Untangling the hedge: Genetic diversity in clonally and sexually transmitted genomes of European wild roses, Rosa L.

While European wild roses are abundant and widely distributed, their morphological taxonomy is complicated and ambiguous. In particular, the polyploid Rosa section Caninae (dogroses) is characterised by its unusual meiosis, causing simultaneous clonal and sexual transmission of sub-genomes. This hemisexual reproduction, which often co-occurs with vegetative reproduction, defies the standard definition of species boundaries. We analysed seven highly polymorphic microsatellite loci, scored for over 2 600 Rosa samples of differing ploidy, collected across Europe within three independent research projects. Based on their morphology, these samples had been identified as belonging to 21 dogrose and five other native rose species. We quantified the degree of clonality within species and at individual sampling sites. We then compared the genetic structure within our data to current rose morpho-systematics and searched for hemisexually co-inherited sets of alleles at individual loci. We found considerably fewer copies of identical multi-locus genotypes in dogroses than in roses with regular meiosis, with some variation recorded among species. While clonality showed no detectable geographic pattern, some genotypes appeared to be more widespread. Microsatellite data confirmed the current classification of subsections, but they did not support most of the generally accepted dogrose microspecies. Under canina meiosis, we found co-inherited sets of alleles as expected, but could not distinguish between sexually and clonally inherited sub-genomes, with only some of the detected allele combinations being lineage-specific.

Power and Weakness of Repetition - Evaluating the Phylogenetic Signal From Repeatomes in the Family Rosaceae With Two Case Studies From Genera Prone to Polyploidy and Hybridization (Rosa and Fragaria).

Plant genomes consist, to a considerable extent, of non-coding repetitive DNA. Several studies showed that phylogenetic signals can be extracted from such repeatome data by using among-species dissimilarities from the RepeatExplorer2 pipeline as distance measures. Here, we advanced this approach by adjusting the read input for comparative clustering indirectly proportional to genome size and by summarizing all clusters into a main distance matrix subjected to Neighbor Joining algorithms and Principal Coordinate Analyses. Thus, our multivariate statistical method works as a "repeatomic fingerprint," and we proved its power and limitations by exemplarily applying it to the family Rosaceae at intrafamilial and, in the genera Fragaria and Rosa, at the intrageneric level. Since both taxa are prone to hybridization events, we wanted to show whether repeatome data are suitable to unravel the origin of natural and synthetic hybrids. In addition, we compared the results based on complete repeatomes with those from ribosomal DNA clusters only, because they represent one of the most widely used barcoding markers. Our results demonstrated that repeatome data contained a clear phylogenetic signal supporting the current subfamilial classification within Rosaceae. Accordingly, the well-accepted major evolutionary lineages within Fragaria were distinguished, and hybrids showed intermediate positions between parental species in data sets retrieved from both complete repeatomes and rDNA clusters. Within the taxonomically more complicated and particularly frequently hybridizing genus Rosa, we detected rather weak phylogenetic signals but surprisingly found a geographic pattern at a population scale. In sum, our method revealed promising results at larger taxonomic scales as well as within taxa with manageable levels of reticulation, but success remained rather taxon specific. Since repeatomes can be technically easy and comparably inexpensively retrieved even from samples of rather poor DNA quality, our phylogenomic method serves as a valuable alternative when high-quality genomes are unavailable, for example, in the case of old museum specimens.

Genetic and chemical diversity of the toxic herb Jacobaea vulgaris Gaertn. (syn. Senecio jacobaea L.) in Northern Germany.

Tansy ragwort, Jacobaea vulgaris Gaertn. (syn. Senecio jacobaea L.), is a common Asteraceae in Europe and Asia and known to be an invasive pest in several regions in the world. Recently it is also spreading immensely in native regions like Northern Germany. Pyrrolizidine alkaloids (PAs), which are found in high amounts in Jacobaea vulgaris, are toxic for humans and potentially lethal for grazing animals. In this study we investigated 27 populations of tansy ragwort in Northern Germany for their PA concentration and composition using liquid chromatography coupled to high resolution mass spectrometry. Furthermore, we investigated the genetic structure of selected populations using amplified length polymorphism markers. We detected 98 different PAs in the samples and considerable differences of PA composition between populations. In contrast, PA content of populations did not differ significantly. Genetic (4%) differentiation among populations was low while average genetic diversity was high (0.35). There was no correlation between genetic and geographic distance. Neither genetic markers nor chemical composition revealed any connection to the geographic pattern. As we could not detect any pattern in genetic or chemical diversity, we suggest that the existence of this diversity is a result of a broad interaction with the environment rather than that of evolutionary constraints in the current selection process driving PA composition in J. vulgaris in certain chemotypes.

Does origin always matter? Evaluating the influence of nonlocal seed provenances for ecological restoration purposes in a widespread and outcrossing plant species.

For restoration purposes, nature conservation generally enforces the use of local seed material based on the "local-is-best" (LIB) approach. However, in some cases recommendations to refrain from this approach have been made. Here we test if a common widespread species with no obvious signs of local adaptation may be a candidate species for abandoning LIB during restoration. Using 10 microsatellite markers we compared population genetic patterns of the generalist species Daucus carota in indigenous and formerly restored sites (nonlocal seed provenances). Gene diversity overall ranged between H e = 0.67 and 0.86 and showed no significant differences between the two groups. Hierarchical AMOVA and principal component analysis revealed very high genetic population admixture and negligible differentiation between indigenous and restored sites (F CT = 0.002). Moreover, differentiation between groups was caused by only one outlier population, where inbreeding effects are presumed. We therefore conclude that the introduction of nonlocal seed provenances in the course of landscape restoration did not jeopardize regional species persistence by contributing to inbreeding or outbreeding depressions, or any measurable adverse population genetic effect. On the basis of these results, we see no obvious objections to the current practice to use the 10-fold cheaper, nonlocal seed material of D. carota for restoration projects.

Formalizing morphologically cryptic biological entities: new insights from DNA taxonomy, hybridization, and biogeography in the leafy liverwort Porella platyphylla (Jungermanniopsida, Porellales).

PREMISE OF THE STUDY: Recognition and formalization of morphologically cryptic species is a major challenge to modern taxonomy. An extreme example in this regard is the Holarctic Porella platyphylla s.l. (P. platyphylla plus P. platyphylloidea). Earlier studies demonstrated the presence of three isozyme groups and two molecular lineages. The present investigation was carried out to elucidate the molecular diversity of P. platyphylla s.l. and the distribution of its main clades, and to evaluate evidence for the presence of one vs. several species. METHODS: We obtained chloroplast (atpB-rbcL, trnL-trnF) and nuclear ribosomal (ITS) DNA sequences from 101 Porella accessions (P. platyphylla s.l., P. × baueri, P. cordaeana, P. bolanderi, plus outgroup species) to estimate the phylogeny using parsimony and likelihood analyses. To facilitate the adoption of Linnean nomenclature for molecular lineages, we chose a DNA voucher as epitype. KEY RESULTS: Phylogenies derived from chloroplast vs. nuclear data were congruent except for P. platyphylla s.l., including a North American lineage that was placed sister to P. cordaeana in the chloroplast DNA phylogeny but sister to the Holarctic P. platyphylla s.str. in the nuclear DNA phylogeny. European and North American accessions of P. cordaeana and P. platyphylla form sister clades. CONCLUSIONS: The genetic structure of P. platyphylla s.l. reflects morphologically cryptic or near cryptic speciation into Holarctic P. platyphylla s.str. and North American P. platyphylloidea. The latter species is possibly an ancient hybrid resulting from crossings of P. cordaeana and P. platyphylla s.str. and comprises several distinct molecular entities.

Metabolic fingerprinting of Leontopodium species (Asteraceae) by means of ¹H NMR and HPLC-ESI-MS.

The genus Leontopodium, mainly distributed in Central and Eastern Asia, consists of ca. 34-58 different species. The European Leontopodium alpinum, commonly known as Edelweiss, has a long tradition in folk medicine. Recent research has resulted in the identification of prior unknown secondary metabolites, some of them with interesting biological activities. Despite this, nearly nothing is known about the Asian species of the genus. In this study, we applied proton nuclear magnetic resonance (¹H NMR) spectroscopy and liquid chromatography-mass spectrometry (LC-MS) metabolic fingerprinting to reveal insights into the metabolic patterns of 11 different Leontopodium species, and to conclude on their taxonomic relationship. Principal component analysis (PCA) of ¹H NMR fingerprints revealed two species groups. Discriminators for these groups were identified as fatty acids and sucrose for group A, and ent-kaurenoic acid and derivatives thereof for group B. Five diterpenes together with one sesquiterpene were isolated from Leontopodium franchetii roots; the compounds were described for the first time for L. franchetii: ent-kaur-16-en-19-oic acid, methyl-15α-angeloyloxy-ent-kaur-16-en-19-oate, methyl-ent-kaur-16-en-19-oate, 8-acetoxymodhephene, 19-acetoxy-ent-kaur-16-ene, methyl-15ß-angeloyloxy-16,17-epoxy-ent-kauran-19-oate. In addition, differences in the metabolic profile between collected and cultivated species could be observed using a partial least squares-discriminant analysis (PLS-DA). PCA of the LC-MS fingerprints revealed three groups. Discriminating signals were compared to literature data and identified as two bisabolane derivatives responsible for discrimination of group A and C, and one ent-kaurenoic acid derivative, discriminating group B. A taxonomic relationship between a previously unidentified species and L. franchetii and Leontopodium sinense could be determined by comparing NMR fingerprints. This finding supports recent molecular data. Furthermore, Leontopodium dedekensii and L. sinense, two closely related species in terms of morphology and DNA-fingerprints, could be distinguished clearly using ¹H NMR and LC-MS metabolic fingerprinting.

To be or not to be the odd one out--allele-specific transcription in pentaploid dogroses (Rosa L. sect. Caninae (DC.) Ser).

BACKGROUND: Multiple hybridization events gave rise to pentaploid dogroses which can reproduce sexually despite their uneven ploidy level by the unique canina meiosis. Two homologous chromosome sets are involved in bivalent formation and are transmitted by the haploid pollen grains and the tetraploid egg cells. In addition the egg cells contain three sets of univalent chromosomes which are excluded from recombination. In this study we investigated whether differential behavior of chromosomes as bivalents or univalents is reflected by sequence divergence or transcription intensity between homeologous alleles of two single copy genes (LEAFY, cGAPDH) and one ribosomal DNA locus (nrITS). RESULTS: We detected a maximum number of four different alleles of all investigated loci in pentaploid dogroses and identified the respective allele with two copies, which is presumably located on bivalent forming chromosomes. For the alleles of the ribosomal DNA locus and cGAPDH only slight, if any, differential transcription was determined, whereas the LEAFY alleles with one copy were found to be significantly stronger expressed than the LEAFY allele with two copies. Moreover, we found for the three marker genes that all alleles have been under similar regimes of purifying selection. CONCLUSIONS: Analyses of both molecular sequence evolution and expression patterns did not support the hypothesis that unique alleles probably located on non-recombining chromosomes are less functional than duplicate alleles presumably located on recombining chromosomes.

Microsatellite analyses of artificial and spontaneous dogrose hybrids reveal the hybridogenic origin of Rosa micrantha by the contribution of unreduced gametes.

Dogroses are characterized by a unique meiosis system, the so-called canina meiosis, which facilitates sexual reproduction at odd-number ploidy. The mostly pentaploid somatic level of dogroses is restored by a merger of haploid sperm cells and tetraploid egg cells. We analyzed experimental hybrids between different dogrose species using microsatellites to determine pollen-transmitted alleles. This information was used to reconstruct the putative hybridogenic origin of Rosa micrantha and R. dumalis and to estimate the frequency of spontaneous hybridization in a natural population. We found no evidence for the hybrid origin of R. dumalis, but our data suggest that R. micrantha presumably arose by hybridization between R. rubiginosa and R. canina or R. corymbifera. We observed only hexaploid individuals of R. micrantha, thus the establishment of this hybridogenic species was favored when unreduced gametes contributed to their origin. We demonstrate that spontaneous hybrids originated infrequently from the parental species in a natural population, but hybridization was often associated with the formation of unreduced gametes. We postulate that unreduced gametes play a major role in the evolutionary success of dogrose hybrids because they provide highly homologous chromosomes crucial for bivalent formation during canina meiosis and thus ensuring this unique form of sexual reproduction.

HybHyp--hybridizing the host: the long reach of parasite genes. A new hypothesis to explain host-parasite interrelationships in plant hybrid complexes.

Ever since existence of sexuality in plants was accepted in around 1700, questions centred about the role and maintenance of sexual reproduction in general, leading to a number of hypotheses like the Vicar of Bray, the Ratchet or the Hitch-hiker theory. Bell (The masterpiece of nature. The evolution and genetics of sexuality. University of California Press, Berkeley, LA, 1982) formulated the Red Queen Hypothesis (RQH) which explains the persistence of sexual reproduction as an outcome of a coevolutionary arms race between hosts and parasites. By sexual recombination and genetic diversification hosts minimize the risk of pathogen infection. Since virulence of pathogens is genetically determined and often species specific, parasites are mostly adapted to common host genotypes, whereas rare and divergent genotypes are less infected and therefore have a selective advantage. Employing Dawkins (The extended phenotype. The long reach of the gene, 1999) central theorem of the extended phenotype to the RQH, mating systems in hosts might be a result of the long reach of the parasites genes. Here now the hypothesis is proposed, that evolution by hybridisation and polyploidy in host plants is an extended phenotype of parasites, a response of hosts triggered by the parasites genes to slow down the effects of the Red Queen strategy of plants. Thus, hybridisation and polyploidy might have evolved by parasite pressure and not by host strategy. This hypothesis is called the "hybridisation-of-the-host-hypothesis".

AFLP markers as a tool to reconstruct complex relationships: A case study in Rosa (Rosaceae).

The genus Rosa has a complex evolutionary history caused by several factors, often in conjunction: extensive hybridization, recent radiation, incomplete lineage sorting, and multiple events of polyploidy. We examined the applicability of AFLP markers for reconstructing (species) relationships in Rosa, using UPGMA clustering, Wagner parsimony, and Bayesian inference. All trees were well resolved, but many of the deeper branches were weakly supported. The cluster analysis showed that the rose cultivars can be separated into a European and an Oriental cluster, each being related to different wild species. The phylogenetic analyses showed that (1) two of the four subgenera (Hulthemia and Platyrhodon) do not deserve subgeneric status; (2) section Carolinae should be merged with sect. Cinnamomeae; (3) subsection Rubigineae is a monophyletic group within sect. Caninae, making sect. Caninae paraphyletic; and (4) there is little support for the distinction of the five other subsections within sect. Caninae. Comparison of the trees with morphological classifications and with previous molecular studies showed that all methods yielded reliable trees. Bayesian inference proved to be a useful alternative to parsimony analysis of AFLP data. Because of their genome-wide sampling, AFLPs are the markers of choice to reconstruct (species) relationships in evolutionary complex groups.

Johannes Reinke (1839-1931) and his "Dominanten" theory--an early concept of gene regulation and morphogenesis.

Johannes Reinke (1839-1931) was one of the most eminent and influential botanists, politician, philosopher and "anti-Haeckelist" of the 19th century. Educated in the mid 19th century he had faced revolutionary changes in the scientific understanding of the origin and evolution of life. Working mainly at the phenotypic level, Reinke was interested in the basic mechanisms and the guiding idea behind all processes which coordinated gene regulation and morphogenesis. What drove Reinke in his search of regulatory mechanisms for evolutionary patterns and processes was his religious belief. For the origin of the very first life, Reinke believed in creation but once life was established, evolution followed natural laws. This led to his early concept of "Dominanten" entities, which regulate gene transcription and morphogenesis, anticipating the ideas of Goldschmidt and v. Uexküll.

Different evolutionary histories of two Phragmidium species infecting the same dog rose hosts.

Rust fungi in the genus Phragmidium are frequent pathogens of both wild and cultivated roses. We investigated the occurrence and relationships of rusts on dog roses, Rosa sect. Caninae (Rosa canina, R. corymbifera and R. rubiginosa) in Germany. Two Phragmidium species, P. mucronatum and P. tuberculatum, were able to infect each of the three dog rose species examined. However, the overall infection of R. rubiginosa was significantly lower, which could be important for rose breeding. Despite overlapping host ranges, the evolutionary background of P. tuberculatum and P. mucronatum is quite distinct. Phylogenetic analyses of the D1/D2 region of the LSU rDNA suggest that P. mucronatum shares a common ancestor with other rose rusts, whereas P. tuberculatum evolved from a Rubus-Sanguisorba rust clade and must have undergone a host shift to Rosa spp.

Evolution by hybridisation. The influence of reticulate evolution on biosymmetrical patterns and processes in plants.

Natural hybridisation in plants is one of the most striking modes of evolution, when looking at the effects reticulation may have on patterns and processes of the evolutionary history of the affected system. Among other functional constraints, symmetry is regarded as a major factor of or for evolution. A loss of symmetry, that means origin of asymmetrical phenomena can in some cases be linked to environmental or genetic stresses, such as hybridisation. It has been shown that the discussion on reticulation and symmetrical evolution is strongly influenced by a zoocentric evolutionary view, but more focus is needed on the differences between the organisms subject to the study. Plant hybridogenic evolution with the subsequent possibility of polyploidisation and the consequent reduced homology is a completely different phenomenon compared to animal evolution. This article examines the role of symmetry for plant evolution by concentration on two topics: (a) spatial asymmetry on multiple levels caused by hybridisation, and (b) temporal asymmetry (changes of rhythms inherent to specific systems) as an effect of reticulation.