Temperature dependence of energy-transducing functions and inhibitor sensitivity in chloroplasts

A comparative analysis of the temperature dependence of energy-transducing reactions in spinach (Spinacia oleracea) chloroplasts and their sensitivity for uncouplers and energy-transfer inhibitors at different temperatures is presented. Arrhenius plots reveal two groups of transitions, around 19°C and around 12°C. Activities that show transitions around 19°C include linear electron flow from water to ferricyanide, its coupled photophosphorylation, the dark-release of the fluorescent probe atebrin, and the slow component of the 515 nm (carotenoid) absorbance decay after a flash. The transitions around 12°C are observed with pyocyanine-mediated cyclic photophosphorylation, light- and dithioerythritol-activated ATP hydrolysis, the dark-release of protons, and the fast 515 nm decay component. It is suggested that both groups of temperature transitions are determined by proton displacements in different domains of the exposed thylakoid membranes. The effects of various uncouplers and an energy-transfer inhibitor are temperature dependent. Some uncouplers also show a different relative inhibition of proton uptake and ATP synthesis at lower temperatures. The efficiency of energy transduction (ATP/e₂) varied with temperature and was optimal around 10°C.     

Study of the temperature effect in three chestnut (Castanea sativa Mill.) cultivars' behaviour

The aim of this work was to analyse the effect of temperature in three chestnut cultivars, Aveleira, Judia and Longal. For this purpose, gas exchange, thylakoid membrane potential, photosynthetic pigment and lipid content data in July, September and October under different temperatures (31, 26 and 18 degrees C) were determined. With respect to gas exchanges, significant changes in photosynthesis rate of Aveleira were observed between July and September (7mumol CO2m(-2)s(-1)). In contrast, Judia and Longal showed a strong increase in this period, 6.1-8.5 and 4.9-6.7 micromol CO2m(-2)s(-1), for Judia and Longal, which represent an increase of about 15% and 43%, respectively. Similar patterns were detected in daylight photosynthesis measurements for Judia and Longal, in which an almost 60% decrease was observed, in contrast to 40% for Aveleira, from morning to midday, when temperatures increased from 27 to 34 degrees C. In addition to high photosynthetic rates in the hottest month, Aveleira was also the sunniest cultivar according its highest value on chlorophyll a/b ratio (3.65). Cultivars also presented maximal thylakoid membrane potential at different temperatures, with their values being 20.8, 17.8 and 17.2 degrees C for Aveleira, Longal and Judia, respectively. These results were also supported by thylakoid fatty acid composition which indicated that the unsaturation index of Aveleira (158) was the lowest in comparison with other two cultivars, 168 and 175, for Longal and Judia, respectively.     

Isolation of chestnut chloroplasts: membrane potentials of chestnut and spinach thylakoids

Typical chestnut thylakoid extracts isolated by mechanical disruption of leaf tissues had an equivalent of 0.28 kg m⁻³ chlorophyll (Chl) which is six times less than in thylakoids obtained from spinach, although Chl content in leaves was only half as small. According to optical microscopy, the vesicles showed a good integrity, exhibiting at 21 °C a high capacity of photon-induced potential membrane generation, which was demonstrated by the almost full 9-amino-6-chloro-2-methoxyacridine fluorescence quenching in a hyper-saline medium containing 150 mM KCl and having osmotic potential of −1.5 MPa. The half-time of the thylakoid potential generation was 11.7 s with the time of dissipation around 8.9 s. In such conditions, spinach thylakoids showed an increased swelling and also differences in the half-time generation which was almost four times faster than was observed in chestnut. However, when spinach thylakoids were incubated in a typical hypo-saline medium without KCl with osmotic potential −0.8 MPa, no additional swelling was observed. Consequently the half-time of potential dissipation was 35 s. Studies with nigericin suggested a chestnut thylakoid ΔpH significantly smaller than that observed in spinach, which was confirmed by the measurements of the ATP driven pumping activity.     

Electric surface charge dynamics of chloroplast thylakoid membranes. Temperature dependence of electrokinetic potential and aminoacridine interaction

Electric surface charge dynamics of unstacked broken chloroplasts at low-ionic strength were studied by free-flow electrophoresis and aminoacridine fluorescence and binding changes over the temperature range 4–36°C. Both illumination and ATP hydrolysis in the dark cause a significant increase of net negative surface charge. The light and dark electrokinetic (ζ) potentials have a broad temperature optimum between 20 and 36°C. The decline at lower temperature shows a transition at about 18°C. The ATP-induced increase of the ζ potential requires preactivation of the ATPase and is dicyclohexylcarbodiimide sensitive. Aminoacridine binding shows a quite different temperature dependence. At lower temperatures there is an increased number of binding sites with a decreased affinity and the binding becomes positively cooperative. It is demonstrated that aminoacridines aggregate to dimers upon binding to the membranes. This phenomenon is stimulated by light and favoured at lower temperatures. The light-dependent extra binding increases sigmoidally with increasing temperature, similar to the increase of ζ potential, but with a less abrupt transition. The different effects of temperature on the electrokinetic and binding data are explained in terms of surface charge screening in the electric double-layer of the thylakoid membrane.     

Leaf Gas Exchange and Water Relations of Grapevines Grown in Three Different Conditions

Diurnal and seasonal changes in the leaf water potential (), stomatal conductance (g _s), net CO₂ assimilation rate (P _N), transpiration rate (E), internal CO₂ concentration (C _i), and intrinsic water use efficiency (P _N/g _s) were studied in grapevines (Vitis vinifera L. cv. Touriga Nacional) growing in low, moderate, and severe summer stress at Vila Real (VR), Pinhão (PI), and Almendra (AL) experimental sites, respectively. In VR and PI site the limitation to photosynthesis was caused more by stomatal limitations, while in AL mesophyll limitations were also responsible for the summer decline in P _N.     

Effect of Chestnut Ink Disease on Photosynthetic Performance

In order to evaluate the evolutionary impact of chestnut ink disease, infected trees (cv. Judia), were compared with non‐infected trees, in three separate months: July, September and October. The aim of this work is to analyse the effects of the infection using parameters related to plant water relations, gas exchange and biometric data of leaves and fruits. In this period, temperatures decreased from 31 to 16°C contrarily to precipitation, which increased from 18 to 178 mm, respectively. In consequence, leaf water potential changed between ?1.6 and ?1.0 MPa while in infected plants the values maintained around ?1.2 MPa over the referred period. Nevertheless, at the gas exchanges level, differences in stomatal conductance, transpiration and photosynthesis were only detected in October. Concerning photosynthesis rate, the infected plants showed, in relation to September, a reduction around 35% whereas in non‐infected plants the decline was 25%. Alterations in the chlorophyll contents were also observed between September and October. In infected plants reduction on total amount of chlorophyll was from 18.6 to 13.4 mg/W_f, while in non‐infected plants values were only decayed from 15.1 to 13.1 mg/W_f. In relation to chlorophyll a/chlorophyll b ratio, plants infected by the oomycete preserved values in the level of 2.6, whereas in healthy plants values changed from 2.5 to 2.3. Leaves and fruits from infected chestnut trees were 13 and 20% smaller, respectively than those from non‐infected. Fruits from infected plants also had less starch but more crude protein.     

Bark stripping in cork oak (Quercus suber L.): effect of an antitranspirant application on gas exchange and water relations of the stripped surface

Quercus suber L. is the primary source of industrial cork, which can be legally collected every 9 years. The main objective of this work was to test the efficiency of an application of an antitranspirant, at three different concentrations, after the bark stripping. For this purpose, several measurements of the gas exchange, water potential, total chlorophylls and the carotenoids contents were determined in cork oak trees, at two times in a day, morning and afternoon. The antitranspirant film was applied immediately after stripping. Transpiration rate showed a significant increase in the afternoon. The parameters, water potential, photosynthetic rates, stomatal conductance and the intrinsic water use efficiency, showed a significant decrease from morning to afternoon. The difference between pigments concentration was not significant throughout the day. Water potential and transpiration rate were high in the treatments with lower antitranspirant concentration. However, the treatment with a higher paraffin concentration showed larger photosynthesis rate. This result suggests that the loss of water observed for the stripping surface can be minimized by a larger concentration of the antitranspirant.