Exploration of intra- and inter-population genetic diversity in <Emphasis Type="Italic">Hedysarum coronarium</Emphasis> 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.     

Assessment of genetic relationships among Pyrus species and cultivars using AFLP and RAPD markers

Twenty-five Pyrus communis L. cultivars including eight traditional Portuguese pears, and four commercial Pyrus pyrifolia (Burm.) Nak. (Japanese pear or `nashi') cultivars were analysed by RAPD and AFLP techniques focusing on their molecular discrimination and the assessment of their genetic relatedness. Twenty-five primers generated 324 RAPD markers, among which 271 (84%) were polymorphic. The AFLP technique, using seven primer combinations, revealed a similar level of molecular polymorphisms (87%), representing 418 polymorphic bands among a total of 478 scored in autoradiographs. The high reproducibility of RAPD and AFLP techniques was confirmed comparing DNA samples from different extractions and different digestions of DNA from the same plant. Three genetic similarity matrices and respective dendrograms were elaborated on using RAPD, AFLP or joint RAPD and AFLP data. Both molecular marker techniques proved their reliability to assess genetic relationships among pear cultivars. P. pyrifolia cultivars exhibit a closer genetic relatedness, clustering apart from P. communis cultivars. Within P. communis, `William's', as well as `Doyenne du Comice', cluster close to their hybrids. Most of the Portuguese cultivars tend to cluster together, indicating to constitute a relatively independent genetic pool, which can be of interest in pear breeding programs.     

Molecular Analysis of Olive Cultivars in the Molise Region of Italy

The Italian region of Molise is characterised by a remarkable variety of microenvironments, where some olive cultivars have settled in the course of many centuries. The study of the varieties cultivated in Molise, and of their relationships with varieties of close regions, is very important in order to clarify the entity of the native olive germplasm, well adapted to the local conditions, and to characterise the oil with regional origin. Eighteen cultivars from Molise have been examined using AFLP markers. They were compared with other cultivars, widely cultivated in Italy and France, in order to evaluate their similarities and to test some hypotheses on their origin and spreading among different areas of cultivation. Numerous varieties showed a high similarity and one case of synonymy (Frantoio-Paesana Bianca) was observed. The results provide useful information in order to distinguish the native germplasm and evaluate the consistency of the olive heritage of the region.     

Applicability of SCAR markers to food genomics: olive oil traceability

DNA analysis with molecular markers has opened a shortcut toward a genomic comprehension of complex organisms. The availability of micro-DNA extraction methods, coupled with selective amplification of the smallest extracted fragments with molecular markers, could equally bring a breakthrough in food genomics: the identification of original components in food. Amplified fragment length polymorphisms (AFLPs) have been instrumental in plant genomics because they may allow rapid and reliable analysis of multiple and potentially polymorphic sites. Nevertheless, their direct application to the analysis of DNA extracted from food matrixes is complicated by the low quality of DNA extracted: its high degradation and the presence of inhibitors of enzymatic reactions. The conversion of an AFLP fragment to a robust and specific single-locus PCR-based marker, therefore, could extend the use of molecular markers to large-scale analysis of complex agro-food matrixes. In the present study is reported the development of sequence characterized amplified regions (SCARs) starting from AFLP profiles of monovarietal olive oils analyzed on agarose gel; one of these was used to identify differences among 56 olive cultivars. All the developed markers were purposefully amplified in olive oils to apply them to olive oil traceability.     

RAPD and ISSR molecular markers in <Emphasis Type="Italic">Olea europaea</Emphasis> L.: Genetic variability and molecular cultivar identification

Thirty Portuguese and eight foreign olive (Olea europaea L.) cultivars were screened using Random Amplified Polymorphic DNA (RAPD) and Inter-Simple Sequence Repeat (ISSR) markers. Twenty RAPD primers amplified 301 reproducible bands of which 262 were polymorphic; and 17 ISSR primers amplified 204 bands of which 180 were polymorphic. The percentage of polymorphic bands detected by ISSR and RAPD was similar (88 and 87%, respectively). The genetic variability observed was similar in the Portuguese and foreign olive cultivars. Seven ISSR and 12 RAPD primers were able to distinguish individually all 38 olive cultivars. Twenty specific molecular markers are now available to be converted into Sequence Characterised Amplified Region (SCAR) markers. Relationships among Portuguese and foreign cultivars is discussed.     

Assessment of genetic diversity in cultivars and wild accessions of Luohanguo (<Emphasis Type="Italic">Siraitia grosvenorii</Emphasis> [Swingle] A. M. Lu et Z. Y. Zhang), a species with edible and medicinal sweet fruits endemic to southern China,using RAPD and AFLP markers

Genetic variation of wild populations and cultivars of Luohanguo (Siraitia grosvenorii), a plant species endemic to southern China, was assessed using random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) markers. Based on the results for 130 individuals from seven populations, a high level of genetic diversity of Luohanguo was observed at the species level. The percentage of polymorphic loci (P) was 89.4%, Nei’s gene diversity (H _e) was 0.239, and Shannon’s information index (H _o) was 0.373 based on the combined AFLP and RAPD data. There was a high degree of genetic differentiation, with 45.1% of the genetic variation attributed to differences between the populations. The genetic diversity of the Luohanguo cultivars is much lower than that of wild populations (P = 41.8%, H _e = 0.141, H _o = 0.211), and a distinct genetic differentiation is observed between the cultivars and wild accessions. The pool of genetic variation in the wild populations provides an excellent gene resource for Luohanguo breeding.     

Total phenolic content,RAPDs, AFLPs and morphological traits for the analysis of variability in <Emphasis Type="Italic">Smallanthus sonchifolius</Emphasis>

Smallanthus sonchifolius is a periennal herb originally cultivated in South America and now grown in several other countries. Recently, greater attention has been focused on this plant due to its agronomical, nutritional and pharmacological characteristics. In this paper the application of RAPDs and AFLPs for the analysis of genetic diversity in a group of 5 Smallanthus sonchifolius landraces is presented. Both methods proceed through the direct analysis of DNA, and their results were compared with the total phenolic content of each landrace and its morphological traits. Using 61 RAPD primers, 85 informative bands were identified, corresponding to 28.7% of polymorphism. In comparison, only six selected AFLP primer pairs produced 84 informative bands, with a similar percentage of polymorphism (23.4%). RAPD and AFLP markers were analyzed separately. Total phenolic content varied twofold among the five landraces analysed, ranging from 3,494 to 6,849 mg/g. Each type of molecular marker resolved two main groups that included the same genotypes, but with different within-group relationships among genotypes. The two groups are consistent with some phenotypic characters but they do not reflect faithfully their geographical origin. Most notably, the two groups comprise landraces with higher and lower total phenolic content, respectively. Dendrograms based on the two molecular data sets graphically depicted the ability of both methods to differentiate all the cultivars studied. Data obtained suggest that the two molecular markers applied are useful to investigate intra-specific genetic variability in Smallanthus sonchifolius, and predict well the total phenolic content of each landrace.     

AFLP Fingerprinting in Pigeonpea (Cajanus cajan (L.) Millsp.) and its Wild Relatives

Detection of DNA polymorphism in cultivated pigeonpea (Cajanus cajan) and two of its wild relatives Cajanus volubilis and Rhynchosia bracteata is reported here for the first time using amplified fragment length polymorphism (AFLP) fingerprinting. For this purpose, two EcoRI (three selective nucleotides) and 14 MseI (three selective nucleotides) primers were used. The two wild species shared only 7.15% bands with the pigeonpea cultivars, whereas 86.71% common bands were seen among cultivars. Similarly, 62.08% bands were polymorphic between C. volubilis and pigeonpea cultivars in comparison to 63.33% polymorphic bands between R. bracteata and pigeonpea cultivars, and 13.28% polymorphic bands among pigeonpea cultivars. The cluster analysis revealed low polymorphism among pigeonpea cultivars and very high polymorphism between cultivated pigeonpea and its wild relatives. The AFLP analysis also indicated that only one primer combination (EcoRI + ACT and MseI + CTG), at the most any four primer pair combinations, are sufficient for obtaining reliable estimation of genetic diversity in closely related cultivars like pigeonpea material analyzed herein. AFLP analysis may prove to be a useful tool for molecular characterization of pigeonpea cultivars and its wild relatives and for possible use in genome mapping.     

Genetic Diversity among Syrian Cultivated and Landraces Wheat Revealed by AFLP Markers

Genetic diversity among some important Syrian wheat cultivars was estimated using Amplified Fragment Length Polymorphism (AFLP) markers. Five Triticum aestivum L. and 10 Triticum turgidum ssp. durum were analyzed with 11 EcoRI–MseI primer pair combinations. Of the approximately 525 detected AFLP markers, only 46.67% were polymorphic. Cluster analysis with the entire AFLP data divided all cultivars into two major groups reflecting their origins. The first one contained T. aestivum L. cultivars, and the T. turgidum ssp. durum cultivars and landraces were grouped in the second. Narrow genetic diversity among all cultivars was detected with an average genetic similarity of 0.884. The lowest similarity index (0.9) was found between Cham5 and Hamary (durum wheat), whereas this value was 0.93 between Salamony and Bouhouth 4 (T. aestivum L.). The narrow genetic diversity level indicates that these genotypes could be originated from the same source. AFLP analysis provides crucial information for studying genetic variation among wheat cultivars and provides important information for plant improvement.     

Intra-specific DNA polymorphism in pineapple (<Emphasis Type="Italic">Ananas comosus</Emphasis> (L.) Merr.) assessed by AFLP markers

Pineapple (Ananas comosus (L.) Merr.) cultivars, often derived from somatic mutations, are propagated vegetatively. It has been suggested by isozyme data that there is little genetic variation among Smooth Cayenne cultivars. A thorough investigation of the genetic variation within the cultivated speciesAnanas comosus, particularly among commercial cultivars, will provide critical information needed for crop improvement and cultivar protection. One-hundred and forty-eight accessions ofA. comosus and 14 accessions of related species were evaluated with AFLP markers. The average genetic similarity ofA. comosus was 0.735 ranging from 0.549 to 0.972, suggesting a high degree of genetic variation within this species. With AFLP markers, discrete DNA fingerprints were detected for each commercial cultivar, breeding line, and intra-specific hybrid. Self-incompatibility, high levels of somatic mutation, and intraspecific hybridization may account for this high degree of variation. However, major cultivar groups of pineapple, such as Cayenne, Spanish, and Queen, could not be distinctively separated. These cultivar groups are based on morphological similarity, and the similar appearance can be caused by a few mutations that occurred on different genetic background. Our results suggest that there is abundant genetic variation within existing pineapple germplasm for selection, and discrete DNA fingerprinting patterns for commercial cultivars can be detected for cultivar protection. The genetic diversity and relationships of fourAnanas species are also discussed.     

Identification and evaluation of <Emphasis Type="Italic">Forsythia</Emphasis> germplasm using molecular markers

This study identifies Forsythia germplasm and evaluated the genetic relationships of F. ×intermedia hybrids, cultivars and their putative parental species. Leaf samples of F. ×intermedia cultivars and species, such as F. koreana and F. suspensa, were collected in the Netherlands, Korea, and USA. Total genomic DNA was extracted and evaluated by randomly amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) analyses. Dendrograms were constructed by using the neighbor-joining (NJ) clustering algorithm applying the interior branch (IB) test or analyzed by STRUCTURE. In the dendrogram generated by RAPD markers, two major clusters were observed. One cluster (CL-I) contained most of the F. ×intermedia cultivars, F. suspensa, and F. koreana. The other cluster (CL-II) included F. europaea, F. ovata, F. densiflora, F. mandshurica, F. japonica, F. viridissima, and cultivars derived from F. ovata. In the AFLP dendrogram, the placement of F. ×intermedia cultivars with F. suspensa was similar, forming cluster CL-A I. The RAPD and AFLP results clearly separated most F. ×intermedia cultivars from F. ovata derived cultivars. The full range of genetic diversity of F. suspensa and F. viridissima should be investigated to verify whether these two species are truly parental taxa for F. ×intermedia. Placement of F. viridissima, F. ovata, and F. japonica in different sub-clusters requires further investigation regarding genetic diversity in the species, and their close relationship with F. koreana, F. mandshurica, and F. saxatalis.     

AFLP and RFLP linkage map in <Emphasis Type="Italic">Coix</Emphasis>

Coix is a genus in the grass family placed in the tribe Maydeae. It is closely related to maize and is also used as a crop plant. Since many valuable traits have been identified recently in Coix, it is considered to be a valuable genetic resource, particularly for maize improvement. In this study, a Coix genetic linkage map was constructed using an F2 population of 131 individuals. Eighty AFLP and 10 RFLP markers were mapped, covering a total length of 1339.5 cM with an average interval of 14.88 cM. The map consisted of 10 linkage groups, were consistent with the chromosome numbers observed cytogenetically. Both AFLP and RFLP markers were used first for genetic analysis in Coix. AFLP markers were generated by two restriction enzyme combinations, EcoRI/MseI and PstI/MseI. A total of 1349 bands were amplified, of which 140 were polymorphic. The polymorphism detection efficiency of the two enzyme combinations was compared, and utility of AFLP markers to construct the linkage map was discussed. Ten RFLP markers detected by three different probes were distributed on eight different linkage groups. The results provide a foundation to map and isolate important genes in Coix, and to investigate its genomic architecture, possible origins, and relationship with maize at the DNA level.     

Genetic Variation and Relationships of Pedigree-Known Oat, Wheat, and Barley Cultivars Releaved by Bulking and Single-Plant Sampling

Applications of bulking procedures have played an increasingly important role in molecular characterization of plant germplasm, but little attention has been made to address the effectiveness of detecting genetic variation and inferring genetic relationships via bulking. An analysis was performed here to compare the genetic variation detected and genetic relationships inferred via bulking and single-plant sampling of five oat (Avena sativa L.), wheat (Triticum aestivum L.), and barley (Hordeum vulgare L.) cultivars with known pedigrees using amplified fragment length polymorphism (AFLP) markers. Three AFLP primer pairs were applied to screen one bulk and eight single-plant samples of each cultivar and up to 140 AFLP bands were scored for each sample. Analyses of these AFLP data showed bulking revealed AFLP variation up to 21.4% less than corresponding single-plant sampling for these crop species and also introduced up to 2.2% upward and 5.1% downward biases in detecting AFLP variations for each cultivar. The genetic relationships inferred by bulking using the Dice's coefficient, the simple matching coefficient, and the Jaccard's coefficient were largely the same, but differed from those in single-plant samples employing average Dice's coefficient, average simple matching coefficient, and AMOVA-based distance method. All of the inferred genetic relationships were not congruent to the known pedigrees. Clearly, substantial biases could exist in detection of AFLP variation and in inference of genetic relationships from bulk samples, even for closely related germplasm, and more efforts to assess the effectiveness of bulking in inferring genetic variation and relationships are needed for more informative molecular characterization of plant germplasm.     

Traceability of plant contribution in olive oil by amplified fragment length polymorphisms

Application of DNA molecular markers to traceability of foods is thought to bring new benefit to consumer's protection. Even in a complex matrix such as olive oil, DNA could be traced with PCR markers such as the amplified fragment length polymorphisms (AFLPs). In this work, fluorescent AFLPs were optimized for the characterization of olive oil DNA, to obtain highly reproducible, high-quality fingerprints, testing different parameters: the concentrations of dNTPs and labeled primer, the kind of Taq DNA polymerase and thermal cycler, and the quantity of DNA employed. It was found that correspondence of fingerprinting by comparing results in oils and in plants was close to 70% and that the DNA extraction from olive oil was the limiting step for the reliability of AFLP profiles, due to the complex matrix analyzed.     

Genetic Diversity and Relationships of Japanese Peach (<Emphasis Type="Italic">Prunus persica</Emphasis> L.) Cultivars Revealed by AFLP and Pedigree Tracing

To evaluate the genetic diversity and to clarify the genetic relationships of Japanese peach cultivars, we analyzed the amplified fragment length polymorphism (AFLP) and traced the pedigree of 17 Japanese commercial peach cultivars and six traditional accessions. Sixteen AFLP primer combinations produced a total of 837 fragments and 146 polymorphic bands with a polymorphism percentage of 17.5%. All of the peach accessions could be identified from differences in at least 10 polymorphic bands. A cluster analysis showed that all the Japanese commercial peach cultivars, except ‘Kiyomi’ and ‘Jichigetsuto’, formed a major group consisting of three sub-groups. Of the six traditional accessions, four were genetically distant from the Japanese commercial peach cultivars while two accessions from China were classified into the Japanese commercial peach cultivars group. Both the AFLP analysis and pedigree tracing suggested that Japanese commercial peach cultivars are mainly derived from ‘Shanhai Suimitsuto’, one of the traditional accessions from China. Although the genetic relationships revealed by AFLP were generally in agreement with those shown by the pedigree information, some contradictions were found. Combining the AFLP results and pedigree information can provide a better understanding of the genetic relationships of Japanese peach cultivars.     

Genetic relationships and diversity among Brazilian cultivars and landraces of common beans (Phaseolus vulgaris L.) revealed by AFLP markers

The genetic variation and relationships among 31 accessions of Phaseolus vulgaris L., and two representatives of Vigna unguiculata L., were evaluated by AFLP analysis. A total of 263 DNA fragments across all materials were scored using nine primer combinations, averaging 32 per primer. More than 95% of the amplification products showed polymorphism, indicating high variation at the DNA level among these accessions. Pair-wise genetic similarity (Jaccard's coefficient) ranged from 0.553 to 0.840, with a mean of 0.765. Twenty-three accessions (70%) clustered into three groups. A majority of the commercial cultivars (91%) clustered within a single group, whereas the landraces were distributed along all the variation. An apparent correlation with phaseolin types was detected. Results of this study suggest that Brazilian landraces truly represent the overall genetic variability of Phaseolus vulgaris, confirming the multiple origins of these materials, and their potential as a source of variation for breeding programs.     

Comparative study of methods for DNA preparation from olive oil samples to identify cultivar SSR alleles in commercial oil samples: possible forensic applications

Virgin olive oil is made from diverse cultivars either mixed or single. Those ensure different tastes and typicity, and these may be also enhanced by the region of production of cultivars. The different olive oil labels correspond to their chemical composition and acidity. Labels also may correspond to a protected origin indication, and thus, such oils contain a given composition in cultivars. To verify the main cultivars used at the source of an olive oil sample, our method is based on DNA technology. DNA is present in all olive oil samples and even in refined oil, but the quantity may depend on the oil processing technology and oil conservation conditions. Thus, several supports were used to retain DNA checking different techniques (silica extraction, hydroxyapatite, magnetic beads, and spun column) to prepare DNA from variable amounts of oil. At this stage, it was usable for amplification through PCR technology and especially with the magnetic beads, and further purification processes were checked. Finally, the final method used magnetic beads. DNA is released from beads in a buffer. Once purified, we showed that it did not contain compounds inhibiting PCR amplification using SSR primers. Aliquot dilution fractions of this solution were successfully routinely used through PCR with different SSR primer sets. This enables confident detection of eventual alien alleles in oil samples. First applied to virgin oil samples of known composition, either single cultivars or mixtures of them, the method was verified working on commercial virgin oil samples using bottles bought in supermarkets. Last, we defined a protocol starting from 2 x 40 mL virgin olive oil, and DNA was prepared routinely in about 5 h. It was convenient to genotype together several loci per sample to check whether alleles were in accordance with those of expected cultivars. Thus, forensic applications of our method are expected. However, the method needs further improvement to work on all oil samples.     

AFLP fingerprinting of sesame (<Emphasis Type="Italic">Sesamum indicum</Emphasis> L.) cultivars: identification,genetic relationship and comparison of AFLP informativeness parameters

Amplified fragments length polymorphism (AFLP) was used to distinguish 20 cultivars of sesame (Sesamum indicum L.) and to elucidate the genetic relationship among these genotypes. The data were also used to estimate the usefulness of parameters currently used to assess the informativeness of molecular markers. A total of 339 markers were obtained using 8 primer combinations. Of the bands, 91% were polymorphic. Five primer combinations were able to distinguish all 20 cultivars used. None of the remaining three primer combinations could distinguish all accessions if used alone, but using all three combinations reduced the probability of a random match to 5 × 10⁻⁵. Polymorphic information content (PIC), resolving power (Rp) and marker index (MI) of each primer combination failed to correlate significantly with the number of genotypes resolved. Jaccard’s similarity coefficients ranged from 0.31 to 0.78. Fifteen cultivars were grouped by four UPGMA-clusters supported by bootstrapping values larger than 0.70. The grouping pattern was similar to the grouping generated by principal coordinate analysis. The results demonstrated that AFLP-based fingerprints can be used to identify unequivocally sesame genotypes, which is needed for cultivar identification and for the assessment of the genetic variability of breeding stocks. We recommend to use the number of cultivars identified by a primer combination instead of PIC, Rp and MI; and to calculate the maximal, instead of average probability of identical match by chance in the assessment of the informativeness of a marker for cultivar identification.     

Analysis of Genetic Diversity and Relationships in Waxy Rice (Oryza sativa L.) using AFLP and ISSR Markers

Opaque endosperm is the main phenotypic indicator for waxy rice, but other phenotypic and genotypic variation among waxy rice accessions has largely been ignored. Previous studies showed that wide diversity in starch physiochemical properties exists in both indica and japonica waxy rices, especially for starch gelatinization temperature (GT) which could be divided into a high- and a low-GT group. In the present study, amplified fragment length polymorphism (AFLP) and inter-simple sequence repeat (ISSR) molecular markers were employed to examine genetic diversity and relationships of 56 waxy rice accessions. A total of 358 AFLP fragments were amplified with five primer combinations, showing a high level of polymorphism (78.3%). A total of 190 ISSR bands were generated with a single primer and a primer pair, showing a very high level of polymorphism (92.2%). The genetic distance matrices obtained from the two sets of markers were significantly correlated (r = 0.731, P = 0.004). The dendrogram generated with combined AFLP and ISSR markers could clearly differentiate the indica and japonica groups. Newly released varieties and breeding lines within each subspecies tended to be clustered together, whereas landraces were more distantly placed in the dendrogram. Only one AFLP band was found specific to the indica type, while no specific bands were found for starch GT. The implications for the conservation and breeding of waxy rice are discussed.     

Analysis of genetic diversity in Tunisian durum wheat cultivars and related wild species by SSR and AFLP markers

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 (Körn) 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 = 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.