Barnabás Wodala

In Vivo Target Sites of Nitric Oxide in Photosynthetic Electron Transport as Studied by Chlorophyll Fluorescence in Pea Leaves

The role of nitric oxide (NO) in photosynthesis is poorly understood as indicated by a number of studies in this field with often conflicting results. As various NO donors may be the primary source of discrepancies, the aim of this study was to apply a set of NO donors and its scavengers, and examine the effect of exogenous NO on photosynthetic electron transport in vivo as determined by chlorophyll fluorescence of pea (Pisum sativum) leaves. Sodium nitroprusside-induced changes were shown to be mediated partly by cyanide, and S-nitroso-N-acetylpenicillinamine provided low yields of NO. However, the effects of S-nitrosoglutathione are inferred exclusively by NO, which made it an ideal choice for this study. QA− reoxidation kinetics show that NO slows down electron transfer between QA and QB, and inhibits charge recombination reactions of QA− with the S2 state of the water-oxidizing complex in photosystem II. Consistent with these results, chlorophyll fluorescence induction suggests that NO also inhibits steady-state photochemical and nonphotochemical quenching processes. NO also appears to modulate reaction-center-associated nonphotochemical quenching.

Monitoring moderate Cu and Cd toxicity by chlorophyll fluorescence and P700 absorbance in pea leaves

We investigated the effect of moderate Cu2+ and Cd2+ stress by applying chlorophyll (Chl) fluorescence and P700 absorbance measurements to monitor the photosynthetic electron transport activity of 3-week-old Pisum sativum L. cv. Petit Provençal plants grown in a modified Hoagland solution containing 50 μM CuSO4 or 5 μM CdCl2. Both heavy metals caused a slight inhibition in PSII photochemistry as indicated by the decrease in the effective quantum efficiency of PSII (ΦPSII), the maximum electron transport capacity (ETRmax), and the maximum quantum yield for electron transport (α). PSI photochemistry was also affected by these heavy metals. Cu2+ and Cd2+ decreased the quantum efficiency of PSI (ΦPSI) as well as the number of electrons in the intersystem chain, and the Cu2+ treatment significantly reduced the number of electrons from stromal donors available for PSI. These results indicate that PSII and PSI photochemistry of pea plants are both sensitive to moderate Cu2+ and Cd2+ stress, which in turn is easily detected and monitored by Chl fluorescence and P700 absorbance measurements. Therefore, monitoring the photochemistry of pea plants with these noninvasive, yet sensitive techniques offers a promising strategy to study heavy metal toxicity in the environment.

Regulation of guard cell photosynthetic electron transport by nitric oxide

Nitric oxide (NO) is one of the key elements in the complex signalling pathway leading to stomatal closure by inducing reversible protein phosphorylation and Ca(2+) release from intracellular stores. As photosynthesis in guard cells also contributes to stomatal function, the aim of this study was to explore the potential role of NO as a photosynthetic regulator. This work provides the first description of the reversible inhibition of the effect of NO on guard cell photosynthetic electron transport. Pulse amplitude modulation (PAM) chlorophyll fluorescence measurements on individual stomata of peeled abaxial epidermal strips indicated that exogenously applied 450nM NO rapidly increases the relative fluorescence yield, followed by a slow and constant decline. It was found that NO instantly decreases photochemical fluorescence quenching coefficients (qP and qL), the operating quantum efficiency of photosystem II (ΦPSII), and non-photochemical quenching (NPQ) to close to zero with different kinetics. NO caused a decrease in NPQ, which is followed by a slow and continuous rise. The removal of NO from the medium surrounding the epidermal strips using a rapid liquid perfusion system showed that the effect of NO on qP and ΦPSII, and thus on the linear electron transport rate through PSII (ETR), is reversible, and the constant rise in NPQ disappears, resulting in a near steady-state value. The reversible inhibition by NO of the ETR could be restored by bicarbonate, a compound known to compete with NO for one of the two coordination sites of the non-haem iron (II) in the QAFe(2+)QB complex.

The cost and risk of using sodium nitroprusside as a NO donor in chlorophyll fluorescence experiments

Sodium nitroprusside (SNP) is a widely used nitric oxide (NO) donor chemical, although it has been reported to release cyanide as well as NO during its photolysis. The aim of this work was to examine this potential side effect of SNP. Chlorophyll fluorescence experiments with pea leaves showed that SNP modifies photosynthetic electron transport. The SNP-induced changes were only partially restored in the presence of a NO-specific scavenger. Moreover, a stoichiometric KCN treatment mimicked the outcome caused by the joint application of the NO donor and NO scavenger. These results confirm the cyanide release of SNP and show that both of its photolytic products reduce the photochemical activity of photosystem II in vivo.

K + outward rectifying channels as targets of phosphatase inhibitor deltamethrin in Vicia faba guard cells

Summary The dominant outward rectifier K + currents were examined in protoplasts from Vicia faba guard cells. In whole-cell patch-clamp recordings, we generally observed that the conductance of the K + inward and the outward rectifier gradually decreases with a half time in the order of 2.3 ± 0.7 min. As a consequence of this rundown, a new steady state was achieved which was 90 ± 5 percnt; lower than that obtained at the beginning of the recording. The rundown of the outward rectifier could be greatly reduced by pre-treating protoplasts either with the membrane permeable drug deltamethrin or by perfusing protoplasts with a pipette solution containing 5 μmol/L cyclosporine A. Furthermore, after the rundown, the conductance of the outward rectifier could be partially restored upon addition of 5 μmol/L deltamethrin to the bath medium. Since deltamethrin and cyclosporine A are established inhibitors of the calcium sensitive phosphatase calcineurin, the data argue for a participation of this type of phosphatase in the control of the activity of K + outward rectifier channels in guard cells.

The roleS of ABA, reactive oxygen species and Nitric Oxide in root growth during osmotic stress in wheat: comparison of a tolerant and a sensitive variety

The effects of PEG 6000-induced osmotic stress (-0.976 MPa) on the root growth of young plants, and the changes in abscisic acid (ABA), reactive oxygen species (ROS) and NO contents were investigated in the root tips of a drought-tolerant and a drought-sensitive wheat cultivar (Triticum aestivum L. cvs. MV Emese and GK Élet, respectively). The root length of cv. MV Emese was more effectively reduced than that of GK Élet by osmotic stress. Concomitantly, the ABA content of the 15-mm apical zone of the roots remained at the control level in GK Élet cultivar, but in MV Emese it decreased significantly after the early phase of the experiment, indicating that the accumulation of ABA is necessary for the maintenance of root growth under osmotic stress. The extent of ROS accumulation relative to the respective control was more pronounced in the elongation zone of roots in MV Emese in the later stages of the experiment, while NO concentrations increased significantly early after PEG exposure, suggesting that high concentrations of ROS and NO were unfavourable for root expansion. In contrast, in cv. Élet, the high NO content in the elongation zone declined to the control level under osmotic stress within 4 days. The changes in root growth due to osmotic stress did not exhibit a correlation with the drought tolerance of the genotypes defined on the basis of the crop yield.

Effect of EDTA-assisted copper uptake on photosynthetic activity and biomass production of sweet sorghum.

Sweet sorghum (Sorghum bicolor L. Moench cv. Rona) is a widely grown sugar crop that is used for bioenergy production. Since sorghum shows increased sensitivity to nutrient deficiency, the objective of this study was to reach an appropriate Cu level in plant tissues using various concentrations of Cu and ethylenediaminetetraacetic acid (EDTA) in order to enhance the photosynthetic activity and biomass production of plants. Copper accumulation increased in the root and stem of plants irrigated for 12 weeks with 0.1 μM CuCl2 both in the presence and absence of 300 μM EDTA and as a consequence, the plant-available Cu concentration in the soil extracts was lower at harvest. Although the copper content of leaves slightly increased, the transport of Fe and Mn, the microelements participating in light reactions of photosynthesis was negatively affected. In spite of this, 0.1 μM CuCl2 alone and with 200 or 300 μM EDTA enhanced the maximal CO2 assimilation rate (Amax) as a function of photon flux density (PPFD) and increased soluble sugar content in all plant parts. The dry mass of plants especially that of stems increased very significantly after 0.1 μM CuCl2 + 300 μM EDTA treatment. These results show that non-toxic concentration of copper in combination with suitable concentration of EDTA can enhance photosynthesis, biomass production, sugar content and the total copper accumulation in the shoot of sweet sorghum plants.