Zouhaier Barhoumi

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.

Potential utilisation of halophytes for the rehabilitation and valorisation of salt-affected areas in Tunisia

In arid and semi-arid regions, irrigation water contributes to salinisation of the upper layer of the soil, where most root activity takes place. Along the path of plant domestication, many crop species have lost resistance mechanisms to various stress conditions [1], including salt stress [2]. Thus, most crop plants do not fully express their original genetic potential for growth, development and yield under salt stress, and their economic value declines as salinity levels increase [3, 4]. Improving salt resistance of crop plants is, therefore, of major concern in agricultural research. A potential genetic resource for the improvement of salt resistance in crop plants resides among wild populations of halophytes [5, 6]. These can be either domesticated into new, salt-resistant crops, or used as a source of genes to be introduced into crop species by classical breeding or molecular methods.

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.

Histochemical Localization of Essential Oils and Bioactive Substances in the Seed Coat of the Halophyte Crithmum maritimum L. (Apiaceae)

Despite seeds and fruits of several halophytes being rich in essential oils and other bioactive substances, the histochemical characterization of these compounds has not received much attention. The aims of the present investigation were to localize the essential oils and the bioactive substances in the seed coat of the oilseed halophyte Crithmum maritimum L. Fruits were collected from the rocky coasts of Tabarka (NW of Tunisia, 36°57′12″ N, 08°45′18″ E). C. maritimum L. seed is shown to be surrounded by two envelopes: The first structure is a secretory envelope, consisting in about 20 canals. The second layer represents the endocarp. As revealed by fluorescence and light microscopy, the essential oils, O-dihydroxyphenols and flavonoids, known as bioactive compounds, were accumulated in the canals. The endocarp layer accumulated polysaccharides, O-dihydroxyphenols, and flavonoids. As a whole, these findings highlight the histochemical features and confirm the valuable quality of C. maritimum L. seeds.

Insights into the physiological responses of the facultative halophyte Aeluropus littoralis to the combined effects of salinity and phosphorus availability.

In this work, we investigate the physiological responses to P deficiency (5μM KH2PO4=D), salt stress (400mM NaCl=C+S), and their combination (D+S) on the facultative halophyte Aeluropus littoralis to understand how plants adapt to these combined stresses. When individually applied, both P deficiency and salinity significantly restricted whole plant growth, with a more marked effect of the latter stress. However, the effects of the two stresses were not additive in plant biomass production since the response of plants to combined salinity and P deficiency was similar to that of plants grown under salt stress alone. In addition the observed features under salinity alone are kept when plants are simultaneously subjected to the combined effects of salinity and P deficiency such as biomass partitioning; the synthesis of proline and the K(+)/Na(+) selectivity ratio. Thus, increasing P availability under saline conditions has no significant effect on salt tolerance in this species. Plants cultivated under the combined effects of salinity and P deficiency exhibited the lowest leaf water potential. This trend was associated with a high accumulation of Na(+), Cl(-) and proline in shoots of salt treated plants suggesting the involvement of these solutes in osmotic adjustment. Proline could be involved in other physiological processes such as free radical scavenging. Furthermore, salinity has no significant effect on phosphorus acquisition when combined with a low P supply and it significantly decreased this parameter when combined with a sufficient P supply. This fact was probably due to salts effect on P transporters. In addition, shoot soluble sugars accumulation under both P deficiency treatments with and without salt likely play an important role in the adaptation of A. littoralis plants to P shortage applied alone or combined with salinity. Moreover, there was a strong correlation between shoot and root intracellular acid phosphatase activity and phosphorus use efficiency which strengthens the assumption that intracellular acid phosphatase enzymes are involved in P remobilization in this species. Finally, our results showed that P availability has no significant effect on salt excretion in A. littorlais which suggests that independently of the P status in the plant, excretion remains priority over other functions requiring energy such as growth. This result could also indicate that salt excretion is not energy-dependent in this species.

The mericarp of the halophyte Crithmum maritimum (Apiaceae): structural features, germination, and salt distribution

At maturation and during seed fall and dispersal, halophyte seeds may be subjected to invasion by salt ions. How these seeds remain viable in such hostile environments is however still unclear, depending for instance on the species and the family. In the Apiaceae, the mericarp (fruit) shows a wide range of morphological and anatomical modifications, many of which may enhance the adaptation to severe environmental conditions. Therefore, structural features, ion accumulation, and long-term floating capacity were investigated in the fruit (mericarp) of the halophyte Crithmum maritimum L. The mericarp was composed of a spongy outer coat, a secretory envelope, a thin endocarp reduced to a unicellular layer delimiting the endosperm, and an embryo. Both of the secretory canals and the endocarp adhered after complete ripening of the mericarp, while the epicarp and much of the mesocarp formed the spongy coat. Assessing long-term floating ability of the fruit under laboratory conditions revealed that even after 60 d, more than 98% of C. maritimum L. mericarps still floated over seawater. Seed germination was delayed and reduced by the spongy coat. The X-ray microanalysis revealed that the spongy coat and the secretory canals contained essentially Cl and Na, while seeds, i.e. endosperm and embryo, accumulated mostly Mg, K and P. In a subsequent experiment designed to simulate salt leaching by rain, most of the salt accumulated in the spongy coat and seeds was released after 2 h imbibition in distilled water. Taken together, these results highlight the protective role of the mericarp and the likely involvement of this structure in the seed dispersal of C. maritimum L. This may ultimately have eco-physiological implications explaining the successful establishment of this halophyte in its native saline biotopes.

Investigation of embryo growth and reserve mobilization of water or salt imbibed seeds of Crithmum maritimum L.

Abstract The embryo growth, endosperm degradation and in situ activity of peroxidase and esterase in seeds imbibed in distilled water or 200mM NaCl were investigated in the halophyte Crithmum maritimum L. Germination was maximal (90%) in distilled water, but was fully inhibited following seed exposure to salt. The completion of the embryo growth (ca. 2mm length) leading to the radicle emergence took 6 d in distilled, but was markedly delayed in 200mM NaCl. The endosperm degradation was markedly delayed by NaCl. Esterase and peroxidase activities were less important in NaCl-imbibed than in H2O-imbibed seeds. The adverse effect of salinity on the embryo growth and the endosperm degradation could partly explain the inhibition of seed germination observed under these circumstances.