AFLP analysis of genetic diversity in leafy kale (<Emphasis Type="Italic">Brassica oleracea</Emphasis> L. convar. <Emphasis Type="Italic">acephala</Emphasis> (DC.) Alef.) landraces,cultivars and wild populations in Europe

AFLP markers were used to characterize diversity and asses the genetic structure among 17 accessions of kale landraces, cultivars and wild populations from Europe. The range of average gene diversity in accessions was 0.11–0.27. Several landraces showed higher levels of diversity than the wild populations and one cultivar had the lowest diversity measures. The landraces that were most genetically diverse were from areas where kales are known to be extensively grown, suggesting in situ conservation in these areas as a supplement to storage of seeds in gene banks. An analysis of molecular variance (AMOVA) showed that 62% of the total variation was found within accessions. For most accessions, genetic distance was not related to geographic distance. Similarities among accessions were probably not caused by recent gene flow since they were widely separated geographically; more likely the relationship among them is due to seed dispersal through human interactions. Our results indicate that a kale population found in a natural habitat in Denmark was probably not truly wild but most likely an escape from a cultivated Danish kale that had subsequently become naturalized.     

Genetic structure and differentiation among grapevines (<Emphasis Type="Italic">Vitis vinifera</Emphasis>) accessions from Maghreb region

Three gene pools representative of Vitis vinifera L. subsp. vinifera (=subsp. sativa Beger) growing in the Maghreb regions (North Africa) from Tunisia (44), Algeria (31) and Morocco (18) and 16 wild grape accessions (Vitis vinifera L. subsp. sylvestris (Gmelin) Beger) from Tunisia were analysed for genetic diversity and differentiation at twenty nuclear microsatellites markers distributed throughout the 19 grape chromosomes. 203 alleles with a mean number of 10.15 alleles per locus were observed in a total of 109 accessions. Genetic diversities were high in all populations with values ranging from 0.6775 (Moroccan cultivars) to 0.7254 (Tunisian cultivars). F _st pairwise values between cultivated grapevine populations were low but found to be significantly different from zero. High F _st pairwise values were shown between wild and cultivated compartments. Two parent offspring relationships, two synonyms and two clones of the same cultivar were detected. The rate of gene flow caused by vegetative dissemination of cultivated grapevine plants was not sufficient to genetically homogenise the pools of cultivars grown in different regions. The Neighbour Joining cluster analysis showed a clear separation according to geographical origins for the cultivated grapevines gene pools and revealed a high dissimilarity between cultivated and wild grapevine. However, three cultivars (Plant d’Ouchtata 1, Plant de Tabarka 3 and Plant d’Ouchtata 3) are very close to wild accessions and may result from a hybridisation between cultivated and wild accessions. The high level of differentiation between cultivated and wild accessions indicates that the cultivated accessions do not derive directly from local wild populations but could mostly correspond to imported materials introduced from others regions during historical times or derived from crossing between them.     

Population structure of the primary gene pool of Oryza sativa in Thailand

The gene pool of cultivated Asian rice consists of wild rice (Oryza rufipogon Griff.), cultivated rice (O. sativa L.) and a weedy form (O. sativa f. spontanea). All three components are widespread in Thailand, frequently co-occurring within fields and providing the opportunity for gene flow and introgression. The purpose to this study is to understand the on-going evolutionary processes that affect the gene pool of rice by analysis of microsatellite variation. Results indicate that O. rufipogon, the wild ancestor of rice, has high levels of genetic variation both within and among populations. Moreover, the variation is structured predominantly by annual and perennial life history. High levels of variation are detected among cultivars indicating Thai cultivated rice has a broad genetic base with only a 20 % reduction in diversity from its wild ancestor. The weedy rice populations reveal varying levels of genetic variation, from nearly as high as wild rice to near zero. Weedy rice is genetically structured into 2 groups. Some populations of invasive weedy rice are the result of hybridization and gene flow between local wild rice and local cultivated rice in the regions of co-occurrence. Other populations of weedy rice are genetically nearly identical to the local cultivated rice. The diversity analysis indicates that the rice gene pool in Thailand is a dynamic genetic system. Gene flow is ongoing among its three main components, first between cultivated and wild rice resulting in weedy rice. Weedy rice in turn crosses with both cultivated varieties and wild rice.     

Phylogenetic position of east Tibetan natural populations in Tartary buckwheat (Fagopyrum tataricum Gaert.) revealed by RAPD analyses

Phylogenetic relationships among cultivated landraces and natural populations of wild subspecies of Tartary buckwheat were investigated by constructing an NJ tree based on RAPD markers focussing on east Tibetan natural populations. Ten plants from three cultivated landraces and 29 plants from five natural populations of wild subspecies in eastern Tibet were used for RAPD analyses. The wild subspecies from eastern Tibet was classified into three types; (1) same type as cultivated landraces; (2) closely related to natural populations of northwestern Yunnan; and (3) an exceptional population, Zhuka, which was closely related to Sichuan populations. Since the type (2) is considered as the wild ancestral type of cultivated Tartary buckwheat, we conclude that eastern Tibet too may be one of the center of origin of this crop.     

Morphologic and Agronomic Diversity of Wild Genetic Resources of Medicago sativa L. Collected in Spain

One hundred and three natural populations of Medicago L. were collected in Spain, mainly from roadsides, non-irrigated or grazed lands. This germplasm was evaluated at Montpellier (France) with control cultivars and Spanish landraces under completely random block design, replicated four times, and observed for 64 qualitative or quantitative characters. Differences between natural populations and cultivated controls are highly significant, but gene flow occurs between wild and cultivated compartment and hybrid populations were identified. On the basis of multivariable analysis, the accessions were grouped into four clusters depending on their proximity with the cultivated pool to facilitate their management and ex situ conservation. The relationship between environment of the site of collection and phenotypic characteristics of the natural populations was also discussed. Different policies of conservation of these genetic resources are suggested to avoid their disappearance even when they disappeared from the other regions of the western Mediterranean. Spanish wild pool of alfalfa, also called ‘Mielga’, appears of great interest for the breeding of alfalfa because it contains a large diversity of characteristics (prostrate habit, rhizomes) linked to tolerance to grazing or drought environment. With the need for more sustainable systems in agriculture, the erosion of natural habitats and the necessity for rehabilitation of degraded ecosystems, the importance of this wild pool is really inestimable.     

Exploration of genetic diversity within Cichoriumendivia and Cichorium intybus with focus on the gene pool of industrial chicory

The present study used 15 simple sequence repeat loci to characterize the genetic diversity of the germplasm that originated the current industrial chicory and to establish the relationships between and inside Cichoriumintybus L. and Cichorium endivia L. Initially we analyzed 19 cultivated C. endivia accessions, 27 wild and 155 cultivated C. intybus accessions distributed among three groups: 83 root chicories, 42 Witloof and 30 leaf chicories. The leaf chicories comprised cultivars corresponding to the Radicchio, Sugarloaf and Catalogne subgroups. The latter has not been previously included in any genetic diversity study. Subsequently, 1297 individuals from the 15 modern root chicory cultivars at the origin of the breeding of the current industrial root chicory cultivars were analyzed. Although the accessions of C. endivia and C. intybus were clearly separated from each other, seven wild C. intybus individuals were genetically closer to C. endivia than to C. intybus, revealing complex genetic interrelationships between these species. The differentiation of C. intybus into three cultivar groups (Witloof, root chicory and leaf chicory) was confirmed. The leaf chicory individuals were divided into three genetic subgroups, corresponding to the Radicchio, Sugarloaf and Catalogne cultivars, thus attesting to the validity of the classification based on morphological factors. Clear differentiation was observed among the Belgian, Polish and Austrian modern industrial root cultivars, but not among the French industrial modern root cultivars. The high phenotypic and genetic variability of the modern industrial root cultivars indicates that this germplasm constitutes a useful gene pool for cultivar improvement and selection.     

Genetic identity and relationships of Iranian apple (Malus?×?domestica Borkh.) cultivars and landraces, wild Malus species and representative old apple cultivars based on simple sequence repeat (SSR) marker analysis

In order to shed light on the role of Iran in apple evolution and domestication, we chose to investigate the relationships of a collection of 159 accessions of wild and domesticated apples including Iranian indigenous apple cultivars and landraces, selected wild species, and old apple scion and rootstock cultivars from different parts of the world. The majority of the wild species belonged to M. sieversii, which is widely believed to be the main maternal wild ancestor of domestic apples, from Kazakhstan and M. orientalis, which is one of the probable minor ancestors of domestic apples, from Turkey and Russia located on the east and west of Iran, respectively. The accessions were assigned into six arbitrary populations for the purpose of generating information on genetic parameters. Nine simple sequence repeat (SSR) loci selected from previous studies in apple were screened over DNA extracted from all the accessions. Results showed that all SSR loci displayed a very high degree of polymorphism with 11–25 alleles per locus. In total, there were 153 alleles across all loci with an average of 17 alleles per locus. The SSR allelic data were then used for estimation of population genetic parameters, including genetic variation statistics, F-statistics, gene flow, genetic identity, genetic distance and then cluster analysis using POPGENE 1.32 software. The F-statistics and gene flow in particular, showed that there was more intra-population than between population variation. The genetic identity and genetic distance estimates, and the dendrogram generated from the un-weighted pair group arithmetic average (UPGMA) method of cluster analysis showed that the Iranian cultivars and landraces were more closely related to M. sieversii from Central Asia (east of Iran) and M. orientalis native to Turkey and Russia than to other accessions of Malus species. Also, the old apple cultivars from different parts of the world have a closer genetic relationship to M. sieversii, M. orientalis and the Iranian apples, than to other wild species. Based on these results, we suggest that the Iranian apples may occupy an intermediate position between the domesticated varieties and wild species. We propose that Iran could be one of the major players in apples’ domestication and transfer from Central Asia to the western countries.     

Origin of cultivated Tatary buckwheat (Fagopyrum tataricum Gaertn.) revealed by RAPD analyses

The phylogenetic relationships among cultivated landraces and natural populations of wild subspecies of Tatary buckwheat were investigated at the individual level by constructing a phylogenetic tree based on RAPD markers. As the PCR templates, DNA of individuals rather than bulked samples, was used. Ten individuals from 10 cultivated landraces, 71 individuals from 21 natural populations of wild subspecies, and 7 individuals from 3 weedy Tatary buckwheat were provided for RAPD analyses. Three groups were recognized: (1) all cultivated landraces and wild subspecies from central Tibet and northern Pakistan; (2) 10 individuals of wild subspecies from northwestern Yunnan; (3) the remaining individuals of wild subspecies from northwestern Yunnan and all individuals of wild subspecies from Sichuan. Group (2) was phylogenetically closely related to group (1). The origin of cultivated Tatary buckwheat, the hybrid origin of weedy Tatary buckwheat and of the wild populations from central Tibet and northern Pakistan are discussed. We arrive at the conclusion that cultivated Tatary buckwheat probably originated in northwestern Yunnan in China.     

Processes affecting genetic structure and conservation: a case study of wild and cultivated <Emphasis Type="Italic">Brassica rapa</Emphasis>

When planning optimal conservation strategies for wild and cultivated types of a plant species, a number of influencing biological and environmental factors should be considered from the outset. In the present study Brassica rapa was used to illustrate this: to develop Scandinavian conservation strategies for wild and cultivated B. rapa, DNA-marker analysis was performed on 15 cultivated and 17 wild accessions of B. rapa plus 8 accessions of the cross compatible B. napus. The B. rapa cultivars were bred in Sweden and Finland in 1944–1997 and the wild B. rapa material was collected from Denmark, Sweden and United Kingdom. The B. napus accessions were bred within the last 20 years in the Scandinavian countries. Results were based on scoring of 131 polymorphic ISSR markers in the total plant material. A Bayesian Markov chain Monte Carlo (MCMC) approach implemented in NewHybrids demonstrated a clear distinction of B. rapa and B. napus individuals except for three individuals that seemed to be backcrosses. The backcrossed hybrids descended from two Swedish populations, one wild and one escaped. The overall pattern of genetic variation and structure in B. rapa showed that cultivated and wild B. rapa accessions formed two almost separated clusters. Geographical origin and breeding history of cultivars were reflected in these genetic relationships. In addition, wild populations from Denmark and Sweden seemed to be closely related, except for a Swedish population, which seemingly was an escaped cultivar. The study point to that many processes, e.g. spontaneous introgression, naturalisation, breeding and agricultural practise affected the genetic structure of wild and cultivated B. rapa populations.     

Genetic differentiation and diversity of phenotypic characters in Chinese wild soybean (<Emphasis Type="Italic">Glycine soja</Emphasis> Sieb. et Zucc.) revealed by nuclear SSR markers and the implication for intraspecies phylogenic relationship of characters

Genetic diversity is reduced from wild soybean to cultivars and from landraces to modern varieties. However, intraspecies genetic diversity loss between characters or phenotypes also existed in wild soybean. We revealed the phylogenic relationship of character types in Chinese wild soybean using 42 SSR markers. We conjectured that white flower, no-seed bloom, grey pubescence, and four seed coat colours were evolutionarily acquired phenotypes. There were a small decrease (∆H = 0.1–6.46%) of gene diversity and a moderate reduction (∆Na = 10.81–53.54%) of number of alleles but a violent loss (80.74–98.59%) of unique alleles in the acquired phenotypes. Our results seemed to suggest that ovoid and elliptic leaves were differentiated at the earliest and subsequently lanceolate leaf appeared before the domestication of soybeans within wild soybean, and that G. gracilis type was another earliest type, maybe emerged since the appearance of soybeans or it was concomitant with the domestication of soybean.     

Characterization of single nucleotide polymorphism in Tunisian grapevine genome and their potential for population genetics and evolutionary studies

In this study, two gene fragments corresponding to the VvMYBA1 and VvMYBA2 loci were sequenced on a sample of grapes including cultivated and wild accessions originating from Tunisia, Germany and France. A total of 42 SNPs were detected in the sequenced fragments giving an average of 1 SNP every 33 bp. High level of polymorphism was observed in the samples either in cultivated or wild accessions. Pattern of nucleotide diversity indicates a non departure from neutrality expectations for wild grapevine sample for gene VvMYBA1 and VvMYBA2 and for cultivated sample for gene VvMYBA1. However, a linkage to a selective sweep was revealed for cultivated grapevine gene pool in gene VvMYBA2. A genetic structure of the studied sample according to accession taxonomic status was revealed by the UPGMA clustering with a considerable overlap. This result was confirmed by significant but low genetic differentiation values between cultivated and wild sample. The number of migrants Nm based on sequence data information between Tunisian cultivars and Tunisian wild accessions showed a low level of gene flow between those germplasms. This finding indicates that Tunisian cultivars do not derive directly from local wild populations but could mostly correspond to imported materials introduced during historical times. However, the possibility that some cultivars derived from ancestral events of local domestication or cross hybridization with native wild plants was not completely excluded for Tunisian grapevine accessions.     

Phenolic Fingerprint of Sicilian Modern Cultivars and Durum Wheat Landraces: A Tool to Assess Biodiversity

Durum wheat landraces have been recognized to have different contents in health‐promoting phytochemicals. These health benefits have been mainly attributed to the presence of phenolic acids and flavonoids existing in this matrix both as soluble free compounds and insoluble bound forms. Sicily is one of the few areas of Southern Europe where it is still possible to find landraces of durum wheat, no longer cropped because of the commercial spread of new high‐yielding varieties. This study aimed to determine the phenolic profile and content of 10 wheat landraces and three genetically improved cultivars to develop an analytical tool for cultivar screening and traceability. Hydroalcoholic extracts from wheat grains were analyzed by means of HPLC with a diode‐array detector and HPLC electrospray ionization MS, revealing 13 metabolites belonging mainly to the classes of hydroxycinnamic acids and flavones C‐glycosides. The quantitative pattern of the 13 phenolic markers, analyzed by applying stepwise linear discriminant analysis, allowed the discrimination between the two groups under study, namely, modern cultivars and durum wheat landraces. Furthermore, all 13 durum wheat varieties were perfectly discriminated in this statistical model, confirming a specific and genotype‐dependent pattern of phenolics concentration.     

Genetic Structure of Wild and Domesticated Populations of Capsicum annuum (Solanaceae) from Northwestern Mexico Analyzed by RAPDs

Levels of genetic variation and genetic structure of 15 wild populations and three domesticated populations of Capsicum annuum were studied by RAPD markers. A total of 166 bands (all of them polymorphic) and 126 bands (125 of them polymorphic) were amplified in wild and domesticated populations, respectively. Mean percentage of polymorphism was 34.2% in wild populations and 34.7% in domesticated populations. Mean and total genetic diversity were 0.069 and 0.165 for wild populations and 0.081 and 0.131 for domesticated populations. Parameters of genetic diversity estimated from 54 bands with frequencies ≥1 − (3/n) (n = sample size) showed that 56.7% of the total variation was within and 43.3% among wild populations, whereas 67.8% of the variation was within and 32.2% among domesticated populations. AMOVA indicated that total genetic diversity was equally distributed within (48.9 and 50.0%) and among (50.0 and 51.1%) populations in both wild and domesticated samples. Wild and domesticated populations were clearly resolved in a UPGMA dendrogram constructed from Jaccard’s distances (average GD = 0.197), as well as by AMOVA (17.2% of variance among populations types, p = 0.001) and by multidimensional scaling analysis. Such differentiation can be associated with domestication as well as different origin of gene pools of the wild (Northwestern Mexico) and cultivated (more probably Central Mexico) samples analyzed. The considerable genetic distances among cultivars (average GD = 0.254) as well as the high number of diagnostic bands per cultivar (33 out of 126 bands), suggest that genetic changes associated with domestication could have resulted from artificial selection intervening in different directions, but the inclusion of more domesticated samples might clarify the nature of distinctions detected here.     

Determining Sources of Diversity in Cultivated and Wild <Emphasis Type="Italic">Lablab purpureus</Emphasis> Related to Provenance of Germplasm by using Amplified Fragment Length Polymorphism

To improve understanding of diversity of Lablab purpureus and establish relationships among 103 germplasm accessions collected from diverse geographic origins, amplified fragment length polymorphism markers were used. Four primer sets selected out of 16 produced 289 clear, repeatable polymorphisms. UPGMA analysis of similarity data clustered the accessions according to their subspecific taxonomic organization, i.e., subsp. purpureus and subsp. uncinatus, as well as to cultivated and wild forms. The well-represented landraces from Africa and Asia, belonging predominantly to subsp. purpureus, displayed moderate genetic diversity. Wild forms from Africa showed far greater levels of diversity that would justify taxonomic re-assessment of the wild subsp. uncinatus. The molecular analysis identified forms that were collected in the wild in India but were genetically placed intermediate between wild and cultivated forms. As these plant types did not exist among the African accessions, it is suggested that they might represent escapes from early attempts of domestication. These results support the suggested pathway of domestication and distribution of L. purpureus from Africa to Asia. Additional members to a previously published core collection of the species are proposed.     

Phylogenetic relationships, interspecific hybridization and origin of some rare characters of wild soybean in the subgenus Glycine soja in China

The Glycine subgenus Soja includes two species, cultivated soybean [(Glycine max (L.) Merr.)] and the progenitor wild soybean (G. soja). However, a morphologically intermediate form, the semi-wild soybean (G. gracilis), exists between the two species, and its taxonomic position is under debate. In this study, we evaluated phylogenetic relationships and occurrence events within the subgenus Soja based on genetic variation of SSR loci using a set of accessions comprising wild soybeans (≤3.0?g 100-seed weight), semi-wild soybeans (>3.0?g) and soybean landraces (≥4.0?g). The results showed that semi-wild soybean accessions collected in natural fields should be treated as a variant of G. soja and not of G. max, and were genetically differentiated from the soybean landraces, even large-seeded semi-wild soybean accessions (6.01–9.0?g) with seed weights overlapping with or exceeding those of soybean landraces. Evolutionary bottleneck analysis indicated that semi-wild soybean is not a transitional form in the domestication of cultivated soybeans from wild soybean. G. soja contained two genetically differentiated forms, small-seeded type (typical, plus 2.01–2.50?g) and a large-seeded type (2.51–3.0?g). Genetically, the large-seeded wild soybean was closer to the semi-wild soybean, although in morphology it resembled the typical wild soybean. Ancestry analysis confirmed that cultivated soybean genes have introgressed into modern wild soybean populations. The green cotyledon character and other rare characters such as white flower, grey pubescence, no-seed bloom, and coloured seed-coats (brown, green, and yellow) in wild soybean were shown to be involved in introgression from cultivated soybeans.     

Using SSR markers to determine the population genetic structure of wild apricot (<Emphasis Type="Italic">Prunus armeniaca</Emphasis> L.) in the Ily Valley of West China

Genetic structure of three wild populations (Xinyuan, Gongliu and Daxigou) of apricot in the Ily Valley, Xinjiang Uygur Autonomous Region of China, was investigated with microsatellite (simple sequence repeat, SSR) markers. The higher polymorphism and greater transportability of these markers between Prunus species proved SSR markers were much efficient for conducting genetic diversity studies in wild apricot. Nei's gene diversity (He) and Shannon's index of diversity (I) were 0.287 and 0.458, respectively. This indicated that the wild apricot in the Ily Valley still maintained a relatively high level of diversity. The Gst of 0.137 and Fst of 0.164 revealed that genetic variation mainly resided among individuals within populations (83.6–86.3%). Population differentiation could also be found according to the distribution of SSR alleles between the populations. Mantel test showed the genetic distance between populations was significantly correlated to the geographical distance. The modest amount of gene flow (2.684) would reduce the disjunction between wild apricots. The long-distance dispersal of pollen by insects was probably the main way of gene flow between populations. Based on the study of population genetic structure, an effective conservation strategy of the species was discussed.     

EST <Emphasis Type="Italic">versus</Emphasis> Genomic Derived Microsatellite Markers for Genotyping Wild and Cultivated Barley

Genetic variation present in wild and cultivated barley populations was investigated using two sources of microsatellite also known as simple sequence repeat (SSR) markers. EST-SSRs are derived from expressed sequences and genomic SSRs are isolated from genomic DNA. Genomic SSR markers detected a higher level of polymorphism than those derived from ESTs. Polymorphism information content was higher in genomic SSRs than EST-derived SSRs. This study showed that the EST-SSR markers developed in cultivated barley are polymorphic in wild and cultivated varieties and produced high quality markers. Ten of these functional markers were polymorphic across the accessions studied. EST markers indicated clearer separation between wild and cultivated barley than genomic SSRs. The EST-SSRs are a valuable source of new polymorphic markers and should be highly applicable to barley genetic resources, providing a direct estimate of functional biodiversity.     

Inferring seed exchange between farmers from population genetic structure of barley landrace Arabi Aswad from Northern Syria

Apart from their significance in genetic conservation, barley landraces are still favoured by local farmers in low-input farming systems. They often perform more predictably under adverse conditions than modern cultivars due to local adaptation. Increased seed exchange between farmers may put adaptation of local populations of barley landraces at risk. Isozyme markers were used to investigate differentiation and infer gene flow between local populations of barley landrace Arabi Aswad in Northern Syria. Inferred gene flow decreased exponentially with geographic distance and may imply that seed exchange between farmers is limited to a regional scale and diminishes over longer distance. Gene flow seems to be a suitable index for estimating seed exchange in highly inbreeding crop species such as barley. In the future, improved mobility of farmers and extension work may facilitate seed exchange over longer distances, and consequently jeopardise preservation of locally adapted landraces in barley.