Characterisation and Variation of Morphological Traits and Storage Proteins in Spanish Emmer Wheat Germplasm (<Emphasis Type="Italic">Triticum Dicoccon</Emphasis>)

Emmer wheat is hulled wheat that was wide cultivated in Spain at the past. Actually, the most of this germplasm is conserved in Germplasm Banks, and only two small populations have been found in Asturias (North of Spain) in a recent collecting mission. In this work, a collection of 31 Spanish emmer lines developed from identical number of accessions of two Germplasm Banks was analysed for morphological spike traits and seed storage protein composition. Up to seven different botanical varieties were detected, which suggest the presence of a wide diversity, although lower than the historically described 10 botanical varieties. At level of seed storage proteins, the lines showed a high diversity, although the new alleles were present with low frequency in materials with scarce agronomic interest for the farmers (var. atratum, var. lagascae or var. pycnurum). This last circumstance could translate in a lost of variability by genetic drift.     

Exploration of intra- and inter-population genetic diversity in <Emphasis Type="Italic">Hedysarum coronarium</Emphasis> L. by AFLP markers

Amplified fragment length polymorphism (AFLP) markers were used to characterize the genetic diversity within and among natural populations and cultivars of Hedysarum coronarium. Twelve populations within Tunisia were evaluated with three AFLP primer combinations. A total of 207 reproducible bands was detected of which 178 (86%) were polymorphic. The great discriminative power of AFLP markers and their ability to represent genetic relationships among Hedysarum plants was demonstrated. Genetic diversity within and among populations was assessed through Principal Component Analysis (PCA) and cluster analysis by using the Neighbor-joining clustering algorithm. AFLP technology has provided evidence of a high degree of intra- and inter-population genetic diversity in H. coronarium. AFLP banding patterns provided molecular markers correlated with the plants’ geotropism. In addition, AFLP markers can differentiate wild accessions from cultivars. Moreover, geographical origins did not correspond to population clustering.     

Characterization of a landrace collection for Tomàtiga de Ramellet (Solanum lycopersicum L.) from the Balearic Islands

Tomàtiga de Ramellet (Ramellet) is a traditional long-storage tomato from the Mediterranean Islands of Mallorca, Menorca, Eivissa (Ibiza), and Formentera (Balearic Islands). To investigate the observed phenotypic and shelf life variability within local varieties, we studied 142 accessions characterized as Ramellet varieties, and 29 accessions representing other local varieties. The 171 accessions constitute a new germplasm collection at the University of Balearic Islands (UIB). The evaluation of the collection demonstrated wide variability in most agronomic, morphological and quality traits of Ramellet accessions and clearly differentiates them from other local varieties with respect to fruit shape, size, fruit composition, and fruit storage. All accessions of Ramellet evaluated have the ripening mutation alcobaça (alc), suggesting an important role for this variant in controlling shelf life. Results of evaluation also showed wide variability between the Ramellet accessions for storage time, suggesting that other traits like fruit size and composition may play a role in fruit conservation. Observed variation within the Ramellet accessions is consistent with a genetically variable population or landrace group. The establishment of this collection will help prevent genetic erosion of this local landrace group and constitute a resource for future investigation and breeding.     

Morphological and chemical variation in 25 collections of the Indian medicinal plant, <Emphasis Type="Italic">Withania somnifera</Emphasis> (L.) Dunal (Solanaceae)

Six phenotypic characters and three withanolide markers were assessed in 25 accessions of Withania somnifera collected from different states of India for studying genetic variability. The variability ranges observed at phenotypic and chemotypic levels were polymorphic. Based on D² values and PCA (Principal Component Analysis) of phenotypic traits like plant height, no. of branches/plant, no. of seeds/berry, root length, root diameter and root yield, these 25 accessions were grouped in five clusters. The relative contribution of each character towards genetic divergence was worked out. Five accessions–AGB002 (Rajasthan), AGB003 (J&K), AGB004 (Madhya Pradesh), AGB006 (J&K) and AGB009 (Punjab) representing clusters 2 and 4 exhibited maximum intra and inter-cluster divergence. Cluster 5 representing accession AGB053 (Andhra Pradesh) was having mixed traits. Chemically most of the accessions in cluster 3 showed uniformity in presence of three marker withanolides Withaferin A, Withanone and Withanolide A in the leaves.     

Determination of genetic variability by RAPD markers in cauliflower,cabbage and kale local cultivars from France

RAPD markers were used to analyse the genetic variability among a cole crop cultivar collection from France and to identify possible duplicate accessions for facilitating the germplasm multiplication. We surveyed 24 cauliflower, 24 cabbage and 48 kale populations using 62, 100 and 89 RAPD markers respectively on 40 seed bulk samples per accession. Genetic distances were calculated on the basis of these markers. Using the phylogenetic package PAUP we compared the genetic clusters obtained for the RAPD markers with the groups based on morphologic and agronomic data. For cauliflowers and cabbages a drastic selection by growers led to an extensive differentiation of morphologic and precocity traits related to geographic origin, which matches the grouping based on RAPD markers. Furthermore, RAPDs detected in some cases misclassification of accessions in the collection. Kales formed an homogeneous group probably due to extensive genetic exchanges leading to a continuous diversity. For the other crops, different horticultural types were separated in several subgroups by the RAPD markers suggesting different genetic origins. RAPD markers provide a rapid and informative approach to analyze the genetic variability in a collection. It can be applied to autogamous (spring cauliflower) or allogamous (winter cauliflower) crops, even in cases where the intra-population diversity is extensive (cabbages).     

Patterns of morphological diversity in relation to geographical origins of wild Lupinus angustifolius from the Aegean region

Summary Patterns of morphological diversity were examined in relation to geographical origins of 157 accessions of wild Lupinus angustifolius from the Aegean region using multivariate techniques. Genetic diversity was extremely large for most morphological traits, with significant variation detected among localities in Greece and within and between collection sites for some traits. Canonical variates and correlation analysis showed that early flowering, tall and large-seeded accessions were associated with warm winters and drier climates in southern Greece. Thirteen groups of accessions were identified by hierarchical cluster analysis of 19 morphological traits, accounting for 81 % of genotype and 41 % of genotype x trait sums of squares. The distinguishing features of these groups were clarified by principal coordinates analysis. Two groups, with very desirable agronomic characteristics, originated from the Dhodhekanisos Islands (Kos, Leros, Patmos) in the south-eastern Aegean: these had rapid and tall growth, prolific podding on the main stem, pods high off the ground, many upper lateral branches, large leaves, pods and seeds, and high seed yield. Accessions from the Kikladhes Islands of the central Aegean were extremely variable and those from Naxos Island were represented in 9 of the 13 groups. Accessions from northern Greece grouped together as later flowering, shorter, and smaller-seeded types, but some accessions from the southern Greek Islands were grouped with the northern mainland types. This study identified regions in Greece, such as the south-eastern islands, where further collection may be warranted for traits of obvious agronomic value for domesticated L. angustifolius. Extreme morphological variability occurs within and between collection sites, and between localities in Greece.     

Relationships among different geographical groups,agro-morphology,fatty acid composition and RAPD marker diversity in Safflower (<Emphasis Type="Italic">Carthamus tinctorius</Emphasis>)

Patterns of geographical diversity, and the relationship between agro-morphological traits and fatty acid composition were assessed for 193 safflower (Carthamus tinctorius) accessions representing forty countries. Accessions were assigned to eight groups based on geographical proximity. Cluster and Principal Component analyses were performed to assess patterns of diversity among the accessions and to select the most distant accessions from each of eight groups for analysis of randomly amplified polymorphic DNA (RAPD) markers. There was a large amount of diversity for agro-morphological traits, fatty acid composition, and RAPD markers. Most correlations among different traits were rather low. Plant height showed a positive correlation with days to flowering (r = 0.63**). Palmitic acid was positively correlated with stearic acid and oleic acid values, and negatively correlated with linoleic acid (P < 0.01). Oleic acid and linoleic acid showed a strong negative correlation (r = −0.89**). The first three principal components together explained 59% of the variation, however, neither principal component analysis (PCA) nor marker analysis revealed a clear relationship between diversity pattern and geographical origin. Accessions from some geographical regions tended to group together, such as accessions from South Western Asia, Central Western Europe, and the Mediterranean region. The correlation between the morphological matrix and the genetic matrix based on RAPD markers was not significant (r = 0.027). Wide diversity in safflower germplasm indicates a considerable potential for improving this crop for both agronomic and quality traits.     

Relationships Among <Emphasis Type="Italic">Brassica napus</Emphasis> (L.) Germplasm from Spain and Great Britain as Determined by RAPD Markers

The genetic diversity and the relationships among a collection of Brassica napus L. European populations were evaluated using random amplified polymorphic DNA markers. The study included 33 accessions of B. napus collected from Galicia (northwestern Spain) and 18 British cultivars, 16 accessions of B. napus and two accessions of Brassica oleracea L. used as controls. DNA from 25 individuals per population was analyzed using 18 decamer primers. One hundred thirty-eight amplification products were scored of which 105 were polymorphic. These bands ranged in size from 350 to 2500 base pairs. Similarity coefficients and cluster analysis were computed and six groups were obtained. Cluster I was the largest and included all the landraces from northwestern Spain, except two accessions that grouped separately into Clusters III and IV, respectively. A low level of genetic variability was detected among the B. napus Spanish genotypes, while considerable diversity was present among the British ones, which grouped into three groups, two main clusters and one group formed by one accession. Cluster II included all commercial varieties grown in Great Britain whereas Cluster V grouped local varieties maintained by the growers for many years. Cluster VI was a singularity formed by one entry. British accessions of B. oleracea had the greatest dissimilarity with all the other populations and grouped separately in Clusters VII and VIII. As conclusion, B. napus landraces used in northwestern Spain as leafy-green vegetable probably have an independent origin from B. napus crops grown in other European regions. Besides, separate domestication in northwestern Spain and Great Britain for a different end use might have led to two distinct gene pools.     

Exploring genetic variation,oil and α-tocopherol content in avocado (<Emphasis Type="Italic">Persea americana</Emphasis>) from northwestern Mexico

The Hass cultivar of avocado is the most widely grown commercial cultivar in Mexico. Unfortunately, this cultivar is poorly adapted to the Mexican low-lands with hot dry climates characteristic of northwestern Mexico. Other well-adapted avocado accessions are available for these regions, but their nutritional traits and genetic diversity have yet to be explored. In this study, we analyze oil content, (α-tocopherol) and genetic variation among five local varieties from northwest Mexico that grow in high temperatures regimes in unfertilized soils and without any agronomic management. We report significant phenotypic variability in oil and α-tocopherol components in different accessions of avocado as determined by HPLC. Interestingly, we find higher α-tocopherol content (45.02–50.66 μg/g of fresh pulp) in some local avocados compared to Hass (32.28 μg/g of fresh pulp). The analyzed accessions represent a moderately polymorphic set of genotypes as measured by microsatellite (10 alleles by locus) and SNP (1 SNP every 164.4 bp) analysis of the VTE3 and VTE4 genes, implied in the biosynthesis of tocopherols. SNP data allowed also identifying differences between the local varieties and controls (Hass and the Mexican race accession). The variation observed at the genetic, morphologic and nutritional levels provide significant new information that may be valuable in selecting and developing avocado genotypes adapted to high-temperature environments.     

Morphological characterization of snake melon (<Emphasis Type="Italic">Cucumis melo</Emphasis> var. <Emphasis Type="Italic">flexuosus</Emphasis>) populations from Palestine

Genetic diversity in 50 snake melon accessions collected from Palestine (West Bank) was assessed by examining variation in 17 phenotypic characters. These accessions belonged to four important landraces of Cucumis melo var. flexuosus: Green “Baladi” (GB), white Baladi (WB), green Sahouri (GS), and white Sahouri (WS). Principal component analysis (PCA) and a dendrogram were performed to determine relationships among populations and to obtain information on the usefulness of those characters for the definition of cultivars. PCA revealed that secondary fruit skin color, flesh color, primary fruit skin color, and secondary skin color pattern were the principal characters to discriminate melon accessions examined in the present study. According to the scatter diagram and dendrogram, landraces of C. melo var. flexuosus: GB, WB, GS, and WS formed different clusters. However, based on Euclidean genetic coefficient distances, GB and WB had the least degree of relatedness with GS and WS, indicating distantly related landraces (Baladi and Sahouri). On the other hand, the highest degree of relatedness was detected between WS on one hand, and both GS and WB on the other indicating closely related cultivars. Fruit traits variability among the different snake melon landraces was evaluated and discussed in this study. This evaluation of fruit trait variability can assist geneticists and breeders to identify populations with desirable characteristics for inclusion in cultivars breeding programs.     

Genetic diversity and phenotypic variation for drought resistance in alfalfa (<Emphasis Type="Italic">Medicago sativa</Emphasis> L.) germplasm collected for drought tolerance

Drought is a major environmental factor hampering alfalfa productivity worldwide. Gene banks provide an array of trait diversity, frequently consisting of specific seed collection projects that focused on acquiring germplasm adapted to specific traits such as drought tolerance. These subsets provide a logical starting point to identify useful breeding resources. In the present study, we investigated the genetic diversity and evaluated 18 agronomic, physiological and quality traits associated with drought tolerance in a subset of 198 alfalfa accessions collected for their potential drought resistance. Significant differences were found among accessions for most of the traits evaluated. Water deficit significantly decreased biomass dry yield by 61.9% compared with well-watered control. A positive correlation was found between relative water content (RWC) and drought susceptible index, while negative correlations were found between RWC and leaf senescence (LS), and between RWC and canopy temperature (CT). Alfalfa accessions with high RWC showed relative lower yield reduction, cooler CT and delayed LS. Genetic variation for forage quality was also investigated and the correlation between agronomic and quality traits was analyzed. Biomass yield under drought was negatively correlated with total protein, relative feed value and relative forage quality, suggesting that the efforts to improve yield under drought tend to negatively affect forage quality. Therefore, maintaining forage quality should be considered for breeding alfalfa with drought resistance. Alfalfa accessions with high drought resistance scores and minimum loss of quality were identified in this study and can be used as genetic resources for developing alfalfa varieties with improved drought resistance and water use efficiency while maintaining forage quality.     

Genetic diversity of wild soybean (<Emphasis Type="Italic">Glycine soja</Emphasis> Sieb. et Zucc.) and Japanese cultivated soybeans [<Emphasis Type="Italic">G. max</Emphasis> (L.) Merr.] based on microsatellite (SSR) analysis and the selection of a core collection

Wild soybeans, Glycine soja, are a source of genetic variation to improve soybeans. To improve the efficiency evaluation of conserved germplasm a core or mini-core collection approach that maximizes allelic diversity in a proportion of the whole collection has frequently been advocated. The genetic diversity of a wild soybean collection (1,305 accessions) plus Japanese cultivated soybeans (53 accessions) were analyzed at 20 SSR marker loci. Higher levels of allelic diversity were found in wild soybeans (28 alleles per locus) than Japanese cultivated soybean (five alleles per locus). The genetic distance between wild soybeans from different regions reflected their proximity. Accessions from Russia consisted of a diverse array of alleles resulting in accessions being spread further apart in a PCA plot than accessions from other regions. Accessions of wild soybean from Korea included many rare alleles and thus had a high representation in the core collection. The two core collections developed here, traditional and mini, consisted of 192 accessions with 97% of the allelic diversity (14% of the whole collection) and 53 accessions with 62.4% of the allelic diversity (5% of the whole collection), respectively.     

Application of isozyme data to the management of the United States national Brassica oleracea L. genetic resources collection

Summary Management of a genetic resources collection is more effective if a curator can accurately identify genotypes and accessions as well as assess intraspecific genetic relationships and the genetic structure of species. Consequently, a study was conducted to determine whether data from starch gel electrophoresis of a specific set of isozymes from plants of Brassica oleracea (cole crops) would be useful in answering four questions the answers to which are essential for effective curatorial activities: Can individual plants be identified? Can specific accessions be identified? What are the genetic relationships among botanical varieties within B. oleracea? What is the genetic structure of the species B. oleracea? Six loci from 4 enzyme systems (LAP, PGD, PGI, and PGM) were analyzed. Individuals and accessions could not usefully be identified using these isozymes, but genetic relationships within and genetic structure of the species were easily determined, resulting in specific recommendations for improving collection management. As expected, highly selected commercial lines exhibited less diversity than average, and so may be of limited value when trying to maximize diversity with a minimum number of accessions. In contrast, landraces and weakly selected lines are more diverse than average, and thus are useful in maximizing per accession diversity. Not only was 93% of the genetic variability in B. oleracea found among accessions and a mere 7% among varieties, but also a cluster analysis showed that accessions of a single botanical variety are often more similar to those of a different variety than to each other. These results suggest that in order to create a genetic resources collection of B. oleracea that faithfully represents the diversity in the species, a curator should assemble a broad array of accessions originating from diverse agroecological niches, having different levels of improvement, and representing all botanical varieties.Abbreviations LAP Leucine amino peptidase - PGD 6-Phosphogluconate dehydrogenase - PGI Phosphoglucoisomerase - PGM Phosphoglucomutase - RAPD Random amplified polymorphic DNA - RFLP Restriction fragment length polymorphism - UPGMA Unweighted pair-group method using arithmetic averages