Diversity among common bean populations from the Abruzzo region (Central Italy): A preliminary inquiry

The Abruzzo region (Central Italy) shows peculiar characteristics: the extension of mountain territory is the highest in mainland Italy and the whole region is less developed as compared to other Central Italy areas. Moreover, the farming systems developed with specific peculiarities parallel to shepherdy. Recently, in the area of the Gole di Popoli, ideally constituting the door of the inner and less developed part of the region, four populations of common bean were collected. Seed morphological traits, phaseolin and lectin profiles were analysed. Three populations appeared heterogeneous in phaseolin and lectin fractions. C, T, and S phaseolin types, C type being predominant, and six lectin patterns were observed. A comparison with common bean populations from the Basilicata region (Southern Italy) showed interesting differences between the two regions. The present contribution, besides the limitation due to the small number of samples, suggests that the Abruzzo, and other Italian regions with similar agro-environmental conditions, might retain highly interesting local populations.     

The common bean populations from Basilicata (Southern Italy). An evaluation of their variation

The variation within a collection constituted by 36 populations of common bean (Phaseolus vulgarisL.) recently collected in Basilicata region (Southern Italy) was studied. These populations are cultivated in marginal areas of the region mainly for farms' self-consumption. An appreciable variation of seed shape, colour and type of pattern was observed; 34 populations are of climbing growth habit. The frequency of the phaseolin types within the collection was investigated. C, T and S phaseolin types were observed, the type C being predominant (18 of 36 landraces); intra-population variation was detected only for two landraces. Local populations were compared with 28 commercial cultivars widely grown in the Basilicata region and accounting for 80–90% of common bean production. Significant divergences in growth habit and frequency of phaseolin types were observed: 22 cultivars had bush growth habit and 20 showed T phaseolin type. These results stress the urgency of actions devoted to the safeguard of this local germplasm. In fact, the diffusion of alloctonous and not typical germplasm is the first step towards the erosion of useful or valuable genes present in those established local populations.     

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.     

Extensive introgression of Middle American germplasm into Chilean common bean cultivars

Summary The genetic diversity of 95. representative Chilean common bean (Phaseolus vulgaris L.) landraces was analyzed using phaseolin seed protein and eight isozyme systems as genetic markers. Four types of phaseolin were found, “C”, “T”, “S” and “H”, in decreasing order of frequency. Each type had a different distribution between the Northern and Southern regions of the country. Nei’s genetic distance based on isozyme diversity indicated that a high percentage of the total variation found in this sample occurred between landraces and only a small percentage of the variation was detected within populations. Cluster analysis based on Nei’s genetic distance and a principal component analysis of isozyme frequencies did not detect a clear association between the geographic distribution of the landraces and their isozyme constitution. However, Nei’s genetic distance analysis clustered the bean landraces into two major groups which had a specific isozyme pattern, seed color, and seed size. The genetic analysis also detected a rare polymorphism for theMdh-2 locus, a null allele at theDiap-2 locus, and polymorphism for theAco-2 locus. The principal component analysis of isozyme frequencies showed that only 30% of the genotypes analyzed were similar to the Andean check and 5% of the samples were similar to Middle American check. This finding suggests a high frequency of hybridization between the Middle America and Andean gene pools in cultivated common bean from Chile.     

Characterization of Some Italian Common Bean (<Emphasis Type="Italic">Phaseolus Vulgaris</Emphasis> L.) Landraces by RAPD,Semi-random and ISSR Molecular Markers

Randomly amplified polymorphic DNA (RAPD), inter-simple sequence repeat (ISSR) and a semi-random PCR system were used to analyze the genetic diversity of 16 Italian common bean landraces and their relationship to four commercial cultivars. Of the primers tested, 8 ISSR, 6 RAPD and 7 semi-random primers produced polymorphic and reproducible DNA fragments. A higher proportion of polymorphic bands were observed using ISSR (85%) and semi-random (90%) primers than RAPD (69%) method. The combination of any two semi-random markers allowed the identification of all 20 bean genotypes. In contrast ISSR (except for primer (CAC)₃GC) and RAPD markers appeared to be less informative as more than two markers were necessary to achieve the same diagnostic level. Moreover, 7 ISSR, 2 RAPD and 8 semi-random exclusive bands were identified as putative population-specific markers. Semi-random and ISSR derived dendrograms showed similar tendencies in terms of genetic relatedness, whereas clustering of genotypes within groups was not similar when compared with the RAPD technique. Despite the different ability to resolve genetic variation among the investigated landraces, two major clusters with less than 60% (ISSR) and 40% (RAPD and semi-random) genetic similarity were formed with all three marker systems. The two groups were correlated with the phaseolin patterns and seed size of the landraces. The analysis showed that the cultivar ȁ8Lingua di Fuocoȁ9 and most of the landraces (13 out of 16) collected in Italy belong to the Andean gene pool, whereas only the three populations from Pratomagno belong to the Middle American gene pool.     

Can Phaseolin Patterns Help Resolve the Phaseolus–Vigna Complex?

Although common bean (Phaseolus vulgaris L.) is cultivated throughout India, the Himalayas hold largest diversity of bean germplasm. No studies on characterization of phaseolin types on this germplasm have been conducted earlier. In order to determine whether the common bean cultivars collected from various areas in the Northern Himalayas represent introductions from the Central American and Andean domestication centers or are local domesticates, we have analyzed the electrophoretic variation (SDS-PAGE) of phaseolin types in several bean accessions. A few species of Vigna were also included in this study to determine whether phaseolin (vignin in Vigna) patterns can be used to resolve the Phaseolus–Vigna complex. The present investigation on phaseolin (globulin) patterns of Phaseolus vulgaris and Vigna spp. clearly shows much variability in globulin patterns. Three new types of phaseolin patterns were recorded. An attempt to resolve phylogenetic problems in this complex was made using the phaseolin data.     

Insight into the origin of common bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.) grown in the state of Jammu and Kashmir of north-western Himalayas

Common bean (Phaseolus vulgaris L.) is considered as one of the principal grain legume crops grown in north-western Himalayan state of Jammu and Kashmir (J&K), India. Huge diversity has been observed for this crop in state Jammu and Kashmir. The hilly regions of the state J&K are famous for producing high quality, tasty and highly flavoured beans. In order to characterise this huge diversity and trace the origin of common bean, systematic efforts have been made for the first time in collection, evaluation and characterization of bean collection from Jammu & Kashmir. A set of 428 common bean lines were initially collected/procured and based on cluster analysis using few qualitative traits/site of collection, a diverse set of 96 lines was selected. The PCR assay for phaseolin locus led to the characterization of 96 lines into Meso-american and Andean types. Out of 96 lines tested, 45 possessed “S” type phaseolin and 51 possess “T” type phaseolin. The ITS region of selected local and exotic lines was Sanger sequenced and the sequence analysis of ~ 800 bp long region revealed the presence of 12 SNPs including one promising SNPs showing significant association with phaseolin patterns. The clustering based on ITS sequence data led to the clear cut separation of common bean lines into two distinct clusters based on their phaseolin types. The results of the present study helped to gain insights into the origin of common bean landraces grown in state of J&K, India.     

Diversity of common bean (Phaseolus vulgaris L.) germplasm from Portugal

Common bean (Phaseolus vulgarisL.) is a traditional crop in Portugal, where farmers growvarieties selected and maintained by themselves. A collection of 88landraces of common bean was evaluated for 17 quantitative andqualitative traits and the biochemical marker phaseolin to displaythe degree of variation of this germplasm. Agronomic data weresubjected to cluster analysis and several groups were identified,with three groups clustering most of the landraces. Regardingphaseolin variation the C and T banding patterns are the mostfrequent ones, so the origin of the Portuguese beans is thus probablythe Andean region of South America. These results give informationabout the origin, diversity and breeding value of the Portuguesegermplasm, that could be useful to widen the genetic base ofcurrently cultivated bean varieties in Europe.     

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

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

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.     

The Sierra Norte of Madrid: an agrobiodiversity refuge for common bean landraces

At the mountain area close to the city of Madrid, common beans were one of the main food crops present in everyday diet until 1960. This paper describes the morphological diversity, for forty-six phenological and morphological traits, found in forty-three traditional varieties of common beans collected in this area and seven commercial varieties used as reference. That comparison suggests that common bean breeding programs have led to later varieties with a higher production of straight pods. Quantitative traits showed also similar values when compared to the Iberian common bean collection, except for the phenological and seed size characters. The phenological differences could suggest an ecological adaptation of the studied landraces to the regional environmental conditions. The different seed size averages of both collections might correspond to the scarce presence of smaller seed-type common beans in Madrid collection. All the accessions collected in the Sierra Norte of Madrid belong to any of the groups included in the Spanish core collection. Madrilenian collection is also composed by indeterminate growth habit varieties, while the presence of bush accessions is relatively uncommon. Most of the Madrilenian landraces have a remarkable fitness for green-pod consumption (42 %) and their seeds are mainly white (30 %), ovate-shape (49 %) and medium-large (40 %). The study of seed storage protein allowed to classify the landraces according to their domesticated gene pools. Most of them (72 %), with T and C phaseolin type, seem to belong to Andean germplasm, while the remaining (28 %), with S and B phaseolin type, to Mesoamerican one. The remarkable morphological diversity of common beans found in this small area is a symptom of a broad genetic base despite genetic erosion, what indicates a widespread crop in the past. Therefore, it is advisable to design agro-environmental policies to promote the production and commercialization of common bean landraces in Sierra Norte of Madrid.     

Collecting maize (<Emphasis Type="Italic">Zea mays</Emphasis> L. convar. <Emphasis Type="Italic">mays</Emphasis>) with potential technological ability for bread making in Portugal

The Portuguese maize bread (“broa”) manufactured from traditional maize landraces still plays an important economic and social role on Central and Northern rural communities of the country. However the traditional maize landraces agricultural systems are changing. Local maize landraces are in risk of disappearing because of the progressive adoption of hybrid varieties not suitable for bread production. These changes are contributing to a major loss of genetic diversity. An expedition took place in the Central region of Portugal (Beira Interior and Beira Litoral) with the purpose of collecting enduring maize landraces with technological ability for bread production and to access the possibility of establishing a participatory plant breeding and conservation program. A total of 51 different maize landraces and 175 other varieties of associated crops were collected. Maize landraces showed to maintain high diversity and potential for improvement. The production relayed on small farms with polycrop, quality oriented, sustainable systems. A participatory plant breeding and conservation program would be possible on this region with the proviso that local authorities would be involved. This program would allow a higher valuation of these maize populations, contributing to halt the current loss of these unique Portuguese maize landraces.     

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.     

Different cucumber melon (<Emphasis Type="Italic">Cucumis melo</Emphasis> L.) races cultivated in Salento (Italy)

South Italy is still a centre of diversity for melons but new cultivars progressively replace its traditional landraces. The area of Salento is important for several typical old crops as the cucumber melon ‘Meloncella’ (Cucumis melo L. var. chate (Hasselq.) Filov) traditionally cultivated for its unripe fruits. Information on cultivation, quality and variation of this Apulian landrace (probably the last relic of a wider cultivation in Europe) is reported. A strategy for its characterization and safeguarding is in progress at the genebank of IGV of Bari (Italy).     

Assessment of genetic variation in common bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.) from Nebrodi mountains (Sicily,Italy)

This study was carried out to estimate the level of diversity existing within some common bean landraces still cultivated in Nebrodi mountains, North-western area of Sicily. The multidisciplinary approach adopted to reach this goal involved the characterisation of collected material through morphological, biochemical and molecular marker analyses. The nutritional quality of seeds was also investigated in view of the proposition of the best landraces as niche products. Results showed that those bean landraces retain a considerable level of heterogeneity. The use of both biochemical and molecular markers showed that all landraces clustered into two main groups, corresponding to the Andean and Mesoamerican gene pools. Our results suggest that the best strategy for preserving the diversity of common bean from a restricted area such as Nebrodi mountains, necessitates of a deep knowledge of germplasm to avoid the loss of precious genetic resources or, on the contrary, the safeguard of populations genetically redundant.     

The evolution of lentil (Lens culinaris Medik.) cultivation in Italy and its effects on the survival of autochthonous populations

At the beginning of the 20th Century Italy was one of the most important producers of lentil in the Mediterranean basin. Presently, this pulse is mainly cultivated in marginal areas of Central and Southern Italy and in some small islands. As a result of the reduced attention devoted to lentil, several autochthonous populations have disappeared together with the traditions related to their cultivation. Moreover, how long the cultivation of those still grown today will continue is unpredictable. Governmental and local associations have a time window for anticipating a possible epoch when to grow these landraces will be no longer profitable. In the last decades the Germplasm Institute of the National Research Council (Bari, Italy) and the Institut für Pflanzengenetik und Kulturpflanzenforschung (Gatersleben, Germany) have collected 63 lentil populations in Italy, presently stored ex situ. The evaluation of this collection has shown the existence of an appreciable morphological, agronomic and genetic diversity that could foster future breeding programmes. It is concluded that there is an urgent need to promote the survival of lentil populations in situ, since this will further support the adaptation and evolution of autochthonous genotypes in their original environments.     

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