Werner C. Antunes

Photosynthesis and photoprotection in coffee leaves is affected by nitrogen and light availabilities in winter conditions.

Coffee is native to shady environments but often grows better and produces higher yields without shade, though at the expense of high fertilization inputs, particularly nitrogen (N). Potted plants were grown under full sunlight and shade (50%) conditions and were fertilized with nutrient solutions containing either 0 or 23 mM N. Measurements were made in southeastern Brazil during winter conditions, when relatively low night temperatures and high diurnal insolation are common. Overall, the net carbon assimilation rate was quite low, which was associated with diffusive, rather than biochemical, constraints. N deficiency led to decreases in the concentrations of chlorophylls (Chl) and total carotenoids as well as in the Chl/N ratio. These conditions also led to qualitative changes in the carotenoid composition, e.g., increased antheraxanthin (A) and zeaxanthin (Z) pools on a Chl basis, particularly at high light, which was linked to increased thermal dissipation of absorbed light. The variable-to-maximum fluorescence ratio at predawn decreased with increasing A+Z pools and decreased linearly with decreasing N. We showed that this ratio was inadequate for assessing photoinhibition under N limitation. Expressed per unit mass, the activities of superoxide dismutase and glutathione reductase were not altered with the treatments. In contrast, ascorbate peroxidase activity was lower in low N plants, particularly under shade, whereas catalase activity was lower in shaded plants than in sun-grown plants, regardless of the N level. Glutamine synthetase activity was greater in sun-grown plants than in shaded individuals at a given N level and decreased with decreasing N application. Our results suggest that the photoprotective and antioxidant capacity per amount of photons absorbed was up-regulated by a low N supply; nevertheless, this capacity, regardless of the light conditions, was not enough to prevent oxidative damage, as judged from the increases in the H(2)O(2) and malondialdehyde concentrations and electrolyte leakage. We demonstrated that N fertilization could adequately protect the coffee plants against photodamage independently of the anticipated positive effects of N on the photosynthetic capacity.

In field-grown coffee trees source-sink manipulation alters photosynthetic rates, independently of carbon metabolism, via alterations in stomatal function

Perturbations of the source-sink balances were performed in field-grown coffee (Coffea arabica) trees to investigate the possible role of carbohydrates in feedback regulation of photosynthesis. Four treatments were applied at the whole-plant level: (i) complete defruiting and maintenance of the full leaf area, (ii) the half crop load and full leaf area, (iii) the full crop load and full leaf area and (iv) the full crop load and half leaf area. Sampling and measurements were performed twice during the phase of dry matter accumulation of fruits. Gas exchange, chlorophyll a fluorescence, carbon isotope labelling and steady-state metabolite measurements were assessed in source leaves. The average rate of net photosynthetic rate (A) and stomatal conductance (g(s)) were larger (> 50%), and carbon isotope composition ratio was lower, in trees with a full crop load and half leaf area than in defruited trees, with individuals of the other two treatments showing intermediate values. However, differences in A seem unlikely to have been caused either by photochemical impairments or a direct end-product-mediated feedback down-regulation of photosynthesis. It is proposed that the decreased A in defruited coffee trees was independent of carbon metabolism and was rather directly related to a lower CO(2) availability coupled to lower g(s).

Changes in stomatal function and water use efficiency in potato plants with altered sucrolytic activity

As water availability for agriculture decreases, breeding or engineering of crops with improved water use efficiency (WUE) will be necessary. As stomata are responsible for controlling gas exchange across the plant epidermis, metabolic processes influencing solute accumulation in guard cells are potential targets for engineering. In addition to its role as an osmoticum, sucrose breakdown may be required for synthesis of other osmotica or generation of the ATP needed for solute uptake. Thus, alterations in partitioning of sucrose between storage and breakdown may affect stomatal function. In agreement with this hypothesis, potato (Solanum tuberosum) plants expressing an antisense construct targeted against sucrose synthase 3 (SuSy3) exhibited decreased stomatal conductance, a slight reduction in CO(2) fixation and increased WUE. Conversely, plants with increased guard cell acid invertase activity caused by the introduction of the SUC2 gene from yeast had increased stomatal conductance, increased CO(2) fixation and decreased WUE. (14)CO(2) feeding experiments indicated that these effects cannot be attributed to alterations in photosynthetic capacity, and most likely reflect alterations in stomatal function. These results highlight the important role that sucrose breakdown may play in guard cell function and indicate the feasibility of manipulating plant WUE through engineering of guard cell sucrose metabolism.

Tobacco guard cells fix CO2 by both Rubisco and PEPcase while sucrose acts as a substrate during light-induced stomatal opening.

Transcriptomic and proteomic studies have improved our knowledge of guard cell function; however, metabolic changes in guard cells remain relatively poorly understood. Here we analysed metabolic changes in guard cell-enriched epidermal fragments from tobacco during light-induced stomatal opening. Increases in sucrose, glucose and fructose were observed during light-induced stomatal opening in the presence of sucrose in the medium while no changes in starch were observed, suggesting that the elevated fructose and glucose levels were a consequence of sucrose rather than starch breakdown. Conversely, reduction in sucrose was observed during light- plus potassium-induced stomatal opening. Concomitant with the decrease in sucrose, we observed an increase in the level as well as in the (13) C enrichment in metabolites of, or associated with, the tricarboxylic acid cycle following incubation of the guard cell-enriched preparations in (13) C-labelled bicarbonate. Collectively, the results obtained support the hypothesis that sucrose is catabolized within guard cells in order to provide carbon skeletons for organic acid production. Furthermore, they provide a qualitative demonstration that CO2 fixation occurs both via ribulose-1,5-biphosphate carboxylase/oxygenase (Rubisco) and phosphoenolpyruvate carboxylase (PEPcase). The combined data are discussed with respect to current models of guard cell metabolism and function.

Resposta fisiológica de clone de café Conilon sensível à deficiência hídrica enxertado em porta-enxerto tolerante

The objective of this work was to determine alterations in physiology and those due to drought tolerance on Conilon coffee (Coffea canephora) contrasting clones regarding the sensitivity to hydric stress. The reciprocal grafting between clones 109A, drought sensitive, and 120, drought tolerant, - 120/109A, 120/120, 109A/120, 109A/109A - along with their ungrafted control plants (109A and 120) were evaluated. Plants were cultivated in 12‑L vases in greenhouse. Six months after grafting, half of the plants was subjected to water deficit, by suspending irrigation until leaves reached the hydric potential of ‑3,0 MPa. When clone 120 was used as rootstock, plants presented deeper roots, although with lower root-biomass, higher ability to postpone leaf dehydration and higher instantaneous water‑use efficiency (WUE). Under severe drought, starch and sucrose contents decreased similarly, regardless of the treatment, whereas leaf concentrations of glucose, fructose, total amino acids and proline were higher in non‑grafted 109A, 109A/109A, and 120/109A plants. These plants showed the lowest WUE values. Solute accumulation was not primarily related to drought tolerance. The use of drought tolerant rootstocks improves to drought tolerance in coffee.

Photosynthetic limitations in coffee plants are chiefly governed by diffusive factors

It has long been held that the regulation of photosynthesis in source leaves may be controlled by carbohydrates. The mechanisms that govern the diurnal fluctuation of photosynthesis and the potential role of feedback regulation by carbohydrates during photosynthesis in coffee (Coffea arabica) leaves were investigated in three independent and complementary experiments. An integrative approach using gas exchange measurements in addition to carbon isotope labelling and steady-state carbohydrate and amino acid analysis was performed. Canonical correlation analysis was also performed. In field-grown plants under naturally fluctuating environmental conditions (Experiment I), the overall pattern of gas exchange was characterised by both low stomatal conductance (gs) and net carbon assimilation rate (A) in the afternoon; no apparent signs of photoinhibition were observed. Under conditions of low air evaporative demand (Experiment II), only slight decreases (~20%) in A were observed at the end of the day, which were associated with a reduction (~35%) in gs. For both conditions, any increase in carbohydrate and amino acid pools over the course of the day was small. In leaves from girdled branches (Experiment III), a remarkable decrease in A and particularly in gs was observed, as were increases in starch but not in hexoses and sucrose pools. Furthermore, the rate of 14CO2 uptake (assessed under saturating CO2 conditions) and the partitioning of recently fixed 14C were not affected by girdling. It is proposed that the diurnal oscillations in A and the differences in A in leaves from girdled and non-girdled branches were merely a consequence of diffusive limitations rather than from photochemical constraints or direct metabolite-mediated down-regulation of photosynthesis.

Arabidopsis gun4 mutant have greater light energy transfer efficiency in photosystem II despite low chlorophyll content

Arabidopsis gun4 mutant is defected in retrograde signaling and is characterized by reduced levels of chlorophyll. However, the impact of this mutation on plant photosynthetic performance remains unclear. We carried out a physiological characterization of gun4 in order to investigate the role of GUN4 for plant photosynthetic performance under light stress. The gun4 plants showed reduced minimal fluorescence in dark-adapted leaves and quantum yield of unregulated energy dissipation of photosystem II (PSII) under non-light-stress condition. The effective quantum yield of the PSII (ΦPSII) and photochemical quenching (qL) were higher in gun4 plants. Higher values of ΦPSII were also observed in gun4 under different light intensities. However, the rate of net carbon assimilation and stomatal conductance were lower in gun4. No differences were detected between the genotypes in the total absorption of 14CO2 as well as in the percentage of 14C flux to basic amino acids, sugars, starch, and cell wall. After light stress, the potential quantum yield of PSII decreased only in wild type and the non-photochemical quenching was higher in gun4. Taken together, the results suggest that gun4 transfers more efficiently the excess of light energy absorbed despite a reduction in chlorophyll and carotenoids content and has greater capacity to dissipate the excess of energy absorbed.