Neila Trifi-Farah

Loss of genetic diversity as a signature of apricot domestication and diffusion into the Mediterranean Basin

BackgroundDomestication generally implies a loss of diversity in crop species relative to their wild ancestors because of genetic drift through bottleneck effects. Compared to native Mediterranean fruit species like olive and grape, the loss of genetic diversity is expected to be more substantial for fruit species introduced into Mediterranean areas such as apricot (Prunus armeniaca L.), which was probably primarily domesticated in China. By comparing genetic diversity among regional apricot gene pools in several Mediterranean areas, we investigated the loss of genetic diversity associated with apricot selection and diffusion into the Mediterranean Basin.ResultsAccording to the geographic origin of apricots and using Bayesian clustering of genotypes, Mediterranean apricot (207 genotypes) was structured into three main gene pools: ‘Irano-Caucasian’, ‘North Mediterranean Basin’ and ‘South Mediterranean Basin’. Among the 25 microsatellite markers used, only one displayed deviations from the frequencies expected under neutrality. Similar genetic diversity parameters were obtained within each of the three main clusters using both all SSR loci and only 24 SSR loci based on the assumption of neutrality. A significant loss of genetic diversity, as assessed by the allelic richness and private allelic richness, was revealed from the ‘Irano-Caucasian’ gene pool, considered as a secondary centre of diversification, to the northern and southwestern Mediterranean Basin. A substantial proportion of shared alleles was specifically detected when comparing gene pools from the ‘North Mediterranean Basin’ and ‘South Mediterranean Basin’ to the secondary centre of diversification.ConclusionsA marked domestication bottleneck was detected with microsatellite markers in the Mediterranean apricot material, depicting a global image of two diffusion routes from the ‘Irano-Caucasian’ gene pool: North Mediterranean and Southwest Mediterranean. This study generated genetic insight that will be useful for management of Mediterranean apricot germplasm as well as genetic selection programs related to adaptive traits.

Genetic diversity in Tunisian perennial ryegrass revealed by ISSR markers

The genetic diversity in Tunisian perennial ryegrass (Lolium perenne) was examined by the help of inter-simple sequence repeats (ISSR). Starting from eighteen accessions, a large number of polymorphic ISSR markers were currently generated using appropriate primers (a total of 136, which average of 12.6 polymorphic bands/primer). These markers were considered to estimate the genetic distance among accessions and to draw phylogenetic trees. Our data provide evidence of a high degree of genetic diversity in Tunisian ryegrass. In addition, both cultivars and wild types present a high degree of divergence suggesting a complex domestication process in this crop. Moreover, spontaneous populations of Tunisian ryegrass have been identified as important ecotypes that are suitable in selection programs to improve grasslands.

Impact of Mapped SSR Markers on the Genetic Diversity of Apricot (Prunus armeniaca L.) in Tunisia

The impact of mapped microsatellites on the study of genetic diversity of Tunisian apricot accessions was assessed. The genetic variability of 47 traditional apricot cultivars originating from several areas in Tunisia was investigated with 32 polymorphic microsatellite loci selected for their location throughout the eight linkage groups of Prunus genome. The higher polymorphism and greater transportability of these markers among Prunus species were proved by the expected heterozygosity (He = 0.56) and Shannon’s index of diversity (I = 1.05), indicating that Tunisian apricot germplasm maintained a substantial level of genetic diversity. According to their geographical origin, the genetic differentiation among groups (north, center, and south; Fst = 0.04) was lower, while the gene flow among groups was consequent (Nm = 4.79), attesting a narrow genetic background of apricot in the country. Both unweighted pair-group method with arithmetic mean dendrogram, based on Nei’s genetic distances and factorial correspondence analysis, separated northern cultivars from central and southern cultivars, revealing the same molecular basis of apricot material in the Center and the South of Tunisia. These results revealed the efficiency of mapped markers for genetic variability measurements compared to randomly ones, however, no advantage was observed considering the genetic relationships among studied accessions.

Use of Microsatellite Polymorphisms to Develop an Identification Key for Tunisian Apricots

Microsatellite polymorphisms in 54 apricot landrace cultivars were identified by using 26 Prunus microsatellite primers. Samples of apricot cultivars were collected in eight growing regions in Tunisia ranging from the north to the south of the country and from the sub-humid to the saharian areas. The primers revealed 103 alleles and 155 different genotypes among the 54 apricot accessions. The most polymorphic microsatellite loci were used to develop an identification key. Five microsatellite primers and the 28 resulting alleles were sufficient to discriminate among all 54 cultivars. These results are discussed in the context of cultivar nomenclature, geographic origins, and the comprehensive fingerprinting of Tunisian apricot collections.

Phylogenetic relationships in the North African genus Hedysarum as inferred from ITS sequences of nuclear ribosomal DNA

Sequences of the internal transcribed spacers (ITS) of nuclear ribosomal DNA were analysed to precise their length (637–643 bp) and resolve phylogenetic relationships among eight Mediterranean species of the genus Hedysarum (Fabaceae). The infra-specific variability levels of the ITS sequences of spontaneous population of H. coronarium proved a lack of polymorphism both in the length and in the sequences examined in this species. Hence, a consensus ITS sequence characterising each Hedysarum species has been investigated for analysis of inter-specific polymorphisms. The level of variation of ITS sequence was high enough to make the ITS1 and ITS2 a useful tool for phylogenetic reconstruction. However, ITS2 seems to be relatively more polymorphic and more informative than ITS1 regarding length or GC percent. The phylogenic relationships in the genus Hedysarum based on ITS1 and ITS2 sequences taken independently or together, are discussed in the context of current work in molecular biosystematics. Results exhibited the distinctiveness of the two H. spinosissimum subspecies (i.e. H. spinosissimum ssp. capitatum and H. spinosissimum ssp. spinosissimum). In addition, the great similarity of the ITS sequences between H. coronarium (the only cultivated species of the genus) and H. carnosum suggests the usefulness of the latter in selection programmes to improve pastoral production in semi-arid areas.

Exploration of intra- and inter-population genetic diversity in Hedysarum coronarium L. by AFLP markers

Amplified fragment length polymorphism (AFLP) markers were used to characterize the genetic diversity within and among natural populations and cultivars of Hedysarum coronarium. Twelve populations within Tunisia were evaluated with three AFLP primer combinations. A total of 207 reproducible bands was detected of which 178 (86%) were polymorphic. The great discriminative power of AFLP markers and their ability to represent genetic relationships among Hedysarum plants was demonstrated. Genetic diversity within and among populations was assessed through Principal Component Analysis (PCA) and cluster analysis by using the Neighbor-joining clustering algorithm. AFLP technology has provided evidence of a high degree of intra- and inter-population genetic diversity in H. coronarium. AFLP banding patterns provided molecular markers correlated with the plants’ geotropism. In addition, AFLP markers can differentiate wild accessions from cultivars. Moreover, geographical origins did not correspond to population clustering.

Grafting versus seed propagated apricot populations: two main gene pools in Tunisia evidenced by SSR markers and model-based Bayesian clustering

Apricot was introduced into the Mediterranean Basin from China and Asian mountains through the Middle-East and the Central Europe. Traditionally present in Tunisia, we were interested in accessing the origin of apricot species in the country, and in particular in the number and the location of its introductions. A set of 82 representative apricot accessions including 49 grafted cultivars and 33 seed propagated ‘Bargougs’ were genotyped using 24 microsatellite loci revealing a total of 135 alleles. The model-based Bayesian clustering analysis using both Structure and InStruct programs as well as the multivariate method revealed five distinct genetic clusters. The genetic differentiation among clusters showed that cluster 1, with only four cultivars, was the most differentiated from the four remaining genetic clusters, which constituted the largest part of the studied germplasm. According to their geographic origin, the five identified groups (north, centre, south, Gafsa oasis and other oases groups) enclosed a similar variation within group, with a low level of differentiation. Overall results highlighted the distinction of two apricot gene pools in Tunisia related to the different mode of propagation of the cultivars: grafted and seed propagated apricot, which enclosed a narrow genetic basis. Our findings support the assumption that grafting and seed propagated apricots shared the same origin.

Relative efficiency of morphological characters and molecular markers in the establishment of an apricot core collection

In order to optimize the management of genetic resources, in most cases a representative sample of the germplasm collections needs to be developed. The establishment of a core collection is thus of major importance either to minimize the cost associated with the management of the associated germplasm or to apply analysis onto representative bases. In order to select a representative core collection among the Tunisian apricot germplasm of 110 accessions large, the Maximization strategy algorithm was used. This algorithm was shown to be the most convenient when using both morphological traits and molecular markers. Three core collections based on morphological characters, molecular markers or the combined data were compared. Our data indicate that both the molecular and the morphological markers have to be considered to obtain a core collection that represents the global diversity of the 110 accessions. Using this method, a subset of 34 selected accessions was found to represent accurately the 110 accessions present in the whole collection (75 to 100% for the morphological characters and 97% of the molecular markers). These results show that the combination of molecular and morphological markers is an efficient way to characterize the apricot core collection and provides an exhaustive coverage for the analyzed diversity on morphological and genetic bases.

Phylogeny of Mediterranean Lathyrus species using Inter Simple Sequence Repeats markers

Abstract The genus Lathyrus constitutes an important phytogenetic patrimony as a source of traditional foodstuffs in Tunisia. An Inter-Simple Sequence Repeats (ISSR) technique was employed to assess genetic diversity and relationships of seven Mediterranean species of the genus Lathyrus belonging to four sections (Lathyrus, Clymenum, Nissolia and Aphaca). The molecular characterization and fingerprint identification of 11 populations representing the Lathyrus species were conducted using 11 ISSR primers (seven 3’ anchored primers and four non-anchored primers). A total of 261 polymorphic markers have been successfully generated, ranging between 400 and 3000 base pairs, with a mean of 23.72 bands/primer. The percentage of polymorphic bands for these species was 98.12%. The ISSR technique was able to characterize and differentiate between the different studied genotypes of the Lathyrus species. Genetic distance within and between populations was estimated by calculating the similarity index. The similarity indices ranged from 0.251–0.741, with a mean of 0.386. Our data provide evidence of high molecular polymorphism, showing that both wild and cultivated species forms constitute an important pool of diversity. Despite the large distant geographical origin site, the accessions per species issued from different geographical origins are relatively related. Moreover, the genetic similarity tree showed that the two species L. sativus and L. cicera are similar showing that L. sativus may be derived from L. cicera, whereas the principal component analysis showed a clear divergence of the L. ochrus population belonging to section Clymenum from the other populations. Furthermore, L. ochrus appears to be closer to L. aphaca and L. nissolia than to the other studied species belonging to section Lathyrus.

Evaluation of genetic diversity in Sulla coronaria from different geographical populations in Tunisia by inter simple sequence repeat (ISSR)

Five oligonucleotides generating 116 markers complementary to simple sequence repeats were used in order to characterize wild and cultivated ecotypes of Sulla coronaria and assess genetic diversity suitable in breeding programs. While analysing populations, a large genetic variability was revealed and supported by the preferentially allogamous mating system of the species. Moreover, the highest level of intra-population variations (H pop /H sp = 69.9) either of wild or cultivated accessions have been strongly evidenced by a significant adaptation to variety of habitats. In addition, the structure of populations was independent from the bioclimatic stages and was not affected by environmental factors as shown by the non correlation between the geographic and the Nei and Li’s genetic distances (r= 0.461 and p=0.068>0.05). The unweighted pair group method with arithmetic mean (UPGMA) genetic relationships showed that some local spontaneous accessions characterised by an orthotropic port (Jebel Zit and Beja) were also molecularly similar to other cultivars. Key words : Sulla coronaria , wild and cultivated forms, intra and inter-populations variability, microsatellites (ISSRs), bioclimatic stages.