The role of NADPH oxidases in regulating leaf gas exchange and ion homeostasis in Arabidopsis plants under cadmium stress.

NADPH oxidase, an enzyme associated with the plasma membrane, constitutes one of the main sources of reactive oxygen species (ROS) which regulate different developmental and adaptive responses in plants. In this work, the involvement of NADPH oxidases in the regulation of photosynthesis and cell ionic homeostasis in response to short cadmium exposure was compared between wild type (WT) and three RBOHs (Respiratory Burst Oxidase Homologues) Arabidopsis mutants (AtrbohC, AtrbohD, and AtrbohF). Plants were grown under hydroponic conditions and supplemented with 50 µM CdCl2 for 24 h. Cadmium treatment differentially affected photosynthesis, stomatal conductance, transpiration, and antioxidative responses in WT and Atrbohs mutants. The loss of function of RBOH isoforms resulted in higher Cd2+ influx, mainly in the elongation zone of roots, which was more evident in AtrbohD and AtrbohF mutants. In the mature zone, the highest Cd2+ influx was observed in rbohC mutant. The lack of functional RBOH isoforms also resulted in altered patterns of net K+ transport across cellular membranes, both in the root epidermis and leaf mesophyll. The analysis of expression of metal transporters by qPCR demonstrated that a loss of functional RBOH isoforms has altered transcript levels for metal NRAMP3, NRAMP6 and IRT1 and the K+ transporters outward-rectifying K+ efflux GORK channel, while RBOHD specifically regulated transcripts for high-affinity K+ transporters KUP8 and HAK5, and IRT1 and RBOHD and F regulated the transcription factors TGA3 and TGA10. It is concluded that RBOH-dependent H2O2 regulation of ion homeostasis and Cd is a highly complex process involving multilevel regulation from transpirational water flow to transcriptional and posttranslational modifications of K/metals transporters.

[Evaluation of different procedures for the extraction of phenolic compounds from a medicinal plant: Verbena officinalis]. / Évaluation de différents procédés d'extraction des composés phénoliques d'une plante médicinale : Verbena officinalis.

Since ancient times, plants have been the main source of bioactive molecules, such as phenolic compounds, capable of remedying various diseases. However, polyphenols' content and efficiency vary greatly as a function of several intrinsic and extrinsic factors. To optimize the procedure for the extraction of active molecules from the medicinal plant Verbena officinalis, effects of the plant origin, selected solvent, and extraction method were assessed. V. officinalis aerial parts were collected in two different regions of Tunisia (Bizerte and Ain Draham), and their bioactive molecules were extracted by maceration, decoction, and by the Soxhlet apparatus, either with water or with ethanol. Significant variability in the extracts' contents of phenolic compounds as well as their antioxidant and antimicrobial capacities were noted depending on the different studied factors. In particular, ethanol extracts were found to generally contain higher concentrations of phenolic compounds and more potent antioxidant capacities than water extracts. However, when tested against various pathogenic bacteria, water extracts were most often at least as active as ethanol extracts to inhibit bacteria growth in vitro. Finally, differences were also observed between V. officinalis samples from Bizerte compared to Ain Draham area. All of these results emphasize the need of adapting various parameters for the optimal extraction of bioactive molecules from a medicinal plant such as V. officinalis.

Title: Évaluation de différents procédés d'extraction des composés phénoliques d'une plante médicinale : Verbena officinalis. Abstract: Depuis la nuit des temps, les plantes ont été la source principale de molécules bioactives, tels les composés phénoliques, capables de remédier à diverses maladies. Cependant, le contenu et l'activité des polyphénols dépendent d'un certain nombre de facteurs intrinsèques et extrinsèques. Dans le but d'optimiser les procédés d'obtention des principes actifs de la verveine (Verbena officinalis), les effets de la provenance de la plante, du solvant et de la méthode d'extraction ont été évalués. Ainsi, la partie aérienne de V. officinalis a été collectée dans deux régions différentes de la Tunisie (Bizerte et Ain Draham). L'extraction a été réalisée par macération, décoction et par l'appareil de Soxhlet tantôt avec de l'eau tantôt avec de l'éthanol pur. Une variabilité significative des teneurs en composés phénoliques ainsi que des capacités antioxydantes et antimicrobiennes des extraits a été observée en fonction des facteurs étudiés. De manière générale, les extraits éthanoliques sont plus riches en composés phénoliques et présentent des activités antioxydantes plus fortes que les extraits aqueux. Cependant, vis-à-vis de différentes souches pathogènes, les extraits aqueux sont souvent au moins aussi puissants que les extraits éthanoliques pour inhiber la croissance bactérienne in vitro. De plus des différences notables sont observées selon que V. officinalis provient de la région de Bizerte ou d'Ain Draham. Ces résultats montrent que la prise en compte de plusieurs paramètres est nécessaire pour optimiser l'efficacité des procédures d'extraction des molécules bioactives de V. officinalis.

Potassium deficiency alters growth, photosynthetic performance, secondary metabolites content, and related antioxidant capacity in Sulla carnosa grown under moderate salinity.

Salinity and K+ deficiency are two environmental constraints that generally occur simultaneously under field conditions, resulting in severe limitation of plant growth and productivity. The present study aimed at investigating the effects of salinity, either separately applied or in combination with K+ deficiency, on growth, photosynthetic performance, secondary metabolites content, and related antioxidant capacity in Sulla carnosa. Seedlings were grown hydroponically under sufficient (6000 µM) or low (60 µM) K+ supply with 100 mM NaCl (C + S and D + S treatments, respectively). Either alone or combined with K+ deficiency, salinity significantly restricted the plant growth. K+ deficiency further increased salt impact on the photosynthetic activity of S. carnosa, but this species displayed mechanisms that play a role in protecting photosynthetic machinery (including non photochemical quenching and antioxidant activity). In contrast to plants subjected to salt stress alone, higher accumulation of phenolic compounds was likely related to antioxidative defence mechanism in plants grown under combined effects of two stresses. As a whole, these data suggest that K+ deficiency increases the deleterious effects of salt stress. The quantitative and qualitative alteration of phenolic composition and the enhancement of related antioxidant capacity may be of crucial significance for S. carnosa plants growing under salinity and K+ deficient conditions.

The effect of salinity on photosynthetic activity in potassium-deficient barley species.

The interactive effects of salinity and potassium deficiency on the growth, mineral elements and photosynthetic performance were investigated in wild (Hordeum maritimum L.) and cultivated barley (Hordeum vulgare L. var. Manel). At 28 d of growth, plants were treated with 3 mM K and 0 mM NaCl (3-0); 3 mM K and 100 mM NaCl (3-100); 0 mM K and 0 mM NaCl (0-0), 0 mM K and 100 mM NaCl (0-100) for 14 d. In both species, biomass production decreased considerably when the two constraints were applied simultaneously. Salinity affected shoots more than roots, whereas for potassium deficiency, the reverse occurred. Generally, potassium uptake was more affected in wild than in cultivated barley and, independent of potassium availability, 100 mM NaCl increased Na+ content in both species, whereas K+ deprivation increased Na(+) content only in H. maritimum shoots (0-0). Potassium-use efficiency (KUE) increased in all treated plants. Potassium deficiency increased the negative effects induced by salt in the photosynthetic process of H. vulgare, and this species seemed to be unable to counteract the negative effects of salinity. H. maritimum showed limitation in CO2 photoassimilation, but this species displayed mechanisms that play a role in avoiding PSII photodamage aimed to dissipate the excess energy.

Implication of phospholipase D in response of Hordeum vulgare root to short-term potassium deprivation.

To verify the possible involvement of lipids and several other compounds including hydrogen peroxide (H(2)O(2)) and glyceraldehyde-3-phosphate dehydrogenase (G3PDH) in the response of Hordeum vulgare to early potassium deprivation, plants were grown in hydroponic conditions for 30d with a modified Hewitt nutrient solution containing 3mM K(+). They were then incubated for increasing periods of time ranging from 2 to 36h in the same medium deprived of K(+). In contrast to leaves, root K(+) concentration showed its greatest decrease after 6h of treatment. The main lipids of the control barley roots were phospholipids (PL), representing more than 50% of the total lipids. PL did not change with treatment, whereas free sterols (FS) decreased following K(+) deprivation, showing a reduction of approximately 17% after 36h. With respect to the individual PL, 30h K(+) deprivation led to a reduction in phosphatidylcholine (PC), phosphatidylserine (PS), and phosphatidylinositol (PI) levels, whereas phosphatidylglycerol (PG), phosphatidylethanolamine (PE), and phosphatidic acid (PA) levels increased. The maximum PA accumulation and the highest phospholipase D (PLD) activation, estimated by an accumulation of phosphatidylbutanol (PtBut), were observed after 24h of K(+) starvation. At the root level, H(2)O(2) showed the maximum value after 6h of incubation in -K solution. In parallel, G3PDH activity reached its minimum. On the basis of a concomitant stimulation of PLD activity and, consequently, PA accumulation, enhancement of H(2)O(2) production, and inhibition of G3PDH activity, we suggest a possible involvement of these three compounds in an early response to K(+) deprivation.

Application of municipal solid waste compost reduces the negative effects of saline water in Hordeum maritimum L.

The efficiency of composted municipal solid wastes (MSW) to reduce the adverse effects of salinity was investigated in Hordeum maritimum under greenhouse conditions. Plants were cultivated in pots filled with soil added with 0 and 40tha(-1) of MSW compost, and irrigated twice a week with tap water at two salinities (0 and 4gl(-1) NaCl). Harvests were achieved at 70 (shoots) and 130 (shoots and roots) days after sowing. At each cutting, dry weight (DW), NPK nutrition, chlorophyll, leaf protein content, Rubisco (ribulose-bisphosphate carboxylase/oxygenase) capacity, and contents of potential toxic elements were determined. Results showed that compost supply increased significantly the biomass production of non salt-treated plants (+80%). This was associated with higher N and P uptake in both shoots (+61% and +80%, respectively) and roots (+48% and +25%, respectively), while lesser impact was observed for K+. In addition, chlorophyll and protein contents as well as Rubisco capacity were significantly improved by the organic amendment. MSW compost mitigated the deleterious effect of salt stress on the plant growth, partly due to improved chlorophyll and protein contents and Rubisco capacity (-15%, -27% and -14%, respectively, in combined treatment, against -45%, -84% and -25%, respectively, in salt-stressed plants without compost addition), which presumably favoured photosynthesis and alleviated salt affect on biomass production by 21%. In addition, plants grown on amended soil showed a general improvement in their heavy metals contents Cu2+, Pb2+, Cd2+, and Zn2+ (in combined treatment: 190%, 53%, 168% and 174% in shoots and 183%, 42%, 42% and 114% in roots, respectively) but remained lower than phytotoxic values. Taken together, these findings suggest that municipal waste compost may be safely applied to salt-affected soils without adverse effects on plant physiology.