Identification and genetic structure of wild Italian <Emphasis Type="Italic">Humulus lupulus</Emphasis> L. and comparison with European and American hop cultivars using nuclear microsatellite markers

Nine genic SSR loci were used to evaluate the genetic diversity and identify accessions in wild Italian Humulus lupulus L., in comparison with widely cultivated European and U.S. commercial cultivars. A collection of 80 wild hop samples from Italy and 43 hop cultivars from Europe and U.S., were characterized. Allelic frequency analysis revealed 65 distinct Italian genotypes and differentiated all the commercial cultivars; moreover, specific alleles were observed for wild and cultivated hops. The number of alleles identified in the wild population were 104 and 123 within all the accessions. The maximum polymorphic information content was evidenced for locus HlGA23 in the Italian wild population and in the whole set of accessions (0.905 and 0.902 respectively). The dendrogram constructed from Euclidean distance with the UPGMA method showed two main clusters, one including commercial American and European accessions and one mostly composed by wild Italian accessions. Model-based clustering (Bayesian method) placed the accessions into five germplasm groups, one of which was characterized by Italian genotypes only. The study showed for the first time the great biodiversity present in Italy, and the remarkable differences with European and American hops. It was also found that within the population of north-central Italy a large genetic variability is present, suited to be studied and exploited; this genetic wealth could be used in future breeding programs in order to develop new hop varieties carrying characteristics useful for brewers.     

Chloroplast Microsatellites to Investigate the Origin of Grapevine

The origin of the grapevine, Vitis vinifera ssp . sativa L., has been investigated with archaeobotanical–archaeological, cultural and historical data indicating a unique domestication centre located in the Caucasian and Middel-East regions about 6–7000 years ago, but, events leading to the domestication of this species are still an open issue. In this work, eight universal chloroplast microsatellites are used to assess genetic relationships among varieties selected as representatives of four distinct geographical groups from Middle-East to Western European regions. Results show that two out of the eight analysed chloroplast loci are polymorphic within the 142 individuals. Allele variants of the cpSSR loci combine in a total of six different haplotypes. The analysis of haplotypes distribution and haplotype diversity (HD) suggest that only three out of the six haplotypes are represented in the Caucasian and Middle-East samples, with 90% of individuals sharing the same haplotype. Moreover, the presence of all six haplotypes in the European accessions, with a high level of haplotype diversity, suggests varietal influx in these areas. Concerning the Western European varieties, especially in Spanish accessions, half of the individuals share haplotype VI which is completely absent in the Caucasian and Middle-East cultivars. This result opens the discussion about the existence of a unique and common domestication centre, located in the Caucasian and Middle-East area, for all the European cultivars. This work suggests the usefulness of chloroplast genome markers to provide information on haplotype distributions that could help to identify further geographical areas for grapevine varietal evolution.     

Genetic structure and differentiation among grapevines (<Emphasis Type="Italic">Vitis vinifera</Emphasis>) accessions from Maghreb region

Three gene pools representative of Vitis vinifera L. subsp. vinifera (=subsp. sativa Beger) growing in the Maghreb regions (North Africa) from Tunisia (44), Algeria (31) and Morocco (18) and 16 wild grape accessions (Vitis vinifera L. subsp. sylvestris (Gmelin) Beger) from Tunisia were analysed for genetic diversity and differentiation at twenty nuclear microsatellites markers distributed throughout the 19 grape chromosomes. 203 alleles with a mean number of 10.15 alleles per locus were observed in a total of 109 accessions. Genetic diversities were high in all populations with values ranging from 0.6775 (Moroccan cultivars) to 0.7254 (Tunisian cultivars). F _st pairwise values between cultivated grapevine populations were low but found to be significantly different from zero. High F _st pairwise values were shown between wild and cultivated compartments. Two parent offspring relationships, two synonyms and two clones of the same cultivar were detected. The rate of gene flow caused by vegetative dissemination of cultivated grapevine plants was not sufficient to genetically homogenise the pools of cultivars grown in different regions. The Neighbour Joining cluster analysis showed a clear separation according to geographical origins for the cultivated grapevines gene pools and revealed a high dissimilarity between cultivated and wild grapevine. However, three cultivars (Plant d’Ouchtata 1, Plant de Tabarka 3 and Plant d’Ouchtata 3) are very close to wild accessions and may result from a hybridisation between cultivated and wild accessions. The high level of differentiation between cultivated and wild accessions indicates that the cultivated accessions do not derive directly from local wild populations but could mostly correspond to imported materials introduced from others regions during historical times or derived from crossing between them.     

Microsatellite DNA Analysis of Wild Hops, Humulus lupulus L.

To study the relationships and genetic diversity among wild hops, Humulus lupulus, we analyzed 133 samples of wild hops collected from Europe, Asia and North America using polymorphism on 11 microsatellite loci. Although only three primers showed bands in Japanese hops, all other samples showed polymorphic bands at most loci. There were no duplicate genotypes among samples of European, Chinese and North American hops, and each individual hop could be distinguished completely. The phylogenetic tree constructed from DA distance with the UPGMA method showed a large cluster comprised of European hops, although Russian hops from the Caucasus and Altai regions were separate from the European cluster. Chinese and North American samples gave distinct clusters suggesting genetic differentiation. This study has indicated that hop microsatellite DNA is differentiated, and is dependent upon the origin in regions of Europe, Asia and North America.     

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.     

Germplasm sources of protective glandular leaf trichomes of hop (<Emphasis Type="Italic">Humulus lupulus</Emphasis>)

We report here that small but numerous foliar glands are present near the base of the abaxial leaf blades of hop (Humulus lupulus L.) at about 2.6 times the density found in the more distal regions, and this concentration of glands appear to protect the basal region from damage by insects. Distinctive wild taxa of H. lupulus are native to distinctive regions of the north temperate world. The much higher presence of protective glands in the three native North American Humulus taxonomic varieties in comparison with the varieties native to Europe and Asia suggests specialized leaf feeders in North America have been responsible for the evolution of greater gland density. Commercial domesticated hop cultivars trace most of their germplasm to the European hop (H. lupulus var. lupulus), which possesses a much lower concentration of leaf glands than the native hops of all other regions of the world, and therefore is likely relatively susceptible to damage. We suggest that breeding for increased density of protective foliage glands may be a non-chemical way of alleviating foliar feeding damage in cultivated hops.     

Genetic diversity in European accessions of the Barley Core Collection as detected by isozyme electrophoresis

Genetic diversity in 79 European accessions of the Barley Core Collections was surveyed using isozyme electrophoresis. A total of 26 alleles were observed at the ten isozyme loci. All loci were polymorphic except Pgd-1 which was monomorphic. The comparison of the results with those of previous studies indicates that most of the alleles occurring in the European Barley are also observed in this set of the European Barley Core Collections. Only five alleles (Est-1 Al; Est-5 Ag, Te; Pgi-1 C and Ndh-2 B) were absent. Nine of 26 alleles were rare alleles, which were detected only in one or two accessions. Moreover, most of rare alleles were detected in 6-rowed winter barley. It is very important to include rare alleles for maximising the genetic variations in core collections. In the set of European Barley Core Collection, 6-rowed barley contained larger diversity than 2-rowed barley; winter type contained larger diversity than spring type. The cluster analysis separated 79 accessions into three major groups. Group I is more complex and comprised 2-rowed spring, 2-rowed winter and 6-rowed winter barley. In this group, 18 accessions in the cluster A and 14 accessions in the cluster B possessed identical genotypes as judged from the ten isozyme data. Principal coordinate analysis could not clearly separate the spring cultivars from the winter barley lines, as well as not separate 2-rowed from 6-rowed barley.     

Chloroplast DNA Variation in the Most Primitive Cultivated Diploid Potato Species Solanum stenotomum Juz. et Buk. and its Putative Wild Ancestral Species using High-Resolution Markers

Solanum stenotomum Juz. et Buk. (2n = 2x = 24) is considered to be the most primitive diploid cultivated species from which all the other Andean cultivated potatoes were originated (Hawkes 1990). To disclose chloroplast DNA (ctDNA) variability and the maternal origin of S. stenotomum, 36 accessions of S. stenotomum and 86 accessions of putative wild ancestral species were determined for ctDNA types and analyzed by high-resolution markers (seven ctDNA microsatellites and an H3 marker). High-resolution markers discriminated 57 different ctDNAs (haplotypes), which were classified into the W-type ctDNA group and C-, S- and A-type ctDNA group, and within the latter group S- and A-type ctDNAs were distinct from each other among many different haplotypes mostly having C-type ctDNA. This ctDNA relationship supported our previous findings obtained for mostly Andean cultivated species (Sukhotu et al. 2004). Compared with other putative ancestral wild species, S. stenotomum showed somewhat limited ctDNA diversity, having two major haplotypes 1 and 2 also found in different wild species in different places. Therefore, the ctDNA in S. stenotomum was of at least dual origins either by successive domestication from different species or else by introgression after initial S. stenotomum arose.     

Characterization of European hazelnut (<Emphasis Type="Italic">Corylus avellana</Emphasis>) cultivars using SSR markers

World hazelnut production is based primarily on selections from the wild. In this study, we used 21 pairs of simple sequence repeat (SSR) primers to investigate genetic diversity in 270 clonal accessions of European hazelnut (Corylus avellana) representing a wide geographic range. Of the 270 accessions, 198 had unique fingerprints while 72 were duplicates. Based on the 198 unique accessions, the number of detected alleles per locus averaged 9.81 and observed heterozygosity (H_o) averaged 0.67. Of the 206 total alleles amplified, 20 were unique to a single accession. A genetic similarity matrix was constructed and the resulting dendrogram revealed four major geographical groups: Central European, Black Sea, English and Spanish-Italian. SSR alleles indicated the parentage of 31 accessions. The fingerprints are publicly available through the Germplasm Resources Information Network (GRIN) database. The identification of duplicate and mislabeled accessions will improve management of hazelnut genebanks, and information on genetic variation in hazelnut will assist the international research community. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

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.     

Using diversity of the chloroplast genome to examine evolutionary history of wheat species

Chloroplast microsatellites (SSRs) are conserved within wheat species, yet are sufficiently polymorphic between and within species to be useful for evolutionary studies. This study describes the relationships among a very large set of accessions of Triticum urartu Thum. ex Gandil., T. dicoccoides (Körn. ex Asch. et Graebn.) Schweinf., T. dicoccon Schrank, T. durum Desf., T. spelta L., and T. aestivum L. s. str. based on their cpSSR genotypes. By characterising the chloroplast diversity in each wheat species in the evolutionary series, the impact on diversity of major evolutionary events such as domestication and polyploidyisation was assessed. We detected bottlenecks associated with domestication, polyploidisation and selection, yet these constrictions were partially offset by mutations in the chloroplast SSR loci that generated new alleles. The discrete cpSSR alleles and haplotypes observed in T. urartu and Aegilops tauschii, combined with other species specific polymorphisms, provide very strong evidence that concur with current opinion that neither species was the maternal and thus cytoplasmic donor for polyploid wheats. Synthetic hexaploid wheats possessed the same chloroplast haplotypes as their tetraploid progenitors demonstrating how the novel synthetic wheat lines have captured chloroplast diversity from the maternal parents, the chloroplast is maternally inherited and novel alleles are not created by genomic rearrangements triggered by the polyploidisation event.     

Polymorphisms Among Chloroplast and Mitochondrial Genomes of <Emphasis Type="Italic">Citrullus</Emphasis> Species and Subspecies

Twelve and six DNA clones representing various parts of chloroplast and mitochondrial genomes, respectively, were used to detect polymorphism among five watermelon cultivars and 21 U.S. Plant Introductions (PIs) collected from diverse geographical locations and representing major groups of Citrullus species. Cluster analysis based on 20 chloroplast DNA (cpDNA) and 10 mitochondrial DNA (mtDNA) restriction fragment length polymorphism (RFLP) markers differentiated the accessions into three major phenetic groups: PIs and watermelon cultivars of Citrullus lanatus subsp. vulgaris (Schrad. ex Eckl. et Zeyh.) Fursa (also designated as C. lanatus var. lanatus) (group I), PIs of C. lanatus var. citroides (of C. lanatus subsp. lanatus Schrad. ex Eckl. et Zeyh.)(group II), and C. colocynthis (L.) Schrad. PIs (group III). The chloroplast and mitochondrial genomes of watermelon cultivars are distinct, but closely related to those of the C. lanatus var. lanatus PIs. On the other hand, the chloroplast and mitochondrial genomes of the wild species C. colocynthis are more similar to those of C. lanatus var. citroides. Polymorphic cpDNA and mtDNA markers identified in this study can complement isozyme and nuclear DNA data used in earlier phylogenetic and phenetic classifications of Citrullus PIs. These cpDNA and mtDNA markers are being used in experiments designed to enhance watermelon cultivars by replacing the chloroplast and mitochondrial genome of cultivated watermelon with those of the wild species C. colocynthis.     

Genetic diversity of <Emphasis Type="Italic">Nelumbo</Emphasis> accessions revealed by RAPD

Total 65 lotus accessions in genus Nelumbo mainly collected from China, were subjected to random amplified polymorphic DNA (RAPD) markers to estimate the genetic diversity and to test the genetic basis of the relationships between morphotypes and molecular markers. Seventeen primers generated a total of 195 highly reproducible and discernible loci, among which 173 were polymorphic. Percent polymorphism varied from 66.7 to 100 with an average of 88.72, and five primers out of them, OPC05, OPG10, OPN20, OPP09 and OPS17, showed 100% polymorphism. A relatively high genetic diversity was detected among all the samples with the similarity coefficient values ranging from 0.45 to 0.85, and Nei’s gene diversity (h) 0.30, and Shannon index (I) 0.46. The UPGMA dendrogram clustered 65 accessions in four clusters and the clustering pattern showed two groups, N. nucifera ssp. nucifera and those accessions related to the American lotus, and some special cultivars, landraces, hybrids and the American lotus. Principal Coordinate Analysis (PCA) further indicated that the genetic diversity of Nelumbo accessions was not evenly distributed, instead, was presented by a clustered distribution pattern. Similar to the results revealed by the dendrogram, two main groups representing the two subspecies of N. nucifera, as well as some special landraces, cultivars of Chinese lotus, the Japanese lotus and hybrids out of the two groups were obtained. Neither the UPGMA dendrogram nor the PCA analysis exhibited strict relationship with geographic distribution and morphotypes among the accessions.     

Genetic diversity of African wild rice (Oryza longistaminata Chev. et Roehr) at the edge of its distribution

Ethiopia is at the edge of the distribution for African wild rice, Oryza longistaminata. Here, chloroplast (cp) and simple sequence repeat (SSR) markers were applied to wild rice accessions in Ethiopia to evaluate how they differ from control O. longistaminata, O. barthii and O. glaberrima accessions which originated from African countries. Based on the cp genomes of African wild rice species, maternally inherited cpINDEL markers were developed. The cp indels helped to elucidate 20 plastid types. African cultivated rice shared a particular plastid type with one of annual O. barthii. Parts of northern wild rice in Ethiopia shared Type 6 with control O. longistaminata. The north group shared another type with parts of the south group. The 16 SSR markers amplified a total of 155 alleles in 215 rice accessions, with mean allelic richness of 9.688 per locus, observed heterozygosity of 0.241, expected heterozygosity of 0.724, polymorphic information content of 0.700, and a significant genetic differentiation of 0.215. Both cpINDEL and nuclear markers analyses suggested that wild rice in Ethiopia belongs to O. longistaminata. However, they carry both a unique plastid type and different population structure from control O. longistaminata collected from other areas in Africa. We concluded that the edge of its distribution maintains unique variation. These populations are regarded as valuable genetic resources for future rice breeding.

     

Genetic diversity of cultivated and wild Ussurian Pear (<Emphasis Type="Italic">Pyrus ussuriensis</Emphasis> Maxim.) in China evaluated with M13-tailed SSR markers

Eight genomic SSR markers with a M13 tail attached were used to assess the genetic diversity of 72 Ussurian Pear accessions (Pyrus ussuriensis Maxim.) in China. The M13-tailed method was effective in discriminating all the 32 wild accessions. All the 40 Ussurian Pear cultivars could be successfully discriminated with the exception of 4 sets of synonymies or spots. A total of 108 alleles were obtained with an average of 13.5 per locus. The expected heterozygosity, observed heterozygosity, and power of discrimination were 0.78, 0.63, and 0.86 respectively. Three triploid cultivars (‘Anli’, ‘Ruan’er’, and ‘Pitaiguo’), and one wild accession, P. ussuriensis ‘Xilin-3’, showed three alleles at some SSRs. The number of alleles and observed heterozygosity per locus for 40 Ussurian Pear cultivars were 9.1 and 0.62, respectively, lower than the values of 32 wild accessions which were 11.3 and 0.65, respectively. A dendrogram based on the SSR genotypes was obtained, showing two major groups corresponding to cultivated group and wild group. All the cultivars fell into the cultivated group. Some subgroups (Nanguoli subgroup, Zhibazi subgroup, Xiangshuili subgroup, Balixiang subgroup, Anli subgroup) could be found in the cultivated group. A very close relationship between ‘Huagaili’ and ‘Miansuan’, and a close relationship between ‘Anli’ and a wild accession, P. ussuriensis ‘Huangshanli’ could be found in Anli subgroup. ‘Nanguoli’ and ‘Xiaowuxiang’ showed a close relationship with at least one identical allele at each locus with the exception of NH015a.     

Identification of Chinese white pear cultivars using SSR markers

109 Pyrus accessions including 92 local Chinese accessions of P. bretschneideri were identified genetically using nine microsatellite loci developed from apple and pear. The nine SSR loci revealed 129 alleles in 109 pear accessions and 114 alleles in 92 Chinese white pears. Among the 92 local Chinese accessions of P. bretschneideri, 70 could be differentiated successfully except for 10 sets of synonymous or mutants. For the 92 accessions, the number of putative alleles per locus ranged from seven to 18, with an average of 12.67; the average values of observed heterozygosity and Shannon’s Information index were 0.60 and 1.85, respectively. A phenogram based on the SSR (simple sequence repeat) genotypes was obtained. The 109 accessions clustered into 11 groups based on geographical origin. The European pears and the Asian pears did not form independent two groups, but three P. communis cultivars grouped together independently. The Japanese P. pyrifolia cultivars mingled together with the Chinese P. bretschneideri cultivars, but four P. ussuriensis cultivars except for one (Jianbali) grouped together independently. The results indicated that the relationship of P. bretschneideri cultivars and P. pyrifolia cultivars was much closer than others.     

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.     

Preliminary screening and selection for cold tolerance in annual wild <Emphasis Type="Italic">Cicer</Emphasis> species

The study was aimed to select cold tolerant accessions of annual wild Cicer species in the highlands in the west Mediterranean area of Turkey. A total of 43 accessions of eight annual wild Cicer species was screened for cold tolerance and compared with the best cold tolerant cultivars, ILC 8262 and FLIP 93-53C. After the sensitive check, ILC 533, was killed, accessions were evaluated using a 1–9 scale. Three accessions of Cicer bijugum, 2 accessions of Cicer reticulatum and 2 accessions of Cicer echinospermum were highly cold tolerant, while 3 accessions of C. bijugum, 9 accessions of C. reticulatum, 2 accessions of C. echinospermum and 1 accession of Cicer pinnatifidum were cold tolerant. All accessions of Cicer judaicum were sensitive to cold. Whereas Cicer chorassanicum and Cicer cuneatum were killed, Cicer yamashitae was recorded as highly sensitive to cold. The cold tolerant accessions of annual wild Cicer species were superior to the best cultivar as far as hardiness is concerned.     

Investigating the origin of hazelnut (<Emphasis Type="Italic">Corylus avellana</Emphasis> L.) cultivars using chloroplast microsatellites

The place and time of European hazel (Corylus avellana L.) domestication is not clear, although it was already cultivated by the Romans. In this study, 75 accessions from Spain, Italy, Turkey, and Iran were analysed using 13 chloroplast microsatellite to investigate the origin and diffusion of hazelnut cultivars. Four loci were polymorphic and identified a total of four different chlorotypes. Their distribution was not uniform in each geographical group. The most frequent chlorotype A was present in all groups. An increase in chlorotype number and diversity from Spain eastward to Italy, Turkey, and Iran was observed. Results suggest that some spread of cultivars occurred from East to West and that hazelnut cultivation was not introduced from the eastern Mediterranean basin into Spain and southern Italy by Greeks or Arabs. Moreover, the results suggest considerable exchange of germplasm between Italy and Spain, probably by the Romans. Hazelnut appears to have been domesticated independently in three areas: the Mediterranean, Turkey, and Iran.