Molecular genetic diversity and association mapping of nut and kernel traits in Slovenian hazelnut (<Emphasis Type="Italic">Corylus avellana</Emphasis>) germplasm

European hazelnut (Corylus avellana L.), cultivated in several areas of the world including Europe, Anatolia, and the USA, is an economically important nut crop due to its high mineral, oleic acid, amino acid, and phenolic compound content and pleasant flavor. This study examined molecular genetic diversity and population structure of 54 wild accessions and 48 cultivars from the Slovenian national hazelnut collection using amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers. Eleven AFLP primer combinations and 49 SSR markers yielded 532 and 504 polymorphic fragments, respectively. As expected for a wind-pollinated, self-incompatible species, levels of genetic diversity were high with cultivars and wild accessions having mean dissimilarity values of 0.50 and 0.60, respectively. In general, cultivars and wild accessions clustered separately in dendrogram, principal coordinate, and population structure analyses with regional clustering of the wild material. The accessions were also characterized for ten nut and seven kernel traits and some wild accessions were shown to have breeding potential. Morphological principal component analysis showed distinct clustering of cultivars and wild accessions. An association mapping panel composed of 64 hazelnut cultivars and wild accessions had considerable variation for the nut and kernel quality traits. Morphological and molecular data were associated to identify markers controlling the traits. In all, 49 SSR markers were significantly associated with nut and kernel traits [P < 0.0001 and LD value (r ²) = 0.15–0.50]. This work is the first use of association mapping in hazelnut and has identified molecular markers associated with important quality parameters in this important nut crop.     

Phenetic and cladistic studies help clarify species assemblages in <Emphasis Type="Italic">Passiflora</Emphasis> section <Emphasis Type="Italic">Dysosmia</Emphasis> (Passifloraceae)

One of the most enigmatic and unusual groups in the passionflower genus, Passiflora section Dysosmia (Passifloraceae), stands out as a group that is notoriously taxonomically complicated. Phenetic and cladistic analyses of Dysosmia were carried out with representatives from all 21 species currently recognized in the section, in order to delineate morphological groups above the species level. The study was based mainly on vegetative morphological characters and included principal coordinates, non-metric multidimensional scaling, cluster, and cladistic analyses. Results from each analysis reveals that three major species assemblages exist within Dysosmia, corresponding largely to vegetative pubescence and fruit color. The results presented here set the stage for future systematic and phylogenetic studies in Passiflora sect. Dysosmia.     

Genetic diversity of some Iranian sweet cherry (<Emphasis Type="Italic">Prunus avium</Emphasis>) cultivars using microsatellite markers and morphological traits

The aim of this study was to characterize 23 important Iranian sweet cherry (Prunus avium) cultivars collected from different provinces of Iran and 1 foreign cultivar, which was used as control, considered for breeding programs by using 21 microsatellite markers and 27 morphological traits. In sweet cherry (Prunus avium) accessions, leaf, fruit, and stone morphological characters were evaluated during two consecutive years. The study revealed a high variability in the set of evaluated sweet cherry accessions. The majority of important correlations were determined among variables representing fruit and leaf size and variables related to color. Cluster analysis distinguished sweet cherry accessions into two distinct groups. Principal component analysis (PCA) of qualitative and quantitative morphological parameters explained over 86.59% of total variability in the first seven axes. In PCA, leaf traits such as leaf length and width, and fruit traits such as length, width, and weight, and fruit flesh and juice color were predominant in the first two components, indicating that they were useful for the assessment of sweet cherry germplasm characterization. Out of 21 SSR markers, 16 were polymorphic, producing 177 alleles that varied from 4 to 16 alleles (9.35 on average) with a mean heterozygosity value of 0.82 that produced successful amplifications and revealed DNA polymorphisms. Allele size varied from 95 to 290 bp. Cluster analyses showed that the studied sweet cherry genotypes were classified into five main groups based mainly on their species characteristics and SSR data. In general, our results did not show a clear structuring of genetic variability within the Iranian diffusion area of sweet cherry, so it was not possible to draw any indications on regions of provenance delimitation. The results of this study contribute to a better understanding of sweet cherry genetic variations in Iran, thus making for more efficient programs aimed at preserving biodiversity and more rational planning of the management of reproductive material.     

Genetic diversity and population structure of pummelo (<Emphasis Type="Italic">Citrus maxima</Emphasis>) germplasm in China

Pummelo (Citrus maxima) is one of the basic species of Citrus. It has been cultivated for about 4000 years in China, and therefore, there are abundant germplasm during the long time of culture. However, there is still a lack of a detailed study of the genetic characteristics of pummelo population. In this study, genetic diversity and population structure among 274 pummelo accessions collected in China were analyzed using 31 nuclear simple sequence repeat (nSSR) markers. The observed heterozygosity was calculated as 0.325 and genetic differentiation Fst as 0.077. Genetic structure analysis divided the whole germplasm into three subpopulations, Pop-a, Pop-b, and Pop-c. Pop-a was composed of accessions mostly from Southeast China, Pop-b was composed of accessions from the central region of South China, and Pop-c was composed of accessions from Southwest China. Meanwhile, the analysis of principal coordinate analysis and neighbor-join tree supported the viewpoint of three subpopulations, and then the possible dispersal routes of pummelos in China were proposed. This study provides an insight into the genetic diversity, facilitates future genome-wide association studies, and promotes the breeding program of pummelo as well.     

Analysis of microsatellite loci variability in rare and endemic species <Emphasis Type="Italic">Allium regelianum</Emphasis> A.K. Becker ex Iljin

SSR analysis of rare and endemic species Allium regelianum, which grows in the south of Russia, was performed for the first time. Variability analysis of 88 accessions of A. regelianum was carried out using four highly polymorphic microsatellite loci (PIC value ranged from 0.55 to 0.72). SSR-analysis made it possible to revealed polymorphism within and among the populations of A. regelianum from Volgograd region. Analysis of Wright’s F-statistics and the analysis of molecular variance showed that more than 90% of total genetic variation of the species was attributed to the differences within the populations and less than 10% of the differences were found among the populations. Cluster analysis of 46 accessions of A. regelianum from three populations of Volgograd region and principal coordinate analysis of all accessions did not reveal differentiation among the populations.     

Determination of morphological variation in subspecies of Isatis glauca Aucher ex Boiss. from Central Anatolia

Isatis glauca Aucher ex Boiss. has four subspecies: glauca, galatica Y?ld?r?ml?, sivasica (Davis) Y?ld?r?ml? and iconia (Boiss.) Davis, naturally growing in Turkey. In this study, 77 accessions from nine I. glauca ssp. populations were characterised in terms of variation in morphological characters, including the number of branches per plant, plant height, diameter of stem base, diameter of middle stem, diameter of apical stem, width and length of fruit, length of fruit stalk, fruit shape, fruit base shape, fruit apices shape, loculus, ondulation, apical process of fruit, fruit weight, basal length of fruit, apical length of fruit, wing width of fruit, loculus length, loculus width, length of seed, dorsal width of seed, lateral width of seed and seed weight. The number of branches had the highest value of coefficient of variation (CV), 0.21–0.86, while the fruit width had the lowest value of CV 0.08–0.20. According to Pearson’s correlation and multiple regression analyses, eco-geographical factors had a substantial effect on morphological variation. A principal component analysis explained 90.61 % of morphological variation through the study of five extracted components. A dendrogram, which was constructed according to the between-group linkages method, based on squared Euclidean distances, represented a hierarchical cluster analysis of 77 accessions. The results of the principal coordinate analysis, which showed differentiation according to morphological characters between nine populations, were consistent with the dendrogram. Consequently, morphological variation of I. glauca ssp. and the significant effects of eco-geographical factors on its morphological variation were discussed.     

Analysis of genetic diversity and population structure in <Emphasis Type="Italic">Capsicum</Emphasis> landraces from North Eastern India using TE-AFLP markers

North eastern (NE) India harbours a precious germplasm repository of Capsicum in the form of various landraces. The present study was undertaken to characterise the extent of genetic variation present in different Capsicum landraces from north eastern India. A set of 171 Capsicum accessions were characterised using three-endonuclease amplified fragment length polymorphism (AFLP) markers. Out of 416 bands obtained from six primer combinations, 254 (61 %) were polymorphic. The pairwise genetic dissimilarity among accessions ranged from 0.03 to 0.97. Cluster analysis based on neighbour joining showed two major clusters. Cluster I contained most of the bhut jolokia accessions whereas cluster II contained all of the Capsicum annuum genotypes. Similar grouping was observed with population STRUCTURE analysis as well as principle coordinate analysis. Analysis of molecular variance (AMOVA) revealed 45 and 54 % variation among and within populations, respectively. This information on population structure analysis and molecular characterisation will be helpful for effective utilisation of this germplasm in Capsicum improvement programs.     

Assessment of genetic diversity in <Emphasis Type="Italic">Mucuna</Emphasis> species of India using randomly amplified polymorphic DNA and inter simple sequence repeat markers

Genus Mucuna which is native to China and Eastern India comprises of perennial climbing legume with long slender branches, trifoliate leaves and bear green or brown pod covered with soft or rigid hairs that cause intense irritation. The plants of this genus are agronomically and economically important and commercially cultivated in India, China and other regions of the world. The high degrees of taxonomical confusions exist in Mucuna species that make authentic identification and classification difficult. In the present study, the genetic diversity among the 59 accessions of six species and three varieties of M. pruriens has been assessed using DNA fingerprinting based molecular markers techniques namely randomly amplified polymorphic DNA (RAPD), inter simple sequence repeats (ISSR) and combined dataset of RAPD and ISSR. Also, genetic relationship among two endemic species of Mucuna namely M. imbricata and M. macrocarpa and two varieties namely IIHR hybrid (MHR) and Dhanwantari (MD) with other species under study was investigated by using cluster analysis and principal coordinate analysis. The cluster analysis of RAPD, ISSR and combined dataset of RAPD and ISSR clearly demonstrated the existence of high interspecific variation than intra-specific variation in genus Mucuna. The utility and efficacy of RAPD and ISSR for the study of intra species and interspecies genetic diversity was evident from AMOVA and PCoA analysis. This study demonstrates the genetic diversity in Mucuna species and indicates that these markers could be successfully used to assess genetic variation among the accessions of Mucuna species.     

Assessment of a non-destructive method to estimate the leaf area of <Emphasis Type="Italic">Armoracia rusticana</Emphasis>

The aim of the study was to analyze horseradish growth for developing a mathematical model to estimate the leaf area based on linear measurements of the leaf surface. Leaf area (LA), number, and morphometric characteristics of the leaves including lamina length (L) and width (W) were evaluated on two horseradish accessions (Cor and Mon) throughout a 2 year growing cycle. In both accessions, increased values of LA and leaf number were found by comparing the second with the first-growing season. Leaf development occurs along with variations in size and not in shape during the plant growth. The leaves are elliptical in shape but tend to be wider and bigger in Cor accession and tapered and similar to narrow ellipses in Mon showing different length/width relationship. Consequently, several regression models relating to the LA and L, W, L², and W² individually or in combination were fitted for each accession based on a set of 1000 leaves. The horseradish LA can be predicted based on either length or width alone. However, the regression linear model LA?=?aLW?+?b (LA?=?0.71LW ??0.27 and LA?=?0.76LW ??3.22 for Cor and Mon, respectively) provided the best LA estimation (R²?>?0.95). The validation of this latter model showed high correlation between LA measured and LA predicted in both accessions (R²?=?0.98). Considering the type of foliage of horseradish, the proposed model can be used to estimate the leaf area throughout the entire crop cycle.     

Assessment of Genetic Diversity in <Emphasis Type="Italic">Secale cereale</Emphasis> Based on SSR Markers

The primary aim of this study was to estimate genetic diversity among Secale cereale L. accessions using 22 previously published simple sequence repeat (SSR) markers. The plant material included 367 rye accessions comprising historical and contemporary cultivars, cultivated materials, landraces, and breeding strains from the Polish breeding company Danko. The studied accessions represented a wide geographical diversity. Several methods were employed to analyze genetic diversity among the Secale cereale L. accessions and to determine population structure: principal coordinate analysis (PCoA), neighbor-joining (NJ), and Bayesian clustering. We also defined a core collection of 25 rye accessions representing over 93 % of SSR alleles. The results of these analyses showed that accessions from the rye gene bank are clearly divergent in comparison with materials received directly from European breeding companies. Our findings suggest also that the genetic pool of current rye cultivars is becoming narrower during breeding processes. The selected panel of SSR markers performed well in detection of genetic diversity patterns and can be recommended for future germplasm characterization studies in rye.     

Morphological variation and evolutionary significance of <Emphasis Type="Italic">Coccinia grandis</Emphasis> (L.) Voigt: an under-exploited cucurbitaceous vegetable crop

Morphological variation was analyzed in wild accessions and cultivars of the vegetatively propagated dioecious Coccinia grandis. Variations of 43 morphological characters, 19 qualitative and 23 quantitative traits, were analyzed among 40 female accessions, including 25 cultivars and 15 wild accessions. Multivariate statistical analyses were used to group accessions according to their morphological similarity. Principal component (PC) analysis revealed that the first three PCs accounted for 50% of the total variance, and differences among the accessions were evidenced principally in relation to fruit characteristics such as fruit weight, fruit length and the number of seeds in each fruit. Analysis of variance carried out in the entire germplasm revealed significant differences within the germplasm, whereas ANOVA carried out between the wild accessions and the cultivars proved the null hypothesis that there are no significant differences between the two groups, and differences were observed only in fruit characters that are targets of human selection. Principal component analysis, UPGMA cluster analysis and discriminant factor analysis revealed strong overlaps between the two groups indicating the ongoing process of evolution and selection in the species.     

Bayesian and Phylogenic Approaches for Studying Relationships among Table Olive Cultivars

To enhance table olive tree authentication, relationship, and productivity, we consider the analysis of 18 worldwide table olive cultivars (Olea europaea L.) based on morphological, biological, and physicochemical markers analyzed by bioinformatic and biostatistic tools. Accordingly, we assess the relationships between the studied varieties, on the one hand, and the potential productivity–quantitative parameter links on the other hand. The bioinformatic analysis based on the graphical representation of the matrix of Euclidean distances, the principal components analysis, unweighted pair group method with arithmetic mean, and principal coordinate analysis (PCoA) revealed three major clusters which were not correlated with the geographic origin. The statistical analysis based on Kendall’s and Spearman correlation coefficients suggests two highly significant associations with both fruit color and pollinization and the productivity character. These results are confirmed by the multiple linear regression prediction models. In fact, based on the coefficient of determination (R ²) value, the best model demonstrated the power of the pollinization on the tree productivity (R ² = 0.846). Moreover, the derived directed acyclic graph showed that only two direct influences are detected: effect of tolerance on fruit and stone symmetry on side and effect of tolerance on stone form and oil content on the other side. This work provides better understanding of the diversity available in worldwide table olive cultivars and supplies an important contribution for olive breeding and authenticity.     

Simple Sequence Repeat and <Emphasis Type="Italic">S</Emphasis>-locus Genotyping to Explore Genetic Variability in Polyploid <Emphasis Type="Italic">Prunus spinosa</Emphasis> and <Emphasis Type="Italic">P. insititia</Emphasis>

Polyploid Prunus spinosa (2n = 4×) and P. insititia (2n = 6×) represent enormous genetic potential in Central Europe, which can be exploited in breeding programmes. In Hungary, 17 cultivar candidates were selected from wild-growing populations including 10 P. spinosa, 4 P. insititia and three P. spinosa × P. domestica hybrids (2n = 5×). Their taxonomic classification was based on their phenotypic characteristics. Six simple sequence repeats (SSRs) and the multiallelic S-locus genotyping were used to characterize genetic variability and reliable identification of the tested accessions. A total of 98 SSR alleles were identified, which presents 19.5 average allele number per locus, and each of the 17 genotypes could be discriminated based on unique SSR fingerprints. A total of 23 S-RNase alleles were identified. The complete and partial S-genotype was determined for 8 and 9 accessions, respectively. The identification of a cross-incompatible pair of cultivar candidates and several semi-compatible combinations help maximize fruit set in commercial orchards. Our results indicate that the S-allele pools of wild-growing P. spinosa and P. insititia are overlapping in Hungary. A phylogenetic and principal component analysis confirmed the high level of diversity and genetic differentiation present within the analysed genotypes and helped clarify doubtful taxonomic identities. Our data confirm that S-locus genotyping is suitable for diversity studies in polyploid Prunus species. The analysed accessions represent huge genetic potential that can be exploited in commercial cultivation.     

A molecular phylogeny of the Old World cypresses (<Emphasis Type="Italic">Cupressus</Emphasis>: Cupressaceae): evidence from nuclear and chloroplast DNA sequences

Studies of phylogenetic relationships among cypresses of the Old World (Cupressus; Cupressaceae) have been plagued by unresolved relationships, poor branch support, and conflict between data sets and methods of analysis. In this study, we combined 5.4 kb of aligned DNA sequence and 157 binary characters with previously published data in examining phylogenetic relationships among Cupressus species. Bayesian and parsimony analysis of the combined data or of the nuclear data alone always recovered three principal clades of Cupressus; however, tests of phylogenetic incongruence could not distinguish between competing relationships among the three principal Cupressus lineages. In contrast, incongruence tests often found statistically significant conflict between the nuclear and plastid data, particularly with respect to the placement of C. chengiana. Consistent with previous studies and prevailing taxonomic opinion, we find C. darjeelingensis more closely related to cypresses of the New World (Hesperocyparis). In contrast, we placed accessions of C. assamica and C. tonkinensis, two putatively Old World species suggested to be misidentified New World taxa by some authors, within well-supported Old World clades. Statistical analysis of genetic distances suggests instances in which taxa recognized as distinct species by some authors are identical or nearly so and may best be considered a single taxon. Conversely, we identify instances in which infraspecific taxa are more distantly related to one another than those traditionally recognized as distinct species. Factors confounding cypress taxonomies, including poor morphological differentiation, misidentification, and the use of accessions of questionable provenance, are discussed.     

Evaluation of Diversity Based on Morphological Variabilities and ISSR Molecular Markers in Iranian <Emphasis Type="Italic">Cynodon dactylon</Emphasis> (L.) Pers. Accessions to Select and Introduce Cold-Tolerant Genotypes

The main goals of the present study were to screen Iranian common bermudagrasses to find cold-tolerant accessions and evaluate their genetic and morphological variabilities. In this study, 49 accessions were collected from 18 provinces of Iran. One foreign cultivar of common bermudagrass was used as control. Morphological variation was evaluated based on 14 morphological traits to give information about taxonomic position of Iranian common bermudagrass. Data from morphological traits were evaluated to categorize all accessions as either cold sensitive or tolerant using hierarchical clustering with Ward’s method in SPSS software. Inter-Simple Sequence Repeat (ISSR) primers were employed to evaluate genetic variability of accessions. The results of our taxonomic investigation support the existence of two varieties of Cynodon dactylon in Iran: var. dactylon (hairless plant) and var. villosous (plant with hairs at leaf underside and/or upper side surfaces or exterior surfaces of sheath). All 15 primers amplified and gave clear and highly reproducible DNA fragments. In total, 152 fragments were produced, of which 144 (94.73%) being polymorphic. The polymorphic information content (PIC) values ranged from 0.700 to 0.928. The average PIC value obtained with 15 ISSR primers was 0.800, which shows that all primers were informative. Probability identity (PI) and discriminating power between all primers ranged from 0.029 to 0.185 and 0.815 to 0.971, respectively. Genetic data were converted into a binary data matrix. NTSYS software was used for data analysis. Clustering was done by the unweighted pair-group method with arithmetic averages and principle coordinate analysis, separated the accessions into six main clusters. According to both morphological and genetic diversity investigations of accessions, they can be clustered into three groups: cold sensitive, cold semi-tolerant, and cold tolerant. The most cold-tolerant accessions were: Taft, Malayear, Gorgan, Safashahr, Naein, Aligoudarz, and the foreign cultivar. This study may provide useful information for further breeding programs on common bermudagrass. Selected genotypes can be evaluated for other abiotic stresses such as drought and salinity.     

Genetic Diversity,Genotype Discrimination,and Population Structure of Mexican <Emphasis Type="Italic">Opuntia</Emphasis> sp., Determined by SSR Markers

The Opuntia (prickly pear) genus, an important horticultural crop in Mexico, is essentially a fruit crop with two variants: sweet (“tunas”) or acid (“xoconostles”) fruits; it is also a source of vegetables “nopalitos” or fodder for livestock, among other uses. Its taxonomical classification has been reported as complex, although few studies on the genetic structure of Mexican Opuntia are available, and genetic differences between the two types of fruits are unknown. Opuntia genotype identification and classification are still mainly based on morphological characters. In this study, the genetic diversity of Mexican Opuntia germplasm with agronomic and economic importance was revealed, using 88 accessions and 13 SSR markers, in an attempt to explore the genetic relationships among them. A total of 159 alleles were detected ranging from 7 to 23 per locus with an average of 12.2. The SSR markers generated unique fingerprints for each Opuntia accession confirming their usefulness for genetic analysis. The accessions’ grouping was defined by several complementary clustering methods, and the moderate incongruences between the different methods did not influence the overall clustering. DAPC and STRUCTURE analyses grouped the accessions into five groups, thus confirming the incorrect delimitation of species in this genus. The following species had no clear boundaries: Opuntia ficus-indica, Opuntia albicarpa, Opuntia megacantha, Opuntia streptacantha, Opuntia lasiacantha, and Opuntia hyptiacantha. However, Opuntia robusta was separated from the rest of the species. Opuntia joconostle and Opuntia matudae, which produce acid fruits, tended to differ from the others. Median-joining simulation classified all genotypes into a complex network, and both linear and reticular ties between Mexican Opuntia genotypes were revealed. The genetic distance revealed in the present study shows the importance of Mexican accessions for conservation and use in breeding programs.