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.     

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 diversity and similarity of pear (<Emphasis Type="Italic">Pyrus</Emphasis> L.) cultivars native to East Asia revealed by SSR (simple sequence repeat) markers

Simple sequence repeat (SSR) markers were used to assess genetic diversity and relationship of Pyrus L. cultivars native mainly to East Asia. A total of 168 putative alleles were generated from six primer-pairs (BGA35, KU10, BGT23b, NH004a, NH011b and NH015a). All the SSR markers showed a high level of genetic polymorphism with a mean of 28 putative alleles per locus and the heterozygosity of 0.63. The Dice’s similarity coefficient between cultivars ranged from 0.02 to 0.98 and Occidental pears generally had low affinities to Asian pears. Ten major groups were generated from all the accessions by UPGMA clusters analysis. Chinese sand pears consisted of four groups with Chinese white pears and Japanese pears, of which Chinese sand pears occurred in all four groups, presenting a large genetic diversity, Chinese white pears were included in three groups, and Japanese pears only fell into one group. In the dendrogram, Chinese sand pears and Chinese white pears did not form discrete group, even subgroups. Some Japanese pear cultivars had high affinities to Chinese sand pear cultivars. These findings supports the authors’ previous viewpoints of Chinese white pears as a variety or an ecotype of Chinese sand pears (P. pyrifolia var. sinensis (Lindley) Y. Teng et K. Tanabe) and the progenitor of Japanese pears coming from China. Cultivars of P. ussuriensis Maxim. were clustered together with one clone of P. hondoensis Nakai et Kikuchi, a relative species of P. ussuriensis. Cultivars of P. communis L. and other Occidental species formed three independent groups and were distant from most Asian pears, except for P. betulaefolia Bge.     

Pear (Pyrus Species) Genetic Resources in Iwate, Japan

Iwateyamanashi (Pyrus ussuriensis Maxim. var. aromatica (Nakai et Kikuchi) Rehd.) is one of the Pyrus species grown wild in Japan. The origin of the Japanese pear (P. pyrifolia) is uncertain but it has been suggested that Iwateyamanashi is the possible progenitor of the modern Japanese cultivar. During the last few decades, the number of Iwateyamanashi trees has been decreasing and therefore, conservation is urgently needed. After 13 explorations in the northern Tohoku region of Japan, 615 pear trees and 30 local names were recognized mainly in Iwate Prefecture. The center of the distribution seemed to be somewhere around Mt. Hayachine to the northern area of Kitakami highlands (from lat. 39 °20′ to 40 °10′N, and from long. 141 °20′ to 141 °50′E). Four morphological characters concerning fruit shape, measured for 85 trees, showed a wide range of continuous variation. For the skin colour of fruit, 51% of trees bore russet type fruits, 22% smooth and 27% intermediate ones. Most of the fruits had five loculi but Sanenashi fruits (seedless pear), one of the old cultivars, had three, and fruits of another two trees had four. More than 80% of trees tended to produce fruits with a calyx but some trees bore fruits without a calyx. These observations indicate a wide range of genetic diversity in Pyrus species which is caused, not only by high heterogeneity in Iwateyamanashi itself, but also by the coexistence of Iwateyamanashi, P. pyrifolia (Burm.) Nakai and hybrid progeny in this area. Already 250 trees have been conserved as genetic resources by grafting at Kobe University.     

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.     

Delimitation of Amaranthus cruentus L. and Amaranthus caudatus L. using micromorphology and AFLP analysis: an application in germplasm identification

The ‘Morelos’ accessions of Amaranthus from Mexico demonstrate taxonomic ambiguity at the basic morphologic level. The main cause is the enormous morphological and genetic variation exhibited by the species in the genus. Although basic morphological criteria can be applied to herbarium specimens or germplasm collections for quick taxonomic identification, the morphological data alone can be misleading. To ascertain the taxonomic identity of the ‘Morelos’ accessions and their hypothesized species affiliation to Amaranthus caudatus or Amaranthus cruentus, we conducted a comparative analysis of phylogenetic relationships among these taxa/accessions using amplified fragment length polymorphism (AFLP) and micromorphology methods. Based on AFLP data, all the controversial ‘Morelos’ accessions can be consistently placed into a single A. cruentus species clade, which is clearly separated from the A. caudatus species clade. The AFLP-based phylogenetic relationship of ‘Morelos’ and delimitation of A. cruentus and A. caudatus are further supported by micromorphology, showing that the combination of these techniques can provide more reliable data for germplasm identification than each method used alone.     

AFLP similarities among historic Danish cultivars of fodder beet (<Emphasis Type="Italic">Beta vulgaris</Emphasis> L. subsp. <Emphasis Type="Italic">vulgaris</Emphasis>var. <Emphasis Type="Italic">rapacea</Emphasis> Koch)

Dead seeds of a fodder beet cultivar ‘Elvetham’ stored under ambient conditions since 1880 were compared to a homonymous sample preserved in an on-farm situation in Denmark. DNA was isolated from single seeds and successfully applied to Amplified Fragment Length Polymorphism (AFLP) analysis of the accessions. Six primer pairs were used to determine the similarity between the two accessions based on 112 polymorphic bands. Furthermore, similarity among seven cultivars of fodder beets representing the main types used in Scandinavia at the end of the 19th century was determined. This analysis was based on 152 polymorphic bands. Differentiation among the seven cultivars was determined to a mean G _ST value of 0.438, while G _ST between the two ‘Elvetham’ accessions was 0.266. A principal coordinate analysis based on jaccards similarity index illustrates that the two ‘Elvetham’ accessions are different from each other. The differentiation is higher than the value found between two separate ‘Eckerndorfer’ accessions. The results indicate that the cultivated accession has changed. Additionally, the value of applying old dead seed material for documentation in gene banks is demonstrated. During the analysis it was found that DNA isolated from seeds and leaves behaved differently in the AFLP process, however, the two fractions assigned to their common accession.     

Relationship of wild and cultivated forms of <Emphasis Type="Italic">Pisum</Emphasis> L. as inferred from an analysis of three markers,of the plastid,mitochondrial and nuclear genomes

Eighty-nine accessions of wild and cultivated peas (12 Pisum fulvum Sibth. et Smith., 7 P. abyssinicum A. Br., 31 wild and 42 cultivated forms of P. sativum L.) were analysed for presence of the variants of three functionally unrelated polymorphic markers referring to different cellular genomes. The plastid gene rbcL either contains or not the recognition site for restriction endonuclease AspLEI (rbcL+ vs. rbcL−); the mitochondrial gene cox1 either contains or not the recognition site for restriction endonuclease PsiI (cox1+ vs. cox1−); the nuclear encoded seed albumin SCA is represented by slow (SCA^S) or fast (SCA^F) variant. Most of the accessions possessed either of two marker combinations: 24 had SCA^F cox1+ rbcL+ (combination A) and 49 accessions had SCA^S cox1− rbcL− (combination B), 16 accessions represented 5 of the rest 6 possible combinations. All accessions of P. fulvum and P. abyssinicum had combination A, the overwhelming majority of cultivated forms of P. sativum had combination B while wild representatives of P. sativum had both combinations A and B, as well as rare combinations. This pattern indicates that combination A is the ancestral state in the genus Pisum L., inherited by P. fulvum and P. abyssinicum, while combination B seems to have arisen in some lineage of wild P. sativum which rapidly fixed mutational transitions of the three markers studied, most probably via a bottleneck effect during the Pleistocene. Then this ‘lineage B’ spread over Mediterranean and also gave rise to cultivated forms of P. sativum. Rare combinations may have resulted from occasional crosses between ‘lineage A’ and ‘lineage B’ in nature or during cultivation, or represent intermediate evolutionary lineages. The latter explanation seems relevant for an Egyptian cultivated form ‘Pisum jomardii Schrank’ (SCA^F cox1− rbcL−) which is here given a subspecies rank. Wild representatives of P. sativum could be subdivided in two subspecies corresponding to ‘lineage A’ and ‘lineage B’ but all available subspecies names seem to belong to lineage B only. Presently all wild forms would better be considered within a fuzzy paraphyletic subspecies P. sativum subsp. elatius (Bieb.) Schmalh. s. l.     

Diversity of aroma patterns in wild and cultivated <Emphasis Type="Italic">Fragaria</Emphasis> accessions

Plant breeders are interested in strawberry species as donors of volatile compounds in breeding programmes because of the diversity and intensity of wild strawberry aroma. Therefore, the topic of this paper is the prospective analysis of four accessions of four wild strawberry accessions in comparison to a standard cultivar of Fragaria × ananassa Duch. by using human sensory, gas chromatography mass spectrometry (GCMS) and gas chromatography–olfactometry (GCO). The wild species have higher aroma intensities compared with the cultivated one. The flavour quality differs significantly. Semiquantitative GC analysis revealed that F. × ananassa cv. ‘Elsanta’ has the lowest content of volatile compounds whereas Fragaria moschata L. ‘Cotta’ has the highest. The aroma impressions, measured by GCO, support the findings of GCMS analyses. The nasal impact frequency (NIF)-profiles of the wild types are more manifold and of higher intensities than those of the cultivated F. × ananassa cv. ‘Elsanta’ which corresponds with the overall flavour impression when tasting the fresh fruits.     

Comparative analysis of organelle DNAs in acid citrus grown in Japan using PCR-RFLP method

PCR-RFLP analyses of three regions for each of chloroplast DNA (cpDNA; rbcL-ORF106, trnD-trnT, trnH-trnK) and mitochondrial DNA (mtDNA; nad7/exon2-exon3, nad7/exon3-exon4, 18S-5S) were performed in 26 cultivars of acid citrus grown in Japan to identify polymorphisms and classify them. The polymorphisms were compared with those of three true Citrus species, i.e., mandarin, pummelo and citron. Ichang papeda (C. ichangensis) was also included in this study to find its relationship with Yuzu. Inter-species cpDNA variation was recognized and the acid citrus were divided into three groups, namely; I (‘Yuzukichi’ and ‘Kinkoyu’), II [sour oranges (‘Kaiseito’, ‘Daidai’ and ‘China daidai’), ‘Nansho daidai’, ‘Kiku daidai’, C. sudachi (‘Mushi yukaku’, ‘Yushi yukaku’ and ‘Yushi mukaku’), C. sphaerocarpa (‘Kabosu’ and ‘Aka kabosu’), C. kizu (‘Taninaka kizu’, ‘Kinosu’ and ‘Kizu’), ‘Zanbo’, ‘Mochiyu’, ‘Jabara’ and ‘Naoshichi’], and III [Yuzu (‘Tetraploid’, ‘Tochikei yuzu’ and ‘Yamanekei yuzu’), ‘Matsuda sudachi’, ‘Zuishoyu’, ‘Hanayu’ and ‘Yuko’]. CpDNA restriction patterns of the three true Citrus species differed from each other as well as from those of ichang papeda. CpDNA restriction patterns of group I of the acid citrus were identical to those of mandarins. Group II showed the same as pummelos. CpDNA restriction patterns of group III were differed from those of the three true Citrus species in the three regions. This group was differed from ichang papeda after digestion of trnH-trnK PCR products with TaqI, HinfI and AluI, while they showed identical restriction patterns in two regions, rbcL-ORF106 and trnD-trnT. Citrons and ichang papeda were placed in groups IV and V, respectively. Based on mtDNA restriction patterns, the acid citrus were divided into three groups; i, ii and iii. In groups i and ii accessions of groups I and II of cpDNA were placed with mandarins and pummelos, respectively. In group iii accessions of group III of cpDNA were placed with ichang papeda. Citrons were placed in a distinct group, iv.     

Genetic Analysis of Egyptian Date (<Emphasis Type="Italic">Phoenix dactylifera</Emphasis> L.) Accessions Using AFLP Markers

Forty-seven samples of date palm (Phoenix dactylifera L.) collected from eight locations in Egypt were studied using four sets of amplified fragment length polymorphism (AFLP) markers with near infrared fluorescence labeled primers. These samples belonged to 21 named accessions and 9 of unknown pedigrees. A total of 350 bands were scored and 233 (66.6%) were polymorphic. Twenty-seven Egyptian accessions and ‘Medjool’and ‘Deglet Noor’accessions from California could beclassified into the major cluster. This major cluster may represent a major group of date palm germplasm in North Africa. There were four other clusters, each containing one or two accessions. The variety ‘Halawy’and one accession of unknown provenance were most likely from hybridization between two clusters. Six groups of accessions of which had the same names, revealed similar but not identical AFLP profiles suggesting these accessions might derive from seedlings rather thanthrough clonal offshoot propagation.     

Genetic diversity and structure of Ethiopian,Yemen and Brazilian C<Emphasis Type="Italic">offea arabica</Emphasis> L. accessions using microsatellites markers

Genetic diversity among 115 coffee accessions from the Coffea Germplasm Collection of IAC was assessed using SSR markers. The germplasm represents 73 accessions of Coffea arabica derived from spontaneous and subspontaneous plants in Ethiopia and Eritrea, species center of origin and diversity, 13 commercial cultivars of C. arabica developed by the Breeding Program of IAC, 1 accession of C. arabica cv. ‘Geisha’, 13 accessions of C. arabica from Yemen, 5 accessions of C. eugenioides, 4 accessions of C. racemosa and 6 accessions of C. canephora. Genetic analysis was performed using average number of alleles per locus (A), proportion of polymorphic loci (P), Shannon’s genetic index (H′ and G′_ST) and clustering analysis. All evaluated species were distinguished by a cluster analysis based on Jaccard’s coefficient. Differentiation between the cultivated plants of C. arabica and accessions derived from spontaneous and subspontaneous plants was observed. Spontaneous and subspontaneous accessions from Ethiopia were separated according to the geographical origin: east and west of the Great Rift Valley. Cultivated plants showed a low genetic diversity with a division in two groups: accessions from Yemen (H′=0,028) and Brazilian commercial cultivars (H′=0,030). The results agreed with previously reported narrow genetic basis of cultivated plants of C. arabica and supported the hypotheses about domestication of the species. This study also showed a significant genetic diversity among accessions from Ethiopia and Eritrea present in the Germplasm Collection of IAC. This diversity is specially observed in accessions from Sidamo (H′=0,143), Kaffa (H′=0,142) and Illubabor (H′=0,147) indicating their importance as source of genetic variability for coffee breeding programs.     

Analysis of the contribution of Mesoamerican and Andean gene pools to European common bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.) germplasm and strategies to establish a core collection

Common bean (Phaseolus vulgaris L.) was introduced in Europe from both Mesoamerican and Andean centres of origin. In this study, a collection including 544 accessions from all European regions showed that the Andean phaseolin types ‘T’ (45.6%) and ‘C’ (30.7%) prevailed over the Mesoamerican ones ‘S’ (23.7%), and accessions with cuboid seed shape (34.9%), maroon coat darker colour seed (44.3%), uniform seed colour (69.6%) were the most frequent. European accessions with phaseolin ‘S’ showed a significantly larger average seed size compared to those from America in the same phaseolin class while those presenting ‘T’ and ‘C’ phaseolin did not. This suggests that, during crop expansion in Europe, sampling or selection favoured the large-seeded races within the Mesoamerican ‘S’ gene pool or, possibly, introgression from Andean germplasm did occur. A core collection was developed using sampling approaches based on the information available in the genebank databases and on phaseolin patterns. Four sampling strategies were used: simple random sampling, and three random-stratified samplings, by logarithm of frequency of accessions by country, by European region, and by phaseolin pattern, respectively. Two sampling strategies resulted in core collections significantly different for phaseolin electrophoretic patterns from the whole collection. Stratification by phaseolin patterns increased the frequency of ‘S’ types (‘C’ type = 33%, ‘T’ type = 5.7% and ‘S’ type = 31.3%). The core collections were validated using seven seed characters, and no significant difference was observed in all strategies. This first developed European bean core collection will help to assess the contribution of the two American gene pools to the European germplasm and their relative importance for breeding purposes.     

Genetic Nature of Yams (Dioscorea sp.) Domesticated by Farmers in Benin (West Africa)

‘Domestication’ is a traditional farmers’ practice reported for yams (Dioscorea sp.) in Benin (West Africa). It involves introducing ‘spontaneous’ (naturally occurring) yams, supposedly wild (D. abyssinica and D. praehensilis), in varieties of the D. cayenensis–D. rotundata cultivated species complex. In this study, we established the genetic nature of ‘predomesticated’ yam plants using the amplified fragment length polymorphism (AFLP) technique. A total of 213 accessions, consisting of 32 predomesticated yams, 70 D. cayenensis–D. rotundata, 86 D. abyssinica and 25 D. praehensilis yams were analysed. Using 91 AFLP markers, three groups of accessions were distinguished, broadly corresponding to the above botanical species. Of the 32 predomesticated accessions, 16% were clustered with D. praehensilis, 37% with D. abyssinica and the remaining 47% with D. cayenensis–D. rotundata. These results demonstrated the use of wild plants by farmers in their domestication process, and suggested that plants derived from intervarietal and interspecific hybridisation may also be subject to this process. This study has shown that through domestication farmers influence and increase the genetic diversity in yam by using sexual reproduction of wild and possibly cultivated yams.     

Genetic Characterization of Brazilian Annual <Emphasis Type="Italic">Arachis</Emphasis> Species from Sections <Emphasis Type="Italic">Arachis</Emphasis> and <Emphasis Type="Italic">Heteranthae</Emphasis> using RAPD Markers

The genus Arachis is divided into nine taxonomic sections. Section Arachis is composed of annual and perennial species, while section Heteranthae has only annual species. The objective of this study was to investigate the genetic relationships among 15 Brazilian annual accessions from Arachis and Heteranthae using RAPD markers. Twenty-seven primers were tested, of which nine produced unique fingerprintings for all the accessions studied. A total of 88 polymorphic fragments were scored and the number of fragments per primer varied from 6 to 17 with a mean of 9.8. Two specific markers were identified for species with 2n = 18 chromosomes. The phenogram derived from the RAPD data corroborated the morphological classification. The bootstrap analysis divided the genotypes into two significant clusters. The first cluster contained all the section Arachis species, and the accessions within it were grouped based upon the presence or absence of the ‘A’ pair and the number of chromosomes. The second cluster grouped all accessions belonging to section Heteranthae.     

A note on the evolution of kabuli chickpeas as shown by induced mutations in <Emphasis Type="Italic">Cicer reticulatum</Emphasis> Ladizinsky

Two distinct chickpeas of the domestic chickpea, C. arietinum L., exist and are referred to as ‘desi’ or microsperma and ‘kabuli’ or macrosperma. Cicer reticulatum Ladiz. is considered to be the wild progenitor of the domestic chickpea. However, the morphological variation in 18 original accessions of C. reticulatum is narrower than those of the domestic chickpeas. The aim of the study is to increase the variability in C. reticulatum. In M₂ generation, a mutant with white flower color was isolated despite of the fact that the parent has the pink flower. Although seed coat color of the parent was dark brown, the mutant was cream like ‘kabuli’ chickpea. It is commonly accepted that the large seeded domestic ‘kabuli’ chickpeas originated from the small seeded ‘desi’ chickpeas, but the induced mutants (white flower and cream seed coat color) of C. reticulatum may suggest an additional path for the evolution of ‘kabuli’ chickpea. ‘Kabuli’ chickpeas could have originated from spontaneous mutants of C. reticulatum. In M₃ generation, multipinnate leaf, erect growth habit, green seed and double-podded chickpeas were isolated. Among these progenies, morphologic variability increased and approached domesticated chickpea. Based on historical records and the induced mutants obtained from this study, the domestic ‘kabuli’ chickpea could have directly emerged from C. reticulatum in ancient Eastern Turkey.     

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.     

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     

The Presence of a New S-RNase Allele (S10) in Asian Pear (Pyrus pyrifolia (Burm; Nakai))

A PCR (polymerase chain reaction) amplification method using newly designed S-RNase primers was carried out in five Korean-bred pear cultivars and ten Japanese-bred pear cultivars. A new S-RNase allele, designated as S₁₀, was discovered from ‘Chengsilri’, containing a 1513 bp and two exons (213 bp in total) that coded for a peptide of 71 amino acids. The S₁₀-RNase allele contained the three conserved cysteine residues peculiar to S-RNase in Japanese pear and one histidine residue essential for RNase activity. We compared nucleotide sequence similarity of the exon regions of ten pear S-RNase alleles. The nucleotide sequence of S₁ showed a high similarity to S₄ (97.4%) and the new S₁₀ shows 77.8% (S₅) to 84.4% (S₄) similarity with the other pear S-RNase alleles. S₁₀ had a unique restriction endonuclease site for ‘HhaI’, with digests yielding fragments of 1235 and 491 bp. The S-genotype of pear cultivar (‘Chengsilri’) was determined to be S₅S₁₀ by PCR–RFLP (restriction fragment length polymorphism). Cluster analysis of 49 known S-RNase alleles of the Rosaceae separated into two divergent groups are as follows: group I: pear and apple, group II: almond, sweet cherry and mume.     

A study of variability in the Sicilian faba bean landrace ‘Larga di Leonforte’

In this paper molecular analysis, physical, technological, and chemical traits were used to estimate the level of variation on five accessions of a locally adapted Sicilian faba bean (Vicia faba L.) landrace named ‘Larga di Leonforte’. DNA analysis was performed using the Amplified Fragment Length Polymorphism (AFLP) molecular marker class and two other faba beans (cv. ‘Aguadulce Samba’—Spain—and landrace ‘Locale di Modica’—Sicily) were used as controls. Although the accessions of ‘Larga di Leonforte’ varied significantly for most of the agronomical and physical traits, this landrace generally had a heavy seed weight, short but large pods, and no more than two seeds per pod. This last characteristic allowed for erect pod angle attitude at maturity. Idratation data showed difference among accessions in seed weight at full hydration and in absorption rate at the very beginning of the hydration process, while any difference among accessions emerged in terms of cooking properties. The six AFLP Eco+3/Mse+3 different primer combinations applied in this research revealed different levels of polymorphism among the faba bean accessions and a total number of 346 amplicons were generated. Around 60% of amplicons displayed a polymorphic pattern among different accessions. Cluster analysis on morphological, technological, chemical and molecular data placed the all five ‘Larga di Leonforte’ accessions into a separated group, and the Sicilian material shares a fairly large amount of similarity with respect to the cultivar ‘Aguadulce Samba’ selected in Spain.