roberto papa

Abstract: Knowledge about the origins and evolution of crop species represents an important prerequisite for efficient conservation and use of existing plant materials. This study was designed to solve the ongoing debate on the origins of the common bean by investigating the nucleotide diversity at five gene loci of a large sample that represents the entire geographical distribution of the wild forms of this species. Our data clearly indicate a Mesoamerican origin of the common bean. They also strongly support the occurrence of a bottleneck during the formation of the Andean gene pool that predates the domestication, which was suggested by recent studies based on multilocus molecular markers. Furthermore, a remarkable result was the genetic structure that was seen for the Mesoamerican accessions, with the identification of four different genetic groups that have different relationships with the sets of wild accessions from the Andes and northern Peru�Ecuador. This finding implies that both of the gene pools from South America originated through different migration events from the Mesoamerican populations that were characteristic of central Mexico.

Abstract: Multilocus digenic linkage disequilibria (LD) and their population structure were investigated in eleven landrace populations of barley (Hordeum vulgare ssp. vulgare L.) in Sardinia, using 134 dominant simple-sequence amplified polymorphism markers. The analysis of molecular variance for these markers indicated that the populations were partially differentiated (F ST = 0.18), and clustered into three geographic areas. Consistent with this population pattern, STRUCTURE analysis allocated individuals from a bulk of all populations into four genetic groups, and these groups also showed geographic patterns. In agreement with other molecular studies in barley, the general level of LD was low (13 % of locus pairs, with P < 0.01) in the bulk of 337 lines, and decayed steeply with map distance between markers. The partitioning of multilocus associations into various components indicated that genetic drift and founder effects played a major role in determining the overall genetic makeup of the diversity in these landrace populations, but that epistatic homogenising or diversifying selection was also present. Notably, the variance of the disequilibrium component was relatively high, which implies caution in the pooling of barley lines for association studies. Finally, we compared the analyses of multilocus structure in barley landrace populations with parallel analyses in both composite crosses of barley on the one hand and in natural populations of wild barley on the other. Neither of these serves as suitable mimics of landraces in barley, which require their own study. Overall, the results suggest that these populations can be exploited for LD mapping if population structure is controlled.

Abstract: Here, we present a brief overview of the main studies conducted on the common bean (Phaseolus vulgaris L.) in Europe and other countries outside its centres of origin. We focus on the proportions of the Andean and Mesoamerican gene pools, and on the inter-gene pool hybridization events. In Europe, for chloroplast microsatellites, 67% of European germplasm is of Andean origin. Within Europe, interesting trends have been seen; indeed, the majority of the Andean type is found in the three macro-areas of the Iberian Peninsula, Italy and central-northern Europe, while, in eastern and south-eastern Europe, the proportion of the Mesoamerican type increased. On a local scale, the contribution of the Mesoamerican type is always low. On other continents, various situations are seen using different markers: in China and Brazil, the Mesoamerican gene pool prevails, while in an African sample, overall, both gene pools are equally represented, with differences in individual countries. The frequency of European bean genotypes deriving from at least one hybridization event was 44% with an uneven distribution. Interestingly, hybrids tend to have intermediate seed size in comparison with �pure� Andean or Mesoamerican types. On other continents, very few hybrids are found, probably because of the different marker systems used.

Abstract: Phaseolus coccineus L. is closely related to P. vulgaris and is the third most important cultivated Phaseolus species. Little is known about the patterns of its diversity. In this work, a representative collection of its worldwide diversity was initially developed. The collection includes 23 wild forms (WFs) and 52 landraces (LRs) from Mesoamerica (the crop domestication area), and 148 LRs from Europe (where the crop was introduced in the XVI century). The collection was studied by using 12 SSR molecular markers that were developed for the P. vulgaris genome. They were proved to be effective and reliable in P. coccineus by this work. Fourteen LRs of P. dumosus (previously identified as a subspecies of P. coccineus) were also studied. The genetic diversity, population structure and phylogenetic relationships were investigated. The results indicate that a) the European and Mesoamerican gene pools are clearly differentiated, b) a certain reduction of diversity occurred with introduction into Europe, and c) the Mesoamerican LRs (P. dumosus included) and WFs are closely related and are connected by a high gene flow. This study provides a picture of the genetic diversity distribution and outcomes with introduction into the Old World which was not available before. It also underlines that the genetic diversity of both WFs and LRs is an important source for Phaseolus spp. breeding programs and deserves to be preserved in situ and ex situ.

Abstract: The olive is considered one of the most important fruit crops of the Mediterranean basin where it shows a wide range of variability, with about 2,000 cultivars. Italy, with about 500 cultivars, plays a fundamental role. The ability to discriminate olive cultivars and estimate genetic variability are important factors in better management of genetic resources and in helping to understand how genetic diversity is partitioned among cultivars. The two main objectives of the present investigation were to evaluate the identity of cultivars grown in Abruzzo region, central Italy, and to study their genetic structure. We applied amplified fragment length polymorphism (AFLP) methodology on 84 genotypes belonging to the most relevant and oldest varieties cultivated in Abruzzo and on six unknown genotypes. The information content of data was evaluated using the Marker Ratio index and the Polymorphic Index Content. Moreover, STRUCTURE software was used to investigate the genetic population structure. The analysis enabled us to clearly distinguish eight cultivars within seven clusters. Additionally, one cluster was found to have various minor cultivars and showed a relatively high level of diversity. The partitioning of genetic diversity showed that the largest amount of molecular variance was within groups. Our data suggest that both sexual and clonal propagation have played an important role in the evolution of olive cultivars. In our hypothesis, some ancestral population spread in central Italy with a relevant role of seed propagation, followed by a selection of superior clones from which more traditional varieties originated. In a few cases, hybridization should be taken into consideration to explain the diffusion of recently developed cultivars.

Abstract: Evolutionary studies in plant and animal breeding are aimed at understanding the structure and organization of genetic variations of species. We have identified and characterized a genomic sequence in Phaseolus vulgaris of 1,200 bp (PvSHP1) that is homologous to SHATTERPROOF-1 (SHP1), a gene involved in control of fruit shattering in Arabidopsis thaliana. The PvSHP1 fragment was mapped to chromosome Pv06 in P. vulgaris and is linked to the flower and seed color gene V. Amplification of the PvSHP1 sequence from the most agronomically important legume species showed a high degree of interspecies diversity in the introns within the Phaseoleae, while the coding region was conserved across distant taxa. Sequencing of the PvSHP1 sequence in a sample of 91 wild and domesticated genotypes that span the geographic distribution of this species in the centers of origin showed that PvSHP1 is highly polymorphic and, therefore, particularly useful to further investigate the origin and domestication history of P. vulgaris. Our data confirm the gene pool structure seen in P. vulgaris along with independent domestication processes in the Andes and Mesoamerica; they provide additional evidence for a single domestication event in Mesoamerica. Moreover, our results support the Mesoamerican origin of this species. Finally, we have developed three indel-spanning markers that will be very useful for bean germplasm characterization, and particularly to trace the distribution of the domesticated Andean and Mesoamerican gene pools.

Abstract: The potential of DNA barcoding was tested as a system for studying genetic diversity and genetic traceability in bean germplasm. This technique was applied to several pure lines of Phaseolus vulgaris L. belonging to wild, domesticated, and cultivated common beans, along with some accessions of Phaseolus coccineus L., Phaseolus lunatus L., and Vigna unguiculata (L.) Walp. A multilocus approach was exploited using three chloroplast genic regions (rbcL, trnL, and matK), four intergenic spacers (rpoB-trnC, atpBrbcL, trnT-trnL, and psbA-trnH), and nuclear ITS1 and ITS2 rDNA sequences. Our main goals were to identify the markers and SNPs that show the best discriminant power at the variety level in common bean germplasm, to examine two methods (tree based versus character based) for biodiversity analysis and traceability assays, and to evaluate the overall utility of chloroplast DNA barcodes for reconstructing the origins of modern Italian varieties. Our results indicate that the neighbor-joining method is a powerful approach for comparing genetic diversity within plant species, but it is relatively uninformative for the genetic traceability of plant varieties. In contrast, the character-based method was able to identify several distinct haplotypes over all target regions corresponding to Mesoamerican or Andean accessions; Italian accessions originated from both gene pools. On the whole, our findings raise some concerns about the use of DNA barcoding for intraspecific genetic diversity studies in common beans and highlights its limitations for resolving genetic relationships between landraces and varieties.

Abstract: Multilocus sequence data collected from domesticated and related wild relatives provides a rich source of information on the effect of human selection on the diversity and adaptability of a species to complex environments. To evaluate the domestication history of common bean (Phaseolus vulgaris L.), multilocus sequence data from landraces representing the various races within the Middle American (MA) and Andean gene pools was evaluated. Across 13 loci, nucleotide diversity was similar between landraces and wild germplasm in both gene pools. The diversity data were evaluated using the approximate Bayesian computation approach to test multiple domestication models and estimate population demographic parameters. A model with a single domestication event coupled with bidirectional migration between wild and domesticated genotypes fitted the data better than models consisting of two or three domestication events in each genepool. The effective bottleneck population size was ~50% of the base population in each genepool. The bottleneck began ~8200 and ~8500 years before present and ended at ~6300 and ~7000 years before present in MA and Andean gene pools respectively. Linkage disequilibrium decayed to a greater extent in the MA genepool. Given the (1) geographical adaptation bottleneck in each wild gene pool, (2) a subsequent domestication bottleneck within each gene pool, (3) differentiation into gene-pool specific races and (4) variable extents of linkage disequilibrium, association mapping experiments for common bean would more appropriately be performed within each genepool.

Abstract: Wild legumes constitute an important component of widespread pastures in the Mediterranean basin. This region is experiencing remarkable effects from climate change, and continuous monitoring of species and population dynamics is important in order to plan and enact valuable conservation programmes. Tripodion tetraphyllum (L.) Fourr. [=Anthyllis tetraphylla L.] (2n = 16), belongs to the tribe Loteae (Fabaceae), and could be very important for soil protection and sward improvement in abandoned or degraded Mediterranean areas. This alternative pasture legume is very closely related to Lotus japonicus and has some important characteristics for survival of the species in difficult and overgrazed Mediterranean areas. In this study, we have investigated the molecular diversity and population structure of T. tetraphyllum from North Africa using ISSR markers and plastidial microsatellites. To date, this is the first study concerning the genetic diversity and geographic differentiation of T. tetraphyllum. Ninety genotypes from three North African countries were analysed according to ISSRs, cpSSRs and one phenotypic trait. T. tetraphyllum shows a clear geographical structure, with differentiation associated with longitudinal differences; moreover, there is a general reduction in genetic diversity from Morocco to Tunisia. With all the markers used, strong differentiation was seen among collection sites. Our data highlight a genetic diversity gradient and cline of distribution, indicating that T. tetraphyllum has extended its area of distribution from Morocco to Tunisia.

Abstract: The objective of this study was to investigate the metabolite variations during industrial pasta processing (from semolina to dried pasta) for five different commercial products. Up to 76 metabolites were detected. Significant differences were observed between wholemeal and refined pasta samples, with the wholemeal pasta richer in many classes of compounds such as phytosterols, policosanols, unsaturated fatty acids, amino acids, carotenoids, minerals, and so on. Significant differences were also observed between samples of refined pasta apparently similar for the actual parameters used for the assessment of pasta quality. The results indicated that a number of metabolites undergo a transformation during the pasta-making process depending on the processing conditions adopted. The approach used in this work shows the high potential of metabolite profiling for food investigations with regard to process-related transformation, safety, and nutrition.

Abstract: The carotenoid concentration of durum wheat is a criterion for the assessment of semolina quality, and it is of particular importance in determining the color of pasta. Thus, the development of a rapid screening method for kernel and semolina color has facilitated increasing the yellow color in durum wheat cultivars. However, the distribution of the pigment might vary within the ear of the wheat. The micromethod for the determination of yellow pigment concentration (YPC) of single wheat kernels now allows the discovery of whether this variation is the same for cultivars with different carotenoid concentrations. All of the cultivars investigated showed similar trends in the arrangements of the kernels within the ears, with the central positions of the ears being the most stable for YPC. Indeed, the best combination of higher YPC and larger kernel size is seen for the basal-central region of the ear, which can be used for the selection during wheat-breeding programs of cultivars with a more intense yellow color.

Abstract: Abstract Molecular linkage maps are an important tool for gene discovery and cloning, crop improvement, further genetic studies, studies on diversity and evolutionary history, and cross-species comparisons. Linkage maps diVer in both the type of marker and type of population used. In this study, gene-based markers were used for mapping in a recombinant inbred (RI) population of Phaseolus vulgaris L. P. vulgaris, common dry bean, is an important food source, economic product, and model organism for the legumes. Gene-based markers were developed that corresponded to genes controlling mutant phenotypes in Arabidopsis thaliana, genes undergoing selection during domestication in maize, and genes that function in a biochemical pathway in A. thaliana. Sequence information, including introns and 3 UTR, was generated for over 550 genes in the two genotypes of P. vulgaris. Over 1,800 single nucleotide polymorphisms and indels were found, 300 of which were screened in the RI population. The resulting LOD 2.0 map is 1,545 cM in length and consists of 275 gene-based and previously mapped core markers. An additional 153 markers that mapped at LOD <1.0 were placed in genetic bins. By screening the parents of other mapping populations, it was determined that the markers were useful for other common Mesoamerican £ Andean mapping populations. The location of the mapped genes relative to their homologs in Arabidopsis thaliana (At), Medicago truncatula (Mt), and Lotus japonicus (Lj) were determine by using a tblastx analysis with the current pseduochromosome builds for each of the species. While only short blocks of synteny were observed with At, large-scale macrosyntenic blocks were observed with Mt and Lj. By using Mt and Lj as bridging species, the syntenic relationship between the common bean and peanut was inferred.

Abstract: This study focuses on the expansion of Phaseolus vulgaris in Europe. The pathways of distribution of beans into and across Europe were very complex, with several introductions from the New World that were combined with direct exchanges between European and other Mediterranean countries. We have analyzed here six chloroplast microsatellite (cpSSR) loci and two unlinked nuclear loci (for phaseolin types and Pv-shatterproof1). We have assessed the genetic structure and level of diversity of a large collection of European landraces of P. vulgaris (307) in comparison to 94 genotypes from the Americas that are representative of the Andean and Mesoamerican gene pools. First, we show that most of the European common bean landraces (67%) are of Andean origin, and that there are no strong differences across European regions for the proportions of the Andean and Mesoamerican gene pools. Moreover, cytoplasmic diversity is evenly distributed across European regions. Secondly, the cytoplasmic bottleneck that was due to the introduction of P. vulgaris into the Old World was very weak or nearly absent. This is in contrast to evidence from nuclear analyses that have suggested a bottleneck of greater intensity. Finally, we estimate that a relatively high proportion of the European bean germplasm (about 44%) was derived from hybridization between the Andean and Mesoamerican gene pools. Moreover, although hybrids are present everywhere in Europe, they show an uneven distribution, with high frequencies in central Europe, and low frequencies in Spain and Italy. On the basis of these data, we suggest that the entire European continent and not only some of the countries therein can be regarded as a secondary diversification center for P. vulgaris. Finally, we outline the relevance of these inter-gene pool hybrids for plant breeding.

Abstract: Background The yellow colour of pasta products is one of the main criteria used by consumers to assess pasta quality. This character is due to the presence of carotenoid pigments in semolina. During pasta processing, oxidative degradation of carotenoid pigments occurs mainly due to lipoxygenase (LOX). In durum wheat (Triticum durum Desf.), two Lpx-1 genes have been identified on chromosome 4B, Lpx-B1.1 and Lpx-B1.2, and evidences have been reported that the deletion of Lpx-B1.1 is associated with a strong reduction in LOX activity in semolina. In the present study, we characterised the Lpx-B1 gene family identified in a durum wheat germplasm collection and related the distribution and expression of the Lpx-B1 genes and alleles to variations in LOX activity in the mature grains. Results In addition to the already known Lpx-B1.1 and Lpx-B1.2 genes, a new gene was identified, Lpx-B1.3, along with three different Lpx-B1.1 alleles, Lpx-B1.1a, Lpx-B1.1b and the partially deleted Lpx-B1.1c. Screening of the germplasm collection showed that all of the genotypes have one of the three Lpx-B1.1 alleles, associated with either Lpx-B1.2 or Lpx-B1.3, thus showing that in this collection the two genes are alternatives. Therefore, based on Lpx-B1 distribution, three different haplotypes were distinguished: haplotype I, carrying Lpx-B1.3 and the Lpx-B1.1b allele; haplotype II carrying Lpx-B1.2 and the Lpx-B1.1a allele; and haplotype III carrying Lpx-B1.2 and the Lpx-B1.1c allele. Determination of Lpx-B1 transcript abundance and total LOX activity in mature grains revealed differences among these three haplotypes: haplotypes I, II and III showed high, intermediate and low levels, respectively, of functional Lpx-B1 transcripts and enzymatic activity. Conclusions In this germplasm collection, the Lpx-B1 gene family accounts for most of the total LOX activity in the mature grains. Information on these Lpx-B1 haplotypes provides significant improvement for prediction of LOX-1 activity levels in mature grains, and will therefore help in breeding programmes aimed at selection of new durum wheat genotypes with higher carotenoid contents in their end products.

Abstract: Background Among the cereal crops, barley is the species with the greatest adaptability to a wide range of environments. To determine the level and structure of genetic diversity in barley (Hordeum vulgare L.) landraces from the central highlands of Ethiopia, we have examined the molecular variation at seven nuclear microsatellite loci. Results A total of 106 landrace populations were sampled in the two growing seasons (Meher and Belg; the long and short rainy seasons, respectively), across three districts (Ankober, Mojanawadera and Tarmaber), and within each district along an altitudinal gradient (from 1,798 to 3,324 m a.s.l). Overall, although significant, the divergence (e.g. FST) is very low between seasons and geographical districts, while it is high between different classes of altitude. Selection for adaptation to different altitudes appears to be the main factor that has determined the observed clinal variation, along with population-size effects. Conclusions Our data show that barley landraces from Ethiopia are constituted by highly variable local populations (farmer's fields) that have large within-population diversity. These landraces are also shown to be locally adapted, with the major driving force that has shaped their population structure being consistent with selection for adaptation along an altitudinal gradient. Overall, our study highlights the potential of such landraces as a source of useful alleles. Furthermore, these landraces also represent an ideal system to study the processes of adaptation and for the identification of genes and genomic regions that have adaptive roles in crop species.

Abstract: Studies of the level and the structure of the genetic diversity of local varieties of Phaseolus vulgaris are of fundamental importance, both for the management of genetic resources and to improve our understanding of the pathways of dissemination and the evolution of this species in Europe. We have here characterized 73 local bean populations from Sardinia (Italy) using seed traits and molecular markers (phaseolins, nuSSRs and cpSSRs). American landraces and commercial varieties were also included for comparison. We see that: 1) the Sardinian material is distinct from the commercial varieties considered; 2) the variation in the seed traits is high and it mostly occurs among populations (95%); 3) compared to the American sample and the commercial varieties, the Sardinian collection has a low level of diversity; 4) the majority (>95%) of the Sardinian individuals belong to the Andean gene pool; 5) the Sardinian material shows a strong genetic structure, both for cpSSRs and nuSSRs; 6) the nuSSRs and cpSSRs concur in differentiating between gene pools, but a lack of congruence between nuclear and chloroplast has been observed within gene pools; and 7) there are three putative hybrids between the Andean and Mesoamerican gene pools. Despite the relatively low level of diversity, which is probably due to a strong founder effect, the Sardinian landraces are worth being conserved and studied further because of their distinctiveness and because hybridization within and between the gene pools could generate variation that will be useful for breeding.

Abstract: Landraces are domesticated local plant varieties that did not experienced a deliberate and intensive selection during a formal breeding programme. In Europe, maize landraces are still cultivated, particularly in marginal areas where traditional farming is often practised. Here, we have studied the evolution of flint maize landraces from central Italy over 50 years of on-farm cultivation, when dent hybrid varieties were introduced and their use was widespread. We have compared an �old� collection, obtained during the 1950s, before the introduction of hybrids, and a recent collection of maize landraces. For comparison, a sample of maize landraces from north Italy, and of improved germplasm, including hybrids and inbred lines were also used. A total of 296 genotypes were analysed using 21 microsatellites. Our results show that the maize landraces collected in the last 5-10 years have evolved directly from the flint landrace gene pool cultivated in central Italy before the introduction of modern hybrids. The population structure, diversity and linkage disequilibrium analyses indicate a significant amount of introgression from hybrid varieties into the recent landrace populations. No evidence of genetic erosion of the maize landraces was seen, suggesting that in-situ conservation of landraces is an efficient strategy for preserving genetic diversity. Finally, the level of introgression detected was very variable among recent landraces, with most of them showing a low level of introgression; this suggests that co-existence between different types of agriculture is possible, with the adoption of correct practices that are aimed at avoiding introgression from undesired genetic sources.

Abstract: In Ethiopia, barley is generally grown in two different planting seasons per year: during the long rainy season (Meher) and the short rainy season (Belg). The aim of the present study was to assess for the first time the role of this 'two-season system' on the structure of the genetic diversity of the Ethiopian barley landraces. We characterised 3,170 individual genotypes from 106 landrace populations using eight morphological spike traits. The diversity within population was higher in the season where barley is more important (Belg), and in general, where its cultivation is in larger plots because of weaker 'competition' with others crops. This indicates that barley diversity has a complex relationship with variations in the surrounding agro-ecosystem. Overall, the divergence between the two seasons was quite small (3.4%), suggesting that seed flow does not occur independently across the years within the two seasons. This would affect the amount of mutations and historic recombination that have accumulated within these populations

Abstract: Chloroplast microsatellites (cpSSRs) provide a powerful tool to study the genetic variation and evolution of plants. We have investigated the usefulness of 39 primer pairs tagging cpSSR loci on a set of eight different genera of Leguminosae (Papilionoideae subfamily) and five species belonging to the Phaseolus genus. Thirty-six �universal� primer pairs were retrieved from the literature, one was re-designed and a further two were designed de novo. The cpSSR loci analysed were highly polymorphic across the individuals examined. Twenty-seven primer pairs were polymorphic in the overall sample, 18 within the Phaseolus genus, and 16 in both P. vulgaris and P. coccineus. The analysis of the plastome sequences of four Leguminosae species (obtained from GenBank) showed that in the loci targeted by universal primer pairs: 1) the originally tagged cpSSRs can be lost; 2) other cpSSRs can be present; and 3) polymorphism arises not only from differences in the numbers of cpSSR repeats, but often from other insertion/ deletion events. Multilocus linkage disequilibrium analysis suggests that homoplasy is not a major problem in our dataset and principal component analysis indicates intelligible relationships among the species considered. Our study demonstrates that this set of chloroplast markers are a useful tool to study the diversity and the evolution of several legumes, and particularly of P. vulgaris and P. coccineus.

Abstract: Together with the knowledge of the population structure, a critical aspect for the planning of association and/or population genomics studies is the level of linkage disequilibrium (LD) that characterizes the species and the population used for such an analysis. We have analyzed the population structure and LD in wild and domesticated populations of Phaseolus vulgaris L. using AFLP markers, most of which were genetically mapped in two recombinant inbred populations. Our results reflect the previous knowledge of the occurrence of two major wild gene pools of P. vulgaris, from which two independent domestication events originated, one in the Andes and one in Mesoamerica. The high level of LD in the whole sample was mostly due to the gene pool structure, with a much higher LD in domesticated compared to wild populations. In relation to association studies, our results also suggest that whole-genome-scan approaches are feasible in the common bean. Interestingly, an excess of inter-chromosomal LD was found in the domesticated populations, which suggests an important role for epistatic selection during domestication. Moreover, our results indicate the occurrence of a strong bottleneck in the Andean wild population before domestication, suggesting a Mesoamerican origin of P. vulgaris. Finally, our data support the occurrence of a single domestication event in Mesoamerica, and the same scenario in the Andes.

Abstract:

Abstract: Genotype-by-environment interactions (GEIs) can affect breeding progress because they often complicate the evaluation and selection of superior genotypes. This drawback can be reduced by gaining insights into GEI processes and genotype adaptation. Here, we have studied the GEIs over a set of 24 barley genotypes that were grown across six environments (location-by-year combinations) in Sardinia, Italy. Three groups of genotypes were analysed: barley landraces (LANs), recombinant inbred lines (RILs), and commercial varieties (VARs). The additive main effects and multiplicative interaction (AMMI) model was used for data analysis, and results evidenced no significant differences in grain yield averages for the 24 genotypes. However, there was a relevant GEI for yield mainly between two of the six environments (one characterised by warm pre-anthesis period and high spring rainfalls, and the other characterised by opposite features) and two groups of genotypes (VAR and LAN). Moreover, a negative trade-off between yield levels of genotypes was seen when the barley genotypes were grown in the contrasting environments. Overall, intermediate GEI levels were seen for RILs in comparison to LANs and VARs, and some of the RILs provided valuable yield levels (e.g. RILs 23 and 52). The results thus show the potential usefulness of LANs as a genetic resource for breeding, e.g. to obtain genotypes adapted to Mediterranean environments, such as the RILs analysed in this study.

Abstract: The study of phenotypic and genetic diversity in landrace collections is important for germplasm conservation. In addition, the characterisation of very diversified materials with molecular markers offers a unique opportunity to define significant marker-trait associations of biological and agronomic interest. Here, 50 tomato landraces (mainly collected in central Italy), nine vintage and modern cultivars, and two wild outgroups were grown at two locations in central Italy and characterised for 15 morpho-physiological traits and 29 simple sequence repeat (SSR) loci. The markers were selected to include a group of loci in regions harbouring reported quantitative trait loci (QTLs) that affect fruit size and/or shape (Q-SSRs) and a group of markers that have not been mapped or shown to have a priori known linkage (NQ-SSRs). As revealed by univariate and multivariate analyses of morphological data, the landraces grouped according to vegetative and reproductive traits, with emphasis on fruit size, shape and final destination of the product. Compared to the low molecular polymorphism reported in tomato modern cultivars, our data reveal a high level of molecular diversity in landraces. Such diversity has allowed the inference of the existence of a genetic structure that was factored into the association analysis. As the proportion of significant associations is higher between the Q-SSR subset of markers and the subset of traits related to fruit size and shape than for all of the other combinations, we conclude that this approach is valid for establishing true-positive marker-trait relationships in tomato.

Abstract: Abstract 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.

Abstract: Background and Aims The main aim of this study was to use an amplified fragment length polymorphism (AFLP)-based, large-scale screening of the whole genome of Phaseolus vulgaris to determine the effects of selection on the structure of the genetic diversity in wild and domesticated populations. Methods Using pooled DNA samples, seven each of wild and domesticated populations of P. vulgaris were studied using 2506 AFLP markers (on average, one every 250 kb). About 10 % of the markers were also analysed on individual genotypes and were used to infer allelic frequencies empirically from bulk data. In both data sets, tests were made to determine the departure from neutral expectation for each marker using an F(ST)-based method. Key Results The most important outcome is that a large fraction of the genome of the common bean (16 %; P <0.01) appears to have been subjected to effects of selection during domestication. Markers obtained in individual genotypes were also mapped and classified according to their proximities to known genes and quantitative trait loci (QTLs) of the domestication syndrome. Most of the markers that were found to be potentially under the effects of selection were located in the proximity of previously mapped genes and QTLs related to the domestication syndrome. Conclusions Overall, the results indicate that in P. vulgaris a large portion of the genome appears to have been subjected to the effects of selection, probably because of linkage to the loci selected during domestication. As most of the markers that are under the effects of selection are linked to known loci related to the domestication syndrome, it is concluded that population genomics approaches are very efficient in detecting QTLs. A method based on bulk DNA samples is presented that is effective in pre-screening for a large number of markers to determine selection signatures.

Abstract: The main aim of this study was to test the patterns of sequence divergence and haplotype structure at the MAT locus of Pyrenophora teres, the causal agent of barley 'net blotch' disease. P. teres is a heterothallic ascomycete that co-occurs in two symptomatological forms, the net form (NF) and the spot form (SF). The mating-type genes MAT1-1-1 and MAT1-2-1 were sequenced from 22 NF isolates (12 MAT1-1-1 and 10 MAT1-2-1 sequences) and 17 SF isolates (10 MAT1-1-1 and seven MAT1-2-1 sequences) collected from Sardinian barley landrace populations and worldwide. On the basis of a parsimony network analysis, the two forms of P. teres are phylogenetically separated. More than 85% of the total nucleotide variation was found between formae speciales. The two forms do not share any polymorphisms. Six diagnostic nucleotide polymorphisms were found in the MAT1-1-1 intron (1) and in the MAT1-1-1 (3) and MAT1-2-1 (2) exons. Three diagnostic non-synonymous mutations were found, one in MAT1-1-1 and two in MAT1-2-1. For comparison with P. teres sequence data, the mating-type genes from Pyrenophora graminea were also isolated and sequenced. Divergence between P. graminea and P. teres is of a similar magnitude to that between NF and SF of P. teres. The MAT genes of P. graminea were closer to those of SF than to NF, with the MAT1-2-1 SF peptide not different from the MAT1-2-1 peptide of P. graminea. Overall, these data suggest long genetic isolation between the two forms of P. teres and that hybridization is rare or absent under field conditions, with each form having some particular niche specialization. This indicates that research on resistance to P. teres should consider the two forms separately, as different species.

Abstract: A deeper understanding of random markers is important if they are to be employed for a range of objectives. The sequence specific amplified polymorphism (S-SAP) technique is a powerful genetic analysis tool which exploits the high copy number of retrotransposon long terminal repeats (LTRs) in the plant genome. The distribution and inheritance of S-SAP bands in the barley genome was studied using the Steptoe � Morex (S � M) double haploid (DH) population. Six S-SAP primer combinations generated 98 polymorphic bands, and map positions were assigned to all but one band. Eight putative co-dominant loci were detected, representing 16 of the mapped markers. Thus at least 81 of the mapped S-SAP loci were dominant. The markers were distributed along all of the seven chromosomes and a tendency to cluster was observed. The distribution of S-SAP markers over the barley genome concurred with the knowledge of the high copy number of retrotransposons in plants. This experiment has demonstrated the potential for the S-SAP technique to be applied in a range of analyses such as genetic fingerprinting, marker assisted breeding, biodiversity assessment and phylogenetic analyses

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Abstract: Efficient assessment of genetic diversity is important for conservation and utilization of genetic resources. We sought to assess the ability of inter simple sequence repeats (ISSRs) as molecular markers to identify genetic diversity both within and among sympatric populations of common bean, Phaseolus vulgaris L. One wild and four domesticated populations originating from the Sierra Norte de Puebla, Mexico, were chosen because they were growing in the same region or field. The ISSR diversity was assessed using four primers that revealed a higher number of strong, polymorphic bands in preliminary analyses. Fifty of these bands could be mapped onto the core linkage map in population BAT93 x Jalo EEP558, whereas four were unlinked. The mapped bands were distributed over nine of the 11 linkage groups, suggesting that they were broadly distributed in the genome. On the basis of a sample of 50 intense bands in these five populations, ISSRs were able to clearly distinguish all populations. Most of the variation was distributed among populations rather than within populations, consistent with the predominantly selfing nature of the species. Differentiation among domesticated populations was much higher (FST {approx} 0.49�0.85) than between the wild and domesticated gene pools (FST = 0.05). Within each population, most loci had achieved near-fixation. Around 7% of individuals showed a lack of correlation between seed type and ISSR fingerprint. Furthermore, each population contained individuals with unusual markers but present in the other populations (frequency < 5 or 10%). Two nonmutually exclusive explanations were discussed�incomplete lineage sorting and introgression�to account for the presence of these unusual individuals. Overall, ISSR markers were very useful to differentiate closely related populations. Further research is necessary to quantify the actual level of outcrossing.

Abstract: Pyrenophora teres f. sp. teres mating-type genes (MAT-1: 1190 bp; MAT-2: 1055 bp) have been identified. Their predicted proteins, measuring 379 and 333 amino acids, respectively, are similar to those of other Pleosporales, such as Pleospora sp., Cochliobolus sp., Alternaria alternata, Leptosphaeria maculans, and Phaeosphaeria nodorum. The structure of the MAT locus is discussed in comparison with those of other fungi. A mating-type PCR assay has also been developed; with this assay we have analyzed 150 isolates that were collected from 6 Sardinian barley landrace populations. Of these, 68 were P. teres f. sp. teres (net form; NF) and 82 were P. teres f. sp. maculata (spot form; SF). Within each mating type, the NF and SF amplification products were of the same length and were highly similar in sequence. The 2 mating types were present in both the NF and the SF populations at the field level, indicating that they have all maintained the potential for sexual reproduction. Despite the 2 forms being sympatric in 5 fields, no intermediate isolates were detected with amplified fragment length polymorphism (AFLP) analysis. These results suggest that the 2 forms are genetically isolated under the field conditions. In all of the samples of P. teres, the ratio of the 2 mating types was consistently in accord with the 1:1 null hypothesis. This ratio is expected when segregation distortion and clonal selection among mating types are absent or asexual reproduction is rare. Overall, sexual reproduction appears to be the major process that equalizes the frequencies of the 2 mating types within populations.

Abstract: Lack of introgression or divergent selection may be responsible for the maintenance of phenotypic differences between sympatric populations of crops and their wild progenitors. To distinguish between these hypotheses, amplified fragment length polymorphism markers were located on a molecular linkage map of Phaseolus vulgaris relative to genes for the domestication syndrome and other traits. Diversity for these same markers was then analyzed in two samples of wild and domesticated populations from Mesoamerica. Differentiation between wild and domesticated populations was significantly higher in parapatric and allopatric populations compared to sympatric populations. It was also significantly higher near genes for domestication compared to those away from these genes. Concurrently, the differences in genetic diversity between wild and domesticated populations were strongest around such genes. These data suggest that selection in the presence of introgression appears to be a major evolutionary factor maintaining the identity of wild and domesticated populations in sympatric situations. Furthermore, alleles from domesticated populations appear to have displaced alleles in sympatric wild populations, thus leading to a reduction in genetic diversity in such populations. These results also provide a possible experimental framework for assessing the long-term risk of transgene escape and the targeting of transgenes inside the genome to minimize the survival of these transgenes into wild populations following introduction by gene flow.

Abstract: The genetic diversity of 66 Phaseolus genotypes was investigated, which included 14 local varieties of Phaseolus vulgaris and nine local varieties of P. coccineus, collected in Marche, central Italy. Their genetic diversity was assessed using three types of molecular marker: inter simple sequence repeats (ISSRs), nuclear gene-tagged simple sequence repeats (SSRs) and chloroplast simple sequence repeats (CpSSRs). Phaseolus vulgaris shows a higher genetic diversity than P. coccineus for the SSRs and CpSSRs, but not for the putative neutral ISSR markers. These data suggest that selection by farmers and adaptation to heterogeneous environments has maintained the diversity in landraces of the common bean. Comparing genetic diversity in Marche with that of the American controls reveals that 71% of the local P. vulgaris varieties in Marche are of Andean origin. The two gene pools of the common bean can be found on the same farm, and there is some evidence of past hybridization events between these two gene pools.

Abstract: A 16S rRNA gene-based fingerprinting method was developed for the identification of Azotobacteraceae and tested onto 48 soil isolates and 28 reference strains belonging to the free-living nitrogen-fixing bacterial group and to the most common species found in soil samples. According to this method, the 16S rRNA gene was amplified using universal primers for Eubacteria and PCR products were subsequently digested with RsaI, HhaI, HpaII, FnuDII, and AluI. The analysis of the restriction profiles obtained showed that the method is able to define a unique species-specific phylotype (SSP) for each of the eight Azotobacteraceae species tested. Cluster analysis was successfully employed for the identification of members of the family Azotobacteraceae, being assignation into species of the isolates confirmed by means of partial 16S rRNA gene sequencing.

Abstract: For the genus Anthyllis (Fam. Fabaceae, tribe Loteae), with few exceptions, little information is available on the genetic variation among and within species. This genus contains 20 species distributed throughout Europe, Africa, and the Mediterranean basin. The most widespread species is A. vulneraria, and over 30 intraspecies taxa have been identified based on plant morphology. To study the molecular phylogeny of the genus, the sequences of the internal transcribed spacers ITS1 and ITS2 of the nuclear ribosomal DNA of 10 Anthyllis species, including 11 subspecies of A. vulneraria and three subspecies of A. montana, were obtained and analysed together with sequences of five other species of the genus obtained from GenBank. Our results suggest that the genus Anthyllis is not monophyletic and is divided in two main clades: the Anthyllis sensu strictu and the "tetraphylla clade". The former includes most of the Anthyllis species, and the latter includes three annual species more closely related to Lotus. All the taxa were also analysed according to seven chloroplast microsatellites, and these data closely confirm the results obtained with the ITS phylogeny.

Abstract: Monoconidial cultures of Pyrenophora teres, the causal agent of barley net blotch, were isolated from leaves collected from six populations of the barley landrace "S'orgiu sardu" growing in five agro-ecological areas of Sardinia, Italy, and genotyped using AFLPs. The 150 isolates were from lesions of either the "net form" (P. teres f. sp. teres) or the "spot form" (P. teres f. sp. maculata) of the disease. Of 121 AFLP markers, 42%, were polymorphic. Cluster analysis resolved the isolates into two strongly divergent groups (F(ST) = 0.79), corresponding to the net (45% of the isolates) and the spot (55% of the isolates) forms (designated the NFR and SFR groups, respectively). The absence of intermediate genotypes and the low number of shared markers between the two groups indicated that hybridization between the two formae is rare or absent under the field condition of Sardinia. Five of the barley populations hosted both forms but in different proportions. The SFR populations were similar in overall polymorphism to the NFR populations. However, compared to the SFR form, the NFR occurred in all fields sampled and showed a higher population divergence (F(ST) = 0.43 versus F(ST) = 0.09 with all isolates; F(ST) = 0.37 versus F(ST) = 0.06 with clone corrected samples) probably due to a lower migration rate. AFLP fingerprints resolved 117 distinct genotypes among the 150 isolates sampled (78%), 87% in SFR and 68% in NFR isolates. Although the absolute numbers may be a function of the number of AFLP markers assayed, the relative difference suggests that clonality is more prevalent among the NFR isolates (with 11 of 46 haplotypes observed more than once), compared with SFR isolates (7 of 71 haplotypes). Both digenic and multilocus linkage disequilibrium analyses suggested that sexual reproduction occurs at significant levels within the NFR and SFR populations, and that the relative contribution of sexual and asexual reproduction varies among different environments.

Abstract: Using amplified fragment length polymorphisms (AFLPs), we analyzed the genetic structure of wild and domesticated common bean (Phaseolus vulgaris L.) from Mesoamerica at different geographical levels to test the hypothesis of asymmetric gene flow and investigate the origin of weedy populations. We showed both by phenetic and admixture population analyses that gene flow is about three- to four-fold higher from domesticated to wild populations than in the reverse direction. This result, combined with other work, points to a displacement of genetic diversity in wild populations due to gene flow from the domesticated populations. The weedy populations appear to be genetically intermediate between domesticated and wild populations, suggesting that they originated by hybridization between wild and domesticated types rather than by escape from cultivation. In addition, the domesticated bean races were genetically similar confirming a single domestication event for the Mesoamerican gene pool. Finally, the genetic diversity of the domesticated bean population showed a lower level of geographic structure in comparison to that of the wild populations.

Abstract: Gene flow is a potential concern associated with the use of transgenic crops because it could affect genetic diversity of related landraces and wild relatives. This concern has taken on added importance with the looming introduction of transgenic crops in centers of crop domestication (Mexico, China) and those producing pharmaceutical compounds. For gene flow to take place among cultivars and their wild relatives, several steps have to be fulfilled, including the presence of cultivars or wild relatives within pollen or seed dispersal range, the ability to produce viable and fertile hybrids, at least partial overlap in flowering time, actual gene flow by pollen or seed, and the establishment of crop genes in the domesticated or wild recipient populations. In contrast with domestication genes, which often make crops less adapted to natural ecosystems, transgenes frequently represent gains of function, which might release wild relatives from constraints that limit their fitness. In most sexually reproducing organisms, the chromosomal region affected by selection of a single gene amounts to a small percentage of the total genome size. Because of gene flow, the level of genetic diversity present in the domesticated gene pool becomes a crucial factor affecting the genetic diversity of the wild gene pool. For some crops, such as cotton and maize, the introduction of transgenic technologies has led to a consolidation of the seed industry and a reduction in the diversity of the elite crop gene pool. Thus, diversity in improved varieties grown by farmers needs to be monitored. Several areas deserve further study, such as the actual magnitude of gene flow and its determinants in different agroecosystems, the long-term effects of gene flow on genetic diversity both across gene pools and within genomes, the expression of transgenes in new genetic backgrounds, and the effects of socio-economic factors on genetic diversity.

Abstract: Amplified fragment length polymorphisms (AFLPs) were used to evaluate genetic relationships within cowpea [ Vigna unguiculata (L.) Walp.] and to assess the organization of its genetic diversity. Nei's genetic distances were estimated for a total of 117 accessions including 47 domesticated cowpea (ssp. unguiculata var. unguiculata), 52 wild and weedy annuals (ssp. unguiculata var. spontanea), as well as 18 perennial accessions of the wild subspecies pubescens, tenuis and alba. AFLP variation was also used to study genetic variation among and within domesticated and wild accessions based on their geographical origin (western, eastern and southern Africa). Wild annual cowpea (var. spontanea) ( H (T)=0.175) was more diverse than domesticated cowpea ( H (T)=0.108). Wild cowpea was more diverse in eastern ( H (S)=0.168) than in western Africa ( H (S)=0.129), suggesting an eastern African origin for the wild taxon. The AFLP data were consistent with earlier findings of a unique domestication event in cowpea in the northern part of the continent and suggested that domestication in eastern or southern Africa was unlikely. It did not allow a more precise localization of domestication due to extensive gene flow between wild and domesticated forms that has led to a large crop-weed complex distributed over the entire African continent. In addition, wild materials from northeastern Africa are still lacking. Overall, the superiority of the AFLP technique over isozymes resided in its ability to uncover variation both within domesticated and wild cowpea, and should be a powerful tool once additional wild material becomes available.

Abstract: One hundred and six landraces belonging to 7 species of the Triticeae tribe were collected in central Italy by DBVBA (Perugia University), DIBIAGA (Ancona University) and ARSSA (Abruzzo Region Agricultural Development Agency) in different individual and joint missions. A few accessions were supplied by private and other public organisations. Triticum dicoccum Schubler is the most widespread species, followed by T. aestivum L., T. monococcum L., T. spelta L., T. turgidum var. durum Desf., Secale cereale L. and Hordeum vulgare L. Besides the presence of landraces reproduced by farmers over generations, information related to on-farm management and to qualitative/organoleptic traits as well as information related to their local names, uses, traditions and social context was gathered during the missions. The majority of the accessions was characterised by morphological and phenological traits and molecular markers. This work shows the presence of morpho-phenologic and genetic differences among landraces and the importance of some species in the agricultural systems and food customs of the investigated area. Particularly for emmer three well distinct landraces are present, "Farro Italia Centrale", "Farro della Garfagnana" and "Farro Italia Meridionale". Other interesting and traditional landraces are the "Solina" common wheat in Abruzzo and the "Orzo mondo" naked barley in Marche. Most of the populations are still cultivated in marginal lands and under low input or organic agronomic conditions; nevertheless, in many cases, they are found near modern varieties in conventional agriculture systems. Moreover, the in situ (on-farm) conservation of Triticeae landraces in central Italy is strictly linked to elderly farmers.

Abstract: Barley is the second most widely cultivated cereal on the island of Sardinia. It is grown for green fodder, grain and straw, all of which are used for animal feed. The cultivation of modern varieties is limited and many farmers grow local populations of the six-row landrace 'S'orgiu sardu'. Twelve local populations (20 strains per population) were evaluated for 13 random amplified polymorphic DNA (RAPD) markers, six isozyme loci and five morphophenological traits. Results showed that Sardinian barley is characterized by a lax six-row spike and is, in general, a spring type. Isozyme polymorphism was detected for only two esterase loci (Estl and Est4). The genetic variation for all types of markers was found located mostly within population (about 84% for isozymes and morphophenological traits and 89% for RAPDs). This finding appears to confinn the hypothesis that the populations studied belong to the same landrace called 'S'orgiu sardu' by farmers. A geographical differentiation between southern and northern populations was found in relation to distinct environmental characteristics. Taking into account all types of marker 209 (90.5%) different genotypes were identified out of 231 individuals. The overall results confirm the high variability that can be found within landrace populations, underlining the value of landraces as a source of genetic variation.

Abstract: Since ancient times, barley has been an important food resource for the people of Sardinia. The oldest traces of its cultivation are from the mid-Neolithic (fourth millennium B.C.). Archaeological, historical and anthropological aspects of barley cultivated in Sardinia are discussed in this paper. We describe the traditional process for making barley bread (orgiathu) in Sardinia, where a special starter called ghimisone was prepared. Today, barley is cultivated only as animal feed, with two uses, grain yield and grazing. Many farmers prefer to grow local populations belonging to landrace locally known as ldquoS' orgiu sardurdquo. Local Sardinian populations of barley evolved in diverse environments, being cultivated from sea-level up to 1000 m elevation, on various soil types at different intensities of abiotic stresses, and with climates and environments associated with various agricultural practices, depending both on production strategies and climatic conditions. These barley materials are thought to be valuable genetic and cultural inheritance which must be preserved and used for both productive and research purposes.

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Abstract: Barley varieties are known to differ in the extent of Na+ and Cl� accumulation in leaves when grown in saline soil or hydroponic culture. In particular, the cv. Chevron accumulates more Na+ than the more salt-tolerant cv. CM67, and has lower leaf K+ concentrations. When salt was applied as a spray to the leaves, CM67 accumulated more Na+ than Chevron, and the selection Sinis 27 (from a landrace collected on the Sinis Peninsula of Sardinia) accumulated more Na+ than Sinis 28. In some cases leaf K+ concentrations decreased in response to high concentrations of salt sprayed on to the leaves. Accumulation of Na+ was greater in the 4th leaf than in the flag leaf. Added CaCI2 had opposite effects when added to the salt applied to the soil or to the saline spray. In the soil, CaCI2 reduced Na+ uptake; applied to the leaf it increased Na+ uptake. Pre-wetting the leaves before the salt spray, or washing the leaves with non-saline water 1 h after the salt spray, reduced the uptake of Na+ and Cl�. It is clear that tolerance to salt applied as salt spray or in the soil are different characteristics.

Abstract: Agriculture is an history of a real symbiosis which developed between crops and man; crops have made it possible to sustain an ever increasing human population. During the nineteenth century a real revolution occurred in agricultural systems, characterized by sharp and significant increases in crop yields and thus in food supply. However, it has been estimated that the global human population will rise substantially in the next 40-50 years. To limit the disruption of natural habitats and the processes of desertification and land degradation, the increases in agricultural production will need to be derived mainly from increases in the productivity of agricultural systems. Increasing crop productivity by increasing agronomic inputs appears not to be feasible in developed countries because given the actual levels of inputs used in most of our agricultural systems, no additional yield return could be obtained. In this context, over the next few decades we will be facing a major challenge: to increase the productivity of agriculture systems in terms of goods and services, while reducing its environmental costs. This idea is the basis of the concept of sustainable agriculture. Sustainable agriculture differs from conventional agriculture in that it aims to maximise our dependence on naturally occurring biological systems. Such systems are less important in conventional agriculture, which is highly dependent on synthetic external inputs, tillage and irrigation. A major consequence of the differences between sustainable and conventional agriculture is that the varieties of plants best suited to the two approaches can be significantly different. As a consequence, breeding strategies that can address these new production circumstances are needed, which means breeding practices that can sustainably enhance agricultural productivity and profitability, while simultaneously addressing food security and biodiversity conservation challenges. On the basis of these considerations, plant breeding for sustainable agriculture has to be implemented considering the following main points: crop and agro-ecosystem diversity, selection environments and genotype x environment interactions, target traits, genomics-assisted breeding and biotechnology, socio-economic context.

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