F. J. Rumsey

Polyploidy, phylogeography and Pleistocene refugia of the rockfern Asplenium ceterach : evidence from chloroplast DNA

Chloroplast DNA sequences were obtained from 331 Asplenium ceterach plants representing 143 populations from throughout the range of the complex in Europe, plus outlying sites in North Africa and the near East. We identified nine distinct haplotypes from a 900 bp fragment of trnL‐trnF gene. Tetraploid populations were encountered throughout Europe and further afield, whereas diploid populations were scarcer and predominated in the Pannonian‐Balkan region. Hexaploids were encountered only in southern Mediterranean populations. Four haplotypes were found among diploid populations of the Pannonian‐Balkans indicating that this region formed a northern Pleistocene refugium. A separate polyploid complex centred on Greece, comprises diploid, tetraploid and hexaploid populations with two endemic haplotypes and suggests long‐term persistence of populations in the southern Mediterranean. Three chloroplast DNA (cpDNA) haplotypes were common among tetraploids in Spain and Italy, with diversity reducing northwards suggesting expansion from the south after the Pleistocene. Our cpDNA and ploidy data indicate at least six independent origins of polyploids.

Chloroplast Phylogeny of Asplenioid Ferns based on rbcL and trnL-F Spacer Sequences (Polypodiidae, Aspleniaceae) and its Implications for Biogeography

Abstract Molecular phylogenies have been generated to investigate relationships among species and putative segregates in Asplenium, one of the largest genera in ferns. Of the ∼700 described taxa, 71 are included in a phylogenetic analysis using the chloroplast rbcL gene and trnL-F spacer. Our results support Hymenasplenium as the sister lineage to all other asplenioid ferns, and all other putative satellite genera are nested within this asplenioid clade. Instead of the classical and well-recognized separation into Old and New World clades, asplenioid ferns reveal a separation of the deeper branches into tropical and temperate clades. Temperate clades have evolved from tropical, more-basal clades and the phylogeny indicates up to six shifts between temperate and tropical preferences in the evolution of this widespread genus. Implications for speciation processes and biogeographic aspects, including the re-colonization of temperate regions after the last glacial period, are discussed and we present a phylogenetic framework from which the historical biogeography of asplenioid ferns can be inferred for Europe and North America.

Does Macaronesia exist? Conflicting signal in the bryophyte and pteridophyte floras

Macaronesia, which includes five mid-Atlantic archipelagos (Azores, Madeira, Selvagems, Canaries, and Cape Verdes), has been traditionally recognized as a distinct biogeographic unit whose circumscription has been intimately associated with the hypothesis that the flora is a relict of a formerly broadly distributed subtropical Tertiary flora. The concept of Macaronesia is revisited here using parsimony and Bayesian analyses of floristic data sets for the moss, liverwort, and pteridophyte floras. All analyses reject the monophyly of Macaronesia s.l., resolving the Cape Verdes with tropical Africa. Of the other Macaronesian archipelagos, the liverwort and pteridophyte analyses support, or could not reject, an Azorean-Madeiran-Canarian clade (hereafter Macaronesia s.s.), but the moss analysis resolves the Canary Islands as sister to North Africa, thus rejecting the concept of Macaronesia s.s. for this group. Dynamic interchange of taxa with neighboring continental areas rather than relictualism best explains the relationships of the Cape Verde cryptogamic flora and the Canary Island moss flora. In contrast, relictualism is consistent with a monophyletic Macaronesia s.s. for liverworts and pteridophytes. However, from the limited information available on relationships of endemic cryptogams, this explanation alone may be unsatisfactory. Spatially congruent patterns may, in fact, conceal a complex mixture of relictual distributions and more recent speciation and dispersal events.

Genetic structure, reproductive biology and ecology of isolated populations of Asplenium csikii (Aspleniaceae, Pteridophyta)

The potential for environmental heterogeneity to generate spatial structuring of genotypes in seed-plant populations that occupy patchy habitats has been demonstrated by several studies, but little is known about the population structure of pteridophytes occupying patchy environments. In this study we have examined the genetic structure of isolated populations of the rock fern Asplenium csikii, an ecological specialist, growing almost exclusively on perpendicular walls of natural rock outcrops. All genetic variation observed in this taxon was partitioned between localities; no allozyme variation was found within a site and each site was colonized by a single multilocus phenotype (MLP). In total, five different MLPs were recorded from the nine localities, with two MLPs present at more than one site. Previous examination of population structure and genetic diversity in another rock fern, A. ruta-muraria, showed that the genetic diversity increases through multiple colonization over time. However, we cannot find any such correlation for A. csikii. All populations are genetically uniform, despite the probably considerable age of the populations and sites. Earlier studies concluded that the ample production of wind-borne propagules would lead to multiple colonization of sites and that reproductive features, such as single-spore colonization and subsequent intragametophytic selfing, would lead to very little genetic structuring of fern populations. In contrast to this prediction, it appears that ecological specialization and the scarcity of the narrowly defined niche contribute strongly to the pronounced partitioning of genetic variability observed in populations of A. csikii.

Host-specific races in the holoparasitic angiosperm Orobanche minor: implications for speciation in parasitic plants

BACKGROUND AND AIMS Orobanche minor is a root-holoparasitic angiosperm that attacks a wide range of host species, including a number of commonly cultivated crops. The extent to which genetic divergence among natural populations of O. minor is influenced by host specificity has not been determined previously. Here, the host specificity of natural populations of O. minor is quantified for the first time, and evidence that this species may comprise distinct physiological races is provided. METHODS A tripartite approach was used to examine the physiological basis for the divergence of populations occurring on different hosts: (1) host-parasite interactions were cultivated in rhizotron bioassays in order to quantify the early stages of the infection and establishment processes; (2) using reciprocal-infection experiments, parasite races were cultivated on their natural and alien hosts, and their fitness determined in terms of biomass; and (3) the anatomy of the host-parasite interface was investigated using histochemical techniques, with a view to comparing the infection process on different hosts. KEY RESULTS Races occurring naturally on red clover (Trifolium pratense) and sea carrot (Daucus carota ssp. gummifer) showed distinct patterns of host specificity: parasites cultivated in cross-infection studies showed a higher fitness on their natural hosts, suggesting that races show local adaptation to specific hosts. In addition, histological evidence suggests that clover and carrot roots vary in their responses to infection. Different root anatomy and responses to infection may underpin a physiological basis for host specificity. CONCLUSIONS It is speculated that host specificity may isolate races of Orobanche on different hosts, accelerating divergence and ultimately speciation in this genus. The rapid life cycle and broad host range of O. minor make this species an ideal model with which to study the interactions of parasitic plants with their host associates.

Host-driven divergence in the parasitic plant Orobanche minor Sm. (Orobanchaceae)

Many parasitic angiosperms have a broad host range and are therefore considered to be host generalists. Orobanche minor is a nonphotosynthetic root parasite that attacks a range of hosts from taxonomically disparate families. In the present study, we show that O. minor sensu lato may comprise distinct, genetically divergent races isolated by the different ecologies of their hosts. Using a three‐pronged approach, we tested the hypothesis that intraspecific taxa O. minor var. minor and O. minor ssp. maritima parasitizing either clover (Trifolium pratense) or sea carrot (Daucus carota ssp. gummifer), respectively, are in allopatric isolation. Morphometric analysis revealed evidence of divergence but this was insufficient to define discrete, host‐specific taxa. Intersimple sequence repeat (ISSR) marker‐based data provided stronger evidence of divergence, suggesting that populations were isolated from gene flow. Phylogenetic analysis, using sequence‐characterized amplified region (SCAR) markers derived from ISSR loci, provided strong evidence for divergence by clearly differentiating sea carrot‐specific clades and mixed‐host clades. Low levels of intrapopulation SCAR marker sequence variation and floral morphology suggest that populations on different hosts are probably selfing and inbreeding. Morphologically cryptic Orobanche taxa may therefore be isolated from gene flow by host ecology. Together, these data suggest that host specificity may be an important driver of allopatric speciation in parasitic plants.

Phylogenetic and biosystematic relationships in four highly disjunct polyploid complexes in the subgenera Ceterach and Phyllitis in Asplenium (Aspleniaceae)

Abstract Phylogenetic studies using DNA sequences of two chloroplast regions, rbc L and trn L-F, demonstrate that the proposed genus Ceterach is a small clade within the large genus Asplenium , and sister to the Phyllitis clade. The Ceterach clade is characterised by irregular anastomosing veins and often densely scaled leaf blades. Its taxonomic status as a group nested within Asplenium is confirmed, and it is accepted here as a subgenus with seven species. The Ceterach clade comprises four lineages that correspond to disjunct polyploid complexes: the A. aureum clade forming a polyploid complex (4×, 6×, 8×) in Macaronesia, the A. ceterach clade forming a polyploid complex (2×, 4×, 6×) in the Mediterranean Basin, the A. paucivenosum clade (4×, 6×) in central Asia, and the A. dalhousiae clade (2×) with a disjunct distribution in the Himalaya, Yemen and Eritrea, and southwestern North America. Asplenium paucivenosum is sister to all other members of the Ceterach clade, whereas A. dalhousiae is sister to the A. aureum clade that includes tetraploid A. aureum , hexaploid A. lolegnamense , and octoploid A. parvifolium . Asplenium ceterach and its variations – including the hexaploid A. ceterach subsp. mediterraneum subsp. nov. first described below – form a monophyletic unit, sister to a clade consisting of A. aureum and A. dalhousiae. Asplenium cordatum from Africa and A. haugthonii from the isolated atlantic island of St. Helena are not members of the Ceterach clade, which suggests that leaf blades with dense indumenta have evolved at least twice within asplenioid ferns. The allotetraploid species A. hybridum has the chloroplast DNA from A. ceterach , and therefore the latter species is the maternal ancestor of the former. The other parent of this hybrid species is A. sagittatum that is nested within the sister clade of Ceterach , the Phyllitis clade comprising A. sagittatum and A. scolopendrium . The findings suggest that the current distribution of Ceterach is either the result of long-distance dispersal or represents fragmented relicts of a previously more widely distributed species.

The origin of the British and Macaronesian endemic Thamnobryum species (Neckeraceae)

Abstract The status and relationships of two British narrow endemic Thamnobryum species (T. angustifolium and T. cataractarum) as well as two Macaronesian endemics (T. fernandesii and T. rudolphianum) were investigated using nuclear (ITS1&2) and plastid (the rps4-trnT-trnL-trnF cluster) markers. Geographic structure present within a monophyletic T. alopecurum containing these narrow endemic taxa, indicates that these submerged multistratose leaved forms in Britain and Madeira have been independently derived from the surrounding T. alopecurum populations and show convergent evolution in response to the extreme rheophilous habitat.

From European Priority Species to Invasive Weed: Marsilea azorica (Marsileaceae) is a Misidentified Alien

Abstract The clover fern Marsilea azorica was described in 1983 from the isolated Azores archipelago in the northern Atlantic, where it is restricted to a single roadside pond. Thought to be an extremely local endemic, it was subsequently listed as a conservation priority species for the Azores, Macaronesia, and Europe, included as ‘critically endangered’ on the IUCN red list, and as ‘strictly protected’ species by the Bern convention and the European Unions habitats directive. However, we present morphological and molecular data (rbcL gene, rps4 gene, rps4-trnS spacer and trnL-trnF spacer sequences), which demonstrate that M. azorica is conspecific with M. hirsuta, a species native to Australia, but widely cultivated and locally invasive in the southern U. S. A. Based on our DNA data, we conclude that these plants are most likely a recent introduction to the Azores from Florida. We recommend removal of Azorean Marsilea from conservation priority lists. While there is no evidence that the small existing population threatens native species, further spread in the Azores should be prevented.

Gene flow between alien and native races of the holoparasitic angiosperm Orobanche minor (Orobanchaceae)

The holoparasitic angiosperm Orobanche minor parasitizes a diverse range of flowering plants from at least 16 orders in both the monocots and eudicots. However, populations of O. minor show host specificity at a local level. Our previous work identified the potential for host specificity to act as a catalyst for genetic divergence among populations of O. minor. Here we have extended this investigation by sampling populations from multiple hosts, across a broad geographic range. Sequence characterised amplified region (SCAR) data identified an exotic host-generalist lineage and a native host-specialist lineage of O. minor, suggesting genetic structure in this species is defined by both host specificity and geography. In addition, host-range overlap, discordant tree topologies, and cryptic morphology indicate the presence of gene flow between alien races and endemic populations. Therefore, repeated introductions of alien races of O. minor from disparate sources leading to introgression with native populations, and cryptic race formation, seem to have contributed to the taxonomic confusion associated with this species. We speculate that radiations associated with broad host range and divergent host ecologies may have promoted the unusually wide geographic distribution and diversification of this species. Finally, evidence of multiple shifts to exotic hosts, coupled with the predicted northward shift in climatic suitability, identify the potential for range expansion in alien races of O. minor, which may threaten nationally scarce native taxa with genetic assimilation. Our phylogenetic analysis provides a framework for identifying host races in Orobanche with a view to setting conservation priorities.