Thirty-four durum wheat cultivars representing the Tunisian durum (Triticum durum Desf.) wheat collection and seven wild species of wheat relatives (Triticum turgidum L., T. dicoccon Schrank., T. dicoccoides (Kö rn) Schweinf., T. araraticum Jakubz., T. monococcum L., Aegilops geniculata Roth, and Aegilops ventricosa Tausch) were analysed with amplified fragment length polymorphism (AFLP) and microsatellite (SSR) markers. Both marker systems used were able to differentiate durum wheat cultivars from the wild relatives and to specifically fingerprint each of the genotypes studied. However, the two marker systems differed in the amount of detected polymorphisms. The 15 SSR markers were highly polymorphic across all the genotypes. The total number of amplified fragments was 156 and the number of alleles per locus ranged from 3 to 24 with an average of 10.4. Two SSR markers alone, Xwms47 and Xwms268, were sufficient to distinguish all 34 durum wheat genotypes. The five AFLP primer pair combinations analysed yielded a total of 293 bands, of which 31% were polymorphic. The highest polymorphic information content (PIC) value was observed for SSRs (0.68) while the highest marker index (MI) value was for AFLPs (7.16) reflecting the hypervariability of the first and the distinctive nature of the second system. For durum wheat cultivars, the genetic similarity values varied between 31.3 and 81% for AFLPs (with an average of 54.2%), and between 3.6 and 72.7% for SSRs (with an average of 19.9%). The rank correlation between the two marker systems was moderate, with r ¼ 0.57, but highly significant. Based on SSR markers, highest genetic similarity (GS) values were observed within the modern cultivars (37.3%), while the old cultivars showed a low level of GS (19.9%). Moreover, the modern cultivars showed low PIC and MI values. UPGMA Cluster analysis based on the combined AFLP and SSR data separated the wild wheat species from the durum wheat cultivars. The modern cultivars were separated from the old cultivars and form a distinct group.
Background
Tunisia is considered a secondary center of diversification of durum wheat and has a large number of abandoned old local landraces. An accurate investigation and characterization of the morphological and genetic features of these landraces would allow their rehabilitation and utilization in wheat breeding programs. Here, we investigated a diverse collection of 304 local accessions of durum wheat collected from five regions and three climate stages of central and southern Tunisia.
Results
Durum wheat accessions were morphologically characterized using 12 spike- and grain-related traits. A mean Shannon-Weaver index (H′) of 0.80 was obtained, indicating high level of polymorphism among accessions. Based on these traits, 11 local landraces including Mahmoudi, Azizi, Jneh Khotifa, Mekki, Biskri, Taganrog, Biada, Badri, Richi, Roussia and Souri were identified. Spike length (H′ = 0.98), spike shape (H′ = 0.86), grain size (H′ = 0.94), grain shape (H′ = 0.87) and grain color (H′ = 0.86) were the most polymorphic morphological traits. The genetic diversity of these accessions was assessed using 10 simple sequence repeat (SSR) markers, with a polymorphic information content (PIC) of 0.69. Levels of genetic diversity were generally high (I = 0.62; He = 0.35). In addition, population structure analysis revealed 11 genetic groups, which were significantly correlated with the morphological characterization. Analysis of molecular variance (AMOVA) showed high genetic variation within regions (81%) and within genetic groups (41%), reflecting a considerable amount of admixture between landraces. The moderate (19%) and high (59%) levels of genetic variation detected among regions and among genetic groups, respectively, highlighted the selection practices of farmers. Furthermore, Mahmoudi accessions showed significant variation in spike density between central Tunisia (compact spikes) and southern Tunisia (loose spikes with open glume), may indicate an adaptation to high temperature in the south.
Conclusion
Overall, this study demonstrates the genetic richness of local durum wheat germplasm for better in situ and ex situ conservation and for the subsequent use of these accessions in wheat breeding programs.
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