Respiratory carbon metabolism following illumination in intact French bean leaves using (13)C/(12)C isotope labeling

The origin of the carbon atoms in the CO(2) respired by French bean (Phaseolus vulgaris) leaves in the dark has been studied using (13)C/(12)C isotopes as tracers. The stable isotope labeling was achieved through a technical device that uses an open gas-exchange system coupled online to an elemental analyzer and linked to an isotope ratio mass spectrometer. The isotopic analysis of the CO(2) respired in the dark after a light period revealed that the CO(2) was labeled, but the labeling level decreased progressively as the dark period increased. The pattern of disappearance depended on the amount of carbon fixed during the labeling and indicated that there were several pools of respiratory metabolites with distinct turnover rates. We demonstrate that the carbon recently assimilated during photosynthesis accounts for less than 50% of the carbon in the CO(2) lost by dark respiration and that the proportion is not influenced by leaf starvation in darkness before the labeling. Therefore, most of the carbon released by dark respiration after illumination does not come from new photosynthates.     

The lack of mitochondrial complex I in a CMSII mutant of Nicotiana sylvestris increases photorespiration through an increased internal resistance to CO2 diffusion

The cytoplasmic male sterile II (CMSII) mutant lacking complex I of the mitochondrial electron transport chain has a lower photosynthetic activity but exhibits higher rates of excess electron transport than the wild type (WT) when grown at high light intensity. In order to examine the cause of the lower photosynthetic activity and to determine whether excess electrons are consumed by photorespiration, light, and intercellular CO(2), molar fraction (c(i)) response curves of carbon assimilation were measured at varying oxygen molar fractions. While oxygen is the major acceptor for excess electrons in CMSII and WT leaves, electron flux to photorespiration is favoured in the mutant as compared with the WT leaves. Isotopic mass spectrometry measurements showed that leaf internal conductance to CO(2) diffusion (g(m)) in mutant leaves was half that of WT leaves, thus decreasing the c(c) and favouring photorespiration in the mutant. The specificity factor of Rubisco did not differ significantly between both types of leaves. Furthermore, carbon assimilation as a function of electrons used for carboxylation processes/electrons used for oxygenation processes (J(C)/J(O)) and as a function of the calculated chloroplastic CO(2) molar fraction (c(c)) values was similar in WT and mutant leaves. Enhanced rates of photorespiration also explain the consumption of excess electrons in CMSII plants and agreed with potential ATP consumption. Furthermore, the lower initial Rubisco activity in CMSII as compared with WT leaves resulted from the lower c(c) in ambient air, since initial Rubisco activity on the basis of equal c(c) values was similar in WT and mutant leaves. The retarded growth and the lower photosynthetic activity of the mutant were largely overcome when plants were grown in high CO(2) concentrations, showing that limiting CO(2) supply for photosynthesis was a major cause of the lower growth rate and photosynthetic activity in CMSII.     

Effect of temperature on net CO2 assimilation and photosystem II quantum yield of electron transfer of French bean (Phaseolus vulgaris L.) leaves during drought stress

The effect of leaf temperature on stomatal conductance and net CO₂ uptake was studied on French bean (Phaseolus vulgaris L.) using either dehydrated attached leaves (25–40% water deficit) or cut leaves supplied with 10^–4 M abscisic acid (ABA) solution to the transpiration stream. Decreasing leaf temperature caused stomatal opening and increased net CO₂ uptake (which was close to zero at around 25° C) to a level identical to that of control leaves (without water deficit) at around 15° C. (i) The ABA effect on stomatal closure was modulated by temperature and, presumably, ABA is at least partly responsible for stomatal closure of french bean submitted to a drought stress. (ii) For leaf temperatures lower than 15° C, net CO₂ uptake was no longer limited by water deficit even on very dehydrated leaves. This shows that dehydrated leaves retain a substantial part of their photosynthetic capacity which can be revealed at normal CO₂ concentrations when stomata open at low temperature. In contrast to leaves fed with ABA, decreasing the O₂ concentration from 21% to 1% O₂ did not increase either the rate of net CO₂ uptake or the thermal optimum for photosynthesis of dehydrated leaves. The quantum yield of PSII electron flow (measured by F/F_m) was lower in 1% O₂ than in 21% O₂ for each leaf pretreatment given (non-dehydrated leaves, dehydrated leaves, and leaves fed with ABA) even within a temperature range in which leaf photosynthesis at normal CO₂ concentration was the same in these two O₂ concentrations. It is concluded that this probably indicates an heterogeneity of photosynthesis, since this difference in quantum yield disappears when using high CO₂ concentrations during measurements.Abbreviations and Symbols ABA abscisic acid - F_m maximum chlorophyll fluorescence - F difference between steady-state chlorophyll fluorescence and F_m - PPFD photosynthetic photon flux density We would like to thank Dr. J.-M. Briantais (Laboratoire d'écologie végétale, Orsay, France) for help during fluorescence measurements and Ms. J. Liebert for technical assistance.     

Effect of dehydration and high light on photosynthesis of two C3 plants (Phaseolus vulgaris L. and Elatostema repens (Lour.) Hall f.)

The effect of drought on the photosynthetic functioning of two C₃ plants, Phaseolus vulgaris and Elatostema repens, has been examined. Leaf net CO₂ uptake measured in normal air was negligible at a leaf water deficit of about 30% while the calculated leaf intercellular CO₂ concentration (Ci) was unchanged. However, both the maximal photosynthetic capacity (CO₂-dependent O₂ evolution) and apparent quantum yield, measured in the presence of saturating CO₂ levels (5 to 14%), only started to decrease within the range of 25 to 30% leaf water deficit. This shows that the drought-induced inhibition seen in normal air is not caused by an inhibition of the photosynthetic mechanism, and that in this case Ci values can be misleading. Both 77 K and room-temperature fluorescence measurements indicate that the functioning of the photosystem-II reaction centre is hardly modified by water shortage. Furthermore, an analysis of photochemical chlorophyll fluorescence quenching shows, in the absence of CO₂, that O₂ can be an efficient acceptor of photosynthetic energy, even in severly dehydrated plants which do not show net CO₂ uptake in normal air. In these plants, O₂ is probably reduced mainly via Mehler-type reactions. High-light treatment given at low O₂ increases photoinhibition as measured by the decrease of apparent quantum yield in dehydrated P. vulgaris, whereas, interestingly, 1% O₂ protects dehydrated E. repens against high-light damage. The two plants could have different protective mechanisms depending upon the O₂ level or different photoinhibitory sites or mechanisms.Abbreviations and symbols Ca, Ci ambient and calculated intercellular CO₂ concentration - Fm, Fo, Fv maximum, initial and variable fluorescence emission - LWD leaf water deficit - PPFD photosynthetic photon flux density - PSII photosystem II - qQ photochemical quenching of chlorophyll fluorescence     

Metabolic origin of carbon isotope composition of leaf dark-respired CO2 in French bean

The carbon isotope composition (delta(13)C) of CO(2) produced in darkness by intact French bean (Phaseolus vulgaris) leaves was investigated for different leaf temperatures and during dark periods of increasing length. The delta(13)C of CO(2) linearly decreased when temperature increased, from -19 per thousand at 10 degrees C to -24 per thousand at 35 degrees C. It also progressively decreased from -21 per thousand to -30 per thousand when leaves were maintained in continuous darkness for several days. Under normal conditions (temperature not exceeding 30 degrees C and normal dark period), the evolved CO(2) was enriched in (13)C compared with carbohydrates, the most (13)C-enriched metabolites. However, at the end of a long dark period (carbohydrate starvation), CO(2) was depleted in (13)C even when compared with the composition of total organic matter. In the two types of experiment, the variations of delta(13)C were linearly related to those of the respiratory quotient. This strongly suggests that the variation of delta(13)C is the direct consequence of a substrate switch that may occur to feed respiration; carbohydrate oxidation producing (13)C-enriched CO(2) and beta-oxidation of fatty acids producing (13)C-depleted CO(2) when compared with total organic matter (-27.5 per thousand). These results are consistent with the assumption that the delta(13)C of dark respired CO(2) is determined by the relative contributions of the two major decarboxylation processes that occur in darkness: pyruvate dehydrogenase activity and the Krebs cycle.     

RELATIONS BETWEEN SUNFLECK SEQUENCES AND PHOTOINHIBITION OF PHOTOSYNTHESIS IN A TROPICAL RAIN FOREST UNDERSTORY HERB

We report the photosynthetic characteristics of a C₃ shade plant native to the tropical rain forest understory. It was shown that Elatostema repens Lour. (Hall) f. (Urticaceae) presents a large light adjustment capacity. The effects of several lightfleck sequences on photoinhibition of photosynthesis and carbon gain are analyzed. Photoinhibition is measured both as a decrease in leaf net CO₂ uptake in limiting light (shown to be linearly correlated to quantum yield of O₂ evolution measured at saturating CO₂) and as a decrease of the ratio of variable fluorescence (Fv) to maximum fluorescence (Fmax) measured in liquid nitrogen. It is shown that lightflecks (from 10 to 30 min in duration) of 700 μmol m^–2 s^–1 (high light) induce photoinhibition, and that the effects of those successive high light periods are additive; there is apparently no recovery from photoinhibition during the low light periods (from 10 to 45 min in duration). In contrast, the Fv/Fmax ratio, though decreasing similarly to quantum yield of net CO₂ uptake on leaves submitted to a continuous illumination of 700 μmol m^–2 s^–1, is only decreased a little on leaves submitted to lightfleck sequences of the same photon flux density. Lightflecks of 250 μmol m^–2 s^–1 are not photoinhibitory. Compared to the control maintained under light growth condition (40 μmol m^–2 s^–1) carbon gain is increased on leaves submitted to lightflecks; this gain remains high throughout the light cycles on leaves submitted to nonphotoinhibitory lightflecks and to the photoinhibitory lightflecks followed by the shortest low light period. In the other cases, carbon gain, higher than that of the control at the beginning of the treatments, decreases and becomes lower than the control carbon gain. Finally, the relevance of photoinhibition in the tropical rain forest understory environment is discussed.     

Stomatal and non-stomatal contribution in the decline in leaf net CO2 uptake during rapid water stress

A simple method is proposed for quantitative evaluation of Stomatal and non-stomatal components of the decline in leaf CO₂ uptake during rapid water stress. The changes in leaf conductance were measured during the stress and were used to calculate the photosynthetic rate which would be observed if Stomatal closure were the only cause of the decline in photosynthesis. Photosynthesis-CO₂ response curves, determined just before the stress, were used for this calculation. The difference between the calculated and the actual rate is a measure of the non-stomatal effect of water stress.
This analysis was tested on Sinapis alba submitted to rapid and severe water stress by excising leaves or roots. Experiments were performed at saturating light conditions under high (61 Pa), normal (34 Pa) or low (11 Pa) ambient CO 2 pressure. The non-stomatal effect on de-rooted plants reaches a maximum at the beginning of the stress and is dependent on the CO 2 pressure: after 45 min its influence is still about 100°, 70° and 8°, respectively, at high, normal and low CO₂. In the excised leaf system in which desiccation was more rapid, the non-stomatal effect accounted for nearly 100° of the assimilation decline whatever the CO₂ pressure.     

Interaction between sublethal pollution by sulphur dioxide and drought stress. The effect on photosynthetic capacity

Five-year-old Picea abies L. plants were grown in growth cabinets in the presence (3.1 μmol m⁻³) or absence of SO₂. After 5 weeks, the photosynthetic capacity of mature needles produced in the year was the same in both conditions. Trees were then submitted in situ to drought stress by withholding water. The decline of leaf photosynthetic capacity was greatest in the presence of SO₂. Chlorophyll decreased only when trees were submitted to dehydration in the presence of SO₂; however, this al-one could not account for the large decline in photosynthetic capacity observed under that condition. Needle water content was the lowest during dehydration in the presence of SO₂. It is concluded that the critical factor in the interaction between pollution by SO₂ and drought stress is the greater dehydration of the tissue found in stressed plants grown in the presence of SO₂. The large decline in photosynthetic capacity under such conditions might be due to this greater dehydration.     

Étude de la pycnogenèse chezPhomopsis mali (Schultz etSacc.)Rob.

Light induces the formation of pycnidia inPhomopsis mali. The induction caused by light can be conserved in darkness. The size and quantity of pycnidia vary with the illuminance. Under low and high illuminances physiologically different pycnidia appear.Phomopsis mali cultivated in darkness in the presence of other microorganisms (fungi or bacteria) can fructify and produces pycnidia similar to those formed under low illuminance and in natural conditions. Our experiments indicate the presence of a pycnogenesis-inducing substance that can exists in different forms and induces the formation of the different pycnidia.