Genetic diversity and population structure of a common bean (Phaseolus vulgaris L.) collection from Calabria (Italy)

Eighty-seven Phaseolus vulgaris landraces, still cultivated in Calabria (Italy), were investigated in order to study the patterns of common bean genetic diversity in this region, to better understand the evolutionary development of beans in Europe and to properly manage these genetic resources. Four American accessions and five Italian varieties were also included. Different markers, such as 12 microsatellites, seed traits, phaseolins and 100-seed weight were combined with different statistical approaches. For each microsatellite, expected (H _e ) and observed (H _o ) heterozygosities, polymorphism information content (PIC), probability of identity (PI) and homozygosity were calculated. Furthermore, in Calabrian group of bean landraces, total (N _a ) and private (N _pa ) number of alleles, observed (H _o ), expected heterozygosities (H _e ) and allelic richness (AR) were calculated. Genetic distances among landraces were estimated using Nei’s coefficient and a cluster analysis using the UPGMA algorithm was performed. The results clearly indicated that: (1) Calabrian germplasm showed a high level of diversity (H _e  = 0.595); (2) Mesoamerican and Andean gene pools were clearly distinguished in Calabrian germplasm, with the Andean gene pool predominating (83 %); (3) Calabrian landraces were largely hybridized within and between the gene pools. A model-based approach, using the STRUCTURE software, was adopted. Six groups, including 4 of Andean origin and one of Mesoamerican origin were identified. Even more interesting, a small group (8 %) showed a distinct genetic structure, in which interspecific hybridizations with runner bean (Phaseolus coccineus L.) could have occurred. Nevertheless, a relatively high proportion of Calabrian bean landraces (12.6 %) was derived from intra and interspecific hybridizations.     

Molecular characterisation of common bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.) accessions from Southwestern Uganda reveal high levels of genetic diversity

Common bean (Phaseolus vulgaris L.) was introduced to East Africa over 400 years ago and is today a fundamental part of food and income security of many smallholder farmers in the region. East Africa is among the top three bean producing and consuming regions of Africa with Uganda, Rwanda and Kenya being leaders in production. The leading common bean producing region in Uganda is the Southwestern (SW) highlands. Production is subsistent and farmers grow mixed varieties (accessions) to minimize losses. The aim of this study was to investigate the impact of farmer selection and production practices on genetic diversity of common bean in SW Uganda. 100 accessions were assembled from SW Uganda and assayed with 6 DNA simple sequence repeat (SSR) markers. A total of 41 alleles were detected giving an overall average gene diversity of 0.299 (30%) in both districts. The accessions clustered into two major gene pools i.e., Mesoamerican and Andean. Within each gene pool there was evidence of clonal populations suggesting wide distribution of certain accessions. Kabale district had a higher average gene diversity (38%) compared to Kisoro district (22%). Detection of Andean phaseolin in a clone set of Mesoamerican gene pool suggests introgession between the two groups.     

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.     

Genetic diversity in sorghum [<Emphasis Type="Italic">Sorghum bicolor</Emphasis> (L.) Moench] germplasm from Southern Africa as revealed by microsatellite markers and agro-morphological traits

Cultivated sorghum [Sorghum bicolor (L.) Moench] is an important food security crop in the semi-arid regions of the world including Asia and Africa. Its genetic diversity is contained mostly in traditional varieties and modern cultivars used by farmers. In this study, agro-morphological traits and molecular markers were used to assess genetic diversity in 22 accessions of cultivated sorghum from five countries (Botswana, Namibia, Swaziland, Zambia and Zimbabwe) in the Southern African Development Community (SADC) region. The study revealed a significant variation among 22 accessions in both qualitative and quantitative morphological traits, indicating the accessions’ promising potential as breeding material. For molecular analysis, 11 microsatellite primer-pairs were used, and generated a total of 70 alleles across 20 accessions. Analysis of molecular variance revealed a high level of genetic variation; 67 % among the accessions and 10 % among the five countries. The patterns of genetic diversity and the relationships observed in this study should provide insights for genetic resource conservation and utilization of sorghum germplasm in the SADC region.     

Water yam (Dioscorea alata L.) diversity pattern in Brazil: an analysis with SSR and morphological markers

Substantial progress was made in the last decade in understanding the diversity in Dioscorea species. However, most of the studies so far concentrated in the ‘yam belt in Africa’. We present a genetic diversity and structure analysis among commercial and local varieties of water yam (Dioscorea alata L.) in Brazil using microsatellite and morphological markers. Twelve microsatellite primers were used to generate DNA profiles of 72 local varieties and 17 commercial accessions of water yam collected in four different regions in Brazil. Also, four morphological traits were evaluated on individual plants under field conditions. The morphological characterization showed considerable diversity. High polymorphism was found with 100 % polymorphism observed for 11 primers and a discriminating power value of 0.92, on average. We did not observe a strong population structure among sampling regions, while most of the genetic diversity was concentrated within regions (95.9 %). Analysis of the relationship between accessions did not clearly separate the local and commercial genotypes. However, the molecular analyses of D. alata showed high intraspecific diversity in local accessions from different regions in Brazil, with the highest Shannon index value (H′ = 0.41) for the Southeast region. These results confirmed an admixture of accessions in all sampling regions, consistent with the lack of a significant correlation between geographic and genetic distances, suggesting that farmers exchanged water yam materials extensively. The genetic diversity can be explained by the result of a continuous exchange of accessions by farmers throughout Brazil.     

Microsatellites based marker molecular analysis of Ghanaian bambara groundnut (Vigna subterranea (L.) Verdc.) landraces alongside morphological characterization

Bambara groundnut an indigenous crop of African origin is drought tolerant and the third most important leguminous crop in Ghana and Sub-Saharan Africa. In order to assess the level of genetic diversity within a small collection of Ghanaian landraces, molecular analysis was performed using microsatellite markers alongside characterization of morphological features. Genetic distance based on Jaccard’s similarity coefficient from the SSR marker analysis ranged from 0.48 to 0.9 among the landraces. 80 individual genotypes were clustered into 17 units with substantial levels of inter- and intra-landrace polymorphism. The analyses of variance from the morphological characterization for all quantitative traits were statistically significant (p < 0.05) except for terminal leaflet width. The first 4 principal components accounted for 41.97, 20.15, 13.39 and 9.81 % respectively of the morphological variations among the landraces. Qualitative traits however accounted for less of these variations. The results of the present study support the availability of high level of polymorphism within the collection of bambara groundnut analysed. This report is useful to crop improvement and germplasm conservation of bambara groundnut in Ghana.     

Target region amplification polymorphism (TRAP) for assessing genetic diversity and marker-trait associations in chickpea (Cicer arietinum L.) germplasm

The effective utilization of crop diversity held in genebanks depends on our knowledge of useful traits and available markers associated with the target traits. Target region amplification polymorphism (TRAP) was used to evaluate the genetic diversity and underlying relationships among 263 accessions of chickpea landraces maintained by the USDA-ARS Western Regional Plant Introduction Station in Pullman, WA, USA. Two-hundred sixty-two TRAP markers were amplified by eight primer combinations. Altogether, 110 (42 %) markers were polymorphic, the other 152 (58 %) displayed no variation. These polymorphic markers revealed important differences among the accessions, with an estimated, mean pair-wise genetic distance of 25.82 %, ranging from 2.8 to 50.0 %. Genetic distance analysis divided the accessions into two major groups, with 113 and 150 accessions each, and substantial association between molecular diversity and geographic origin was evident. Bayesian analysis of population structure revealed two groups (K = 2) with evidence for six sub-groups. Additionally, the population structure of a subset of 110 lines was determined (K = 3) for testing marker-trait associations (MTAs). Phenotypic traits included the concentrations of protein and nine mineral elements in the seeds. Two MTAs were significant (p < 0.01) for concentrations of Ca and K, and three MTAs were significant for Cu and Ni concentrations. The results indicate that this population is useful for genome-wide association studies on other economic traits given the level of genetic diversity uncovered and the marker-trait associations in seed minerals discovered.     

Italian common bean landraces: diversity and population structure

Phaseolus vulgaris L. is an important species that originated in Mesoamerica. A Mesoamerican and an Andean gene pool are usually distinguished in the domesticated forms. Many bean landraces are still cultivated in Italy and the Department of Applied Biology maintains an ex situ collection of 146 landraces. Although protection schemes are being developed in Italy, most landraces are extinct or at risk of extinction. To facilitate their conservation and use, geographical, morphological, biochemical and molecular (SSR) data were collected and analysed to estimate the diversity and the genetic structure of the collection. Data confirmed that both the Mesoamerican and the Andean gene pools were introduced in Italy and, although a distinction between the two gene pools exists, the Italian landrace diversity is clearly structured in three clusters that are not simply ascribable to the original gene pools. The observed structure appears also to be due to adaptation to the different environmental conditions determined by altitude. This was confirmed by assessing the presence of selective effects for some of the SSR used in this study. Finally, a certain extent of admixture in Italian landrace diversity suggests past (or recurring) hybridisation events among gene pools. The combined use of morphological, biochemical and molecular data clearly distinguished almost all the landraces. The data gathered here can assist landrace in situ protection schemes that are being developed in Italy, be used to register landraces in the European common catalogue of ‘conservation varieties’ for seed commercialisation and contribute to a better use of Italian common bean diversity in breeding for organic and conventional production systems.     

利用SSR分析小豆种质遗传多样性

摘要：小豆是一类重要的食用豆,本研究利用45对SSR引物对小豆基因组DNA进行了SSR标记的筛选鉴定,共筛选出多态性良好、扩增效果稳定的SSR引物18对。利用筛选出的18个SSR标记,分析了来自我国栽培小豆优异种质53份和日本引进种质27份,旨在阐明其遗传多样性特点,为育种利用提供理论依据。结果表明,在所有参试的80份小豆种质中共鉴定出等位变异92个,平均每个位点为5.1个;其中53份国内小豆和27份日本小豆的等位变异数分别为89个和74个,平均每个位点分别为4.9个和4.1个。所有供试小豆平均每个位点的多态信息含量（PIC）为0.64,变化范围为0.23～0.83,其中国内小豆的平均PIC值为0.63,变化范围为0.23~0.86;日本小豆平均PIC值为0.61,变化范围为0.20~0.81。国内栽培小豆和日本小豆在等位变异数、多态信息含量（PIC）、遗传相似性系数均存在差异,UPGMA聚类分析将参试的80份小豆明显分为五大类,聚类结果与小豆种质地理起源呈现出一定的相关性。试验表明,在小豆遗传育种中,可以通过种质资源相互利用来拓宽育成品种的遗传基础,同时这些SSR标记对于小豆资源DNA指纹图谱构建、遗传作图、基因型鉴定及分子标记辅助育种具有重要意义。     

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.     

Evaluation of genetic diversity in a Camelina sativa (L.) Crantz collection using microsatellite markers and biochemical traits

A genomic DNA library enriched with GA/TC repeats from Camelina sativa variety Calena has been analysed. After sequencing of about 200 randomly selected clones, approximately 60 % of them showed to contain simple or compound microsatellites with a high number of repeats. Among all microsatellite markers analysed 15 primer pairs amplified polymorphic fragments. Forty C. sativa accessions of different origin were genotyped with 15 microsatellite markers that generated 134 alleles with an average of 8.93 alleles per locus. The observed heterozygosity (Ho) among the accessions ranged from 0.0 to 0.15 with an average of 0.0370, whereas the average of expected heterozygosity (He) among accessions was 0.2769. The analysis of the average total heterozygosity (H_T = 0.651), the intrapopulation genetic diversity (H_S = 0.260), the interpopulation genetic diversity (D_ST = 0.391) and the coefficient of genetic differentiation among populations (G_ST = 0.574) demonstrated that 57.4 % of the genetic diversity is among the accessions, while 42.6 % resides within them. Phylogenetic tree of the 40 C. sativa accessions was constructed based on Nei’s genetic distance. The unweighted pair group method with arithmetic mean (UPGMA) dendrogram shows, except for CAM108 and CAM170, a clear discrimination among C. sativa accessions grouping them in five subgroups. ANOVA analysis indicates significant differences in some biochemical and agronomic parameters among the C. sativa accessions grouped according to Nei’s genetic distance. The result of the Tukey HSD test demonstrated that the A4 subgroup showed a significant higher TWS and linoleic acid (LA) content, while the subgroup A1 showed a significant higher linolenic and lower LA content compared to the remaining groups.     

Population structure and genetic differentiation among the USDA common bean (Phaseolus vulgaris L.) core collection

Genetic diversity data were collected from a large population of common bean (Phaseolus vulgaris L.) landraces representing the United States Department of Agriculture core collection. The data were based on microsatellite data from all linkage groups. A procedure was developed to determine if we collected sufficient marker data to adequately estimated pairwise diversity. The diversity data were used to define populations using distance and model-based approaches. Genetic differentiation and genetic isolation by distance data were collected. Diversity was also compared for markers linked and unlinked to domestication loci. Using a model-based approach, the landraces were divided into the traditional Middle American and Andean gene pools. Diversity was greater for the Middle American gene pool. Six Middle American and three Andean subpopulations were defined, and the Middle American subpopulations exhibited strong geographic identity. Unlike other studies, seed size varied considerably with subpopulations, and a number of the subpopulations contained landraces from multiple common bean races. All of the subpopulations were highly differentiated, with the Middle American subpopulations showing the greatest differentiation. Genetic isolation by distance was observed among the Middle American and Andean subpopulations but not among subpopulations within a gene pool. Within each gene pool, diversity was lower for markers linked to domestication loci.     

Agro-morphological and molecular variability in <Emphasis Type="Italic">Triticum boeoticum</Emphasis> accessions from Zagros Mountains,Iran

Genetic erosion in cultivated wheat provides a good reason for studying genetic diversity in crop wild relatives. In the present study, genetic diversity of 32 accessions belonging to T. boeoticum species collected from different parts of Iran were evaluated using 13 morphological traits as well as ten inter-simple sequence repeat primers. Statistical analysis for morphological traits showed significant differences among accessions (except number of fertile tillers and total tillers per plant). In principal component analysis, the first three PCA showed 82.65 % of the total morphological variation. Based on the morphological traits, accessions were separated into two main groups by cluster analysis. In molecular analysis, polymerase chain reactions amplified 105 DNA fragments, out of which, 95 (90.47 %) were polymorphic. From geographic perspective, the accessions sampled from western and southwestern of Iran showed the highest and lowest polymorphism, respectively. However, the maximum values of effective number of alleles (Ne), Nei’s gene diversity (He) and Shannon’s information index (I) was related to accessions collected from NW regions. Also, according to cluster analysis and PCoA plot genetic diversity was not related to geographical distribution. Overall, our results revealed a remarkable level of genetic diversity among studied Iranian T. boeoticum accessions; especially accessions collected from Kermanshah and Lorestan provinces, which can be of interest for future breeding programs. So, conservation of germplasm of these areas is recommended.     

Structure of genetic diversity among common bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.) varieties of Mesoamerican and Andean origins using new developed microsatellite markers

A common bean genomic library was constructed using the ‘IAC-UNA’ variety enriched for (CT) and (GT) for microsatellite motifs. From 1,209 sequenced clones, 714 showed microsatellites distributed over 471 simple and 243 compound motifs. GA/CT and GT/CA were the most frequent motifs found among these sequences. A total of 123 microsatellites has been characterized. Out of these, 87 were polymorphic (73.7%), 33 monomorphic (26.8%), and 3 (2.4%) did not amplify at all. In a sample of 20 common bean materials selected from the Agronomic Institute Germplasm Bank, the number of alleles per locus varied 2–9, with an average of 2.82. The polymorphic information content (PIC) of each marker varied from 0.05 to 0.83, with a 0.45 average value. Cluster and principal coordinate analysis of the microsatellite data were consistent with the original assignment of the germplasm accessions into the Andean and Mesoamerican gene pools of common bean. Low polymorphism levels detected could be associated with the domestication process. These microsatellites could be a valuable resource for the bean community because of their use as new markers for genetic studies.     

A note on the variation of phaseolin among common bean landraces collected in Eastern Asia

Summary Phaseolin seed protein was used as a marker to reveal the origin, Mesoamerican or Andean, of common bean (Phaseolus vulgaris) accessions cultivated in some countries (Korea, China, and Japan) of eastern Asia. Andean cultivars (T and C phaseolin patterns) are predominant in this geographical area. Introductions from Middle America, represented by cultivars with S phaseolin type, also occurred at lower rate. In all cases, genotypes with larger seeds have been favoured.