Spanish melons (Cucumis melo L.) of the Madrid provenance: a unique germplasm reservoir

Melon (Cucumis melo L.) landraces of the Madrid provenance, Spain, have received national distinction for their high fruit quality and sensorial attributes. More specifically, a unique array of Group Inodorus landraces have been continuously cultivated and conserved by farmers in the municipality of Villaconejos since the 19th century. Their genetic relationships to other Group Inodorus and Flexuous melon market classes is not known, and, thus, a study was designed to determine their genetic relationships using 52 simple sequence repeat (SSR) markers, and then make genetic comparisons between these accessions and a previously published “Standard Reference Germplasm Array” (RA) containing Group Inodorus (14 Spanish and one USA), Flexuosus (1 Spanish), and Cantalupensis ( 2 USA) melon accessions. This subset consisted of 15 Spanish Group Inodorus landraces that circumscribed the genetic variation of major Spanish melon market classes (Groups Inodorus and Flexuosus), and USA commercial varieties (Groups Cantalupensis and Inodorus). Based on genetic distances, Villaconejos (Madrid) genotypes differed substantially from RA subset accessions, thus defining their genetic uniqueness. Principal component analysis (PCA) partitioned the accessions examined into four distinct groups revealing that Villaconejos black epidermis melons (landraces ‘Largo’, ‘Largo Negro Escrito’ and ‘Puchero’) were distinctly different from all other accessions examined, as cluster analysis separated Rochet market type Villaconejos’ accessions (landraces ‘Mochuelo’, ‘Mochuelo Tradicional’ and ‘Melón de Villaconejos’) from RA of the same market type. Genetic assessment of principal Spanish market classes revealed comparatively low intra-market heterogeneity in Piel de Sapo type accessions and high heterogeneity in Black and Yellow market type accessions. While a relatively high level of genetic introgression was detected between Yellow and Green market types, black epidermis market types were genetically unique. Given the uniqueness and high genetic diversity resident in Villaconejos landraces, this germplasm pool should be considered as a genetic source for broadening the comparatively narrow genetic base of Group Cantalupensis and Inodorus melon market types, especially standard commercial Spanish Group Inodorus market types (e.g., Piel de Sapo, Rochet, and Canari).     

Assessment of genetic diversity among African cassava Manihot esculenta Grantz accessions resistant to the cassava mosaic virus disease using SSR markers

A study was conducted to determine the extent of genetic diversity among African cassava (Manihot esculenta Crantz) accessions resistant to the cassava mosaic virus disease (CMD), using simple sequence repeat (SSR) markers. The accessions included a breeding stock (clone 58308), five improved lines, 62 CMD resistant and 10 CMD susceptible landraces. Genetic diversity was assessed among accessions in five cluster groups derived from UPGMA analysis on data from 18 SSR primer pairs. Average gene diversity, He, was high in all cluster groups, with an average heterozygosity of 0.591 ± 0.061. The estimator of inbreeding Fis revealed a low level of inbreeding within groups and averaged −0.262 ± 0.142. Gene diversity among all accessions was 51.4% and gene diversity within cluster groups was 46.6%, while 4.8% was due to diversity between the different cluster groups. The amount of genetic differentiation measured by Gst and Fst were 9.6% and 12.1% respectively, indicating a weak genetic structure.     

Genic SSRs for European and North American hop (Humulus lupulus L.)

Eight genic SSR loci were evaluated for genetic diversity assessment and genotype identification in Humulus lupulus L. from Europe and North America. Genetic diversity, as measured by three diversity indices, was significantly lower in European cultivars than in North American wild accessions. Neighbor Joining cluster analysis separated the hop genotypes into European and North American groups. These eight SSRs were useful in uniquely identifying each accession with the exception of two sets of European landraces and a pair of Japanese cultivars, ‘Shinshuwase’ and ‘Kirin II’. An accession from Manitoba grouped with the European (EU) cluster reflecting the group’s genetic similarity to older Manitoba germplasm used to develop ‘Brewer's Gold’ and the gene pool arising from this cultivar. Cultivars grouped closely with one of their immediate parents. ‘Perle’ grouped with its parent ‘Northern Brewer and ‘Willamette’ grouped with its parent ‘Fuggle H’. Wild American accessions were divided into two subgroups: a North Central group containing mostly H. lupulus var. lupuloides and a Southwestern group containing H. lupulus var. neomexicanus accessions. These eight SSRs will be valuable for genotype identification in European and wild American germplasm and may potentially prove useful for marker-assisted selection in hop. PCR products from four previously reported primer pairs that amplify the same intronic SSR regions as do the genic SSRs in this study were compared in eight common cultivars. Different primer pairs generated robust markers at the chs2 and chi loci. However, only the HLC-004B and HLC-006 primer pairs amplified successfully at the chs3 and chs4 loci. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

AFLP analysis of genetic diversity in leafy kale (<Emphasis Type="Italic">Brassica oleracea</Emphasis> L. convar. <Emphasis Type="Italic">acephala</Emphasis> (DC.) Alef.) landraces,cultivars and wild populations in Europe

AFLP markers were used to characterize diversity and asses the genetic structure among 17 accessions of kale landraces, cultivars and wild populations from Europe. The range of average gene diversity in accessions was 0.11–0.27. Several landraces showed higher levels of diversity than the wild populations and one cultivar had the lowest diversity measures. The landraces that were most genetically diverse were from areas where kales are known to be extensively grown, suggesting in situ conservation in these areas as a supplement to storage of seeds in gene banks. An analysis of molecular variance (AMOVA) showed that 62% of the total variation was found within accessions. For most accessions, genetic distance was not related to geographic distance. Similarities among accessions were probably not caused by recent gene flow since they were widely separated geographically; more likely the relationship among them is due to seed dispersal through human interactions. Our results indicate that a kale population found in a natural habitat in Denmark was probably not truly wild but most likely an escape from a cultivated Danish kale that had subsequently become naturalized.     

Molecular diversity of Omani wheat revealed by microsatellites: I. Tetraploid landraces

Results of archaeological studies indicate a millennia-old cultivation history for wheat (Triticum spp.) in Oman. However, in spite of numerous collection surveys and efforts for phenotypic characterization of Omani wheat landraces, no attempts have been made using molecular tools to characterize this germplasm. To fill this gap, 29 microsatellite markers revealing 30 loci were used to study the genetic diversity of 38 tetraploid wheat landrace accessions comprising the species T. dicoccon, T. durum and T. aethiopicum. A total of 219 alleles were detected whereby the number of alleles per locus ranged from 2 to 16 with an average number of 7.1 alleles per locus. The highest number of alleles occurred in the B genome with on average 7.9 alleles per locus as compared to the A genome with 6.5 alleles per locus. Heterogeneity was detected for all microsatellites except for GWM 312, GWM 601 and GWM 192B with an average heterogeneity over all primers and lines of 14.4%. Approximately 10% of the accessions contained rare alleles with an average allele frequency <4%. Gene diversity across microsatellite loci ranged from 0.26 to 0.85. The pairwise comparison of genetic similarity ranged from 0.03 to 0.91 with an average of 0.2. Cluster analysis revealed a clear separation of the two species groups T. dicoccon versus T. durum and T. aethiopicum. Within the species clusters regional patterns of subclustering were observed. Overall, this study confirmed the existence of a surprisingly high amount of genetic diversity in Omani wheat landraces as already concluded from previous morphological analyses and showed that SSR markers can be used for landraces’ analysis and a more detailed diversity evaluation.     

A wide range of genetic diversity in pear (<Emphasis Type="Italic">Pyrus ussuriensis</Emphasis> var. <Emphasis Type="Italic">aromatica</Emphasis>) genetic resources from Iwate,Japan revealed by SSR and chloroplast DNA markers

Iwateyamanashi (Pyrus ussuriensis var. aromatica) is one of the Pyrus species which grows wild in Japan. The number of Iwateyamanashi trees has been decreasing, so conservation and evaluation is urgently needed. Over 500 accessions of Pyrus species collected from Iwate in northern Tohoku region are maintained at Kobe University as an Iwateyamanashi germplasm collection. In order to investigate the genetic diversity, five SSR (simple sequence repeat) markers, developed from Japanese and European pear were examined for 86 Pyrus individuals including 58 accessions from Iwate. These SSR loci could discriminate between all the Iwate accessions except for 10 that bear seedless fruit, as well as determine the genetic diversity in Iwateyamanashi germplasms. High levels of variation were detected in 41 alleles and the mean observed heterozygosity across 5 loci was 0.50 for the Iwate accessions. Seedless accessions sharing identical SSR genotype with the local pear variety “Iwatetanenashi” were supposed to have been propagated vegetatively via grafting. In an UPGMA phenogram, Japanese pear varieties (P. pyrifolia) were clustered into two groups with some Iwate accessions including seedless ones. Another 38 Iwate accessions were not clustered clearly, and there was no clear relationship between these accessions and geographical distribution or morphological characters. Allele frequency revealed that the Iwate accessions were genetically more divergent than the Japanese pear varieties. Most Japanese pears possessed a 219 bp deletion at a spacer region between the accD and psaI genes in the chloroplast DNA (cpDNA), but other Pyrus species and two Iwateyamanashi trees did not. In the Iwate accessions, 79.3% had a deletion type cpDNA and others had a standard type cpDNA without deletion. These results are indicative of the wide range of genetic diversity in the Iwate accessions which include Japanese pear varieties. A combination of SSR and cpDNA analyses revealed high heterogeneity in Iwateyamanashi and coexistence of Iwateyamanashi and hybrid progeny with P. pyrifolia. These could be reasons for the wide range of continuous morphological variation described previously.     

Genetic diversity and spatial structure in a new distinct <Emphasis Type="Italic">Theobroma cacao</Emphasis> L. population in Bolivia

Cacao (Theobroma cacao L.) is an important economic crop in the Bolivian Amazon. Bolivian farmers both cultivate cacao, and extract fruits from wild stands in the Beni River region and in valleys of the Andes foothills. The germplasm group traditionally used is presently referred to as “Cacao Nacional Boliviano” (CNB). Using DNA fingerprinting technology based on microsatellite markers, we genotyped 164 Bolivian cacao accessions, including both cultivated and wild CNB accessions sampled from the Amazonian regions of La Paz and Beni, and compared their SSR profiles with 78 reference Forastero accessions from Amazonian cacao populations, including germplasm from the Ucayali region of Peru. Results of multivariate ordination and analysis of molecular variance show that CNB cacao has a unique genetic profile that is significantly different from the known cacao germplasm groups in South America. The results also show that cultivated CNB and wild CNB populations in the Beni River share a similar genetic profile, suggesting that the cultivated CNB is of indigenous origin in Bolivia. The level of genetic diversity, measured by allele richness and gene diversity in the Bolivian cacao, is moderately high, but was significantly lower than gene diversity in the other Amazonian cacao populations. Significant spatial genetic structure was detected in the wild CNB population, using analysis of autocorrelation (rc = 0.232; P < 0.001) and Mantel tests (Rxy = 0.276; P < 0.001). This finding is also highly valuable to support in situ conservation and sustainable use of CNB genetic diversity in Bolivia.     

Rice Diversity Collection,Conservation and Management in Northeastern India

Rice is the principal crop of northeastern region of India, where a considerable range of diversity exists. The germplasm collection has also unfold the occurrence of large number of rice landraces in the region. Between 1985 and 2002, a total 2639 accessions of rice germplasms including their wild relatives have been collected. Most of these germplasm material have been conserved in the National Gene Bank, New Delhi following their necessary characterization and evaluation. With the establishment of the Regional Gene Bank Module with medium-term storage facilities (6–7°C with 40–45% relative humidity) at Barapani, Meghalaya, the rice germplasm conservation (ex situ) have been initiated in the region.     

Genetic diversity in wild and cultivated black raspberry (Rubus occidentalis L.) evaluated by simple sequence repeat markers

Breeding progress in black raspberry (Rubus occidentalis L.) has been limited by a lack of genetic diversity in elite germplasm. Black raspberry cultivars have been noted for showing very few phenotypic differences and seedlings from crosses between cultivars for a lack of segregation for important traits. Despite these challenges, little molecular work has been done to explore genetic diversity and relationships in wild and cultivated black raspberry germplasm. Microsatellite, or simple sequence repeat (SSR), markers are highly polymorphic codominant markers useful for studying genetic diversity, population genetics, genetic fingerprinting and other applications. We examined genetic diversity in 148 wild and cultivated black raspberry accessions using 21 polymorphic SSR markers. Black raspberry cultivars clustered tightly and showed higher than expected heterozygosity while that of wild accessions was low. Relationships between wild black raspberry accessions were poorly resolved and regional clusters were mostly absent from our analysis. Our results indicated that wild black raspberry germplasm is a relatively untapped resource available for future breeding.     

Characterization of Seashore Paspalum (Paspalum vaginatum Swartz) Germplasm by Transferred SSRs from Wheat, Maize and Sorghum

One hundred and thirty SSR markers from wheat, maize and sorghum were screened for the transferability to Paspalum. The transfer rate was 67.5, 49.0 and 66.8% respectively. This would be a very efficient approach for DNA marker development for species which are not well studied molecularly. The polymorphism level for transferred SSR markers was 51.5% within species (Paspalum vaginatum) and 87.1% among Paspalum species. The high level of polymorphism is directly related to the high degree of heterozygosity maintained by its way of reproduction, i.e. self-incompatibility. Forty transferred polymorphic SSR markers were selected and used for characterization and evaluation of seventy-three Paspalum accessions. In total, 209 polymorphic bands were detected from these 40 SSR markers, with an average of five polymorphic bands per marker. The Paspalum accessions clustered into three major groups. Two very similar dendrograms can be generated from either 109 or 209 polymorphic bands. This led us to determine that 18 of the transferred SSR markers were sufficient for genetically differentiating the investigated germplasm accessions. The number of SSR markers required for germplasm characterization and evaluation is discussed. This is the first report of the transfer of SSR markers from major field crops to newly emerged environmental turfgrasses.     

Molecular evidence of genetic diversity changes in pea (<Emphasis Type="Italic">Pisum sativum</Emphasis> L.) germplasm after long-term maintenance

Maintaining germplasm genetic integrity is a key objective of long-term ex situ conservation. Periodic regeneration, performed on limited plots with small number of individuals, increases the risk of genetic drift and genetic diversity changes. In this study, six accessions of white flowered, dry seed pea varieties (Pisum sativum L. subsp. sativum var. sativum): Bohatyr, Klatovsky zeleny, Hanák, Moravsky hrotovicky krajovy, Raman and Viktoria-75 and four accessions of colour flowered, fodder pea (P. sativum subsp. sativum var. arvense (L.) Poiret: Arvika, Česky banán, Moravská krajová and Niké, representing Czechoslovak varieties and landraces, bred over the last 40–80 years, were analyzed using ten microsatellite locus specific markers. Each accession was represented by 20 individual seeds of two temporally different samples, spanning the period of 20 or 40 years. Together with intra-accession variation (except of cv. Hanák), evidence of genetic changes, e.g. differences in allele frequencies as well as genetic composition of sample, was detected in six out of ten accessions (Arvika, Bohatyr, Česky banán, Moravsky hrotovicky krajovy, Moravská krajová and Raman). Evidence of genetic erosion was found in three accessions (Česky banán, Moravsky hrotovicky krajovy and Raman), while in another three (Arvika, Bohatyr and Moravská krajová) the level of diversity was found to have increased. Moreover in three samples of Bohatyr (2004) and Klatovsky zeleny (1963 and 2004), low levels of heterozygosity was detected. These results demonstrate that in pea, a self-pollinating and highly homozygous plant, the danger of the loss of genetic integrity exists. These findings are significant for long-term ex situ germplasm management.     

Variation in melon (<Emphasis Type="Italic">Cucumis melo</Emphasis>) landraces adapted to the humid tropics of southern India

We present here the first comprehensive genetic characterization of melon landraces from the humid tropics of southern India. The genetic diversity among 50 melon landraces collected from 3 agro-ecological regions of southern India (6 agro-ecological sub-regions) was assessed by variation at 17 SSR loci, morphological traits of plant habit and fruit, 2 yield-associated traits, pest and disease resistance, biochemical composition (ascorbic acid, carotenoids, titrable acidity) and mineral content (P, K, Fe, Zn). Differences among accessions were observed in plant and fruit traits. Melon germplasm with high titrable acidity, higher than average amounts of mineral content and resistance to Cucumber mosaic virus, Zucchini yellow mosaic virus, powdery mildew (races 1, 2, 3, 5), Fusarium wilt (races 1, 2), Aphis gossypii and leafminer was recorded in the collection. A high level of genetic variability in melon germplasm was suggested by the SSR analysis. Comparative analysis using SSRs of the genetic variability between Indian melons from north, south, and east regions and reference accessions of melon from Spain, France, Japan, Korea, Iraq, Zambia showed regional differentiation between Indian melon accessions and that Indian germplasm was weakly related to the melon accessions from other parts of the world, suggesting that an important portion of the genetic variability found within this melon collection has not been used yet for the development of new cultivars. Additional collections of acidulus melon germplasm should be made in southern India and adequate management of this important genetic resource is clearly a necessity.     

Morpho-agronomic and simple sequence repeat-based diversity in colored rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) germplasm from peninsular India

Knowledge of the genetic diversity in the germplasm accessions is important for the efficient germplasm management. We studied 45 colored rice accessions, which had been earlier collected from peninsular India. The accessions were evaluated at two diverse locations for 12 morpho-agronomic traits and genotyped using 50 simple sequence repeat (SSR) markers. Significant genotypic variability was observed for all morpho-agronomic traits studied. Dendrogram and principal component analysis based on morpho-agronomic traits separated the accessions into three clusters. The first two principal components accounted for 82% of the total variation. The most discriminatory traits were number of grains per panicle, biomass yield, and days to flowering and maturity. The SSR analysis revealed high polymorphic information content value of 0.84. Though, Mantel test did not show a significant correlation, yet the classification based on phenotyping and genotyping data showed good agreement for the expression of morpho-agronomic traits in the formation of clusters. Five accessions had high concentration (> 100 μg/g) of Fe and one of Zn. Our study revealed the presence of large genetic variation among the colored rice accessions evaluated and moderate agreement between morpho-agronomic and SSR–based classifications and of these with geographic diversity.     

Assessment of genetic diversity in emmer (Triticum dicoccon Schrank) × durum wheat (Triticum durum Desf.) derived lines and their parents using mapped and unmapped molecular markers

In the last few years, the renewed interest for emmer wheat (Triticum dicoccon Schrank) in Italy has stimulated breeding programs for this crop releasing improved genotypes obtained not only by selection from landraces, but even by crosses with durum wheat (Triticum durum Desf.) varieties. The purpose of this work has been to uncover the genetic make-up of some emmer × durum derivatives, specifically by comparing the differences from their parents. Genetic diversity of advanced breeding lines and varieties derived from a durum × emmer cross has been evaluated on the basis of AFLP and SSR markers in comparison with the corresponding emmer and durum wheat parent for addressing the seminal question of how much ‘wild’ variation remains after selection for agronomic type.     

Genetic Variation Among Portuguese Landraces of ‘Arrancada’ Wheat and <Emphasis Type="Italic">Triticum petropavlovskyi</Emphasis> by AFLP-based Assessment

Portuguese wheat landraces, ‘Arrancada’ were collected from the Aveiro region, Portugal before the 1950s. We found in eight accessions of `Arrancada' hexaploid wheat with the long glume phenotype. We assessed the comparative genetic diversity among Portuguese `Arrancada' wheat and Triticum petropavlovskyi Udacz. et Migusch. using AFLP assays and discuss the origin of long glumed `Arrancada' wheat. With the four primer pairs a total of 4885 visible bands were scored corresponding to 99 AFLP markers as putative loci, of which 55 markers (54%) were polymorphic. UPGMA clustering and PCO grouping showed that long glumed ‘Arrancada’ wheat and T. petropavlovskyi were genetically diverse. Long glumed ‘Arrancada’ hexaploid wheat separated into two clusters (groups) in both the UPGMA dendrogram and in PCO analysis. Four long glumed accessions fell in the cluster of tetraploid wheat. A similar argument could be made for another four accessions which belong to the cluster of hexaploid wheat. The substantial level of genetic variation indicated that long glumed ‘Arrancada’ wheat and T. petropavlovskyi originated independently. It is most likely that the P-gene of long glumed ‘Arrancada’ hexaploid wheat was introduced from T. turgidum ssp. polonicum (L.) Thell. to T. aestivum via natural introgression or breeding. We suggest that the long glumed ‘Arrancada’ hexaploid wheat did not originate from T. aestivum through spontaneous mutation at the P locus     

Genetic identity and relationships of Iranian apple (Malus?×?domestica Borkh.) cultivars and landraces, wild Malus species and representative old apple cultivars based on simple sequence repeat (SSR) marker analysis

In order to shed light on the role of Iran in apple evolution and domestication, we chose to investigate the relationships of a collection of 159 accessions of wild and domesticated apples including Iranian indigenous apple cultivars and landraces, selected wild species, and old apple scion and rootstock cultivars from different parts of the world. The majority of the wild species belonged to M. sieversii, which is widely believed to be the main maternal wild ancestor of domestic apples, from Kazakhstan and M. orientalis, which is one of the probable minor ancestors of domestic apples, from Turkey and Russia located on the east and west of Iran, respectively. The accessions were assigned into six arbitrary populations for the purpose of generating information on genetic parameters. Nine simple sequence repeat (SSR) loci selected from previous studies in apple were screened over DNA extracted from all the accessions. Results showed that all SSR loci displayed a very high degree of polymorphism with 11–25 alleles per locus. In total, there were 153 alleles across all loci with an average of 17 alleles per locus. The SSR allelic data were then used for estimation of population genetic parameters, including genetic variation statistics, F-statistics, gene flow, genetic identity, genetic distance and then cluster analysis using POPGENE 1.32 software. The F-statistics and gene flow in particular, showed that there was more intra-population than between population variation. The genetic identity and genetic distance estimates, and the dendrogram generated from the un-weighted pair group arithmetic average (UPGMA) method of cluster analysis showed that the Iranian cultivars and landraces were more closely related to M. sieversii from Central Asia (east of Iran) and M. orientalis native to Turkey and Russia than to other accessions of Malus species. Also, the old apple cultivars from different parts of the world have a closer genetic relationship to M. sieversii, M. orientalis and the Iranian apples, than to other wild species. Based on these results, we suggest that the Iranian apples may occupy an intermediate position between the domesticated varieties and wild species. We propose that Iran could be one of the major players in apples’ domestication and transfer from Central Asia to the western countries.     

Extent and Pattern of Genetic Diversity for Morpho-agronomic Traits in Ethiopian Highland Pulse Landraces II. Faba Bean (<Emphasis Type="Italic">Vicia faba</Emphasis> L.)

A field experiment was conducted in 2001 at Holetta and Kulumsa, Ethiopia, to study genetic diversity in Ethiopian faba bean (Vicia faba L.) landraces. One hundred sixty random germplasm accessions were grown in an alpha lattice design with two replications. Data on 12 traits were collected and analyzed. Significant differences were observed among the accessions for most of the traits (except number of pods/podding nodes) at each location even though differences pooled over location were mostly non-significant. Cluster analysis distinguished seven diversity classes of different sizes. Accessions from the northern half of the country (North and South Wello, North Gonder and North Shewa) were closely related while those from the southern part of the country (Arsi) were highly diverse. Cumulative effects of a number of characters dictated differentiation of the accessions into clusters. Some overlapping were encountered between accessions from the northern and those from the southern parts of the country. The study revealed that accessions from different regions might have similar genetic background and those from the same origin might also have different genetic background. Therefore, geographic diversity should not necessarily be used as an index of genetic diversity and parental selection should be based on a systematic study of genetic diversity in a specific population. Genetic distances between most of the clusters were significant that crosses between parents selected out of them are expected to generate desirable progenies. Future germplasm collection, conservation and utilization strategies should put more focus not only on inter-regional diversity in the country as a whole but also on intra-regional diversity in Arsi.     

Collection of Lupinus angustifolius L. Germplasm and Characterisation of Morphological and Molecular Diversity

Lupinus angustifolius L. is a Mediterranean species, domesticated in the 20th century, representing an important grain legume crop in Australia and other countries. This work is focused on the collection of wild germplasm and on the characterisation of morphological and molecular diversity of germplasm accessions. It reports the collection of 81 wild L. angustifolius accessions from the South and Centre of Portugal, available at the ‘Instituto Superior de Agronomia Gene Bank’, with subsequent morphological and molecular characterisation of a selection of these and other accessions. A multivariate analysis of morphological traits on 88 L. angustifolius accessions (including 59 wild Portuguese accessions, 15 cultivars and 14 breeding lines) showed a cline of variation on wild germplasm, with plants from Southern Portugal characterised by earlier flowering, higher vegetative development and larger seeds. AFLP and ISSR molecular markers grouped modern cultivars as sub-clusters within the wider diversity of wild germplasm, revealing the narrow pool of genetic diversity on which domesticated accessions are based. The importance of preserving, characterising and using wild genetic resources for L. angustifolius crop improvement is outlined by the results obtained.