation in low N leaves was caused by the coordinated decreases in the activities of key enzymes involved in CO₂ assimilation as a result of direct N limitation, not by the indirect feedback repression of CO₂ assimilation via sugar accumulation.

Own-rooted 1-year-old `Concord' grapevines (Vitis labruscana Bailey) were fertigated twice weekly for 11 weeks with 1, 10, 20, 50, or 100 μm iron (Fe) from ferric ethylenediamine di (o-hydroxyphenylacetic) acid (Fe-EDDHA) in a complete nutrient solution. As Fe supply increased, leaf total Fe content did not show a significant change, whereas active Fe (extracted by 2,2′-dipyridyl) content increased curvilinearly. Chlorophyll (Chl) content increased as Fe supply increased, with a greater response at the lower Fe rates. Chl a: b ratio remained relatively constant over the range of Fe supply, except for a slight increase at the lowest Fe treatment. Both CO₂ assimilation and stomatal conductance increased curvilinearly with increasing leaf active Fe, whereas intercellular CO₂ concentrations decreased linearly. Activities of key enzymes in the Calvin cycle, ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), NADP-glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoribulokinase (PRK), stromal fructose-1,6-bisphosphatase (FBPase), and a key enzyme in sucrose synthesis, cytosolic FBPase, all increased linearly with increasing leaf active Fe. No significant difference was found in the activities of ADP-glucose pyrophosphorylase (AGPase) and sucrose phosphate synthase (SPS) of leaves between the lowest and the highest Fe treatments, whereas slightly lower activities of AGPase and SPS were observed in the other three Fe treatments. Content of 3-phosphoglycerate (PGA) increased curvilinearly with increasing leaf active Fe, whereas glucose-6-phosphate (G6P), fructose-6-phosphate (F6P), and the ratio of G6P: F6P remained unchanged over the range of Fe supply. Concentrations of glucose, fructose, sucrose, starch, and total nonstructural carbohydrates (TNC) at both dusk and predawn increased with increasing leaf active Fe. Concentrations of starch and TNC at any given leaf active Fe content were higher at dusk than at predawn, but both glucose and fructose showed the opposite trend. No difference in sucrose concentration was found at dusk or predawn. The export of carbon from starch breakdown during the night, calculated as the difference between dusk and predawn measurements, increased as leaf active Fe content increased. The ratio of starch to sucrose at both dusk and predawn also increased with increasing leaf active Fe. In conclusion, Fe limitation reduces the activities of Rubisco and other photosynthetic enzymes, and hence CO₂ assimilation capacity. Fe-deficient grapevines have lower concentrations of nonstructural carbohydrates in source leaves and, therefore, are source limited.

Bench-grafted `Fuji' apple [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.] trees on Malling 26 (M.26) rootstocks were fertigated for 6 weeks with N concentrations ranging from 0 to 20 mm. These treatments produced levels of leaf N ranging from 0.9 to 4.3 g·m^-2. Over this range, leaf absorptance increased curvilinearly from 74.8% to 92.5%. The light saturation point for CO₂ assimilation expressed on the basis of absorbed light increased linearly at first with increasing leaf N, then reached a plateau at a leaf N content of ≈3 g·m^-2. Under high light conditions (photosynthetic photon flux of 1500 μmol·m^-2·s^-1), the amount of absorbed light in excess of that required to saturate CO₂ assimilation decreased with increasing leaf N. Chlorophyll fluorescence measurements revealed that the maximum photosystem II (PSII) efficiency of dark-adapted leaves was relatively constant over the leaf N range, except for a slight decrease at the lower end. As leaf N increased, nonphotochemical quenching declined under high light, and there was an increase in the efficiency with which the absorbed photons were delivered to open PSII centers. The photochemical quenching coefficient remained high except for a decrease at the lower end of the leaf N range. Actual PSII efficiency increased curvilinearly with increasing leaf N, and was highly correlated with light-saturated CO₂ assimilation. The fraction of absorbed light potentially going into singlet oxygen formation was estimated to be ≈10%, regardless of leaf N status. It was concluded that there was more excess absorbed light in low N leaves than in high N leaves under high light conditions. Nonphotochemical quenching was enhanced with decreasing leaf N to reduce both the PSII efficiency and the probability of damage from photooxidation by excess absorbed light.