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.     

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.     

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

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

Genetic 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.     

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.     

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.     

Level and Transmission of Genetic Heterozygosity in Apricot (Prunus armeniaca L.) Explored Using Simple Sequence Repeat Markers

In this study, 17 peach simple sequence repeat (SSR) sequences were used in the exploration of the genetic heterozygosity level of several apricot cultivars from Spain, France, Greece, and the USA, and 23 descendants. The genotypes can be classified in three groups as a function of their genetic heterozygosity (1) local cultivars from Murcia (Spain) (‘Gitanos’ and ‘Pepito del Rubio’) and several descendants from crosses among these cultivars, with very low genetic heterozygosities (less than 0.30); (2) cultivars from France and Spain (‘Moniquí’, ‘Currot’ and ‘Bergeron’) and several descendants, with intermediate levels of genetic heterozygosity (around 0.45); and (3) cultivars ‘Orange Red’ and ‘Goldrich’ from North America and ‘Lito’ from Greece, with the remaining descendants, having genetic heterozygosities higher than 0.50. The results showed the high increase of genetic heterozygosity in the case of descendants from complementary crosses. The use of cultivars from North America could increase greatly the genetic heterozygosity in the Spanish apricot breeding programs, enlarging the genetic variability of the local cultivars. On the other hand, in the case of transgressive crosses among local Spanish cultivars, the increase of genetic heterozygosity was much lower.     

Genetic diversity of cowpea [<Emphasis Type="Italic">Vigna unguiculata</Emphasis> (L.) Walp.] in four West African and USA breeding programs as determined by AFLP analysis

Cowpea is an important grain legume and hay crop of many tropical and subtropical regions, especially in the dry savanna region of West Africa. The cowpea gene pool may be narrow because of a genetic bottleneck during domestication. Genetic variation within specific breeding programs may be further restricted due to breeding methods, ‘founder effects’ and limited exchange of germplasm between breeding programs. Genetic relationships among 60 advanced breeding lines from six breeding programs in West Africa and USA, and 27 landrace accessions from Africa, Asia, and South America were examined using amplified fragment length polymorphism (AFLP) markers with six near infrared fluorescence labeled EcoRI + 3/1bases/MseI + 3/1bases primer sets. A total of 382 bands were scored among the accessions with 207 polymorphic bands (54.2%). Despite a diverse origin, the 87 cowpea accessions shared a minimum 86% genetic similarity. Principal coordinates analysis showed clustering of breeding lines by program origin, indicating lack of genetic diversity compared to potential diversity. Accessions from Asia and the Americas overlapped and were distinct from West African breeding lines, indicating that germplasm from Asia and the Americas have common origins outside West Africa. US and Asian breeding programs could increase genetic variability in their programs substantially by incorporating germplasm from West Africa, while national programs in West Africa should consider introgression of Asian germplasm and germplasm from other parts of Africa into their programs to ensure long-term gains from selection.     

Quantitative and Qualitative Traits of Natural Ecotypes of Perennial Grasses (Dactylis glomerata L., Festuca arundinacea Schreb., Phalaris tuberosa L., Brachypodium rupestre (Host) R. et S.) Collected in Southern Italy

In 1999, natural populations of Dactylis glomerata L., Festuca arundinacea Schreb., Phalaris tuberosa L. and the macrotherm species Brachypodium rupestre (Host) R. et S., were collected as seed in Southern Italy (Apulia, Basilicata and Campania regions) to evaluate the potential as forage of native germplasm under dry conditions, using available commercial cultivars as controls. The persistence of accessions, biometric, phenologic, productive and qualitative parameters of forage have been studied for a 3-year period (2000–2002) in a typical Mediterranean environment. D. glomerata is widely present in the studied area with two subspecies (subsp. glomerata L., subsp. hispanica (Roth) Nyman). The research pointed out significant differences in plant size, earliness and single plant dry matter (DM) production. All the natural ecotypes belonging to this species showed higher persistence than the control cultivars. Some natural ecotypes appeared to have potential for improving DM yield (subsp. glomerata ecotypes) and reducing neutral detergent fibre (NDF) content (subsp. hispanica ecotypes). Natural ecotypes belonging to F. arundinacea, P. tuberosa, and B. rupestre are sporadically present in the studied area. Among them, two ecotypes of P. tuberosa, showing higher winter growth and earliness compared to cv. ‘Holdfast’, seem to be interesting for a future breeding programme.     

Identification of sweet cherry cultivars (Prunus avium L.) and analysis of their genetic relationships by chloroplast sequence-characterised amplified regions (cpSCAR)

Ten cpSCAR markers that show polymorphism in Prunus avium were used to fingerprint sweet cherry cultivars. The purpose of the study was also to contribute to identification and to help determine their genetic interrelationships. Samples of ‘0900 Ziraat’, a superior Turkish variety, which were collected in several locations all over Turkey, had identical cpSCAR patterns, and they resembled a common European haplotype, A. ‘Sweetheart’, ‘Summit’ and ‘Canada Giant’ and their haplotype are intermediate between the previously described haplotypes A and B, which were originally found in Central and Eastern European sweet and wild cherries, and those from Northern Turkey, respectively. The data therefore suggests a local maternal descent (within Europe and Asia Minor) of the cultivars analysed. Our results show that chloroplast DNA analysis is a straightforward way to classify cherry cultivars. We compare our results to others previously reported for sweet cherry cultivars, and conclude that cpSCAR diversity data could be considered for phylogenetic studies in this group.     

Evaluation ofFusarium wilt-resistance in primitive races ofGossypium hirsutum L.

Seven primitive races ofGossypium hirsutum L.punctatum, marie-galante, richmondi, latifolium, morrilli, palmeri, mexicanum and five F₁ crosses were inoculated withFusarium f. sp.vasinfectum physiological strain I. in the wilt field of Cixi Cotton Institute in 1987. There were highly significant differences among these primitive races.Punctatum andpalmeri were highly resistant. The resistances ofmarie-galante andmexicanum was similar with that of cultivar ‘Simian2’;richmondi, latifolium andmorrilli were susceptible. The disease incidence of F₁ in five primitive races hybridized with cultivar ‘Simian2’ was generally higher than that of mid-parent value. Inheritance of wilt resistance was partially dominant. In addition, 178 accessions of eight primitive races were evaluated for their resistance as before in 1989. There were significant differences in resistance among races and intra-race accessions. There were wilt resistance accessions among primitive races in which disease incidence was below 10%–25%. Inpalmeri, marie-galante, richmondi andpunctatum, one accession was immune with 0% disease incidence, respectively; 22.7–77.4% accessions were susceptible with 50% disease incidence. It showed that primitive races were mixed populations. They should be evaluated and screened for their utilization in cotton breeding program. Some best plants were selected from F₂ of two crosses, ‘Simian2’ ×palmeri and ‘Simian2’ ×mexicanum, based on the resistance toFusarium wilt and the response to photoperiod. 11 lines had been obtained with good fiber quality and relatively high potential production by backcrossing improvement. These lines are now being tested.     

Variation in morphology and seed production of snow clover [<Emphasis Type="Italic">Trifolium pratense</Emphasis> L. subsp. <Emphasis Type="Italic">nivale</Emphasis> (Koch) Arcang.] germplasm from the Rhaetian Alps,Italy

A recent approach to restoration of degraded mountain landscapes (e.g. ski slopes) recommend to sow seed mixtures including ‘site-specific’ species, i.e. native germplasm adapted to the prevailing conditions and ecologically compatible with the area of intervention. To better exploit the potential of these species and enhance their spread in revegetation projects, deeper knowledge is needed, however, on the existing variation in their germplasm, particularly for economically useful traits such as seed yield. Snow clover is one of the recommended ‘site-specific’ legumes. This study evaluated 12 populations of snow clover, collected in three valleys of the Rhaetian Alps, Italy, to assess their variation and highlight any promising material. The evaluation encompassed a mountain and a lowland site, to verify the behaviour of the germplasm also in one environment markedly different from that of origin. Differences among valleys were observed for flower colour, growth habit and type, flowering time and susceptibility to mildew, although the overall pattern of phenotypic diversity was similar among valleys. Some effect of the evaluation site on the expression of morphological and seed production traits, and on the overall diversity, was also evident. Furthermore, the lowland site experienced very high plant mortality towards the end of the first growing season. Individual populations proved a valuable source of variation for seed production. Some populations, in particular, were worth of attention for a possible selection of high-seed-yielding snow clover cultivars. An assessment of the mating system suggested a lack of self-tripping or an almost absolute self-incompatibility in snow clover.     

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.     

Sources of resistance to downy mildew in narbon (Vicia narbonensis) and common (Vicia sativa) vetches

Downy mildew (Peronospora viciae (Berk) de Bary) is an important disease of vetches (Vicia spp.) in the Mediterranean Region. Narbon and common vetch germplasm accessions originating from different countries, and advanced breeding narbon vetch genotypes, were evaluated for reaction to downy mildew for two seasons under field conditions. In narbon vetch, there were resistant sources from both germplasm accessions and advanced genotypes. From the germplasm accessions, 25 accessions were resistant while from the advanced breeding genotypes, 10 had resistance, with 2–3 ratings. In common vetch, many accessions were highly resistant to downy mildew. These were widely distributed in different countries, mainly Turkey, Italy, Syria and Iran. In general, common vetch had higher sources of resistance than the narbon vetch accessions. These new sources of resistance to downy mildew will be incorporated into the forage legume improvement project in a continuous effort to identify cultivars suitable for replacing fallow in the cereal-based cropping systems.     

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.     

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.     

Vigna vexillata (L.) A. Rich. Cultivated as a Root Crop in Bali and Timor

Vigna vexillata is considered as a pantropical distributed wild species closely related to the cowpea (Vigna uniculata) and adapted to infertile soils. The species is occasionally used for its storage roots as well as forage and erosion control plant. The objective of this study was to pursue personal communications that V. vexillata has be transformed into a cultivar in Indonesia. Seven Indonesian islands were visited (Java, Bali, Sumba, Flores, Timor, Kalimantan and Sulawesi). Wild V. vexillata was found in Java, Bali, Sumba, Flores and Timor, occasionally used as wild ‘forest food’, and cultivated V. vexillata was found in Bali and Timor, Seven cultivars were collected and two of these were made available for the National Botanic Garden of Belgium. The cultivars were primarily cultivated for their storage roots, propagated by seeds, required no scarified seeds for good germination and formed non-dehiscent pods. On-farm root yields of 18–30 t ha⁻¹ and seed yields of 0.7–1.2 t ha⁻¹ were estimated. A brief discussion about the common names of wild V. vexillata is given and it is proposed to use the name ‘tuber cowpea’ for V. vexillata accessions which are cultivated for their storage roots. The material may be of interest for regions, where the growing season is too short or the rainfall too low for sweet potato and cassava as well as to incorporate cultivar characteristics into wild V. vexillata accessions which are used for their storage roots.