Macaronesia: a source of hidden genetic diversity for post‐glacial recolonization of western Europe in the leafy liverwort Radula lindenbergiana

Aim Bryophytes exhibit apparently low rates of endemism in Macaronesia and differ from angiosperms in their diversity patterns by the widespread occurrence of endemics within and among archipelagos. This paper investigates the phylogeography of the leafy liverwort Radula lindenbergiana to determine: (1) whether or not morphologically cryptic diversification has occurred in Macaronesia, and (2) the relationships between Macaronesian and continental populations. Location Macaronesia, Europe, Africa. Methods Eighty‐four samples were collected across the species’ distribution range and sequenced at four chloroplast DNA (cpDNA) loci (atpB–rbcL, trnG, trnL and rps4). Phylogenetic reconstructions and Bayesian ancestral area reconstructions were used in combination with population genetics statistics (H, N_ST, F_ST) to describe the pattern of present genetic diversity in R. lindenbergiana and infer its biogeographic history. Results Patterns of genetic diversity in R. lindenbergiana exhibit a striking westwards gradient, wherein haplotype (0.90) and nucleotide (0.0038 ± 0.0019) diversity peak in Macaronesia, with a substantial endemic component. We found 20.9% of the genetic variance between biogeographic regions, and most pairwise F_ST comparisons between regions are significantly different from zero. The global N_ST (0.78) is significantly higher than the global F_ST (0.20), providing evidence for the presence of phylogeographic signal in the data. Ancestral area reconstructions suggest that the haplotypes currently found in western Europe share a Macaronesian common ancestor. Main conclusions The haplotype diversification exhibited by R. lindenbergiana in Macaronesia is comparable to that reported for many angiosperm groups at the species level. The apparent lack of radiation among Macaronesian bryophytes may thus reflect the reduced morphology of bryophytes in comparison with angiosperms. The high diversity found among Macaronesian haplotypes, especially in Madeira and the Canary Islands, and the significant N_ST/F_ST ratio between Macaronesia and all the other biogeographic regions (an indication that mutation rate exceeds dispersal rates) suggest that Macaronesian archipelagos could have served as a refugium during the Quaternary glaciations. Many haplotypes currently found in Europe share a Macaronesian common ancestor, and this further suggests that Macaronesia might have played a key role in the back‐colonization of the continent.     

Novel bibenzyl derivatives and ent-cuparene-type sesquiterpenoids from Radula species

Four novel prenyl bibenzyls, perrottetin A–D were isolated from the liverwort Radula perrottetii together with a new sesquiterpene phenol, ent-2, 3-dihydroxycuparene and a prenyl dihydrochalcone, and their structures were determined by spectral methods and chemical transformations. Further investigation of the French species R. complanata resulted in the isolation of four new bibenzyls, 3, 4′-dimethoxybibenzyl, 3,5-dihydroxy-4-(2, 3-epoxy-3-methylbutyl)bibenzyl, radulanin H and radulanolide. R. buccinifera was chemically close to R. complanata. The composition of bibenzyls found in R. perrottetii and R. oyamensis were different from that of the other Radula species referred to in the section Radula. Some prenyl bibenzyls showed antimicrobial activity against Streptococcus aureus (20 μg/ml). The allergenic activity of R. complanata is due to (+)-frullanolide from the liverwort Frullania dilatata which is intermingled in R. complanata.     

Isothecium myosuroides var. brachythecioides (Dixon) Braithw. reinstated as a species,I. interludens Stirt.

The taxon currently known as Isothecium myosuroides var. brachythecioides (Dixon) Braithw. is reinstated as a species, I. interludens Stirt., the name originally assigned by Stirton when it was first described. The morphological characters separating I. myosuroides s.s. and I. interludens are summarised. The sporophytes of I. interludens, described here for the first time, differ in shape, and pattern of exothecial cells from those of I. myosuroides. A molecular analysis based on variation of the ITS region shows that I. holtii Kindb. and I. alopecuroides (Lam. ex Dubois) Isov. are not monophyletic, calling for a re-assessment of these species, whereas the monophyly of the other species, including I. interludens, cannot be rejected. The analysis failed, however, to yield molecular synapomorphies for I. interludens and other species within I. myosuroides s.l., which we interpret as the incomplete sorting of alleles among recently evolved species. Isothecium interludens is a European endemic with a hyperoceanic temperate distribution. Its nested position within a clade including the two Macaronesian endemics I. prolixum (Mitt.) M.Stech and I. montanum Draper, Hedenäs, M.Stech, T.Lopes & Sim-Sim is suggestive of a Macaronesian origin, in line with the idea that the European Atlantic fringe flora assembled during the course of the last interglacial from Macaronesian ancestors. Isothecium therefore offers a model of prime importance for the study of ongoing speciation in mosses.     

Integrative taxonomy reveals too extensive lumping and a new species in the moss genus Amphidium (Bryophyta)

An integrative taxonomic approach, including molecular phylogenetic reconstructions based on plastid rps4-trnF and nuclear ITS sequences, statistical analysis of morphological-anatomical characters, and classical taxonomy, indicates that the reduction of 13 Amphidium species to three in a recent morphological revision represents a case of too extensive lumping. Instead, six Amphidium species can be distinguished based on molecular and morphological data, the widespread Amphidium lapponicum, A. mougeotii, and A. tortuosum, as well as the Macaronesian endemic A. curvipes, the North American endemic A. californicum, and a newly discovered species from Central Asia (southern Siberia and northern Mongolia), A. asiaticum sp. nov. Diagnostic morphological characters for all six species are discussed. The present data confirm that species diversity of Amphidium is highest in the Holarctic, where all six species occur.     

Karyological evolution and molecular phylogeny in Macaronesian dendroid spurges (Euphorbia subsect. Pachycladae)

 Euphorbia subsect. Pachycladae is a taxon of primarily Macaronesian distribution, defined by morphological and biogeographical criteria. On the basis of morphological data, it is a heterogeneous group within which at least three complexes of species can be distinguished. To ascertain whether it is a natural group and discover its phylogenetic relations, we performed a cladistic analysis of the sequences of ribosomal nuclear DNA and a karyological study. The results of the two studies are concordant and show that the sub-section is polyphyletic and includes three different groups. The first monophyletic group is made up of the Macaronesian endemics E. atropurpurea complex and E. lamarckii complex, which form a polytomy with E. dendroides as the basal species. The lauroid species E. longifolia and E. stygiana represent the second monophyletic group, which derive from Mediterranean forms of E. sect. Helioscopia Dumort. Both species are paleopolyploid (2n=44) with highly symmetrical karyotypes. Finally, E. balsamifera, with a Canarian, African and Arabian distribution, remains isolated in a basal position. Its karyotype, with 2n=20 chromosomes, differs from the Macaronesian model and displays analogies with African cactiform spurges. On the basis of the results, some hypotheses are formulated about speciation processes in the three groups. Received March 3, 2001 Accepted October 28, 2001     

Genetic structure of two endangered pitcher plants,Sarracenia jonesii and Sarracenia oreophila (sarraceniaceae)

Sarracenia jonesii and S. oreophila arc insectivorous perennial plants of the southeastern United States. Both pitcher plant taxa arc rare and endangered. Allozyme diversity was assessed for eight of the ten extant populations of S. jonesii and 14 of the 35 remaining S. oreophila populations. Genetic diversity was low and comparable for both species (H_es = 0.086 and 0.082 for S. jonesii and S. oreophila, respectively). Mean population genic diversity (H_ep) was 0.061 for S. jonesii and 0.060 for S. oreophila. Estimates of genetic diversity were typical of those commonly associated with endemic species. Small populations of each species and geographically disjunct populations tended to maintain less genetic diversity. Indirect estimates of gene flow were comparable for S. oreophila (Nm = 1.62) and S. jonesii (Nm = 1.07).     

Zonal plant communities of the Ribesalbes‐Alcora Basin (La Rinconada mine,eastern Spain) during the early Miocene

Changes in lobule morphology in Radula subgenus Cladoradula show liverworts have the capacity for dramatic, relatively rapid morphological change by heterochrony. In individuals of R. bipinnata, R. boryana and R. tenax, lobules on secondary and tertiary shoots are progenetic with respect to lobules on primary shoots, in that the slope of the relationship between growth duration and shape does not change. However, in R. campanigera, lobules on secondary and tertiary branches exhibit different slopes from primary branches, but have the same growth duration, a pattern consistent with neoteny. The trajectory of allometric growth is extended or truncated in different species compared with outgroup and ancestral nodes. Changes in duration of lobule growth explain 85% of variation in lobule shape between species. Species are related by relatively shallow nodes in the crown of the Radula subgenus Cladoradula clade, suggesting that divergence and associated heterochronic changes have occurred relatively recently. The rapid morphological diversification in the crown contrasts with the relative stasis between the ancestral node and R. brunnea, the outgroup used in this analysis. A robust primary axis may be required to hold shoots away from vertical surfaces to maximize light interception, and hypermorphosis in lobule ontogeny could be a by‐product of the longer growth durations required to build axes sufficiently large to perform this structural role. Alternatively, the large auriculate lobules could function in external water transport systems by providing continuity of surfaces for solute transport via capillary action. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173 , 153–175.     

How many species of Isothecium (Lembophyllaceae,Bryophyta) are there in Macaronesia? A survey using integrative taxonomy

Species boundaries are sometimes difficult to assess, especially when molecular data do not neatly match morphologically defined units. This study investigates the moss genus Isothecium, with special emphasis on Macaronesian populations. Morphological studies are combined with the analysis of three rapidly evolving markers: nuclear internal transcribed spacer and plastid trnG and trnL‐trnF. The results of the morphological studies suggest that Isothecium is represented by five species in Macaronesia, including a new endemic species from Madeira, I sothecium montanum sp. nov. , which is described here. The molecular results are less conclusive than the morphology results in delimiting species of this genus, even when indels are included as informative. Once possible methodological shortcomings have been discarded, the results can be interpreted as having been caused by incomplete lineage sorting, probably as a consequence of recent speciation. The molecular results also suggest that the origin of the Macaronesian endemics may be explained by at least two independent colonization events. Finally, the delimitation of a new endemic species of Isothecium in Macaronesia indicates that current knowledge on the taxonomy of spore‐producing plants may be far from complete in this hotspot of biodiversity. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 00 , 000–000.     

Phylogeography and molecular phylogeny of Macaronesian island Tarphius (Coleoptera: Zopheridae): why are there so few species in the Azores?

Aim We used a phylogenetic framework to examine island colonization and predictions pertaining to differentiation within Macaronesian Tarphius (Insecta, Coleoptera, Zopheridae), and explain the paucity of endemics in the Azores compared with other Macaronesian archipelagos. Specifically, we test whether low diversity in the Azores could be due to recent colonization (phylogenetic lineage youth), cryptic speciation (distinct phylogenetic entities within species) or the young geological age of the archipelago. Location Macaronesian archipelagos (Azores, Madeira and the Canary Islands), northern Portugal and Morocco. Methods Phylogenetic analyses of mitochondrial and nuclear genes of Tarphius beetles of the Azores, other Macaronesian islands and neighbouring continental areas were used to investigate the origin of island biodiversity and to compare patterns of colonization and differentiation. A comparative nucleotide substitution rate test was used to select the appropriate substitution rate to infer clade divergence times. Results Madeiran and Canarian Tarphius species were found to be more closely related to each other, while Azorean taxa grouped separately. Azorean taxa showed concordance between species and phylogenetic clades, except for species that occur on multiple islands, which segregated by island of origin. Divergence time estimates revealed that Azorean Tarphius are an old group and that the most recent intra‐island speciation event on Santa Maria, the oldest island, occurred between 3.7 and 6.1 Ma. Main conclusions Our phylogenetic approach provides new evidence to understand the impoverishment of Azorean endemics: (1) Tarphius have had a long evolutionary history within the Azores, which does not support the hypothesis of fewer radiation events due to recent colonization; (2) the current taxonomy of Azorean Tarphius does not reflect common ancestry and cryptic speciation is responsible for the underestimation of endemics; (3) intra‐island differentiation in the Azores was found only in the oldest island, supporting the idea that young geological age of the archipelago limits the number of endemics; and (4) the lack of evidence for recent intra‐island diversification in Santa Maria could also explain the paucity of Azorean endemics. Phylogenetic reconstructions of other species‐rich taxa that occur on multiple Macaronesian archipelagos will reveal whether our conclusions are taxon specific, or of a more general nature.     

Recent origin,active speciation and dispersal for the lichen genus Nephroma (Peltigerales) in Macaronesia

Aim We reconstructed the phylogeny of the lichen genus Nephroma (Peltigerales) to assess the relationships of species endemic to Macaronesia. We estimated dates of divergences to test the hypothesis that the species arose in Macaronesia (neo‐endemism) versus the oceanic archipelagos serving as refugia for formerly widespread taxa (palaeo‐endemism). Location Cosmopolitan with a special focus on the archipelagos of the Azores, Madeira and the Canary Islands. Methods DNA sequences were obtained from 18 species for three loci and analysed using maximum parsimony, maximum likelihood and Bayesian inferences. Divergence dates were estimated for the internal transcribed spacer (ITS)‐based phylogeny using a relaxed molecular clock. Reconstruction of the ancestral geographical range was conducted using the Bayesian 50% majority rule consensus tree under a parsimony method. Results The backbone phylogenetic tree was fully supported, with Nephroma plumbeum as sister to all other species. Four strongly supported clades were detected: the Nephroma helveticum, the N. bellum, the N. laevigatum and the N. parile clades. The latter two share a common ancestor and each includes a widespread Holarctic species (N. laevigatum and N. parile, respectively) and all species endemic to Macaronesia. The data suggest a neo‐endemic origin of Macaronesian taxa, a recent range expansion from Macaronesia of both widespread species, a range expansion limited to the Mediteranean Basin and south‐western Europe for another taxon, and a long dispersal event that resulted in a speciation event in the western parts of North America. Main conclusions The Macaronesian endemic species belong to two sister clades and originated from a most recent common ancestor (MRCA) shared with one widely distributed taxon, either N. parile or N. laevigatum. Estimates of the mean divergence dates suggest that the endemics originated in the archipelagos after the rise of the volcanic islands, along with the ancestor to the now widespread species, which probably expanded their range beyond Macaronesia via long‐distance dispersal. This study provides the first phylogenetic evidence of Macaronesian neo‐endemism in lichenized fungi and provides support for the hypothesis that oceanic islands may serve as a source for the colonization of continents. However, further data are needed to properly assess the alternative hypothesis, namely colonization from western North America.     

Intercontinental disjunctions in Cryptotaenia (Apiaceae, Oenantheae): an appraisal using molecular data

Aim The angiosperm genus Cryptotaenia (family Apiaceae, tribe Oenantheae) exhibits an anomalous distribution pattern, with five of its eight species being narrow endemics geographically isolated from their presumed relatives. We examined the monophyly of the genus and ascertained the phylogenetic placements of its constituent members in order to explain their distribution patterns. Location Eastern North America, eastern Asia, the Caucasus, southern Italy, Macaronesia and Africa. Methods In total, 173 accessions were examined for nuclear rDNA ITS sequence variation, representing nearly all major lineages of Apiaceae subfamily Apioideae and seven species of Cryptotaenia. Sampling of tribes Oenantheae, Scandiceae and Pimpinelleae was comprehensive. Phylogenetic analyses included Bayesian, maximum parsimony and neighbour‐joining methods; biogeographical scenarios were inferred using dispersal–vicariance analysis (diva ). Results Cryptotaenia is polyphyletic and includes three distant lineages. (1) Cryptotaenia sensu stricto (C. canadensis, C. japonica, C. flahaultii and C. thomasii) is maintained within tribe Oenantheae; C. canadensis and C. japonica, representing an eastern North American–eastern Asian disjunction pattern, are confirmed to be sister species. (2) Cryptotaenia elegans, endemic to the Canary Islands, is placed within Scandiceae subtribe Daucinae along with two woody endemics of Madeira, Monizia edulis and Melanoselinum decipiens. The phylogeny of these Canarian and Madeiran endemics is unresolved. Either they constitute a monophyletic sister group to a clade comprising some Mediterranean and African species of Daucus and their relatives, or they are paraphyletic to this clade. The herbaceous/woody genus Tornabenea from Cape Verde, once included in Melanoselinum, is not closely related to the other Macaronesian endemics but to Daucus carota. (3) The African members of Cryptotaenia (C. africana, C. calycina and possibly C. polygama) comprise a clade with some African and Madagascan umbellifers; this entire clade is sister group to Eurasian Pimpinella. Main conclusions Elucidating the phylogeny of the biogeographically anomalous Cryptotaenia sensu lato enabled hypotheses on the biogeography of its constituent lineages. Cryptotaenia sensu stricto exhibits a holarctic distribution pattern, with its members occurring in regions that were important glacial refugia. The genus probably originated in eastern Asia and from there dispersed to Europe and North America. For the Macaronesian endemic species –C. elegans, M. edulis and M. decipiens–diva reconstructs either a single dispersal event to Macaronesia from the Mediterranean/African region, or a single dispersal followed by a back‐dispersal to the mainland. The radiation of Tornabenea from Cape Verde followed a second dispersal of Daucinae to Macaronesia. Woodiness in Melanoselinum/Monizia and Tornabenea, therefore, is a derived and independently acquired trait. The African members of Cryptotaenia are derived from an ancestor arriving from the Middle East.     

Characterization of the Contents of Oil Bodies from the Liverwort Radula complanata1

Abstract: With a combination of enzymatic treatment and micromanipulation, the contents of oil bodies of Radula complanata were obtained. A GC‐MS analysis revealed that 3‐methoxybibenzyl is the main constituent of these compartments. Its structure was elucidated by NMR spectroscopy.     

Genetic introgression of hybrid Rhododendron x intermedium Tausch is habitat mediated: Evidences from south-eastern Alps (Italy)

Hybridization between Rhododendron ferrugineum L. and R. hirsutum L. in south-eastern Alps was examined in order to (i) evaluate the breeding direction and the extent of backcrossing between hybrids and the parental species, (ii) define which processes facilitate speciation and maintain species identities and (iii) clarify the role of rock geochemistry in hybridization events. Individuals of three hybrid populations were analysed by morphological and molecular markers. The internal transcribed spacer and trnH–psbA distinguished the parental species and F₁ hybrids while only the simple sequence repeat markers recognized genotype classes: F, H, F₁, F₂, BxF (backcross to R. ferrugineum) and BxH (backcross to R. hirsutum). Combining morphological and molecular data, we found that the tested populations had complex genetic structure: the F₁ individuals produce F₂ hybrids and backcross to parental species. Due to R. hirsutum phenology, most backcrossing events were with this parental species (asymmetric hybridization). Geochemical analyses indicate that alkaline soil conditions linked to calcareous dolomitic rocks promoted the genetic assimilation of R. hirsutum. In addition, R. x intermedium shows a higher edaphic adaptation than R. hirsutum as it can be found on a wide range of calcareous-dolomitic rocks as well as on weakly acidic soils of natural or anthropogenic origin.     

Adaptive radiation and genetic differentiation in the woodySonchus alliance (Asteraceae:Sonchinae) in the Canary Islands

The woodySonchus alliance consists of 19 species ofSonchus subg.Dendrosonchus, one species ofSonchus subg.Sonchus and species of five genera (i.e.Babcockia, Sventenia, Taeckholmia, Lactucosonchus, Prenanthes), and is restricted primarily to the archipelago of the Canaries in the Macaronesian phytogeographical region. An enzyme electrophoretic study, including 13 loci, was conducted to assess genetic diversity within and divergence among species of the alliance. Nei's genetic identities (distances) between genera and/or subgenera range from 0.490 (0.714) to 0.980 (0.013), and pairwise comparisons of all populations show relatively high genetic identities, with a mean of 0.804. The high identities further support the genetic cohesiveness of the alliance and its single origin on the Macaronesian islands. Species in the alliance also show about 50%; higher total genetic diversity (H_T) than the mean for other oceanic endemics. There is greater divergence between endemics or species on older islands compared to those on younger islands, which suggests that time is a factor for divergence at allozyme loci. Furthermore, populations on older islands have higher total genetic diversities and lower identities than conspecific populations on younger islands. These results imply early colonization, radiation, and divergence of the woodySonchus alliance on older islands followed by subsequent colonization to younger islands. The taxonomic distribution of alleles in the alliance indicates lineage sorting also played a role in divergence among species. Lineage sorting may also produce nonconcordance with either taxonomic designation or the pattern of variation obtained from other molecular markers such as ITS sequences of nrDNA. Timing for the origin and radiation of the alliance agrees with the estimate based on ITS sequences, and suggests that the early divergence and rapid radiation took place during the Late Tertiary on either Gran Canaria or Tenerife.     

Plant functional types and their ecological responses to salinization in saline grasslands, Northeastern China

Photosynthetic pathways (C₃, C₄, and CAM) and morphological functional types (e.g. shrubs, high perennial grasses, short perennial graminaceous plants, annual grasses, annual forbs, perennial forbs, halophytes, and hydrophytes) were identified for the species from salinity grasslands in Northeastern China, using the data from both stable carbon isotope ratios (¹³C) and from the references published between 1993 and 2002. 150 species, in 99 genera and 37 families, were found with C₃ photosynthesis, and most of these species are dominants [e.g. Leymus chinensis (Trin.) Tzvel., Calamagrostis epigeios (L.), Suaeda corniculata (C.A. Mey.) Bunge]. 40 species in 25 genera and 8 families were identified with C₄ photosynthesis [e.g. Chloris virgata Sw., Aeluropus littoralis (Gouan) Parlat] and 1 species with CAM photosynthesis. Gramineae is the leading family with C₄ photosynthesis (27 species), Chenopodiaceae ranks the second (5 species). The significant increase of C₄ proportions with intense salinity suggested this type plant is remarkable response to the grassland salinization in the region. 191 species were classified into eight morphological functional types and the changes of most of these types (e.g. PEF, HAL, and HPG) were consistent with habitats and vegetation dynamics in the saline grassland. My findings suggest that the photosynthetic pathways, combined with morphological functional types, are efficient means for studying the linkage between species and ecosystems in this type of saline grassland in Northeastern China.This revised version was published online in March 2005 with corrections to the page numbers.     

新疆饼干衣属地衣分类学研究

该研究以采自新疆的100余份饼干衣属(Rinodina)地衣标本为研究材料,通过形态解剖特征观察、地衣化学成分分析以及分子生物学鉴定方法鉴定出9个种,包括2个中国新记录种——阿富汗饼干衣(Rinodina afghanica)和古氏饼干衣(Rinodina guzzinii),7个常见种分别是：包氏饼干衣(R. bohlinii)、毕氏饼干衣(R. bischoffii)、代谢饼干衣(R. metaboliza)、密果饼干衣(R. pycnocarpa)、特雷氏饼干衣(R. trevisanii)、甘肃饼干衣(R. straussii)和地生饼干衣(R. terrestris)。并提供了新疆饼干衣属地衣的分种检索表、每个物种的详细描述、新记录种的特征图片以及系统发育分析。