Genetic and phenotypic diversity in a germplasm working collection of cultivated tropical yams (Dioscorea spp.)

A working collection of yam (Dioscorea spp.) comprising 53 landraces and seven improved cultivars of four species (D. alata L., D. cayenensis Lam., D. dumetorum (Kunth) and D. rotundata Poir.) was evaluated for phenotypic and genetic diversity. The evaluation involved field assessment of 24 morphological traits and DNA analysis with 32 Simple sequence repeat (SSR) polymorphic markers. Diversity was greater between species than within species; highest in D. rotundata and lowest in D. alata and D. cayenensis. Analysis based on combined phenotypic and SSR marker data sets revealed a close relationship between D. rotundata and D. cayenensis, but D. alata and D. dumetorum remained as distinct species. D. alata was related genetically to D. rotundata and D. cayenensis, but phenotypically to D. dumetorum. The study showed that cultivars obtained from different farmers may bear the same name but be genetically different. Polymorphic SSR markers were identified that may be used for genetic analysis across yam species. The working collection assessed in this study represents a good gene pool for intra- and inter-specific hybridization in yam genetic improvement.     

Genetic diversity and species delimitation in the cultivated and wild Guinea yams (Dioscorea spp.) from Southwest Ethiopia as determined by AFLP (amplified fragment length polymorphism) markers

Yams (Dioscorea spp.) rank as the fourth most important root and tuber crop after potatoes, cassava and sweet potatoes. They are an economic crop in most of the tropics especially in West Africa, which produces over 95 % of the world output. Despite their cultural and economic importance there is taxonomic confusion regarding Guinea yams. The currently used classification scheme, which relies on vegetative and inflorescence characters, does not consistently delimit species boundaries between members of Guinea yams (D. cayenensis Lam.–D. rotundata Poir. complex), their wild relatives (D. abyssinica Kunth and D. praehensilis Benth.,) and D. sagittifolia Pax. Establishing the taxonomic identity of the germplasm and understanding the systematic relationships among crops is vital to the management of genetic resources and the utilization of accessions. In this study, amplified fragment length polymorphism (AFLP) genetic fingerprinting was used to evaluate and characterize 43 individual plants, belonging to different populations of wild and cultivated Guinea yams. The three primer combinations used in the AFLP analyses generated 158 scorable bands, with an overall polymorphism of 78 %. Ordination and cluster analyses of AFLP data failed to produce any clear species boundary between either the Guinea yam accessions or between them and their wild relatives. The average genetic similarity between the study individuals of Guinea yams and their wild relatives ranged from 60 to 100 %. The first, second and third principal coordinates axes cumulatively account for 77.45 % of the total variation. AFLP analyses also revealed a higher genetic divergence among cultivated Guinea yam accessions of the Sheko cultivars. Ordination and cluster analysis using UPGMA revealed no clear species boundaries between members of the complex. Thus, the taxonomy of these “species” needs to be revisited using other markers.     

Assessment of genetic diversity in Galega officinalis L. using ISSR and SRAP markers

Goat’s rue is well known for its traditional medicinal importance. In the present study, inter-simple sequence repeat (ISSR) and sequence related amplified polymorphism (SRAP) molecular markers were employed for the first time to access the genetic diversity and relationships of 35 wild goat’s rue accessions obtained from Russia and part of Europe countries. A total of 100 bands were amplified by ten ISSR primers, of which 77(77%) were polymorphic, and 59 polymorphic bands (67.1%) were observed in 88 bands amplified by seven SRAP primers. Polymorphic information content (PIC?=?0.426), Shannon’s information index (I?=?0.432), and average expected heterozygosity (He?=?0.292) generated by ISSR primer were higher than that of SRAP analysis (PIC?=?0.422, I?=?0.380, and He?=?0.257,). The study indicated that ISSR were more effective than SRAP markers for assessing the degree of genetic variation of goat’s rue. UPGMA cluster analysis revealed inconsistencies in the clustering patterns, as the Mantel’s test between the dendrograms for ISSR and SRAP data indicated a poor fit for the ISSR and SRAP data types (r?=?0.358). Whereas the pattern of clustering of the genotypes remained more or less the same in SRAP and combined data of ISSR and SRAP. The results of principal coordinates analysis also supports their UPGMA clustering. These results have an important implication for goat’s rue germplasm characterization, improvement, and conservation.     

Genetic diversity of cultivated flax (Linum usitatissimum L.) and its wild progenitor pale flax (Linum bienne Mill.) as revealed by ISSR markers

Little is known about the genetic diversity of pale flax (Linum bienne Mill.), the wild progenitor of cultivated flax (L. usitatissimum L.), and ex situ germplasm of pale flax was scarce. Effort was made to collect 34 pale flax accessions and five landrace accessions of cultivated flax in Turkey. The inter simple sequence repeat (ISSR) technique was applied to characterize this set of flax germplasm, along with one Turkish cultivar, one Russian cultivar, five winter and four dehiscent type accessions of cultivated flax. Twenty-four ISSR primer pairs detected a total of 311 DNA fragments, of which 298 bands were polymorphic across 493 flax samples (roughly 10 samples per accession). These polymorphic bands had frequencies ranging from 0.002 to 0.998 and averaging 0.38. Accession-specific ISSR variation (Fst values) ranged from 0.469 to 0.514 and averaged 0.493. There was 49.3% ISSR variation resided among these 50 accessions, 35.9% harbored among landrace, winter, dehiscent types of cultivated flax and pale flax, and 38.2% present among 34 pale flax accessions. Pale flax displayed more ISSR variation than landraces and dehiscent type, but less than winter type, of cultivated flax. Clustering 493 individual plants revealed that these assayed plants were largely grouped according to their plant types and that pale flax was genetically more close to the dehiscent type, followed by the winter type and landrace, of cultivated flax. Pale flax collected within the geographic range of 180 km displayed a significant spatial genetic autocorrelation. Genetic distances among the pale flax accessions were significantly associated with their geographic distances and elevation differences. These findings are significant for understanding flax domestication and its primary gene pool.     

Genetic diversity of anchote (Coccinia abyssinica (Lam.) Cogn.) from Ethiopia as revealed by ISSR markers

Anchote (Coccinia abyssinica) is plant endemic in Ethiopia with a high calcium content grown for its edible tuberous roots. In spite of its importance as food security crop, there is no information available on molecular genetic diversity of this crop. Therefore, the aim of this study was to assess genetic diversity within and among 12 populations of anchote using ISSR markers. Using nine ISSR primers, a total of 87 scorable bands was generated of which 74 were polymorphic. Within population diversity based on polymorphic bands ranged from 13.8 to 43.53 % with a mean of 33.05 %, Nei’s genetic diversity of 0.04–0.156 with a mean of 0.12, Shannon information index of 0.07–0.23 with a mean of 0.175 and analysis of molecular variation (AMOVA) of 51.4 % were detected. With all diversity parameters, the highest diversity was obtained from Gimbi, Bedele and Ale populations, whilst the lowest was from Manna. AMOVA showed a 48.56 % between populations variability and significantly lower than that of within population variation. Population differentiation with F_ST was 48.56 %. From Jaccard’s pairwise similarity coefficient, Decha and Nedjo were most related populations exhibiting 0.76 similarity and Manna and Nedjo were the most distantly related populations with similarity of 0.52. The only pentanucliotide primer used in the study, Primer 880 (GGAGA)3, showed a unique band in some individuals that appeared to be associated with morphological quantitative traits (lowest seed number, high protein content, largest fruit size and smallest vine length). Illubabor and Gimbi populations exhibited highest genetic diversity so that the populations should be considered as the primary sites in designing conservation areas for this crop.     

Genetic relationship and species identification of Dioscorea polystachya Turcz. in Yams determined by ISSR,ISAP, SRAP and SCAR markers

Genetic Resources and Crop Evolution - Dioscorea polystachya Turcz. is the authentic herb Yam in Chinese traditional medicine. In the medicinal marketplace, some related Dioscorea species are...     

Spatially structured genetic diversity of the Amerindian yam (Dioscorea trifida L.) assessed by SSR and ISSR markers in Southern Brazil

Dioscorea trifida L. (Dioscoreaceae) is among the economically most important cultivated Amerindian yam species, whose origin and domestication are still unresolved issues. In order to estimate the genetic diversity maintained by traditional farmers in Brazil, 53 accessions of D. trifida from 11 municipalities in the states of São Paulo, Santa Catarina, Mato Grosso and Amazonas were characterized on the basis of eight Simple Sequence Repeats (SSR) and 16 Inter Simple Sequence Repeats (ISSR) markers. The level of polymorphism among the accessions was high, 95 % for SSR and 75.8 % for ISSR. The SSR marker showed higher discrimination power among accessions compared to ISSR, with D parameter values of 0.79 and 0.44, respectively. Although SSR and ISSR markers led to dendrograms with different topologies, both separated the accessions into three main groups: I—Ubatuba-SP; II—Iguape-SP and Santa Catarina; and III—Mato Grosso. The accessions from Amazonas State were classified in group II with SSR and in a separate group with ISSR. Bayesian and principal coordinate analyzes conducted with both molecular markers corroborated the classification into three main groups. Higher variation was found within groups in the AMOVA analysis for both markers (66.5 and 60.6 % for ISSR and SSR, respectively), and higher Shannon diversity index was found for group II with SSR. Significant but low correlations were found between genetic and geographic distances (r = 0.08; p = 0.0007 for SSR and r = 0.16; p = 0.0002 for ISSR). Therefore, results from both markers showed a slight spatially structured genetic diversity in D. trifida accessions maintained by small traditional farmers in Brazil.     

Genetic diversity in Tunisian perennial ryegrass revealed by ISSR markers

The genetic diversity in Tunisian perennial ryegrass (Lolium perenne) was examined by the help of inter-simple sequence repeats (ISSR). Starting from eighteen accessions, a large number of polymorphic ISSR markers were currently generated using appropriate primers (a total of 136, which average of 12.6 polymorphic bands/primer). These markers were considered to estimate the genetic distance among accessions and to draw phylogenetic trees. Our data provide evidence of a high degree of genetic diversity in Tunisian ryegrass. In addition, both cultivars and wild types present a high degree of divergence suggesting a complex domestication process in this crop. Moreover, spontaneous populations of Tunisian ryegrass have been identified as important ecotypes that are suitable in selection programs to improve grasslands.     

Analysis of genetic diversity of Ruthenia Medic (Medicago ruthenica (L.) Trautv.) in Inner Mongolia using ISSR and SSR markers

Ruthenia Medic is tolerant to drought, cold, high salinity, resistance to trampling and high quality features. Inter-simple sequence repeat (ISSR) and simple sequence repeat (SSR) molecular markers were employed for the first time to access the genetic diversity and relationships of 30 wild Ruthenia Medic accessions obtained from Inner Mongolia in the present study. A total of 94 bands were amplified by ten ISSR primers, of which 83 (88.5 %) were polymorphic, and 57 polymorphic bands (80.4 %) were observed in 69 bands amplified by ten SSR primers. Shannon’s information index (I = 0.487), and average expected heterozygosis (He = 0.329) generated by ISSR primer were higher than that of SSR analysis (I = 0.372, He = 0.231). The study indicated that ISSR were more effective than SSR markers for assessing the degree of genetic variation of Ruthenia Medic. UPGMA cluster analysis revealed inconsistencies in the clustering patterns, as the Mantel’s test between the dendrograms for ISSR and SSR data indicated a poor fit for the ISSR and SSR data types (r = 0.0970). Whereas the pattern of clustering of the genotypes remained relatively the same in ISSR and combined data of ISSR and SSR. The results of principal components analysis also supports their UPGMA clustering. These results have an important implication for Ruthenia Medic germplasm characterization, improvement, and conservation.     

Genetic diversity in wild and cultivated black raspberry (Rubus occidentalis L.) evaluated by simple sequence repeat markers

Breeding progress in black raspberry (Rubus occidentalis L.) has been limited by a lack of genetic diversity in elite germplasm. Black raspberry cultivars have been noted for showing very few phenotypic differences and seedlings from crosses between cultivars for a lack of segregation for important traits. Despite these challenges, little molecular work has been done to explore genetic diversity and relationships in wild and cultivated black raspberry germplasm. Microsatellite, or simple sequence repeat (SSR), markers are highly polymorphic codominant markers useful for studying genetic diversity, population genetics, genetic fingerprinting and other applications. We examined genetic diversity in 148 wild and cultivated black raspberry accessions using 21 polymorphic SSR markers. Black raspberry cultivars clustered tightly and showed higher than expected heterozygosity while that of wild accessions was low. Relationships between wild black raspberry accessions were poorly resolved and regional clusters were mostly absent from our analysis. Our results indicated that wild black raspberry germplasm is a relatively untapped resource available for future breeding.     

Genetic diversity,population structure and genetic relationships in apricot (Prunus armeniaca L.) germplasm of Jammu and Kashmir,India using ISSR markers

Genetic Resources and Crop Evolution - The collection and characterization of apricot germplasm is an important step for its utilization, conservation, and breeding programs. In the present study,...     

Comparison between phenetic characterisation using RAPD and ISSR markers and phenotypic data of cultivated chestnut (Castanea sativa Mill.)

Patterns of phenotypic and phenetic variability in sixPortuguese cultivars of chestnut (Castanea sativa Mill.) are evaluated. Morphological characterisationwas based on the quantification of seventeen traits. Varianceanalysis showed significant differences among cultivars, and cultivar× year for all the traits studied, and trees within cultivarsshowed also some significant differences for some of themorphological variables. A significant correlation was obtainedbetween length of the leaf blade and the percentage of unisexual andandrogynic inflorescence with the effective thermal index,accumulated rainfall from April to October and from July to October,or the accumulated temperature below seven during the dormant period.Principal Component and cluster analysis were performed to group thecultivars, according to their similarity coefficients. For molecularcharacterisation, 125 RAPD and 157 ISSR polymorphic markers wereamplified using 28 and 7 primers respectively. High level ofcongruence among the two marker systems (r =90.5%) was obtained from comparison of pheneticsimilarities based on the percentage of shared fragments. ISSRmarkers revealed important advantages over RAPDs, due to a higheffective multiplex ratio (12.5 for ISSR compared with 2.2 forRAPD analysis) and reproducibility. Although morphological andmolecular results are comparable, slight differences are showed incluster analysis UPGMA dendrograms. Molecular analysis explainedhomonym situations among 'Martainha' and'Longal' cultivars in Portugal.     

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.     

Genetic diversity in the Olive tree (Olea europaea L. subsp. europaea) cultivated in Portugal revealed by RAPD and ISSR markers

To assess the genetic diversity of the most important olive cultivars used in Portugal, a base collection was established with two hundred and one accessions of eleven cultivars from the different agro-ecological-regions (AER) of olive oil production. Inter-cultivar diversity was evaluated using seven RAPD primers producing fifty-nine polymorphic markers that enable cultivar distinction. Discriminant analysis according to fruit use and AER revealed a genetic structure associated with local selection both for fruit exploitation and agro-ecological adaptation. Intra-cultivar diversity of the ancient cultivar Galega was also investigated. Three RAPD and five ISSR primers produced ninety-three polymorphic markers upon seventy-seven accessions from five AERs. Total accession discrimination was achieved. UPGMA clustering and discriminant analysis revealed that the genetic diversity was predominantly structured according to accessions origin. The within and among AER variation revealed by AMOVA supported this genetic structure and showed a high proportion of intra-AER variability. These evidences suggest that Galega is composed by a mixture of different genotypes adapted to local conditions, indicating that this cultivar is in an early stage of domestication and should be treated as a landrace instead of a uniform cultivar. The assessment of Galega genetic diversity within each of the five AERs indicated the highest significant level (H_g = 6.23 at p< 0.001) in Ribatejo-Santarém. This finding associated with the distinctiveness of Galega in relation to other Portuguese cultivars and with the recent insights of olive tree domestication allowed us to hypothesize that Ribatejo-Santarém was the ecological region of origin and dispersion of this ancient cultivar.