Abderrazak Smaoui

Influence of biological, environmental and technical factors on phenolic content and antioxidant activities of Tunisian halophytes

Halophyte ability to withstand salt-triggered oxidative stress is governed by multiple biochemical mechanisms that facilitate retention and/or acquisition of water, protect chloroplast functioning, and maintain ion homeostasis. Most essential traits include the synthesis of osmolytes, specific proteins, and antioxidant molecules. This might explain the utilization of some halophytes as traditional medicinal and dietary plants. The present study aimed at assessing the phenolic content and antioxidant activities of some Tunisian halophytes (Cakile maritima, Limoniastrum monopetalum, Mesembryanthemum crystallinum, M. edule, Salsola kali, and Tamarix gallica), depending on biological (species, organ and developmental stage), environmental, and technical (extraction solvent) factors. The total polyphenol contents and antioxidant activities (DPPH and superoxide radicals scavenging activities, and iron chelating and reducing powers) were strongly affected by the above-cited factors. Such variability might be of great importance in terms of valorising these halophytes as a source of naturally secondary metabolites, and the methods for phenolic and antioxidant production.

Phytodesalination of a salt-affected soil with the halophyte Sesuvium portulacastrum L. to arrange in advance the requirements for the successful growth of a glycophytic crop.

In the present work, we studied the potential of the obligate halophyte, Sesuvium portulacastrum L., to desalinize an experimentally-salinized soil after the following criteria: (i) decrease in soil salinity and sodicity, (ii) plant biomass capacity to accumulate sodium ions, and (iii) phytodesalinized soil quality (equivalent to growth of a glycophytic test culture of Hordeum vulgare L.). The cultivation of the halophyte on the salinized soil (phytodesalination culture) led to a marked absorption of Na(+) ions by S. portulacastrum roots and their accumulation in the above-ground biomass up to 872 mg plant(-1) and 4.36 g pot(-1) (about 1 tha(-1)). The decrease in salinity and sodicity of the phytodesalinized soil significantly reduced the negative effects on growth of the test culture of H. vulgare. Furthermore, the phytodesalination enabled H. vulgare plants to keep a high water content and to develop a higher biomass with relatively high K and low Na contents.

ABA, GA3, and nitrate may control seed germination of Crithmum maritimum (Apiaceae) under saline conditions

Impaired germination is common among halophyte seeds exposed to salt stress, partly resulting from the salt-induced reduction of the growth regulator contents in seeds. Thus, the understanding of hormonal regulation during the germination process is a main key: (i) to overcome the mechanisms by which NaCl-salinity inhibit germination; and (ii) to improve the germination of these species when challenged with NaCl. In the present investigation, the effects of ABA, GA(3), NO(-)(3), and NH(+)(4) on the germination of the oilseed halophyte Crithmum maritimum (Apiaceae) were assessed under NaCl-salinity (up to 200 mM NaCl). Seeds were collected from Tabarka rocky coasts (N-W of Tunisia). The exogenous application of GA(3), nitrate (either as NaNO(3) or KNO(3)), and NH(4)Cl enhanced germination under NaCl salinity. The beneficial impact of KNO(3) on germination upon seed exposure to NaCl salinity was rather due to NO(-)(3) than to K(+), since KCl failed to significantly stimulate germination. Under optimal conditions for germination (0 mM NaCl), ABA inhibited germination over time in a dose dependent manner, but KNO(3) completely restored the germination parameters. Under NaCl salinity, the application of fluridone (FLU) an inhibitor of ABA biosynthesis, stimulated substantially seed germination. Taken together, our results point out that NO(-)(3) and GA(3) mitigate the NaCl-induced reduction of seed germination, and that NO(-)(3) counteracts the inhibitory effect of ABA on germination of C. maritimum.

Localization of potential ion transport pathways in vesicular trichome cells of Atriplex halimus L.

The secreting glandular trichomes are recognized as an efficient structure that alleviates salt effects on Atriplex halimus. They are found on buds, young green stems, and leaves. They occupy both the leaf surfaces and give them a whitish color. Their histogenesis and ultrastructure were investigated in the third young leaves. They appear in early stage of plant development and its initiation continuous until just the leaf final development state. Each trichome contains two parts; a stalk which has high electron opacity, embedded in epidermal cells, and bears a second one which is unicellular, called bladder cell and has a low electron density. The bladder cell appears as a huge vacuole and the well-reduced cytoplasm which is pushed close to the wall, contains only a few organelles. Concurrently, the use of silver chloride precipitation technique shows that, in secretion process, salt follows a symplasmatic pathway which is consolidated by the presence of numerous plasmodesmata between the stalk cell(s), and the bladder one and the neighboring mesophyll cells. In addition, according to lanthanum-tracer study, salt can be excreted apoplastically. In fact, the heavy element can be transported via endocytosis vesicles, and by Golgi, endoplasmic reticulum, and lysosome (G.E.R.L.) network toward the storage vacuoles.

PHYTODESALINATION OF A MODERATELY-SALT-AFFECTED SOIL BY SULLA CARNOSA

The aim of this investigation was to evaluate the ability of the indifferent halophyte Sulla carnosa Desf. to desalinize a moderately-salt-affected soil. Seeds were sown on a fertile soil added or not with 1.5 g NaCl. kg−1. Analogous treatments without plantation (control and salinized) were also used. Plant culture was performed under greenhouse conditions in non-perforated pots containing 10 kg soil each and irrigated with non-saline tap water. After 80 days of treatment, shoots were harvested. Soil samples were also collected after division of soil column in each pot into two horizons. Our results showed that salt addition increased electrical conductivity of saturation paste extract (ECe) from 3.3 to 8.4 dS. m−1 and soluble sodium concentration from 0.32 to 1.15 g. kg−1 soil in the upper horizon. In the lower horizon however, Na+ concentration was quasi-constant and then ECe was less increased. Plant culture inversed this pattern of sodium accumulation and salinity. Its productivity and phytodesalination capacity in 80 days were 5.0 t DW. ha−1 and 0.3 t Na+. ha−1 (24% of the added quantity), respectively. Interestingly, sodium dilution within biomass (41.5–45.6 mg. g−1 DW) and the non-altered nutrition make this plant suitable for forage as second use after phytodesalination.

Sesuvium portulacastrum maintains adequate gas exchange, pigment composition, and thylakoid proteins under moderate and high salinity.

Cuttings of Sesuvium portulacastrum L. (Aizoaceae) were taken from plants cultivated under severe saline conditions. The obtained seedlings were grown on sand and irrigated with nutrient solution over 5 weeks under no (0 mM NaCl), moderate (200 mM NaCl), or high (400 mM NaCl) salinity conditions. A follow-up of gas exchange was performed weekly and pigment levels and patterns of fully expanded leaves were determined after 3 and 5 weeks of treatment. At the end of the 5-week period, immunoblot analysis of the main polypeptides of photosystem I and II was performed with the aim to investigate salt-induced variations in photosystem composition. Net CO2 assimilation rate (Pn) increased under salinity up to 3 weeks of treatment then decreased to reach the value of 0mM-treated plants at the end of the experiment. For stomatal conductance (gs) and intercellular CO2 concentration (Ci), the opposite occurred. These results were concomitant with an increase in practically all pigment levels, mainly under high salinity, with the exception of zeaxanthin. The de-epoxidation index (DEPS index) was much lower under saline than non-saline conditions in the 3rd week, indicating light stress in 0mM-treated plants. At the end of the experiment, this index showed much lower values with no significant differences between treatments, which coincided with no significant differences in gas exchange as well. Protein amounts of D1, CP47, and CP43 did not show noticeable variations with salt treatment, whereas LHCII underwent a slight but significant decrease (-15%) at the highest NaCl concentration. LHCI polypeptides were unaffected by the salt treatments, where conversely, the highest concentration induced a significant decrease in PsaA/B amount (-18%).

In Vitro Antiproliferative Effect of Arthrocnemum indicum Extracts on Caco-2 Cancer Cells through Cell Cycle Control and Related Phenol LC-TOF-MS Identification

This study aimed to determinate phenolic contents and antioxidant activities of the halophyte Arthrocnemum indicum shoot extracts. Moreover, the anticancer effect of this plant on human colon cancer cells and the likely underlying mechanisms were also investigated, and the major phenols were identified by LC-ESI-TOF-MS. Results showed that shoot extracts had an antiproliferative effect of about 55% as compared to the control and were characterised by substantial total polyphenol content (19 mg GAE/g DW) and high antioxidant activity (IC50 = 40 μg/mL for DPPH test). DAPI staining revealed that these extracts decrease DNA synthesis and reduce the proliferation of Caco-2 cells which were stopped at the G2/M phase. The changes in the cell-cycle-associated proteins (cyclin B1, p38, Erk1/2, Chk1, and Chk2) correlate with the changes in cell cycle distribution. Eight phenolic compounds were also identified. In conclusion, A. indicum showed interesting antioxidant capacities associated with a significant antiproliferative effect explained by a cell cycle blocking at the G2/M phase. Taken together, these data suggest that A. indicum could be a promising candidate species as a source of anticancer molecules.

Effects of magnesium deficiency on photosynthesis and carbohydrate partitioning

Magnesium nutrition is often forgotten, while its absence adversely affects numerous functions in plants. Magnesium deficiency is a growing concern for crop production frequently observed in lateritic and leached acid soils. Competition with other cations (Ca2+, Na+, and K+) is also found to be an essential factor, inducing magnesium deficiency in plants. This nutrient is required for chlorophyll formation and plays a key role in photosynthetic activity. Moreover, it is involved in carbohydrate transport from source-to-sink organs. Hence, sugar accumulation in leaves that results from the impairment of their transport in phloem is considered as an early response to Mg deficiency. The most visible effect is often recorded in root growth, resulting in a significant reduction of root/shoot ratio. Carbohydrate accumulation in source leaves is attributed to the unique chemical proprieties of magnesium. As magnesium is a nutrient with high mobility in plants, it is preferentially transported to source leaves to prevent severe declines in photosynthetic activity. In addition, Mg is involved in the source-to-sink transport of carbohydrates. Hence, an inverse relationship between Mg shortage and sugar accumulation in leaves is often observed. We hereby review all these aspects with a special emphasis on the role of Mg in photosynthesis and the structural and functional effects of its deficiency on the photosynthetic apparatus.

Selection of a halophyte that could be used in the bioreclamation of salt-affected soils in arid and semi-arid regions

Vegetative bioremediation or bioreclamation of salt-affected soils is an economic solution mainly for developing countries since chemical additions are becoming increasingly expensive. However, to be efficient, this approach needs sufficient irrigation. In this investigation, we evaluated the ability of some halophytes to desalinize a saline soil under non-leaching conditions with the aim of selecting appropriate species that could be used for this purpose in arid and semi-arid regions where precipitation is too low to leach salts from the rhizosphere. Three perennial species were used in this experiment: Arthrocnemum indicum (Willd.) Moq., Suaeda fruticosa Forsk., and Sesuvium portulacastrum L. Seedlings were grown on a saline soil under greenhouse conditions and irrigated with tap water for 170 days. Irrigations were carried out with almost no leaching. Soil salinity was significantly reduced in halophyte-grown soil as compared to the control. Plants were able to decrease the soil electrical conductivity by absorbing soluble salts, mainly sodium ions. Among the three studied species, Sesuvium portulacastrum L. was the most productive and was able to accumulate in shoots nearly 30% of the sodium content of each pot over the 170 days. Thus, Sesuvium portulacastrum L. seems to be the most promising species for saline soil desalination in arid and semi-arid regions.

Physiological and leaf metabolome changes in the xerohalophyte species Atriplex halimus induced by salinity.

Atriplex halimus is a xerohalophyte plant, which could be used as cash crops. This plant was integrated in Tunisian government programs the aim of which is to rehabilitate saline areas and desert. To investigate its strategies involved in salt tolerance, A. halimus was grown hydroponically under controlled conditions with increasing salinity. Plants were harvested and analyzed after 60 days of treatment. The biomass of A. halimus increased by moderate salinity and decreased significantly at high salinity compared to control plants at 400 mM. Despite of the large amounts of Na(+) observed in the leaves of Atriplex plants, leaf water contents and leaf succulence kept on increasing in treated plants and decreased over 150 mM NaCl. This confirmed the compartmentation and the efficient contribution of Na(+) in the osmotic adjustment. Analysis of the metabolic profiles showed an accumulation of carbohydrates and amino acids. The leaf tissues preferentially stored proline, α alanine and sucrose. Increasing NaCl levels were also accompanied by a significant accumulation of malate in leaves. Involvement of these solutes in osmotic adjustment was considered low. Nevertheless, they seemed to have an important role in controlling photosynthesis which capacity was enhanced by low salinity and decreased with increasing salinity (evaluated by actual photochemical efficiency of photosystem II and chlorophyll contents). The unchanged maximum photochemical efficiency of photosystem II accompanied by the increase of the non-photochemical quenching, the enhancement of the total antioxidant activity and the decrease of the malondialdehyde contents in leaves showed efficient protection of membranes and photosystem II from photo-oxidative damage. This protection seemed to be attributed to proline and sucrose largely accumulated in leaves treated with salt.