Medicago sativa and M. tunetana reveal contrasting physiological and metabolic responses to drought.

Alfalfa production is frequently constrained by drought, indicating the importance of assessing species biodiversity in endemic close relatives to enhance forage production under future global change conditions. In the present study, plants of two ecotypes of M. tunetana, native to Tunisia, and four commercial cultivars of M. sativa were subjected to two water regimes (control vs drought [15% field capacity]). Physiological, isotopic and metabolic analyses were used to characterize leaf and nodule profiles of the plants. Biomass, gas exchange and the maximum carboxylation rate (Vcmax) indicated significant decreases in photosynthetic capacity under drought in M. sativa cultivars. However, M. tunetana ecotypes maintained photosynthetic performance and aboveground biomass under drought conditions. Furthermore, nitrogen isotope composition (δ15N) in nodules and leaves was significantly decreased, which reveals a reduction in the N2 fixing activity of nodules under drought conditions that was not translated into lower leaf N content but was probably due to lower N demand. Analyses of starch, soluble sugar, and amino acid content in leaves and nodules have clearly proven the ability of Medicago spp. cultivars to increase the accumulation of osmo-protectors under drought. This study demonstrated the genetic variability of the strategy adopted among the studied cultivars in response to drought. In this sense, M. tunetana, and in part the M. sativa cultivar adapted to Mediterranean conditions, seem capable of maintaining adequate biomass, photosynthesis and biological N2 fixation in comparison to the other M. sativa cultivars.

Eco-Geographical, Morphological and Molecular Characterization of a Collection of the Perennial Endemic Species Medicago tunetana (Murb.) A.W. Hill (Fabaceae) from Tunisia.

In order to characterize and conserve the endemic pastoral species Medicago tunetana, many prospecting missions were carried out in mountainous regions of the Tunisian ridge. Twenty-seven eco-geographical and morphological traits were studied for six M. tunetana accessions and followed by molecular analysis using seven Simple Sequence Repeat (SSR). Only five markers were polymorphic and reproductible in the six M. tunetana populations. A total of 54 alleles were observed with an average of 10.8 bands/primer/genotype. Mean Polymorphism Information Content (PIC), Nei gene diversity (h) Shannon's information index (I) indicated the high level of polymorphism. The generated dendrogram with hierarchical UPGMA cluster analysis grouped accessions into two main groups with various degree of subclustring. All the studied accessions shared 57% of genetic similarity. Analysis of variance showed high significant difference between morphological traits among M. tunetana populations where MT3 from Kesra showed different morphological patterns regarding leaf, pod and seeds traits. Canonical correspondence analysis (CCA) showed two principal groups of M. tunetana populations based on potassium, total and active lime contents in soil. Our results suggest that SSR markers developed in M. truncatula could be a valuable tool to detect polymorphism in M. tunetana. Furthermore, the studied morphological markers showed a large genetic diversity among M. tunetana populations. This approach may be applicable for the analysis of intra specific variability in M. tunetana accessions. Our study could help in the implementation of an effective and integrated conservation programs of perennial endemic Medicago.

Responses to Salinity in Four Plantago Species from Tunisia.

The genus Plantago is particularly interesting for studying the mechanisms of salt tolerance in plants, as it includes both halophytes and glycophytes, as well as species adapted to xeric environments. In this study, the salt stress responses of two halophytes, P. crassifolia and P. coronopus, were compared with those of two glycophytes, P. ovata and P. afra. Plants obtained by seed germination of the four species, collected in different regions of Tunisia, were subjected to increasing salinity treatments for one month under greenhouse conditions. Morphological traits and biochemical parameters, such as ion accumulation and the leaf contents of photosynthetic pigments, osmolytes, oxidative stress markers and antioxidant metabolites, were measured after the treatments. Salt-induced growth inhibition was more pronounced in P. afra, and only plants subjected to the lowest applied NaCl concentration (200 mM) survived until the end of the treatments. The biochemical responses were different in the two groups of plants; the halophytes accumulated higher Na+ and proline concentrations, whereas MDA levels in their leaves decreased, indicating a lower level of oxidative stress. Overall, the results showed that P. coronopus and P. crassifolia are the most tolerant to salt stress, and P. afra is the most susceptible of the four species. Plantago ovata is also quite resistant, apparently by using specific mechanisms of tolerance that are more efficient than in the halophytes, such as a less pronounced inhibition of photosynthesis, the accumulation of higher levels of Cl- ions in the leaves, or the activation of K+ uptake and transport to the aerial part under high salinity conditions.

Genetic diversity and population structure of six species of Capparis in Tunisia using AFLP markers.

In order to study the genetic diversity, the phylogeographic pattern and hybridization between six Tunisian Capparis species, 213 accessions of Caper were genotyped with three primer combinations of amplified fragment length polymorphism (AFLP) markers. Out of 750 fragments generated, 636 were polymorphic and 407 of them were restricted to a single species. STRUCTURE and PCoA analyses clearly separated morphologically different populations into six distinct genetic ones. The UPGMA analysis grouped the species into three main clusters: G1 grouped C. spinosa subsp. spinosa var. spinosa and C. sicula subsp. sicula; G2 grouped C. ovata subsp. ovata and C. orientalis and G3 clustered C. zoharyi and C. aegyptia. Populations from G1, G2 and G3 were mainly distributed in arid, subhumid, and semi-arid bioclimates, respectively. Additional genetic studies on Capparis could help to identify genes underlying speciation events and local adaptation to geographic areas leading to the development of breeding programs.