Insight into mechanisms of multiple stresses tolerance in a halophyte Aster tripolium subjected to salinity and heavy metal stress.

The study aimed at comparing metabolic reactions of a halophyte Aster tripolium to abiotic stresses. Profiling of endogenous phytohormones, soluble carbohydrates and stress-related amino acids was conducted in plants exposed to moderate and high salinity (150 and 300 mM NaCl), and heavy metal salts CdCl2 or PbCl2 (100 and 200 µM). High NaCl and Pb doses inhibited growth of A. tripolium (Stress Tolerance Index STI) of 37% and 32-35%, respectively. The plants tolerated moderate salinity and Cd (STI = 91% and STI = 83-96%, respectively). Toxic metals accumulated mainly in the roots but Cd translocation to the shoots was also observed. The stressors did not affect total concentrations of the main growth promoting phytohormones but we observed enhanced deactivation of auxins and gibberellins, and reduced accumulation of jasmonate precursor. ABA content increased under stress except for moderate salinity. A common reaction was also activation of osmotic adjustment, however it was disparately manifested under salinity and metallic stress. The distinct responses to salinity and metallic stresses involved changes in carbohydrate profile and altered interplay between salicylic acid content and the pool of active gibberellins. The content of active jasmonates diversified A. tripolium reactions to salt excess and each of the heavy metals. This parameter was linked to the accumulation of ethylene precursor. The results of the study can be used to decipher potential co-tolerance mechanism of this halophyte species to multiple environmental stresses.

Disentangling the photochemical salinity tolerance in Aster tripolium L.: connecting biophysical traits with changes in fatty acid composition.

A profound analysis of A. tripolium photochemical traits under salinity exposure is lacking in the literature, with very few references focusing on its fatty acid profile role in photophysiology. To address this, the deep photochemical processes were evaluated by Pulse Amplitude Modulated (PAM) Fluorometry coupled with a discrimination of its leaf fatty acid profile. Plants exposed to 125-250 mm NaCl showed higher photochemical light harvesting efficiencies and lower energy dissipation rates. under higher NaCl exposure, there is evident damage of the oxygen evolving complexes (OECs). On the other hand, Reaction Centre (RC) closure net rate and density increased, improving the energy fluxes entering the PS II, in spite of the high amounts of energy dissipated and the loss of PS II antennae connectivity. Energy dissipation was mainly achieved through the auroxanthin pathway. Total fatty acid content displayed a similar trend, being also higher under 125-250 mm NaCl with high levels of omega-3 and omega-6 fatty acids. The increase in oleic acid and palmitic acid allows the maintenance of the good functioning of the PS II. Also relevant was the high concentration of chloroplastic C16:1t in the individuals subjected to 125-250 mm NaCl, related with a higher electron transport activity and with the organization of the Light Harvesting Complexes (LHC) and thus reducing the activation of energy dissipation mechanisms. All these new insights shed some light not only on the photophysiology of this potential cash-crop, but also highlight its important saline agriculture applications of this species as forage and potential source of essential fatty acids.

In vitro plant regeneration of Aster scaber via somatic embryogenesis.

We established an in vitro plant regeneration system via somatic embryogenesis of Aster scaber, an important source of various biologically active phytochemicals. We examined the callus induction and embryogenic capacities of three explants, including leaves, petioles, and roots, on 25 different media containing different combinations of α-naphthalene acetic acid (NAA) and 6-benzyladenine (BA). The optimum concentrations of NAA and BA for the production of embryogenic calli were 5.0 µM and 0.05 µM, respectively. Media containing higher concentrations of auxin and cytokinin (such as 25 µM NAA and 25 µM BA) were suitable for shoot regeneration, especially for leaf-derived calli, which are the most readily available calli and are highly competent. For root induction from regenerated shoots, supplemental auxin and/or cytokinin did not improve rooting, but instead caused unwanted callus induction or retarded growth of regenerated plants. Therefore, plant growth regulator-free medium was preferable for root induction. Normal plants were successfully obtained from calli under the optimized conditions described above. This is the first report of the complete process of in vitro plant regeneration of A. scaber via somatic embryogenesis.

Elevated atmospheric CO2 concentration enhances salinity tolerance in Aster tripolium L.

Our study aimed at investigating the influence of elevated atmospheric CO(2) concentration on the salinity tolerance of the cash crop halophyte Aster tripolium L., thereby focussing on protein expression and enzyme activities. The plants were grown in hydroponics using a nutrient solution with or without addition of NaCl (75% seawater salinity), under ambient (380 ppm) and elevated (520 ppm) CO(2). Under ambient CO(2) concentration enhanced expressions and activities of the antioxidant enzymes superoxide dismutase, ascorbate peroxidase, and glutathione-S-transferase in the salt-treatments were recorded as a reaction to oxidative stress. Elevated CO(2) led to significantly higher enzyme expressions and activities in the salt-treatments, so that reactive oxygen species could be detoxified more effectively. Furthermore, the expression of a protective heat shock protein (class 20) increased under salinity and was even further enhanced under elevated CO(2) concentration. Additional energy had to be provided for the mechanisms mentioned above, which was indicated by the increased expression of a beta ATPase subunit and higher v-, p- and f-ATPase activities under salinity. The higher ATPase expression and activities also enable a more efficient ion transport and compartmentation for the maintenance of ion homeostasis. We conclude that elevated CO(2) concentration is able to improve the survival of A. tripolium under salinity because more energy is provided for the synthesis and enhanced activity of enzymes and proteins which enable a more efficient ROS detoxification and ion compartmentation/transport.

Aster tripolium L. and Sesuvium portulacastrum L.: two halophytes, two strategies to survive in saline habitats.

Aster tripolium L. (Dollart, Germany) and Sesuvium portulacastrum L. (Dakhla, Morocco) are potential halophytic vegetables, fodder plants, and ornamentals for re-vegetating saline land. To compare their strategies involved in salt tolerance both plants were grown with 0%, 1.5%, and 3% (Aster) or 0%, 2.5%, and 5% (Sesuvium) NaCl in the watering solution. The growth rate was reduced in both species with increasing NaCl concentrations. The quotient of Na(+)/K(+) indicates that Aster accumulates more K(+) in comparison to Na(+) while the reverse is true for Sesuvium. Osmolality of the leaf sap increased with increasing NaCl concentration in both Aster and Sesuvium. Transpiration rate was severely reduced in both Aster (3%) and Sesuvium (5%) plants after 10 d of NaCl watering. The CO(2) assimilation rate decreased in Aster (3%) and Sesuvium (5%) NaCl-treated plants from day 5 to day 10. The most important results from chlorophyll fluorescence measurements were derived from the non-photochemical quenching analysis (NPQ). First, both plants had linearly increasing levels of NPQ with increasing NaCl concentrations. Second, Sesuvium had almost half the NPQ value when compared to Aster under increased soil salinity. In Aster P-ATPase activities were decreased in plants treated with 3% NaCl after three days of treatment, F-ATPase activities increased with increasing NaCl concentrations and no clear changes were measured in V-ATPase activities. In Sesuvium any changes could be observed in the three ATPase activities determined. To conclude, Aster and Sesuvium use different strategies in adaptation to soil salinity.

Quantification and fatty acid profiles of sulfolipids in two halophytes and a glycophyte grown under different salt concentrations.

This study was aimed at understanding the role of sulfolipids in salt tolerance mechanisms of the halophytes Aster tripolium L., Compositae, and Sesuvium portulacastrum L., Aizoaceae, and of the glycophyte Arabidopsis thaliana (L.) Heynh., Brassicaceae. In Aster and Sesuvium the sulfolipid contents increased significantly under salt stress conditions (517 mM or 864 mM). In Arabidopsis, changes in sulfolipid contents were not observed (NaCl up to 100 mM). The fatty acid profile of sulfoquinovosyldiacylglycerol (SQDG) in Aster was modified with increasing NaCl concentrations. LC-MS analyses of sulfolipids from Aster and Sesuvium revealed the presence of 18:3/18:3 and 16:0/18:3 molecules. Obviously, the function of sulfolipids during salt stress differs between halophytic species and between halophytes and glycophytes where sulfolipid accumulation was not observed.

Photosynthetic limitations of a halophyte sea aster (Aster tripolium L) under water stress and NaCl stress.

To understand the mechanisms of salt tolerance in a halophyte, sea aster ( Aster tripolium L.), we studied the changes of water relation and the factors of photosynthetic limitation under water stress and 300 mM NaCl stress. The contents of Na(+) and Cl(-) were highest in NaCl-stressed leaves. Leaf osmotic potentials ( Psi(s)) were decreased by both stress treatments, whereas leaf turgor pressure ( Psi(t)) was maintained under NaCl stress. Decrease in Psi(s) without any loss of Psi(t) accounted for osmotic adjustment using Na(+) and Cl(-) accumulated under NaCl stress. Stress treatments affected photosynthesis, and stomatal limitation was higher under water stress than under NaCl stress. Additionally, maximum CO(2) fixation rate and O(2) evolution rate decreased only under water stress, indicating irreversible damage to photosynthetic systems, mainly by dehydration. Water stress severely affected the water relation and photosynthetic capacity. On the other hand, turgid leaves under NaCl stress have dehydration tolerance due to maintenance of Psi(t) and photosynthetic activity. These results show that sea aster might not suffer from tissue dehydration in highly salinized environments. We conclude that the adaptation of sea aster to salinity may be accomplished by osmotic adjustment using accumulated Na(+) and Cl(-), and that this plant has typical halophyte characteristics, but not drought tolerance.