Pyrrolizidine alkaloids in two endemic capeverdian Echium species

Echium hypertropicum Webb and Echium stenosiphon Webb subsp. stenosiphon are capeverdian shrubs used in folk medicine for the treatment of cough and gastrointestinal diseases. Acid-base extraction was used to obtain alkaloid-rich fractions from both species. GC–MS and ESI-MS/MS analysis indicated the presence of pyrrolizidine alkaloids (PAs) and purified substances were also analyzed by 1D and 2D NMR experiments. A total of ten alkaloids were detected, eight of which were identified by comparing their molecular masses and mass fragmentation patterns with the NIST database and data in the literature. The hepatotoxic diesters echimidine and 7-(2-methylbutyryl)-9-echimidinylretronecine were identified in both species. Echimidine was the major component in the diethyl ether fraction from leaves of E. hypertropicum, whereas the 7-(2-methylbutyryl)-9-echimidinylretronecine was the major component in the dichloromethane fraction from leaves of E. stenosiphon. To our knowledge, this is the first report on the occurrence of pyrrolizidine alkaloids in capeverdian species of Echium. This study on Echium led to the identification of constituent pyrrolizidine alkaloids, serving to assist taxonomists with the complex taxonomy of this genus.     

Molecular diversity and relationships of North American Elymus trachycaulus and the Eurasian E. caninus species

The morphological similarity of Elymus trachycaulus to the Eurasian E. caninus has often been noted. This has lead to controversial and contradicting taxonomic treatments. Nevertheless, there has been no systematic investigation on molecular genetic similarity between E. trachycaulus and E. caninus. In this study, random amplified polymorphic DNA (RAPD) analysis was used to study the similarity between the two species. RAPD analysis of 38 samples representing E. caninus and E. trachycaulus complex yielded 111 interpretable RAPD bands. The Jaccard’s similarity values for E. caninus ranged from 0.38 between accessions H10345 and H10353 to 0.97 between accessions H8745 and H10096, with an average of 0.67. The Jaccard’s similarity values for E. trachycaulus complex ranged from 0.09 between E. trachycaulus ssp. subsecundus (PI 537321) and E. trachycaulus ssp. violaceus (PI 272612) to 0.78 between accessions PI 315368 and PI 372644, with an average of 0.43. The results from different analyses (NJ and PCA) were similar but not identical. The molecular genetic separation between E. caninus and E. trachycaulus was consistent. The PCA analysis clearly separated all E. caninus accessions from E. trachycaulus and its subspecies. The NJ analysis also showed separation between most accessions of E. caninus and E. trachycaulus. Further analysis excluding E. trachycaulus ssp. subsecundus and ssp. violaceus revealed that E. caninus species and E. trachycaulus species were clearly separated into two distinct groups. The RAPD data thus support the treatment of E. caninus and E. trachycaulus as distinct species. The analyses further indicate that E. violaceus is nested within E. trachycaulus, and more related to E. trachycaulus complex rather than to E. caninus.     

Isozyme diversity in some Canarian woody endemisms of the genus Echium L. (Boraginaceae)

Oceanic archipelagos are considered hot-spots of biodiversity because they harbor unique biota with a high level of endemicity. However, the endemic biodiversity of oceanic islands is very vulnerable to extinction. In recent decades, intensive exploitation of these territories and human-mediated introduction of alien species have posed unprecedented threats to the long-term survival of the endemic contingent. The very limited population genetic information available until now for the 28 Canarian endemic Echium taxa has hindered the development of conservation strategies for the ca. 25% of them that are under threat. In this paper, we analyze the levels and distribution of genetic isozyme diversity in 23 natural populations of three endangered endemics of restricted distribution (E. acanthocarpum, CR; E. onosmifolium ssp. spectabile, EN; and E. callithyrsum, VU), and two endemics of wide distribution and in principle free of threat (E. decaisnei and E. onosmifolium). Our results reveal high levels of genetic variability in all these taxa that have plausibly been reached despite a predominance of selfing. They also point out a high incidence of inbreeding in the reproductive dynamics of populations and suggest the potential value of hybridisation processes in shaping the genetic makeup of these species. Among-population differentiation, as estimated by Gst, and genetic distances within taxa are low overall, but they do support the current taxonomic separation between the two subspecies of E. onosmifolium and do not furnish evidence that the current status of the endangered species may be attributed to genetic factors. We use the genetic parameters to suggest some guidelines to help implement a conservation strategy of these taxa.     

Conservation of genetic diversity in British populations of the diploid endemic Coincya monensis ssp monensis (Isle of Man Cabbage): the risk of hybridisation with the tetraploid alien, Coincya monensis ssp cheiranthos

Coincya monensis is represented in the British flora by two, cytologically distinct subspecies. Coincya monensis ssp monensis is an endemic diploid with a coastal sand dune distribution that includes a number of isolated populations. Coincya monensis ssp cheiranthos is a tetraploid alien, well established in South Wales in early successional habitats. Both subspecies share similar life form traits, flowering times and pollinators. Cluster analysis and phylogenetic reconstruction based on sequences of the mitochondrial nad4 gene confirmed the distinction between alien and endemic taxa. Tetraploid populations carry more polymorphic RAPDs loci and their genetic diversity is partitioned more within than among populations. In contrast, C. monensis ssp monensis has a distinct population genetic structure. Analysis of the multilocus genetic data confirmed a structure of genetically isolated, endemic population clusters in Scotland, Arran, the Isle of Man and South Wales. Experimental hybridisation showed the two subspecies are interfertile. Multivariate analysis of RAPDs data resolved hybrids between alien and endemic clusters and hybrids contained a proportion of alien-specific polymorphic loci. Hybrids of alien maternal parentage contained the mitochondrial nad4 sequence characteristic of the alien subspecies. Since the alien subspecies can invade mobile sand dune communities from urban sites and compete for pollinators, there is a risk that alien and endemic populations will mix and introgress. Conservation of endemic genetic diversity in Britain will require protection for all C. monensis ssp monensis populations. Currently, the most disjunct endemic population in South Wales is most at risk from introgression.     

Analysis of the genetic diversity and relationships among and within species of Hippophae (Elaeagnaceae) based on RAPD markers

RAPD markers were used to assess the genetic diversity and inter- and intra-specific relationships of the genus Hippophae L. and to study the correlation between genetic distances and geographic distances among populations of H. rhamnoides ssp. sinensis. The results analyzed by the percentage of polymorphic loci and Shannon information index indicated that a high level of genetic diversity existed both among and within species of the genus Hippophae. In the UPGMA dendrogram, the species or subspecies were clustered into two main groups but not strictly grouped according to sect. Hippophae and sect. Gyantsensis Lian. The multiple regression analysis and Mantel test both indicated a significant correlation between genetic distance and altitude distance among populations of H. rhamnoides ssp. sinensis, and the cluster analysis suggested that the genetic variation among populations of H. rhamnoides ssp. sinensis was linked to their monophyletic origin. Moreover, some degree of genetic differentiation was found among samples collected at different times.     

Genetic Variation in Hippophae rhamnoides ssp. sinensis (Elaeagnaceae) Revealed by RAPD Markers

Hippophae rhamnoides ssp. sinensis is endemic to China, and it is a dioecious, outcrossing plant. Although many studies have been undertaken mainly on its agricultural, nutritional, medical, and ornamental value, little is known about its population genetics. This study uses random amplified polymorphic DNA to investigate the genetic diversity and population genetic structure of 13 natural populations of the subspecies sinensis. Fifteen primers amplified 107 reproducible bands, with 95 (88.79%) being polymorphic. The gene diversity within population was 0.168, considerably lower than that of tree species and most perennial, outcrossing species, but higher than that of annual or short-lived, selfing species. The Gst value showed that 18.3% of the total genetic variation resided among populations, a little lower than that of outcrossing species. The present results are quite similar to those previously reported in another subspecies, H ssp. . rhamnoides rhamnoides. The low genetic differentiation among populations in ssp. sinensis may be attributed to the long-distance dispersal of seeds facilitated by birds, in addition to its characteristics of outcrossing, wind pollination, and widespread distribution. No association between genetic distance and geographical distribution was found. The population relationships revealed by the UPGMA dendrogram parallel this result, in that genetic distance did not increase with geographic separation. This pattern of population differentiation may imply the adaptation of ssp. s populations to the local environment, given that its habitats vary greatly across its distribution.     

Development of SCAR molecular markers for early and late differentiation of Eucalyptus globulus ssp globulus from E. globulus ssp maidenii

The genus Eucalyptus includes over 700 species, some of which are the most widely planted hardwoods worldwide. Each species and subspecies of Eucalyptus present different characteristics regarding its wood quality and yield. This fact makes it very important to work with known species/subspecies so as to optimize handling and conservation of forest resources. Some of them are morphologically very similar, making it difficult to differentiate by simple observation. This is the case with Euclyptus globulus ssp globulus and E. globulus ssp maidenii, which can only be distinguished in the adult tree. These issues can be avoided using well-characterized seeds. This can be quite expensive, especially for the small growers. An alternative approach is to develop simple methods for the differentiation of the two subspecies. In this work, we develop a quick method, based on SCAR molecular markers derived from RAPD molecular markers, for the differentiation of the subspecies of E. globulus, in particular E. globulus ssp globulus and E. globulus ssp maidenii.     

Taxonomic Relationship Among Seven Species of Groupers (Genus <Emphasis Type="Italic">Epinephelus</Emphasis>; Family Serranidae) as Revealed by RAPD Fingerprinting

Phenotypic identification of groupers of the genus Epinephelus is based on color patterns and a suite of morphologic characters. However, these characters often show intraspecific variations and differences between juveniles and adults of the same species. The present work is an attempt to study and ratify the status of Epinephelus spp. using random amplified polymorphic DNA (RAPD) analysis based on samples drawn from southeast and southwest coasts of India. The RAPD fingerprints generated in Epinephelus diacanthus, E. areolatus, E. chlorostigma, E. bleekeri, E. coioides, E. tauvina, and E. malabaricus with 4 primers (OPA 01, OPA 07, OPF 08, and OPF 10) were consistent, reproducible, and yielded species-specific diagnostic markers in all the species. A total of 59 RAPD loci in the size range of 70 to 4500 bp were produced from all 4 arbitrary primers. An unweighted pair-group method with arithmetic mean (UPGMA) dendogram was constructed based on genetic distance values to show the genetic relationships among the 7 species. All the individuals of each species formed monophyletic species clusters. The mean intraspecies genetic distance value (0.305) was significantly lower than the interspecies value (0.365). Epinephelus malabaricus was most distantly related to E. diacanthus and E. bleekeri. The genetic relationship was very close among E. coioides, E. tauvina, and E. malabaricus and also between E. chlorostigma and E. bleekeri. Within-species genetic polymorphism was highest in E. chlorostigma (49.15%) and lowest in E. tauvina (25.42%).     

Genetic relationships in Lens species and parentage determination of their interspecific hybrids using RAPD markers

Broadening of the genetic base and systematic exploitation of heterosis in cultivated lentils requires reliable information on genetic diversity in the germplasm. The ability of random amplified polymorphic DNA (RAPD) to distinguish among different taxa of Lens was evaluated for several geographically dispersed accessions/cultivars of four diploid Lens species. This study was carried out to assess whether RAPD data can provide additional evidence about the origin of the cultivated lentil and to measure genetic variability in lentil germplasm. Three cultivars of Lens culinaris ssp. culinaris, including one microsperma, and two macrosperma types, and four wild species (L. culinaris ssp. orientalis, L. odemensis and L. nigricans) were evaluated for genetic variability using a set of 1 11-mer and 14 random 10-mer primers. One hundred and fifty-eight reproducible and scorable DNA bands were observed from these primers. Genetic distances between each of the accessions were calculated from simple matching coefficients. Split decomposition analysis of the RAPD data allowed construction of an unrooted tree. This study revealed that (1) the level of intraspecific genetic variation in cultivated lentils is narrower than that in some wild species. (2) L. culinaris ssp. orientalis is the most likely candidate as a progenitor of the cultivated species, (3) L. nigricans accession W6 3222 (unknown) and L. c. ssp. orientalis W6 3244 (Turkey) can be reclassified as species of L. odemensis and (4) transmission of genetic material in Lens interspecific hybrids is genotypically specific, as identified by the RAPD markers in our study.     

RAPD Variation Among North Vietnamese <Emphasis Type="Italic">Flemingia macrophylla</Emphasis> (Willd.) Kuntze ex Merr. accessions

Flemingia macrophylla (Willd.) Kuntze ex Merr., a multi-purpose legume with potential as dry-season forage crop, mainly occurs in subhumid to humid environments of tropical and subtropical Asia. Despite increasing interest in conservation of germplasm suitable for low-input production systems information on the genetic diversity of F. macrophylla is extremely scarce. The creation of baseline data is supposed to contribute to more efficient conservation management and to identify collecting strategies of novel germplasm. Random amplified polymorphic (RAPD) markers were used to investigate the genetic variation among 37 F. macrophylla accessions. Germplasm analysed in this study originated from Bac Kan province, Northeast Vietnam. Eight primers generated a total of 47 amplified RAPD loci of which 38 were polymorphic. Jaccard’s similarity coefficients among accessions ranged from 0.069 to 1 with a mean of 0.67. The UPGMA dendrogram revealed three clusters along with three outliers. No correspondence between geographic and genetic distance was found (Mantel test: R = 0.21; P = 0.016). Analysis of molecular variance (AMOVA) revealed significant (P < 0.001) differentiation between accessions collected in lowland and upland regions. Results of UPGMA clustering were confirmed by the pattern of principle coordinates analysis (PCO) plotting. Future collecting strategies should target populations at large distances and along the altitudinal range. Ex situ conservation should encompass those accessions that showed genetic divergence. In situ conservation may consist of establishing a system of interconnected population fragments to guarantee continuing genetic exchange via corridors and of rehabilitating degraded habitats.     

RAPD Profiling in Detecting Genetic Variation in Endemic Coelonema (Brassicaceae) of Qinghai-Tibet Plateau of China

Random amplified polymorphic DNA (RAPD) markers were used to measure genetic diversity of Coelonema draboides (Brassicaceae), a genus endemic to the Qilian Mountains of the Qinghai-Tibet Plateau. We sampled 90 individuals in 30 populations of Coelonema draboides from Datong and Huzhu counties of Qinghai Province in P.R. China. A total of 186 amplified bands were scored from the 14 RAPD primers, with a mean of 13.3 amplified bands per primer, and 87% (161 bands) polymorphic bands (PPB) was found. Analysis of molecular variance (AMOVA) shows that a large proportion of genetic variation (84.2%) resides among individuals within populations, while only 15.8% resides among populations. The species shows higher genetic diversity between individuals than other endemic and endangered plants. The RAPDs provide a useful tool for assessing genetic diversity of rare, endemic species and for resolving relationships among populations. The results show that the genetic diversity of this species is high, possibly allowing it to adapt more easily to environmental variations. The main factor responsible for the high level of differentiation within populations and the low level of diversity among populations is probably the outcrossing and long-lived nature of this species. Some long-distance dispersal, even among far separated populations, is also a crucial determinant for the pattern of genetic variation in the species. This distributive pattern of genetic variation of C. draboides populations provides important baseline data for conservation and collection strategies for the species. It is suggested that only populations in different habitats should be studied and protected, not all populations, so as to retain as much genetic diversity as possible.     

A survey of equid mitochondrial DNA: Implications for the evolution, genetic diversity and conservation of Equus

The evolution, taxonomy and conservation of the genus Equuswere investigated by examining the mitochondrial DNA sequences of thecontrol region and 12S rRNA gene. The phylogenetic analysis of thesesequences provides further evidence that the deepest node in thephylogeny of the extant species is a divergence between twolineages; one leading to the ancestor of modern horses (E.ferus, domestic and przewalskii) and the other to thezebra and ass ancestor, with the later speciation events of the zebrasand asses occurring either as one or more rapid radiations, or withextensive secondary contact after speciation. Examination of the geneticdiversity within species suggested that two of the E. hemionussubspecies (E. h. onager and E. h. kulan) onlyrecently diverged, and perhaps, are insufficiently different to beclassified as separate subspecies. The genetic divergence betweendomestic and wild forms of E. ferus (horse) and E.africanus (African ass) was no greater than expected within anequid species. In E. burchelli (plains zebra) there was anindication of mtDNA divergence between populations increasing withdistance. The implications of these results for equid conservation arediscussed and recommendations are made for conservation action.     

Phylogeography of silver‐studded blue,Plebejus subsolanus (Lepidoptera: Lycaenidae), in the central mountainous regions of Japan

The silver‐studded blue, Plebejus subsolanus, is widely distributed in the Russian Altai mountains, northeastern China, the Korean Peninsula, and the Japanese archipelago. In Japan, the species is distributed across wide elevation ranges from the lowlands of Hokkaido to the subalpine zone of Honshu. Current subspecies classification in Japan is as follows: ssp. iburiensis, occurring in lowland grasslands in Hokkaido; ssp. yaginus in lower mountain grasslands in Honshu; and ssp. yarigadakeanus in higher mountain grasslands in Honshu. The habitat of this species has been markedly reduced due to recent habitat destruction and land‐use changes. Here, we undertook phylogeographic analyses of two subspecies, ssp. yaginus and yarigadakeanus in the central mountainous regions of Japan, based on two mitochondrial gene sequences, in order to collect information for establishing effective conservation strategies. From 57 samples from the four mountain ranges, we obtained a haplotype network comprised of 12 haplotypes. Because of the haplotype network topology, the geographic distribution of haplotypes and the correspondence of haplotype divergence to subspecies taxonomy, we provisionally divided the haplotypes into three haplogroups: YR1 and YR2, which comprised ssp. yarigadakeanus, and YG, which comprised ssp. yaginus. Mitochondrial DNA genetic differentiation generally agreed with morphological subspecies classification. The haplotype network suggested that ssp. yarigadakeanus populations had multiple origins, and the subspecies character of “bright blue of the male's wings” was assumed to have evolved independently in each subalpine meadow. We found that P. subsolanus was genetically differentiated depending upon the elevation at each mountain region, suggesting that each haplogroup should be a conservation unit.     

Genetics,taxonomy and the conservation of British <Emphasis Type="Italic">Euphrasia</Emphasis>

In Britain the genus Euphrasia comprises ca 20 diploid and tetraploid plant species, including several endemics. However, their conservation is impeded by taxonomic uncertainty. Analysis of cpDNA and AFLP variation was used to assess their taxonomic status and establish the extent of barriers to gene exchange among them. Differences in ploidy level constitute a very strong barrier to genetic exchange, although this is not absolute. The diploid endemics E. vigursii and E. rivularis form morphologically and genetically definable units which show some level of reproductive isolation. Within tetraploid Euphrasia, the species showed varying degrees of distinctness. Analysis of geographically paired samples from two widespread outcrossing taxa E. arctica and E. nemorosa provides evidence for extensive genetic exchange between them. However AFLP data indicate that this outbreeding species complex possesses a gene pool distinct from that of the widespread inbreeding tetraploids. The widespread and endemic inbreeding tetraploids contain examples of morphologically and genetically definable taxa, but also species whose distinctness is more equivocal. The conservation implications of this study are that species-based action plans are potentially suitable for conservation of the diploid endemics E. vigursii and E. rivularis. In contrast we contend that a species-based conservation framework, developed with reproductively isolated and genetically distinct groups in mind, requires modification for conservation of the complex and dynamic diversity found within the tetraploids. The adoption of ‘taxonomic’ action plans, designed to protect the evolutionary processes generating Euphrasia diversity, may provide a supplementary solution for conserving this type of variation.     

Nine microsatellite DNA primers for <Emphasis Type="Italic">Hippophae rhamnoides</Emphasis> ssp. <Emphasis Type="Italic">sinensis</Emphasis> (Elaeagnaceae)

Hippophae rhamnoides ssp. sinensis occurs mainly in the arid regions of northwest China. The wood stands of this subspecies play an important role in maintaining the local ecosystems in these regions. In addition, the genetic characteristics are essential to understand the historical range changes of this subspecies and its morphological differentiation with other subspecies. In this study, we developed nine microsatellite loci for this subspecies for the first time. We used the combining biotin capture method to enrich AG/CT/AC/GT/CG/GTG/CCA microsatellites. Twenty-six microsatellites were isolated from the enriching library and nine of them were found to be polymorphic through screening 12 distantly distributed individuals. The number of alleles per locus ranged from three to twelve and expected heterozygosity from 0.2659 to 0.4767, respectively. We further performed cross-priming tests in another subspecies and two congeneric species. These firstly isolated loci will provide a useful tool to investigate the genetic structure of this subspecies and its morphological differentiation from the other subspecies.     

Assessment of genetic diversity of native species in Izu Islands for a discriminate choice of source populations: Implications for revegetation of volcanically devastated sites

In using native species for revegetation, it is necessary choose source populations carefully to reduce the risk of planting suboptimal germplasm. To make preliminary recommendations for native species to use in the revegetation of a volcanically devastated area on Miyake Is., Japan, we investigated the genetic variation of Alnus sieboldiana, Miscanthus sinensis ssp. condensatus, and Polygonum cuspidatum var. terminalis in the Izu Islands and on the Izu Peninsula based on chloroplast DNA (cpDNA) sequence variations and amplified fragment length polymorphisms (AFLPs). The amount and pattern of differentiation differ between organelle and nuclear markers, suggesting the necessity of evaluation based on both types of markers. Within-population diversity did not vary among populations, suggesting that it does not need to be considered in the choice of a source population. The pattern and degree of differentiation varied among species, and geographical proximity did not necessarily accord with genetic similarity, suggesting that the site of an appropriate source population varies among species and should be determined empirically rather than by assuming that close proximity predicts genetic similarity. The Izu Peninsula populations deviated from the island populations in all species. Comparison of cpDNA sequences with those of related species indicates the possibility of hybridization with related species on the Izu Peninsula, suggesting that seeds collected from populations where related species live sympatrically should not be used for revegetation. These findings indicate the need to assess the genetic diversity empirically by using organelle and nuclear markers to avoid unintended consequences of genetic mixing associated with revegetation.     

Comparative analysis of genetic relationship and diagnostic markers of several taxa of Guizotia Cass. (Asteraceae) as revealed by AFLPs and RAPDs

Amplified fragment length polymorphism (AFLP) and random amplified polymorphic DNA (RAPD) were employed to examine the genetic relationship between Guizotia taxa, to suggest the taxonomic status of some of these taxa, and to identify their diagnostic markers. Results from AFLPs and RAPDs share some features in common, both revealing G. scabra ssp. schimperi as the most closely related taxon to G. abyssinica, and indicating that G. arborescens and G. zavattarii are the most divergent taxa. Most of the diagnostic markers revealed in this study were specific to G. arborescens and G. zavattarii. Our analysis suggests that G. scabra ssp. scabra, G. scabra ssp. schimperi, Chelelu and Ketcha are separate species. In this study, AFLP was found to be superior to RAPD in detecting genetic variation, in internal consistency of the data and in the fitness of its clusters to genetic similarity data. AFLPs revealed genetic relationship between Guizotia taxa that is more inline with the cytogenetic and hybridization studies than that revealed by RAPDs.     

Variability for restriction fragment lengths and phylogenies in lentil

Summary Thirty accessions of domesticated (Lens culinaris ssp. culinaris) and wild (L. culinaris ssp. orientalis, L. culinaris ssp. odemensis, L. nigricans ssp. ervoides and L. nigricans ssp. nigricans) lentil were evaluated for restriction fragment length polymorphisms (RFLPs) using ten relative low-copy-number probes selected from partial genomic and cDNA libraries of lentil. Nei's average gene diversity was used as a measure of genetic variability for restriction fragment lengths within subspecies and a dendrogram was constructed from genetic distance estimates between subspecies. The wild lentils L. culinaris ssp. orientalis and L. culinaris ssp. odemensis showed the greatest variability for restriction fragment lengths and were closely positioned to domesticated lentil in the dendrogram. Little variability for restriction fragment lengths was observed within accessions of L. nigricans ssp. ervoides and L. nigricans ssp. nigricans. This observation is consistent with a previously published proposal that nigricans may have been independently domesticated. Estimates of genetic variability based on RFLPs tended to be greater than estimates from isozymes.     

Intra- and inter-specific variations in Lens revealed by RAPD markers

Randomly amplified polymorphic DNA (RAPD) markers were used to estimate intra- and interspecific variations in the genus Lens (lentil). Twenty cultivars of L. culinaris ssp. culinaris, including 11 microsperma (small-seeded) and nine macrosperma (large-seeded) types, and 16 wild relatives (four accessions each of L. culinaris ssp. orientalis, L. odemensis, L. nigricans and L. ervoides), were evaluated for genetic variability using a set of 40 random 10-mer primers. Fifty reproducibly scorable DNA bands were observed from ten of the primers, 90% of which were polymorphic. Genetic distances between each of the accessions were calculated from simple matching coefficients. A dendrogram showing genetic relationships between them was constructed by an unweighted pair-group method with arithmetical averages (UPGMA). This study revealed that (1) expect for L. ervoides, the level of intraspecific variation in cultivated lentil is lower than that in wild species, (2) L. culinaris ssp. orientalis is the most likely candidate for a progenitor of the cultivated species, and (3) microsperma and macrosperma cultivars were indistinguishable by the RAPD markers identified here.     

Genetic differentiation of metallicolous and non-metallicolous Armeria maritima (Mill.) Willd. taxa (Plumbaginaceae) in Central Europe

This study investigates the genetic differentiation within the Central European Armeria maritima (Mill.) Willd. complex with special reference to the metallicolous populations using AFLP markers. Our sampling comprised all metallicolous (ssp. halleri, hornburgensis, bottendorfensis, eifeliaca, calaminaria), and non-metallicolous taxa (ssp. maritima, elongata, alpina). Geographical and genetic distances between populations were moderately positively correlated. Genetic variability of metallicolous and non-metallicolous populations was not significantly different. Lowland populations were clearly differentiated from the alpine populations. Within the lowland group metallicolous and non-metallicolous populations were not genetically differentiated. All lowland populations show a regional differentiation and close relationships to ssp. elongata. Thus, the metallicolous taxa should not be maintained as subspecies. Likewise, their treatment as varieties of a ssp. halleri s.l. is critical because this taxon cannot be consistently characterized throughout its geographical range and may be an artefact itself. If a taxonomical recognition should be considered necessary it is advisable to treat the microendemics as varieties of ssp. elongata.