evidence for radiations of cheilanthoid ferns in the greater cape floristic region

1269

Eiserhardt amp al bull Cheilanthoid fern radiations in southern AfricaTAXON 60 (5) bull October 2011 1269ndash1283

IntroductIon

The winter-rainfall area of the south-western tip of Africa is characterised by a highly distinct and diverse flora (Gold-blatt amp Manning 2002 Linder 2003 2005 Born amp al 2007) and harbours two of the worldrsquos biodiversity hotspots (Myers amp al 2000) the Cape Floristic Region (CFR) and the Succu-lent Karoo (SK) The two regions differ in climate soils and vegetation (Cowling amp al 1997 Milton amp al 1997) but are flo-ristically closely related (Born amp al 2007) and together called the Greater Cape Floristic Region (GCFR Juumlrgens 1997) The GCFR shows an exceptionally high level of plant endemism (7152 at species level) and much higher plant species richness (asymp10000 spp Born amp al 2007) than predicted by global mod-els of vascular plant diversity (Kreft amp Jetz 2007) A unique feature of the GCFR is that much of its diversity is concentrated in relatively few hyper-diverse endemic clades (the so-called ldquoCape Cladesrdquo Linder 2003) The origin of this exceptional flora has been extensively studied and today it is perhaps the best-founded model system to explore the evolutionary and ecological history of biodiversity hotspots (Linder 2005 2006 2008 Forest amp al 2007 Verboom amp al 2009a) However the major drivers of plant diversification in the GCFR are still elusive (Linder amp al 2010 Schnitzler amp al 2011)

Discussed drivers include factors causing non-adaptive or adaptive speciation (Linder 2003 Linder amp al 2010) as well as factors reducing extinction (Verboom amp al 2009b Schnitzler amp al 2011) High topographic heterogeneity possibly in con-junction with climate change or sea level fluctuations might have promoted population fragmentation (Linder 2003 Ver-boom amp al 2009b) It has also been suggested that climatic os-cillations caused repeated shifts in the geographic ranges of the biomes resulting in fragmentation of species ranges (Midgley amp al 2005) Genetic differentiation might be aided by excep-tionally short-ranged pollen and seed dispersal (Ellis amp al 2006 Linder amp al 2010) Steep climatic and edaphic gradients bush fires and a high diversity of narrowly endemic pollinators have been suggested to increase the rate of adaptive plant speciation in the GCFR (Verboom amp al 2004 Ellis amp al 2006 Johnson 2010 Linder amp al 2010 Schnitzler amp al 2011) A recent genetic study found evidence in favour of adptive speciation with simul-taneous divergence in habitat preferences and pollinators (Gladi-olus Rymer amp al 2010 Linder 2010) Adaptive changes in phenology might have played an important role (Linder 2003) especially during the formation of summer-dry climate from the Mid-Miocene onwards (Warren amp al 2011) The idea that rapid adaptive radiations in response to aridification are the pri-mary cause of high plant diversity in the GCFR (Levyns 1964

Evidence for radiations of cheilanthoid ferns in the Greater Cape Floristic Region

Wolf L Eiserhardt12 Jens G Rohwer1 Stephen J Russell3 Jovita C Yesilyurt3 amp Harald Schneider3

1 Biocenter Klein Flottbek University of Hamburg Ohnhorststrasse 18 22609 Hamburg Germany2 Department of Bioscience Aarhus University Ny Munkegade 114 8000 Aarhus C Denmark3 Department of Botany Natural History Museum London SW7 5BD UKAuthor for correspondence Harald Schneider hschneidernhmacuk

Abstract The Greater Cape Floristic Region (GCFR) of southern Africa is characterised by large endemic radiations of flowering plants the so-called lsquoCape Cladesrsquo but it is unknown whether such radiations are also found in non-angiosperms We hypoth-esise that GCFR-endemic lineages exist in the xeric adapted cheilanthoid ferns To test this hypothesis with special emphasis on the alternative hypothesis of frequent colonization the phylogenetic relations divergence times and ancestral areas of the cheilanthoid ferns of the GCFR and adjacent regions are investigated The dataset includes 22 cheilanthoid fern species occur-ring in the GCFR With two exceptions all GCFR-endemics are part of two clades that diversified in the Afro-Madagascan region The GCFR-endemics are further concentrated in three high-endemism subclades that did not originate simultaneously but within the timeframe of angiosperm Cape Clades diversification According to ancestral area reconstructions the ancestors of the two larger Afro-Madagascan clades were likely GCFR-endemic and a substantial part of the diversification history of these clades took place in the GCFR The high diversity of cheilanthoids in the GCFR appears to stem primarily from in situ diversification not from immigration This pattern resembles the numerous radiations of angiosperm clades in the GCFR and may be caused by the same factors We did not find evidence for rapid synchronised radiations as documented for some albeit not all angiosperm Cape Clades High diversification within and dispersal out of the GCFR has likely enriched cheilanthoid diversity in the Afro-Madagascan region

Keywords biogeography Cape flora Cheilanthes chloroplast phylogeny Pellaea pteridophytes

Supplementary Material Table S1 and Figures S1ndashS3 (all in the Electronic Supplement) and the alignment are available in the Supplementary Data section of the online version of this article (httpwwwingentaconnectcomcontentiapttax)

1270

TAXON 60 (5) bull October 2011 1269ndash1283Eiserhardt amp al bull Cheilanthoid fern radiations in southern Africa

Richardson amp al 2001) is increasingly challenged by molecular age estimation studies (Verboom amp al 2009b Schnitzler amp al 2011) Many cape clades are too old for this explanation some dating back to the early Cenozoic (Verboom amp al 2009b) There is evidence that it applies to radiations in the SK and lowlands of the CFR (eg Klak amp al 2004) whereas montane lineages are older indicating that the Cape Fold Mountains have served as a refuge for Fynbos vegetation and associated taxa (Verboom amp al 2009b Schnitzler amp al 2011)

Several factors have likely acted in concert to cause the exceptional diversity of the GCFR The origin of this biodiver-sity hotspot may thus be best addressed by reconstructing the history of a diverse range of lineages with substantial differ-ences in their biology Many angiosperm Cape Clades are well studied whereas comparable studies are lacking for species-rich lineages of non-angiosperms such as ferns liverworts and mosses Understanding the origin of these lineages will help elucidate how the mechanisms underlying the diversification of the Cape Clades depend on particular life history aspects such as dispersal ability and reproductive biology

The Cape region also harbours a high diversity of ferns (Burrows 1990 Jacobsen 1983 Roux 2001 Schelpe amp Anthony 1986) but relatively little attention has been given to the diversification of ferns in this or other biodiversity hot-spots Studies on fern diversification found remarkable simi-larities to angiosperm diversification eg the colonization and radiation of the Diellia clade (Aspleniaceae) in Hawailsquoi (Schneider amp al 2005) the rapid radiation of tree ferns in the Madagascan biodiversity hotspot (Janssen amp al 2008 Sch-neider amp al 2010a) and the diversification of derived ferns in New Zealand (Perrie amp Brownsey 2007) In these examples the

high dispersal capacity of some ferns plays a role but patterns of local diversification were also found (Wolf amp al 2001 Schnei-der amp al 2005 Janssen amp al 2007 2008 Perrie amp Brownsey 2007) The majority of extant fern species belong to lineages that diversified in the shadow of angiosperms (Schneider amp al 2004 Schuettpelz amp Pryer 2009) and spatial diversity patterns are explained by the same environmental variables in ferns and angiosperms although humidity has a stronger impact on ferns (Kreft amp al 2010) However the two groups differ strongly in size with only about 9000 species of ferns (Smith amp al 2006) but at least 220000 angiosperm species (Scotland amp Wortley 2003) The importance of local radiation events needs to be seen in relation to the total diversity of the group for example a local radiation resulting in 10 fern species contributes about 01 to the global fern diversity and corresponds roughly to an angiosperm radiation generating about 220 new species The lower number of fern species coincides with the lower number of detected local fern radiations (Schneider amp al 2010b)

Cheilanthoid ferns (Pteridaceae subfam Cheilanthoideae) form a group of 278 or more species that generally have a prefer-ence for xeric and semi-xeric seasonal climates (Tryon amp al 1990 Schuettpelz amp al 2007) This is a less common adaption since ferns are most diverse in regions with abundant rainfall and equable temperatures (Barrington 1993) Cheilanthoids are an important element of the southern African fern flora (Fig 1) and show a higher degree of endemism than other ferns within the GCFR including many narrow-ranged endemics (Jacobsen 1983 Anthony 1984 Schelpe amp Anthony 1986 Burrows 1990) Based on this Anthony (1984) hypothesised that the south-western tip of Africa has been a centre of chei-lanthoid speciation resulting in the high number of endemics in the GCFR similar to the high endemism levels of several angiosperm groups in the Cape flora In situ diversification as such does not appear to be unusual as globally six major centres of cheilanthoid diversity have been described (Tryon amp Tryon 1973) one of which is South and East Africa together with Madagascar (henceforth Afro-Madagascar) These centres were proposed to be ldquomajor centres of autochthonous evolutionrdquo (Tryon amp Tryon 1973) which has since been corroborated with phylogenetic evidence (Gastony amp Rollo 1995 1998 Kirkpat-rick 2007 Prado amp al 2007 Rothfels amp al 2008 Windham amp al 2009) These studies support the scenario of a limited impact of long-distance dispersal on the diversification history of cheilanthoids Evidence for frequent long-distance dispersal was found in other fern groups such as the Afro-Madagascan polygrammoid ferns (Janssen amp al 2007) but not in the Mala-gasy scaly tree ferns (Janssen amp al 2008) and African mar-sileaceous ferns (Nagalingum amp al 2007) Previous studies on cheilanthoids included too few Afro-Madagascan species to test the phylogenetic coherence of this particular region or explore the relationships of the GCFR-endemics Here we aim to explore evidence for one or several in situ radiations of these xeric ferns in the GCFR We build a phylogeny from five plas-tid DNA loci that have previously been used with success to reconstruct relationships among cheilanthoid ferns (Gastony amp Rollo 1995 1998 Kirkpartrick 2007 Rothfels amp al 2008) We employ ancestral range reconstruction methods and explore

Fig 1 Distribution and species richness of cheilanthoid ferns in south-ern Africa based on distribution data provided in Burrows (1990) Cir-cle size scales with the number of taxa found in a 05 times 05deg grid cell centred on the circle midpoint Maximum circle size equals 16 spe-cies Circles with solid fill represent roughly the extent of the Greater Cape Floristic Region

0 1000km

1271


the respective contributions of local diversification events and colonization by long-distance dispersal to the assembly and maintenance of fern diversity in the GCFR We also estimate the timing of diversification events and compare these estimates with the diversification history of angiosperm Cape Clades

MaterIals and Methods

Taxon sampling mdash Our analysis included 175 of the 278+ species of cheilanthoid ferns (Tryon amp al 1990) To generate a global phylogeny of cheilanthoid ferns we downloaded all DNA sequences of these ferns accessible from GenBank in June 2009 and only retained loci with 10 or more sequences (Table S1 in the Electronic Supplement) We retained the protein coding regions atpA rbcL and rps4 as well as three non-coding regions the rps4-trnS intergenic spacer (IGS) and the trnL-trnF and trnG-trnR regions which include both an intron and an IGS These regions have previously been used with success to reconstruct the phylogeny of smaller samples of cheilanthoids (Gastony amp Rollo 1995 1998 Kirkpartrick 2007 Prado amp al 2007 Zhang amp al 2007 Rothfels amp al 2008 Windham amp al 2009) We generated sequences of previously un-sampled species in order to get 100 coverage of the species present in the GCFR and as many species as possible from the rest of southern Africa to explore relationships with the GCFR species

The first author obtained silica material of South African cheilanthoids during extensive fieldwork in the Richtersveld area Northern Cape Province and surroundings of Cape Town Western Cape Province in 2006 and 2007 Voucher specimens of silica-dried material were carefully identified by comparison with herbarium specimens and deposited in HBG Material of two species was provided by JP Roux (Compton Herbarium Cape Town) Additional samples were obtained from herbarium specimens kept in BM We included new sequences of three cheilanthoid species that occur from the Macaronesian islands throughout the European Mediterranean to Asia Minor and of five species from the Brazilian region These sequences were included to check for potential relationships with southern African cheilanthoids The Appendix provides all information concerning taxa and specimens used to generate new DNA sequences including the GenBank accession numbers of these sequences

Datasets mdash DNA was extracted using a modified CTAB protocol (Doyle amp Doyle 1987 Trewick amp al 2002) DNA am-plification and cycle sequencing was performed using standard protocols described in previous studies (eg Trewick amp al 2002 Schneider amp al 2004) A ball mill (Retsch Duumlsseldorf Germany) was used to pulverize samples for extraction and a NanoDrop 1000 Photometer (PeqLab Erlangen Germany) was used to measure the DNA content of extractions and PCR reactions Some PCR reactions were performed with PuReTaq Ready-To-Go PCR Beads (GE Heathcare Waukesha Wiscon-sin USA) All employed primers were published in previ-ous studies and are accessible at httpwwwpryerlabnet with the exception of the primer sets for atpA (Schuettpelz amp al 2006) trnL-trnF (Trewick amp al 2002) and rps4 + rps4-trnS

(Kirkpatrick 2007) The sequences were generated with the ABI PRISM Bigdye Terminator Cycle Sequencing Ready Reac-tion Kit (Applied Biosystems Foster City California USA) All sequences were assembled and edited using Sequencher v48 (Gene Codes Corporation Ann Arbor Michigan USA) and manually aligned using Mesquite v26 (Maddison amp Mad-dison 2007) Ambiguously aligned regions were excluded from all phylogenetic analyses

Given the heterogeneity of DNA regions employed in previous phylogenetic studies on cheilanthoid ferns we as-sembled eight datasets Five datasets include a single cpDNA region ie atpA rbcL rps4 + rps4-trnS IGS trnL-trnF or trnG-trnR These datasets are available for download in the Supplementary Data section of the online version of this article We also assembled four lsquosupermatrixrsquo datasets (1) all taxa for which at least one region is present (ldquoallTaxardquo 204 taxa) (2) all taxa for which rbcL is present (ldquorbcL-backbonerdquo 159 taxa) (3) selected taxa for which rbcL is present (ldquodatingrdquo 90 taxa) (4) only taxa with all five regions present (ldquonoGapsrdquo 27 taxa)

Phylogenetic analyses mdash Each of the eight datasets was analysed using maximum parsimony (MP) maximum likeli-hood (ML) and Bayesian inference of phylogeny (BPP) For MP all datasets were analysed in PAUP v40b10 (Swofford 2003) using a two-step search strategy In the first step we performed 1000 random-addition replicates with branch swap-ping employing the subtree pruning and regrafting algorithm (SPR) and a maximum of 10 most parsimonious trees kept in memory for each replicate In the second step the sampled trees were subjected to branch swapping using the tree bisec-tion reconnection algorithm (TBR) and a maximum number of 10000 most parsimonious trees Strict-consensus trees were calculated from the obtained trees Internal branch support was assessed by 1000 bootstrap replicates performed in PAUP for each dataset Each bootstrap replicate was calculated using one starting tree constructed with the simple addition algorithm and subsequently swapped with TBR holding a maximum of five trees in memory (Salamin amp al 2003)

ML analyses were carried out in GARLI v096 (Zwickl 2006) employing a general time reversible model with gamma-distribution rates and a proportion of invariable sites (GTR + I + Γ) This model was chosen because it covers a wide range of simpler nested models That way choosing a model with too few parameters is avoided while simpler models can emerge from parameter inference if they apply (Crisp amp al 2010) We assigned a single model to the entire dataset also in the case of supermatrices as GARLI does not allow partitioned analysis All parameters eg base frequencies gamma rates and proportion of invariable sites were estimated simultane-ously with the search for the optimal tree Ten search replicates were carried out on each dataset using the default settings of GARLI One hundred ML bootstrap replicates were run to assess the support for internal nodes using the same model and settings as above with two search replicates per bootstrap replicate Bootstrap values were computed using the SumTrees script (Sukumaran amp Holder 2008)

BPP were calculated using BayesianMarkov Chain Monte Carlo analyses as implemented in MrBayes v3122

1272


(Huelsenbeck amp Ronquist 2001) As in ML the GTR + I +Γ model was chosen However we specified separate models for the coding and non-coding parts of the alignment For the single-region datasets MCMC-analyses were carried out for 3 million generations with samples every 100th generation For the supermatrix datasets the number of generations was not defined beforehand but the analyses were terminated once a standard deviation of split frequencies of 001 had been reached Here we sampled every 1000th generation Posterior probabil-ity values were obtained with SumTrees omitting an appropri-ate number of burn-in trees as determined by visual inspection of the LogL traces Likelihood and Bayesian analyses were carried out on a cluster infrastructure via the CIPRES Portal v21 (Miller amp al 2009)

Divergence time analyses mdash Divergence time estimates were performed on the ldquodatingrdquo dataset using the BEAST v154 package (Drummond amp Rambaut 2007) The dataset was ana-lysed using the GTR + I + Γ model with up to 10 gamma catego-ries simultaneously estimating the parameters of the substitution model and a relaxed clock employing uncorrelated lognormal distribution We used separate models of nucleotide substitution for coding and non-coding parts of the dataset Analyses were done assuming either the Yule process or Birth-death model of diversification The topology was fixed to the topology in-ferred from the ldquoallTaxardquo dataset using maximum likelihood The chronogram was calibrated by divergence time estimates from an independent study on fern diversification by Schuett-pelz amp Pryer (2009) Cheilanthoid ferns are nearly absent in the fossil record and the interpretation of the available fossils is ambiguous given the low number of characters preserved and the degree of homoplasy in cheilanthoids We calibrated the crown age with 527 Myr (see Schuettpelz amp Pryer 2009) with a normal standard deviation of 10 Default conditions were used for all other parameters Three runs of 10 million generations each were generated sampling every 1000th generation The results were carefully studied for convergence using Tracer v14 (Rambaut amp Drummond 2007) The first 1000 sampled trees of each run were discarded as burn-in and the remainder summarized using TreeAnnotator (part of the BEAST package) and visualized with FigTree v13 (httptreebioedacuksoftwarefigtree)

Biogeographic analyses mdash Based on various floristic treatments we built a database with distributional information for all cheilanthoid species in our analysis Where necessary we used IPNI (wwwipniorg) and GBIF (wwwgbifnet) as an additional source of information mainly to guide our search for relevant floras Information on southern African species was obtained from Jacobsen (1983) Anthony (1984) and Burrows (1990) We scored the occurrence of each species for the fol-lowing regions (1) the GCFR (2) Afro-Madagascar excluding the GCFR (3) Australasia (4) Asia north of the Wallacersquos Line (5) Asia Minor Mediterranean Europe and North Africa to Macaronesia (6) North and Central America north of the Pana-manian Isthmus (7) Northern and Western South America and (8) Eastern Brazil These regions reflect distinct centres of cheilanthoid distributions (Tryon amp Tryon 1973)

Likelihood Analysis of Geographic Range Evolution (LAGRangE Ree amp Smith 2008) was used to reconstruct

ancestral ranges on the chronogram obtained from BEAST anal-ysis of the ldquodatingrdquo dataset The analysis was configured using the LAGRangE configurator available online (httpwwwreelabnetlagrange) We used neither range nor dispersal constraints and estimated the rate parameters during analysis Because of the limited number of ranges that can be analysed in LAGRangE we combined the Asian region with the Mediterranean and the two South American regions Preliminary analyses indicated that this would not have any bearing on the results for the GCFR species As a complementary approach we employed Dispersal-Vicari-ance Analysis (DIVA Ronquist 1996) using the hemionitid and notholaenid clades of the ML phylogeny (Fig 2) In contrast to LAGRangE this method allowed using the full set of regions moreover it is independent of the uncertainties associated with molecular age estimation and the most comprehensively sam-pled phylogeny could be used instead of the chronogram which includes only a subset of taxa We checked carefully for evidence suggesting migration into the GCFR

In order to explore biogeographic relationships within the GCFR we also performed a LAGRangE analysis on a subset of taxa with a more fine-scale definition of regions CFR SK Namibia and eastern South AfricaEast Africa The last two regions were distinguished because some species are endemic to Namibia north of the GCFR (Burrows amp al 1990)

results

The phylogenetic analyses of the various datasets recov-ered highly similar topologies We did not encounter any con-flicting signals among the bootstrap trees of the five utilized cpDNA regions the differences among these regions are re-stricted to the information content Dataset statistics are given in Table 1 while Table 2 summarizes the results in the context of the relationships of cheilanthoids found in the GCFR We focus on the results of the combined analyses as these subsume the signals of the five single cpDNA regions Figure 2 shows the phylogram obtained by ML analysis of the ldquoallTaxardquo dataset which is representative of our phylogenetic findings

The two biogeographic analyses ie LAGRangE and DIVA yielded similar results The LAGRangE reconstructions mapped on the chronogram are shown in Fig 3 The majority of Afro-Madagascan species (including the GCFR-endemics) were clustered in three clades These clades diversified entirely within in the region (clades 1 2 and 3 in Fig 2 and 3) with the exception of Cheilanthes viridis Pellaea calomelanos and P boivinii which originated in Afro-Madagascar but expanded their ranges subsequently into Asia or the Mediterranean All three Afro-Madagascan clades belong to the hemionitid clade (Fig 2B) sensu Windham amp al (2009)

All GCFR-endemics except for Cheilanthes rawsonii and Pellaea rufa belong to either clade 1 or clade 3 Three strongly supported groups of GCFR-endemics were found in clade 1 a group of five GCFR-endemics (clade 1a C kunzei C del-toidea C capensis C hastata C robusta Fig 4) was sister to the rest of the clade According to the fine-scale biogeographic analysis (Fig 4) this clade originated and diversified in the SK

1273


Table 1 Dataset statistics nTax number of terminal taxa nChar number of included characters nPI number of parsimony-informative characters CI consistency index RI retention index RC rescaled consistency index NMPT number of most parsimonious trees used for consensus LMPT length of the most parsimonious trees lnL log likelihood of the best tree found in GARLI

nTax nChar nPI CI RI RC NMPT LMPT lnLatpA 80 1515 275 0436 0715 0312 10000 1215 ndash875746rbcL 147 1381 328 0330 0724 0239 10000 1848 ndash1246870rps4 + rps4-trnS IGS 103 989 424 0480 0827 0397 10000 1783 ndash1137488trnL-trnF 114 804 388 0486 0753 0365 10000 1779 ndash1024720trnG-trnR 66 1005 419 0458 0676 0309 216 2071 ndash1194111noGaps 27 5646 927 0655 0699 0458 2 3104 ndash2488271allTaxa 204 5620 1829 0430 0747 0321 10000 8713 ndash5578240rbcL-backbone 159 5620 1703 0443 0716 0317 10000 7980 ndash5131125

Table 2 Phylogenetic evidence for the existence of Cape and South African radiations of cheilanthoid ferns Clade abbreviations as given in Fig 2 Bootstrap valuesposterior probabilities are given for clades with bootstrap gt 50 or posterior probability gt 05 NA stands for clades represented with less than two taxa in the given dataset ndash indicates a conflicting topology in the given dataset ~ indicates that the clade is unresolved but not conflicted in the given dataset

CladeDataset 1 1a 1b 1c 2 2a 3 3a 3bMaximum parsimony

atpA ndash 59 ndash 65 93 NA 62 68 97rbcL ~ lt50 ~ 78 71 68 ndash 55 ~rps4 + rps4-trnS lt50 82 lt50 99 100 58 ~ ndash ndashtrnL-F ~ 67 67 99 93 NA ~ 55 97trnG-R lt50 82 lt50 100 100 100 78 ~ 62noGaps 96 99 98 100 NA NA 100 65 100allTaxa lt50 99 61 100 79 ~ lt50 61 94rbcL-backbone 52 99 69 100 81 73 ~ 61 97

Maximum likelihoodatpA ndash 61 77 ndash 95 NA 69 73 95rbcL ndash lt50 lt50 ndash 59 75 ndash 51 ndashrps4 + rps4-trnS lt50 87 lt50 100 99 61 64 lt50 77trnL-F 69 89 77 96 97 NA 65 84 100trnG-R lt50 90 lt50 100 100 100 94 54 72noGaps 100 100 100 100 NA NA 100 99 100allTaxa 88 98 93 97 93 lt50 72 87 98rbcL-backbone 87 99 95 100 86 76 73 93 97

Bayesian inferenceatpA ~ 97 ~ ~ 100 NA 87 97 100rbcL ~ 93 73 ~ 89 94 ~ 95 64rps4 + rps4-trnS 97 100 75 100 100 79 96 ~ 100trnL-F 84 98 99 100 100 NA 93 100 100trnG-R 97 100 96 100 100 100 100 93 96noGaps 100 100 100 100 NA NA 100 100 100allTaxa 100 100 100 100 100 ~ 99 100 100rbcL-backbone 100 100 100 100 100 100 93 100 100

1274


with subsequent independent expansion of C capensis and C hastata into the CFR A second group (clade 1c) contained a species (C parviloba) whose range is centred on the CFR but extends to other parts of southern Africa nested within a group of two CFR endemics (C depauperata C contracta)

It is likely (Figs 3ndash4) that this clade has diversified within the CFR and the current widespread distribution of C parviloba is of recent origin Within clade 3 one widespread species (C multifida) was nested within two GCFR-endemics (C in-duta Pellaea pteroides clade 3a) A third GCFR-endemic

Fig 2A Maximum likelihood phylogeny of the allTaxa dataset basal part The species in bold face occur in the Greater Cape Floristic Region (GCFR) Species marked with an asterisk () are endemic in the GCFR Branch lengths are proportional to the number of substitutions per site The branch leading to the outgroup Calciphilopteris ludens is not true to scale (dashed line) Abbreviations (clade names following Windham amp al (2009) B bommeriid clade M myriopterid clade N notholaenid clade P pellaeid clade S Cheilanthes skinneri clade The inlay figure shows the entire phylogram the arrow indicates where it has been split into Fig 2A and Fig 2B

002

Pellaea brachyptera

Notholaena californica

Pellaea breweri

Cheilanthes aurea

Argyrochosma delicatula

Notholaena schaffneri

Paragymnopteris bipinnata

Notholaena rosei

Cheilanthes eatoniiCheilanthes tomentosa

Notholaena candida

Paragymnopteris marantae

Paragymnopteris sargentii

Cheilanthes skinneri

Notholaena trichomano

Astrolepis windhamii

Notholaena aliena

Cheilanthes fendleri

Notholaena aschenborniana

Argyrochosma limitanea

Pellaea ternifolia

Pellaea rufa

Astrolepis cochisensis

Pellaea truncata

Pellaea notabilis

Cheilanthes leucopoda

Cheilanthes wrightii

Notholaena copelandii

Paraceterach muelleri

Notholaena bryopoda

Pellaea cordifolia

Cheilanthes lendigera

Notholaena grayi

Pellaea ovata

Cheilanthes allosuroides

Bommeria hispida

Pellaea pringlei

Platyloma rotundifolia

Notholaena sulphurea

Pellaea wrightiana

Cheilanthes rawsonii

Cheilanthes lanosa

Cheilanthes myriophylla

Cheilanthes covillei

Cheilanthes brachypus

Platyloma falcataPlatyloma nana

Pellaea mucronata

Cheilanthes notholaenoides

Calciphilopteris ludens

Notholaena lemmonii

Paragymnopteris vestita

Astrolepis sinuata

Pellaea sagittata

Notholaena greggii

Notholaena neglecta

Argyrochosma jonesii

Notholaena affinis

Cheilanthes parryiCheilanthes feei

Pellaea andromedifolia

Cheilanthes newberryi

Paragymnopteris delavayi

Pellaea glabella

Pellaea atropurpurea

Pellaea times glaciogena

Cheilanthes gracillima

Cheilanthes aurantiaca

Cheiloplecton rigidum

Cheilanthes bonariens

Notholaena standleyi

Argyrochosma nivea

Cheilanthes horridula

Argyrochosma fendleri

Bommeria ehrenbergiana

Cheilanthes alabamensis

Notholaena rigida

Pellaea bridgesii

Pellaea intermedia

Argyrochosma incana

B

P

S

N

M

Fig 2B

1275


Fig 2B Maximum likelihood phylogeny of the allTaxa data-set crown group The species in bold face occur in the Greater Cape Floristic Region (GCFR) Species marked with an asterisk () are endemic in the GCFR Branch lengths are proportional to the number of substitutions per site Abbreviations A Aleuritopteris clade C Chei-lanthes clade D Doryopteris clade H hemionitid clade sensu Windham amp al (2009) Clades 1 2 and 3 occur in the Afro-Madagascan region

Doryopteris pilosa

Aleuritopteris albomarginata

Cheilanthes bergiana

Cheilanthes deltoidea

Doryopteris paradoxa

Cheilanthes decora

Cheilanthes contracta

Pellaea patula

Pellaea dura

Pellaea doniana

Cheilanthes acrostica

Cheilanthes arizonica

Cheilanthes pulchella

Aleuritopteris formosana

Sinopteris albfusca

Ormopteris crenataOrmopteris pinnata

Doryopteris lorentzii

Aspidotis carlotta-halliae

Pellaea calomelanos

Doryopteris pedata

Hemionitis tomentosa

Cheilanthes involuta

Cheilanthes eckloniana

Cheilanthes insignis

Doryopteris triphylla

Cheilanthes kunzei

Doryopteris collina

Adiantopsis radiata

Cheilanthes multifida

Cheilanthes buchtienii

Hemionitis rufa

Pentagramma triangularis

Pellaea trichophylla

Pellaea paupercula

Cheilanthes distans

Doryopteris nobilis

Doryopteris sagittifolia

Cheilanthes flexuosa

Aleuritopteris niphobola

Ormopteris cymbiformis

Notholaena chinensis

Pellaea pteroides

Doryopteris ornithopus

Cheilanthes dinteri

Aleuritopteris squamosa

Hemionitis (Parahemionitis ) arifolia

Cheilanthes robusta

Cheilanthes pentagona

Ormopteris gleichenioides

Aleuritopteris argentea

Cheilanthes marlothii

Leptolepidium dalhousiae

Doryopteris pentagona

Doryopteris rediviva

Pellaea maxima

Cheilanthes induta

Cheilanthes namaquensis

Doryopteris lomariacea

Aleuritopteris tamburii

Cheilanthes leachii

Pellaea nitidula

Pellaea longipilosa

Cheilanthes hirta

Cheilanthes micropteris

Pellaea pectiniformis

Cheilanthes kuhnii

Cheilanthes cf kunzei

Cheilanthes viridis

Cheilanthes parviloba

Pellaea nitidula

Trachypteris pinnata

Pellaea boivinii

Aleuritopteris cf farinosa

Hemionitis palmata

Doryopteris concolor

Aleuritopteris likiangensis

Sinopteris grevilleoides

Cheilanthes hastata

Aleuritopteris duclouxii

Cheilanthes capensis

Aspidotis densa

Ormopteris riedelii

Pellaea cf dura

Cheilanthes quadripinnata

Cheilanthes venusta

Doryopteris pedatoides

Aspidotis californica

Cheilanthes depauperata

Hemionitis levyi

Cheilanthes goyazensis

Cheilanthes maderensis

Adiantopsis chlorophylla

Aleuritopteris grisea

D

C

A

1

1b

1a

2

2a

3

3a

3b

Cheilanthes intramarginalis

002

1c

HFig 2A

1276


(C namaquensis) potentially groups with this clade but its positsion within clade 3 differed between analysesdatasets Anthony (1984) divided C multifida into two subspecies with the typical subspecies (C multifida subsp multifida) endemic to the GCFR This evidence and the ancestral area reconstruc-tion results suggest strongly that clade 3a potentially together with C namaquensis diversified at the Cape

The LAGRangE results also indicate that the ancestors of clade 1 and possibly clade 3 were GCFR-endemic While the likelihood for an ancestral GCFR distribution of clade 3 was found to be 100 (Fig 3) the ancestral distribution of clade 1 was more ambiguous The global biogeographic analy-sis (where the GCFR was treated as a single region) yielded moderate support for GCFR-endemic ancestor of this group

Fig 3 Subtree of a chronogram obtained from analysis of a dataset including 90 taxa of cheilanthoids with BEAST showing the basal portion of the hemionitid clade (cf Fig 2B) Scale axis = million years Values in square brackets are 95 confidence intervals of age estimates Letters at internal nodes indicate the initial geographic distributions of the descendant lineages of each speciation event (LAGRangE reconstruction) Splits without annotation have the same range reconstruction as the previous split Where LAGRangE produced several possible reconstructions only the one with the highest likelihood is shown Pie-charts indicate the likelihood of the shown reconstruction [] Reconstructions without pie-chart have 100 likelihood Ranges a North America b South America c Mediterranean and Asia d Australasia e Afro-Madagascar excluding the GCFR f Greater Cape Floristic Region (GCFR) all regions Bold branches illustrate the parts of the phylogeny where the most likely reconstruction is GCFR-endemic (f)

ee

00100200300

[1147]

[881762]

[78171]

[175265]

[119194]

[109204]

[41123]

[209296]

[126200]

[187278][99204]

[73147]

[48115]

[44132]

[84187]

[95168][86159]

[130270]

[114207]

[85156]

[1866]

[157247]

[208294]

[64163]

[155234]

[130214]

[172254]

[37104]

[67134]

[154244]

[841905]

[55143]

[48114]

[179265]

[2266]

[49143]

Ch parvilobaCh contractaCh depauperataCh hirtaCh ecklonianaCh marlothiiCh dinteriCh deltoideaCh capensisCh hastataCh kunzeiCh robustaCh micropterisO crenataD paradoxaD sagittifoliaD collinaD lorentziiD concolorCh quadripinnP calomelanosCh viridis glCh viridisP pteroidesCh multifidaCh indutaCh namaquensAd radiataCh flexuosaT pinnataH tomentosaH levyiH arifoliaD pilosaP boiviniiP duraAsp californicaCh arizonicaPgr triangularisCh intramarg

[215305]

[233332]

[213302]

[254363]

baab

cabce

deff

f

bf

bb

bb

fef

efe

efe

efe

ff

fb

aa

babc

abca

cabc

Asian clade (A)

aa

cefffefefeefffffbbbbbbefcef

cefe

fefffabbbbaceceeaabb

1

3

2

1277


(P = 065 Fig 3) while an ancestral distribution in the SK and Namibia north of the GCFR was recovered in the fine-scale analysis (P = 07 Fig 4) According to LAGRangE a large pro-portion of the diversification of clade 3 took also place in the GCFR but likelihoods for this were only around 50 (Fig 3) DIVA was ambiguous with respect to these patterns

Of the three clades containing GCFR-endemics clade 1a appears to be the oldest (Fig 3) It started to accumulate diversity around 15 Ma (95 of the posterior probability dis-tribution between 99 and 204 Ma) A relatively young crown age of about 4 Myr [22 66] was found for the diversification of clade 1c which consists of C contracta C depauperata and C parviloba Clade 3a comprising two species and one subspecies endemic to the GCFR has a crown age of about 13 Myr [95 168]

While clades 1 and 3 contain the large majority of GCFR-endemics (9 of 11) in three well-defined groups (1a 1c 3a) two endemic species were found in very isolated positions Pellaea rufa emerged as member of the pellaeid (P) clade and Cheilanthes rawsonii as member of the myriopterid (M) clade and both were deeply nested within taxa that occur in Central and Northern America

Besides these two species and species of clades 1 and 3 only Doryopteris concolor occurs in the GCFR This species has a pantropical distribution and was found nested within the core Doryopteris clade that is centred in the Brazilian region

The geographic relationships of the three Afro-Madagas-can clades were partially resolved in the ancestral area analysis

Clade 1 formed a group with a clade consisting of the Brazilian C micropteris and the Australian C distans DIVA recon-structed the ancestral distribution of this group to definitely contain the GCFR and the Australasian region plus possibly South America andor Afro-Madagascar LAGRangE was very ambiguous for this node suggesting an ancestral South Ameri-can distribution with a low likelihood value

Clade 3 is part of the D clade that is named according to the genus Doryopteris Clade D consists of the core of Doryopteris and Neotropical genera such as Adiantopsis and Ormopteris (= Pellaea sect Ormopteris) The ancestor of clade 3 and its closest relative had a disjunct distribution in the Brazilian re-gion and the GCFR andor remaining Afro-Madagascar ac-cording to DIVA but a South American distribution according to LAGRangE (Fig 3) This indicates that clade 3 originated in and dispersed out of the Brazilian region with subsequent speciation in the African region Clade 2 is most closely related to the genus Hemionitis and their common ancestor must have been present in the African region and potentially also in South America

The newly sequenced species of Cheilanthes originat-ing from the Macaronesian-Mediterranean floristic region (C acrostica C maderensis C pulchella) and form a well-supported clade nested within clade A They are most closely related to species occurring in East Asia and thus only distantly related to sub-Saharan cheilanthoids

dIscussIon

Phylogenetic framework mdash The backbone of the recov-ered phylogeny is congruent with previously reported results (Gastony amp Rollo 1995 1998 Kirkpartrick 2007 Prado amp al 2007 Zhang amp al 2007 Rothfels amp al 2008 Windham amp al 2009) suggesting that a reliable phylogenetic framework for studying the diversification of GCFR cheilanthoids was cre-ated Only few of the analysed taxa had been sequenced for all five DNA regions thus we had to reconstruct phylogenetic relationships from matrices with considerable gaps for most species It is controversial if large proportions of missing data as encountered in our dataset have a detrimental effect on the accuracy of phylogenetic inference (Wiens amp Moen 2008 Lemmon amp al 2009) A variety of different sampling strate-gies and phylogenetic methods were used to explore the effect of missing data but largely congruent results were recovered We also explored supertree approaches (results not shown) de-signed to recover clades supported in different datasets (Baum 1992 Ragan 1992) Largely concordant results were found in both supermatrix and supertree approaches each with good internal support again indicating that an accurate phylogenetic signal was recovered from the data This phylogeny therefore appears to be well-suited for addressing the diversification history of cheilanthoids in the GCFR

Radiations in the GCFR mdash This study recovered strong evidence not only for single speciation events in the GCFR but also for local radiations ie the accumulation of several species originating from a common ancestor from the region

Fig 4 LAGrangE reconstruction of ancestral ranges in clade 1 treat-ing the Fynbos (CFR) and Succulent Karoo (SK) biomes as separate areas Letters at internal nodes indicate the initial geographic dis-tributions of the descendant lineages of each speciation event (LA-GRangE reconstruction) E eastern South Africa to East Africa N Namibia north of the GCFR Splits without annotation have the same range reconstruction as the previous split Where LAGRangE pro-duced several possible reconstructions only the one with the highest likelihood is shown Numbers at nodes are probabilities [] of the shown reconstruction Branch lengths are proportional to time scale axis in million years

100200

Ch parviloba

Ch contracta

Ch depauperata

Ch hirta

Ch eckloniana

Ch marlothii

Ch dinteri

Ch deltoidea

Ch capensis

Ch hastata

Ch kunzei

Ch robusta

CFRNE

CFR

CFR

CFRNE

CFRNE

N

N

SK

SKCFR

SKCFR

SK

SK

5466

28

69

88

84

9399

93

00

87 55

CFRCFRN

NSK

N

SK

(84)

1278


We found three core ldquoCape Cladesrdquo that likely diversified in the GCFR resulting in relatively large groups of range-restricted species (clades 1a 1c and 3a in Figs 2ndash3) This pattern resem-bles that of several angiosperm groups (Linder 2003 2008 Verboom amp al 2009a) although these clades are significantly less speciose than the large angiosperm Cape Clades To de-tect an evolutionary response to factors that are special for the GCFR Cape and non-Cape lineages should be compared (Barraclough 2006) In cheilanthoids more narrow-ranged endemic species are found in the GCFR than in the rest of Af-rica (Anthony 1984 also Schelpe 1970 1977 Jacobsen 1983 Burrows 1990 Roux 2001) and these endemic GCFR species originated likely from in situ speciation Thus the hypothesis of a ldquocentre of cheilanthoid fern speciationrdquo analogous to the angiosperm speciation in the Cape flora (Anthony 1984) is here substantiated with phylogenetic information for the first time

The timing of diversification has featured prominently in explanations of angiosperm radiation in the GCFR (eg Richardson amp al 2001 Goldblatt amp al 2002 Linder amp Hardy 2004 Linder 2008 Sauquet amp al 2009 Verboom amp al 2009b) Our divergence time estimates did not indicate elevated diversification rates over a certain period of time which con-stitute a radiation sensu Linder (2003) but we did not deem our taxon sampling to be comprehensive enough for a formal test (eg lineage-through-time plots) Nor do the cheilanthoid GCFR clades appear to have originated simultaneously How-ever angiosperm GCFR clades also diversified at different time periods (Linder 2008 Verboom amp al 2009b) and the hypothesis of a coincident burst of speciation giving rise to the GCFR biomes is increasingly being challenged (Linder amp Hardy 2004 Hopper amp al 2009 Sauquet amp al 2009 Schnitzler amp al 2011) The crown ages of clades 1a 1c and 3a fall well within the range of ages documented for angiosperm Cape Clades (Verboom amp al 2009b) This holds even if the entire clades 1 and 3 are interpreted as Cape Clades (ancestrally Cape endemic) that showed a high degree of lineage dispersal into tropical Africa It must however be kept in mind that our divergence time estimates are based on a single secondary cali-bration point (Schuettpelz amp Pryer 2009) At present this is the only available option to date our phylogeny as reliable fossils are not available Thus our results may be deemed uncertain however cheilanthoids as a whole would have to be much older than indicated by all existing divergence time estimates (Pryer amp al 2004 Schneider amp al 2004 Schuettpelz amp Pryer 2009) in order to invalidate our conclusions

The factors that caused the diversification of angiosperms in the Cape may also have impacted the diversification of the ferns of the region Our results are most compatible with a scenario in which the high species diversity of the Cape chei-lanthoids is the result of relatively long-term stability and isola-tion of the region combined with a mosaic of steep ecological gradients (Goldblatt amp Manning 2002 Linder 2003 Cowling amp al 2009 Schnitzler amp al 2011) We did not test to what extent speciation is associated with allopatry or ecological divergence like it has been done for some angiosperm Cape Clades (eg Schnitzler amp al 2011) However the finding that groups with very different reproductive biology have radiated

in the GCFR informs the discussion on the factors that promote in situ diversification in this region First animal pollination has been suggested to play a role in the diversification of Cape plants both in the form of divergent adaptation to pollinators and generally short dispersal distance of pollinators causing genetic isolation (Johnson 2010 Linder amp al 2010) In the Afromontane region some groups with specialised plant-pollinator interactions have diversified in situ in contrast to wind-pollinated groups (Galley amp al 2007) At the Cape however both groups with highly specialised insect pollina-tion (eg in Iridaceae Orchidaceae) and groups that do not de-pend on pollinators (eg Cheilanthoideae Restionaceae) have radiated indicating that pollination is relevant for some Cape radiations at best Second limited seed dispersal has been suggested to promote plant diversification in the GCFR by restricting gene flow and increasing the incidence of allopatric speciation (Linder 2003 Ellis amp al 2006 Linder amp al 2010) Ferns are known as good dispersers (cf Wolf amp al 2001) accordingly the GCFR-endemic cheilanthoids have relatively large distributions compared to endemic angiosperms (Figs S1ndashS3) Our findings illustrate that not only groups with short dispersal distances have radiated at the Cape Interestingly cheilanthoids do not seem to have allopatric distributions in the GCFR with the caveat that distribution data is available only at relatively coarse resolution (05 times 05deg Burrows 1990) Meanwhile at least some of the GCFR species are clearly specialised in certain soils or microhabitats (Burrows amp al 1990) There is evidence that speciation in some angiosperm Cape Clades is associated with ecological divergence (Ellis amp al 2006 Schnitzler amp al 2011) and the same might ap-ply to cheilanthoids At present it is not possible to test this formally because habitat preferences have not been recorded consistently for all species

Although most cheilanthoid GCFR-endemics are rela-tively widespread within the region species tend to be con-fined to either of the two biomes indicating that specialisation to the different environments of CFR and SK may have played a role in cheilanthoid fern diversification Biome association is not randomly distributed with respect to phylogeny clades 1a and 1c appear to have diversified within the SK and CFR respectively This parallels the situation in angiosperms where SK and CFR clades can often be distinguished (Verboom amp al 2009b) Clade 1a matches a pattern inferred for some angio-sperm Cape Clades that are thought to originate from the more arid northern parts of the GCFR (Barker amp al 2004 Bellstedt amp al 2008 Schrire amp al 2009) The same could apply to clade 1 as a whole for which an ancestral distribution in the SKNamibia was reconstructed It seems that this ancestor split into a SK lineage (clade 1a) and a clade that first diver-sified in the Namibian uplands and subsequently expanded eastward (clade 1b) From the last lineage the CFR endemic clade 1c was derived relatively recently However this sce-nario depends crucially on the monophyly of clade 1 Except for P pteroides the species of clade 3a are all widespread in the GCFR indicating that the biomes did not play a ma-jor role for the diversification of this group Phytogeographic centres within the CFR biome (Weimarck 1941 Goldblatt

1279


amp Manning 2002) have provided insight into the fine-scale biogeography of some angiosperm Cape Clades suggesting both diversification within and vicariance between centres (eg Leucadendron Barker amp al 2004) However cheilan-thoids are generally more widespread and not restricted to these regions The different scale of endemism and geographic phylogenetic structure might be caused by differences in dis-persal limitation while environmental gradients equally im-pact cheilanthoid and angiosperm diversification The high degree of GCFR endemism on the species as well as clade level suggests that rainfall seasonality is an important fac-tor for cheilanthoid ecology The GCFR is largely congruent with the winter-rainfall zone of southern Africa (Born amp al 2007) and the transition to summer-rainfall tropical Africa can be extremely steep resulting in abrupt floristic turnover (N Juumlrgens pers comm) GCFR-endemics are likely adapted to winter-rainfall but cheilanthoid species outside this region may have preferences for summer-rainfall regimes (Burrows 1990) The observed clustering of GCFR-endemics may be due to the phylogenetic conservation of adaptation to winter-rainfall regimes However under such a scenario the fact that some cheilanthoid species that mainly occur in summer-rainfall regions have small outlier populations in the GCFR needs to be explained This could be due to one of the following factors (1) intra-specific variation in the speciesrsquo phenology (2) these plants grow in azonal habitats eg around springs or creeks with all-year moisture where rainfall seasonality is less important (Goldblatt amp Manning 2002) or (3) the GCFR populations occur due to a mass effect (Shmida amp Whittaker 1981) ie species establish temporarily in a suboptimal habi-tat if propagule pressure is high With the possible exception of these few outlier populations of summer-rainfall species just mentioned the overall distinctness of the GCFR is likely the result of phylogenetic conservatism in tolerances towards rainfall seasonalityaridity these niche characters appear to drive the distinctness of the cheilanthoid flora in the region

Geographic relationships of GCFR clades mdash The GCFR clades recovered here are clearly related to the cheilanthoid flora of the remaining Afro-Madagascar region However ancestral area reconstructions suggest that the GCFR clades may not have originated from a pre-existing Afro-Madagascan flora with the exception of clade 1c which derives from a lineage that diversified in the Namibian mountains (Fig 4) All three Afro-Madagascan clades and particularly clade 3 show relationships with species occurring in the Americas The provenance of clade 1 however is problematic to reconstruct from our data This clade appears to be related to C distans the only Australasian Cheilanthes species included here Aus-tralasia in particular the diversity centre of south-western Australia (cf Tryon amp Tryon 1973) is the only region that is severely under-sampled and underrepresented in our phylog-eny There are clear relationships between the angiosperm Cape flora and the flora of Australia (Linder 2005) a relation between clade 1 and the Australian cheilanthoids cannot be rejected although an ancestral South American distribution is suggested both by DIVA and LAGRangE The situation for clade 2 is much clearer the ancestor of this group arrived in

Africa from South America These relationships could eas-ily be misinterpreted as support for a southern Gondwana origin of these clades However the diversification of the cheilanthoids occurred much later than the break-up of the southern Gondwanan continent in the Upper Cretaceous and early Palaeocene all existing divergence time estimates agree with the lack of fossil evidence for a pre-Eocene origin of the clade (Schneider amp al 2004 Schuettpelz amp Pryer 2009) Frequent exchange between Africa and the Americas is now widely accepted for many groups of plants and animals (eg Givnish amp Renner 2004 Renner 2004) but only few cases are documented for the Cape flora (Galley amp Linder 2006) Ancestral area reconstructions suggest that South American cheilanthoid lineages might initially have reached the GCFR after which the Afro-Madagascan (non-GCFR) lineages would have diverged from initial GCFR populations while clade 1a and 3a continued to diversify within the region Interestingly Cheilanthes rawsonii and Pellaea rufa provide two more ex-amples of species that reached the GCFR from the Neotropics Relationships between New World and African ferns are not uncommon and dispersal appears to occur more frequently in the west-east direction (Moran amp Smith 2001) Our re-sults corroborate this trend and suggest a potential role of the GCFR as a beachhead for colonisation of Africa by New World ferns In contrast the GCFR cheilanthoids show very limited exchange with the floras of Asia and the Mediterra-nean only some widespread Afro-Madagascan species that occur in the Cape also displayed ldquoout of Africardquo expansions but migrations in the other direction are not evident This is perhaps unexpected because cheilanthoid ferns are expected to be less affected by deserts as geographical barriers given their greater tolerance against limited access to water Never-theless the Sahara or the wet rainforests of the Congo appear to be strong barriers segregating sub-Saharan and Mediter-ranean cheilanthoids which show little relatedness to each other More exhaustive sampling of cheilanthoid ferns in Asia Australia and the Americas will provide the necessary resolu-tion to confirm some of the patterns observed in this study

A considerable part of the Afro-Madagascan cheilanthoid diversity (as sampled by us) appears to have originated in the GCFR or from lineages that once were GCFR-endemic Ex-pansion of GCFR lineages into other parts of sub-Saharan Africa has also been documented in angiosperms (eg Galley amp Linder 2006 Galley amp al 2007) In particular many Cape lineages have dispersed northwards along the temperate peaks of the Great Escarpment (Galley amp al 2007) Few cheilanthoid species might represent this track such as C quadripinnata with a distribution range mostly restricted to the eastern es-carpment of southern Africa (Burrows 1990) Other Afro-Madagascan species that seem to derive from Cape lineages are remarkably widespread and variable in their ecological preferences (eg P calomelanos C viridis C multifida) Evolution of a broad ecological niche probably allowed these species to lsquoescapersquo the GCFR The provenance of the tropical African species in clade 1b remains unclear According to the global biogeographic reconstruction the ancestor of clade 1 was GCFR-endemic indicating that this lineage dispersed from

1280


the GCFR northwards into Namibia However this reconstruc-tion might be an artefact of treating SK and CFR as a single region The fine-scale biogeographic analysis (Fig 4) indicates that the ancestor of clade 1 occurred both in the SK and the Namibian uplands rendering vicariant speciation more likely Divergent adaptation to summer and winter rainfall might be a possible explanation for this split

Notes on the classification of southern African cheilan-thoids mdash This study contributes to a growing body of knowl-edge on the phylogenetic history of cheilanthoid ferns (Gastony amp Rollo 1995 1998 Zhang amp al 2003 2007 Kirkpatrick 2007 Prado amp al 2007 Schuettpelz amp al 2007 Rothfels amp al 2008 Windham amp al 2009) A comprehensive phylogenetic study of the subfamily Cheilanthoideae is needed and generic boundaries have to be redefined since the current classifica-tion is unnatural and cannot be used to adequately address biogeographic questions The classification within cheilanthoid ferns has always been a matter of debate because morphologi-cal similarities among the species may be more often the result of convergent evolution rather than one of common ancestry (Windham amp al 2009) More recently the majority of southern African cheilanthoid ferns have been classified as either Chei-lanthes or Pellaea (Anthony 1984 Burrows 1990) but both of these genera are polyphyletic Clade C includes the type of Cheilanthes (C micropteris from the Brazilian region) together with species occurring in the Cape region and an Australian species (Fig 2) This tentatively suggests a pseudo-Gondwanan distribution for the genus Cheilanthes although our sampling outside the GCFR is insufficient to exhaustively explore this hypothesis and we are looking forward to the results of a broader study as outlined by Windham amp al (2009) While species belonging to clade 1 may be kept within Cheilanthes but species belonging to clades 2 and 3 may be members of different genera (Fig 2) Importantly among the species from the GCFR only Pellaea rufa belongs to the genus Pellaea A new genus classification of southern Africa cheilanthoid ferns is thus very necessary though this classification must be based on a global and not on a regional study

acknowledgeMents

We thank all colleagues that deposited their DNA sequences to GenBank various colleagues eg Norbert Juumlrgens Koos Roux Tassilo Feuerer Alison Paul Barbara Rudolph and Julia Llewellyn-Hughes and her team for supporting this project during fieldwork laboratory work and with access to herbarium specimens and silica samples Royal Botanic Garden Edinburgh for providing access to living collections BIOTA Southern Africa for making the collec-tion of cheilanthoids in the Northern Cape Province possible and Cape Nature for issuing the collection permits AAA005-00058-0028 and AAA005-00086-0028 WLE received support from the SYN-THESYS Project httpwwwsynthesysinfo which is financed by European Community Research Infrastructure Action under the FP6 ldquoStructuring the European Research Areardquo Programme We also thank four anonymous reviewers for comments on previous versions of the manuscript

Anthony NC 1984 A revision of the southern African species of Chei-lanthes Swartz and Pellaea Link Contr Bolus Herb 11 1ndash293

Barker NP Vanderpoorten A Morton CM amp Rourke JP 2004 Phylogeny biogeography and the evolution of life-history traits in Leucadendron (Proteaceae) Molec Phylog Evol 33 845ndash860

Barraclough TG 2006 What can phylogenetics tell us about specia-tion in the Cape flora Diversity amp Distrib 12 21ndash26

Barrington DS 1993 Ecological and historical factors in fern bioge-ography J Biogeogr 20 275ndash279

Baum BR 1992 Combining trees as a way of combining data sets for phylogenetic inference and the desirability of combining gene trees Taxon 41 3ndash10

Bellstedt DU Van Zyl L Marais EM Bytebier B de Vil-liers CA Makwarela AM amp Dreyer LL 2008 Phylogenetic relationships character evolution and biogeography of southern African members of Zygophyllum (Zygophyllaceae) based on three plastid regions Molec Phylog Evol 47 932ndash949

Born J Linder HP amp Desmet P 2007 The greater Cape floristic region J Biogeogr 34 147ndash162

Burrows JE 1990 Southern African ferns and fern allies Sandton Frandsen

Cowling RM Proches S amp Partridge TC 2009 Explaining the uniqueness of the Cape flora Incorporating geomorphic evolu-tion as a factor for explaining its diversification Molec Phylog Evol 51 64ndash74

Cowling RM Richardson DM amp Mustart PJ 1997 Fynbos Pp 99ndash130 in Cowling RM Richardson DM amp Pierce SM (eds) Vegetation of Southern Africa Cambridge Cambridge Uni-versity Press

Crisp MD Isagi Y Kato Y Cook LG amp Bowman DM 2010 Livistona palms in Australia Ancient relics or opportunistic im-migrants Molec Phylog Evol 54 512ndash523

Doyle JJ amp Doyle JA 1987 A rapid DNA isolation procedure for small quantities of fresh tissue Phytochem Bull 19 11ndash15

Drummond AJ amp Rambaut A 2007 BEAST Bayesian evolution-ary analysis by sampling trees BMC Evol Biol 7 214 DOI 1011861471-2148-7-214

Ellis AG Weis AE amp Gaut BS 2006 Evolutionary radiation of ldquostone plantsrdquo in the genus Argyroderma (Aizoaceae) Unraveling the effects of landscape habitat and flowering time Evolution 60 39ndash55

Forest F Grenyer R Rouget M Davies TJ Cowling RM Faith DP Balmford A Manning JC Proches S Van der Bank M Reeves G Hedderson TAJ amp Savolainen V 2007 Preserving the evolutionary potential of floras in biodiversity hotspots Nature 445 757ndash760

Galley C amp Linder HP 2006 Geographical affinities of the Cape flora South Africa J Biogeogr 33 236ndash250

Galley C Bytebier B Bellstedt DU amp Linder HP 2007 The cape element in the Afrotemperate flora From Cape to Cairo Proc Roy Soc London Ser B Biol Sci 274 535ndash543

Gastony GJ amp Rollo DR 1995 Phylogeny and generic circumscrip-tions of cheilanthoid ferns (Pteridaceae Cheilanthoideae) inferred from rbcL nucleotide sequences Amer Fern J 85 341ndash360

Gastony GJ amp Rollo DR 1998 Cheilanthoid ferns (Pteridaceae Cheilanthoideae) in the southwestern United States and adjacent MexicomdashA molecular phylogenetic reassessment of generic lin-eages Aliso 17 131ndash144

Givnish TJ amp Renner SS 2004 Tropical intercontinental disjunc-tions Gondwana breakup immigration from the boreotropics and transoceanic dispersal Int J Pl Sci 165 S1ndashS6

Goldblatt P amp Manning JC 2002 Plant diversity of the Cape region of southern Africa Ann Missouri Bot Gard 89 281ndash302

Goldblatt P Savolainen V Porteous O Sostaric I Powell M Reeves G Manning JC Barraclough TG amp Chase MW

lIterature cIted

1281


2002 Radiation in the Cape flora and the phylogeny of peacock irises Moraea (Iridaceae) based on four plastid DNA regions Molec Phylog Evol 25 341ndash360

Hopper SD Smith RJ Fay MF Manning JC amp Chase MW 2009 Molecular phylogenetics of Haemodoraceae in the Greater Cape and Southwest Australian Floristic Regions Molec Phylog Evol 51 19ndash30

Huelsenbeck JP amp Ronquist F 2001 MRBAYES Bayesian infer-ence of phylogeny Bioinformatics 17 754ndash755

Jacobsen WBG 1983 The ferns and fern allies of southern Africa Durban Butterworth

Janssen T Bystriakova N Rakotondrainibe F Coomes D La-bat JN amp Schneider H 2008 Neoendemism in Madagascan scaly tree ferns results from recent coincident diversification bursts Evolution 62 1876ndash1889

Janssen T Kreier H-P amp Schneider H 2007 Origin and diversi-fication of African ferns with special emphasis on Polypodiaceae Brittonia 59 159ndash181

Johnson SD 2010 The pollination niche and its role in the diversifica-tion and maintenance of the southern African flora Philos Trans Ser B 365 499ndash516

Juumlrgens N 1997 Floristic biodiversity and history of African arid regions Biodivers amp Conservation 6 495ndash514

Kirkpatrick REB 2007 Investigating the monophyly of Pellaea (Pteridaceae) in the context of a phylogenetic analysis of cheilan-thoid ferns Syst Bot 32 504ndash518

Klak C Reeves G amp Hedderson T 2004 Unmatched tempo of evolution in Southern African semi-desert ice plants Nature 427 63ndash65

Kreft H amp Jetz W 2007 Global patterns and determinants of vascu-lar plant diversity Proc Natl Acad Sci USA 104 5925ndash5930

Kreft H Jetz W Mutke J amp Barthlott W 2010 Contrasting environmental and regional effects on global pteridophyte and seed plant diversity Ecography 33 408ndash419

Lemmon AR Brown JM Stanger-Hall K amp Lemmon EM 2009 The effect of ambiguous data on phylogenetic estimates obtained by maximum likelihood and Bayesian inference Syst Biol 58 130ndash145

Levyns MR 1964 Migrations and the origins of the Cape flora Trans Roy Soc South Africa 37 85ndash107

Linder HP 2003 The radiation of the Cape flora southern Africa Biol Rev Cambridge Philos Soc 78 597ndash638

Linder HP 2005 Evolution of diversity The Cape flora Trends Pl Sci 10 536ndash541

Linder HP 2006 Investigating the evolution of floras Problems and progressmdashAn introduction Diversity amp Distrib 12 3ndash5

Linder HP 2008 Plant species radiations Where when why Philos Trans Ser B 363 3097ndash3105

Linder HP 2010 Gradual speciation in a global hotspot of plant di-versity Molec Ecol 194583ndash4585

Linder HP amp Hardy CR 2004 Evolution of the species-rich Cape flora Philos Trans Ser B 359 1623ndash1632

Linder HP Johnson SD Kuhlmann M Matthee CA Nyffeler R amp Swartz ER 2010 Biotic diversity in the South-ern African winter-rainfall region Curr Opin Environm Sustain 2 109ndash116

Maddison WP amp Maddison DR 2007 Mesquite A modular system for evolutionary analysis version 20 httpmesquiteprojectorg (accessed 11 May 2009)

Midgley GF Reeves G amp Klak C 2005 Late Tertiary and Quater-nary climate change and centres of endemism in the southern Af-rican flora Pp 230ndash242 in Purvis A Gittleman JL amp Brooks T (eds) Phylogeny and conservation Cambridge Cambridge University Press

Miller MA Holder MT Vos R Midford PE Liebowitz T Chan L Hoover P amp Warnow T 2009 The CIPRES Portals CIPRES 2009-08-04 httpwwwphyloorgsub_sectionsportal

(accessed 4 Aug 2009) (Archived by WebCite(r) at httpwww webcitationorg5imQlJeQa)

Milton SJ Yeaton RI Dean WRJ amp Vlok JHH 1997 Suc-culent karoo Pp 131ndash166 in Cowling RM Richardson DM amp Pierce SM (eds) Vegetation of Southern Africa Cambridge Cambridge University Press

Moran R amp Smith A 2001 Phytogeographic relationships between Neotropical and African-Madagascan pteridophytes Brittonia 53 304ndash351

Myers N Mittermeier RA Mittermeier CG da Fonseca GAB amp Kent J 2000 Biodiversity hotspots for conservation priorities Nature 403 853ndash858

Nagalingum NS Schneider H amp Pryer KM 2007 Molecular phylogenetic relationships and morphological evolution in the het-erosporous fern genus Marsilea Syst Bot 32 16ndash25

Perrie L amp Brownsey P 2007 Molecular evidence for long-distance dispersal in the New Zealand pteridophyte flora J Biogeogr 34 2028ndash2038

Prado J Rodrigues CD Salatino A amp Salatino MLF 2007 Phylogenetic relationships among Pteridaceae including Brazilian species inferred from rbcL sequences Taxon 56 355ndash368

Pryer KM Schuettpelz E Wolf PG Schneider H Smith AR amp Cranfill R 2004 Phylogeny and evolution of ferns (monilo-phytes) with a focus on the early leptosporangiate divergences Amer J Bot 91 1582ndash1598

Ragan MA 1992 Phylogenetic inference based on matrix representa-tion of trees Molec Phylog Evol 1 53ndash58

Rambaut A amp Drummond A 2007 Tracer version 15 httpbeast bioedacukTracer

Ree RH amp Smith SA 2008 Maximum likelihood inference of geographic range evolution by dispersal local extinction and cladogenesis Syst Biol 57 4ndash14

Renner S 2004 Plant dispersal across the tropical Atlantic by wind and sea currents Int J Pl Sci 165 S23ndashS33

Richardson JE Weitz FM Fay MF Cronk QCB Linder HP Reeves G amp Chase MW 2001 Rapid and recent origin of species richness in the Cape flora of South Africa Nature 412 181ndash183

Ronquist F 1996 DIVA version 11 ftpuuse or ftpsystbotuuseRothfels CJ Windham MD Grusz AL Gastony GJ amp Pryer

KM 2008 Toward a monophyletic Notholaena (Pteridaceae) Resolving patterns of evolutionary convergence in xeric-adapted ferns Taxon 57 712ndash724

Roux JP 2001 Conspectus of southern African Pteridophyta Pre-toria Sabonet

Rymer PD Manning JC Goldblatt P Powell MP amp Savolainen V 2010 Evidence of recent and continuous specia-tion in a biodiversity hotspot A population genetic approach in southern African gladioli (Gladiolus Iridaceae) Molec Ecol 194765ndash4782

Salamin N Chase MW Hodkinson TR amp Savolainen V 2003 Assessing internal support with large phylogenetic DNA matrices Molec Phylog Evol 27 528ndash539

Sauquet H Weston PH Barker NP Anderson CL Cantrill DJ amp Savolainen V 2009 Using fossils and molecular data to reveal the origins of the Cape proteas (subfamily Proteoideae) Molec Phylog Evol 51 31ndash43

Schelpe EACLE 1970 Pteridophyta Pp 1ndash254 in Exell AW amp Launert E (eds) Flora Zambesiaca London The Crown Agents for Overseas Governments and Administrations

Schelpe EACLE 1977 Pteridophyta Pp 1ndash197 in Fernandez RB Launert E amp Mendes EJ (eds) Conspectus Florae Angolensis Lisboa Junta de Investigacoes Cientiacuteficas do Ultramar

Schelpe EACLE amp Anthony NC 1986 Pteridophyta In Leist-ner OA (ed) Flora of Southern Africa Cryptogam volumes Pretoria Department of Agriculture and Water Supply

Schneider H Janssen T Bysrtiakova N Heinrichs H Hen-nequin S amp Rakotondrainibe F 2010a Rapid radiations and

1282


neoendemism in the Madgascan biodiversity hotspot Pp 3ndash16 in Glaubrecht M (ed) Evolution in action Berlin Springer

Schneider H Kreier H-P Janssen T Otto E Muth H amp Hein-richs J 2010b Key innovations versus key opportunities Iden-tifying causes of rapid radiations in derived ferns Pp 61ndash76 in Glaubrecht M (ed) Evolution in action Berlin Springer

Schneider H Ranker TA Russell SJ Cranfill R Geiger JMO Aguraiuja R Wood KR Grundmann M Klober-danz K amp Vogel JC 2005 Origin of the endemic fern genus Diellia coincides with the renewal of Hawaiian terrestrial life in the Miocene Proc Roy Soc London Ser B Biol Sci 72 455ndash460

Schneider H Schuettpelz E Pryer KM Cranfill R Magallon S amp Lupia R 2004 Ferns diversified in the shadow of angio-sperms Nature 428 553ndash557

Schnitzler J Barraclough TG Boatwright JS Goldblatt P Manning JC Powell MP Rebelo T amp Savolainen V 2011 Causes of plant diversification in the Cape biodiversity hotspot of South Africa Syst Biol 60 343ndash357

Schrire BD Lavin M Barker NP amp Forest F 2009 Phylogeny of the tribe Indigofereae (Leguminosae-Papilionoideae) Geographi-cally structured more in succulent-rich and temperate settings than in grass-rich environments Amer J Bot 96 816ndash852

Schuettpelz E amp Pryer KM 2009 Evidence for a Cenozoic radiation of ferns in an angiosperm-dominated canopy Proc Natl Acad Sci USA 106 11200ndash11205

Schuettpelz E Korall P amp Pryer KM 2006 Plastid atpA data provide improved support for deep relationships among ferns Taxon 55 897ndash906

Schuettpelz E Schneider H Huiet L Windham MD amp Pryer KM 2007 A molecular phylogeny of the fern family Pteridaceae Assessing overall relationships and the affinities of previously unsampled genera Molec Phylog Evol 44 1172ndash1185

Scotland RW amp Wortley AH 2003 How many species of seed plants are there Taxon 52 101ndash104

Shmida A amp Whittaker RH 1981 Pattern and biological microsite effects in two shrub communities southern California Ecology 62 234ndash251

Smith AR Pryer KM Schuettpelz E Korall P Schneider H amp Wolf PG 2006 A classification for extant ferns Taxon 55 705ndash731

Sukumaran J amp Holder MT 2008 SumTrees Summarization of split support on phylogenetic trees version 102 Part of the Den-droPy phylogenetic computation library version 213 httppypi pythonorgpypiDendropy

Swofford DL 2003 PAUP Phylogenetic analysis using parsimony (and other methods) version 4 Sunderland Massachusetts Sinauer

Trewick SA Morgan-Richards M Russell SJ Henderson S Rumsey FJ Pinter I Barrett JA Gibby M amp Vogel JC 2002 Polyploidy phylogeography and Pleistocene refugia of the rockfern Asplenium ceterach Evidence from chloroplast DNA Molec Ecol 11 2003ndash2012

Tryon RM amp Tryon AF 1973 Geography spores and evolutionary relations in the cheilanthoid ferns Bot J Linn Soc 67 S145ndashS153

Tryon RM Tryon AF amp Kramer KU 1990 Pteridaceae Pp 230ndash256 in Kubitzki K Kramer KU amp Green PS (eds) The families and genera of vascular plants vol 1 Pteridophytes and gymnosperms New York Springer

Verboom GA Archibald JK Bakker FT Bellstedt DU Conrad F Dreyer LL Forest F Galley C Goldblatt P Henning JF Mummenhoff K Linder HP Muasya AM Oberlander KC Savolainen V Snijman DA Van der Niet T amp Nowell TL 2009b Origin and diversification of the Greater Cape flora Ancient species repository hot-bed of recent radiation or both Molec Phylog Evol 51 44ndash53

Verboom GA Dreyer LL amp Savolainen V 2009a Understanding the origins and evolution of the worldrsquos biodiversity hotspots The biota of the African lsquoCape Floristic Regionrsquo as a case study Molec Phylog Evol 51 1ndash4

Verboom GA Linder HP amp Stock WD 2004 Testing the adaptive nature of radiation Growth form and life history divergence in the African grass genus Ehrharta (Poaceae Ehrhartoideae) Amer J Bot 91 1364ndash1370

Warren BH Bakker FT Bellstedt DU Bytebier B Classen-Bockhoff R Dreyer LL Edwards D Forest F Galley C Hardy CR Linder HP Muasya AM Mummenhoff K Oberlander KC Quint M Richardson JE Savolainen V Schrire BD Van der Niet T Verboom GA Yesson C amp Hawkins JA 2011 Consistent phenological shifts in the making of a biodiversity hotspot The Cape flora BMC Evol Biol 11 39 DOI 1011861471-2148-11-39

Weimarck H 1941 Phytogeographical groups centres and intervals within the Cape flora Acta Univ Lund 37 3ndash143

Wiens JJ amp Moen DS 2008 Missing data and the accuracy of Bayesian phylogenetics J Syst Evol 46 307ndash314

Windham MD Huiet L Schuettpelz E Grusz AL Rothfels CJ Beck J Yatskievych G amp Pryer KM 2009 Using plastid and nuclear DNA sequences to redraw generic boundaries and demystify species complexes in cheilanthoid ferns Amer Fern J 99 128ndash132

Wolf PG Schneider H amp Ranker TA 2001 Geographic distribu-tions of homosporous ferns Does dispersal obscure evidence of vicariance J Biogeogr 28 263ndash270

Zhang GM Zhang XC amp Chen ZD 2003 Phylogeny of crypto-grammoid ferns and related taxa based on rbcL sequences Nordic J Bot 23 485ndash493

Zhang GM Zhang XC Chen ZD Liu HM amp Yang WL 2007 First insights in the phylogeny of Asian cheilanthoid ferns based on sequences of two chloroplast markers Taxon 56 369ndash378

Zwickl DJ 2006 Genetic algorithm approaches for the phyloge-netic analysis of large biological sequence datasets under the maximum likelihood criterion Dissertation The University of Texas Austin

Appendix Taxa and vouchers for species sampled

Species locality voucher number (herbarium) name in molecular dataset GenBank accession numbers for atpA rbcL rps4-trnS trnG-trnR trnL-trnF (resp)

Cheilanthes acrostica (Balb) Tod Spain Andalusia W Eiserhardt AND2-2 (HBG) Cheil_sp_medi_2 GU935460 ndash GU935524 GU935555 GU935581 Cheilanthes bergiana Schltdl South Africa Natal Schelpe 4414 (BM) Ch_bergiana GU935471 ndash GU935530 ndash ndash Cheilanthes capensis (Thunb) Sw South Africa Western Cape Paarl W Eiserhardt WE-062d (HBG) Ch_capensis ndash GU935511 GU935536 ndash ndash Cheilanthes cf kunzei Mett South Africa Richtersveld Vioolsdrif BIOTA 127273 (HBG) Ch_cf_kunz_1 GU935470 GU935508 GU935539 GU935575 GU935613 South Africa Richtersveld Numees BIOTA 127321 (HBG) Ch_cf_kunz_2 ndash ndash ndash ndash GU935614 South Africa Richtersveld Vioolsdrif BIOTA 127502 (HBG) Ch_cf_kunz_3 ndash ndash ndash ndash GU935604 Cheilanthes contracta (Kunze) Mett ex Kuhn South Africa Western Cape Matjiesfontein W Eiserhardt WE-200732 (HBG) Ch_con-trac_1 GU935477 GU935518 ndash ndash GU935592 South Africa Western Cape Paarl W Eiserhardt WE-065 (HBG) Ch_contrac_2 GU935478 GU935519 GU935533 GU935570 GU935593 Cheilanthes deltoidea Kunze South Africa Namaqualand Umdaus BIOTA 127270 (HBG) Ch_deltoid_1 GU935467 GU935512 GU935537 GU935572 GU935605 South Africa Richtersveld Tatasberg BIOTA 127466 (HBG) Ch_deltoid_2 ndash ndash ndash ndash GU935608 South

1283


Africa Namaqualand Vandersterrberg BIOTA 127483 (HBG) Ch_deltoid_3 ndash ndash ndash ndash GU935606 South Africa Namaqualand Umdaus BIOTA 127272 (HBG) Ch_deltoid_4 ndash ndash ndash ndash GU935607 Cheilanthes depauperata Baker South Africa Western Cape Matjiesfontein W Eiserhardt WE-200734 (HBG) Ch_depaupera GU935476 GU935516 GU935532 ndash GU935590 Cheilanthes dinteri Brause Namibia Fyndraai BIOTA sn (HBG) Ch_dinteri GU935461 GU935506 GU935527 GU935558 GU935582 Cheilanthes eckloniana (Kunze) Mett South Africa Transvaal BJ Turner 620 (BM) Ch_ecklonian GU935473 GU935513 GU935540 ndash GU935585 Cheilanthes goyazensis (Taub) Domin Brazil Minas Gerais JC Yesliyurt amp J Prado 554 (BM) Ch_goyazensi ndash ndash ndash ndash JN122018 Cheilanthes guanchica Bolle Spain Andalusia W Eiserhardt AND2-1 (HBG) Ch_sp_medi_1 GU935459 GU935504 GU935525 GU935554 GU935580 Cheilanthes hastata (L f) Kunze South Africa Western Cape Paarl W Eiserhardt WE-063 (HBG) Ch_hastata_1 GU935469 GU935510 GU935538 GU935574 GU935612 South Africa Western Cape Matjiesfontein W Eiserhardt WE-200733 (HBG) Ch_hastata_2 GU935468 GU935509 ndash ndash GU935611 Cheilanthes hirta Sw South Africa Drakensberg Schelpe 3120 (BM) Ch_hirta_1 GU935474 GU935515 GU935531 ndash GU935589 Zimbabwe Great Zimbabwe J Liedtke sn (HBG) Ch_hirta_2 ndash ndash ndash ndash GU935588 Cheilanthes induta Kunze South Africa Western Cape Algeria W Eiserhardt WE-200731 (HBG) Ch_induta GU935465 GU935501 GU935542 GU935563 GU935597 Cheilanthes involuta (Sw) Schelpe amp NC Anthony ndash Mugg sn (BM) P_involuta ndash ndash GU935546 ndash ndash Cheilanthes leachii (Schelpe) Schelpe ndash H J Benson 127 (BM) Ch_leachii GU935457 ndash GU935522 ndash GU935578 Cheilanthes maderensis Lowe Italy Pantelleria J Vogel CHEI-23 (BM) Ch_maderensi ndash GU935505 GU935526 ndash ndash Cheilanthes marlothii (Hieron) Domin Namibia Fyndraai BIOTA sn (HBG) Ch_marloth_1 GU935472 GU935514 GU935541 GU935569 GU935586 Zimbabwe Great Zimbabwe J Liedtke sn Ch_marloth_2 ndash ndash ndash ndash GU935587 Cheilanthes multifida (Sw) Sw South Africa Western Cape Paarl W Eiserhardt WE-064 (HBG) Ch_multifi_1 GU935464 GU935500 GU935544 GU935562 GU935596 South Africa Western Cape Koue Bokkeveld W Eiserhardt WE-200730 (HBG) Ch_multifi_2 GU935463 GU935499 ndash ndash GU935595 Cheilanthes multifida subsp lacerata NC Anthony amp Schelpe Tanzania Raino Lampinen sn (BM) Ch_multifida GU935462 GU935498 GU935543 GU935561 GU935594 Cheilanthes namaquensis (Baker) Schelpe amp NC Anthony South Africa Western Cape Matjiesfontein W Eiserhardt WE-200735 (HBG) Ch_sp_SA006 GU935480 GU935492 GU935528 ndash GU935583 South Africa Western Cape Hoek se Berg W Eiserhardt WE-200737 (HBG) Ch_sp_SA008 GU935481 GU935493 ndash GU935559 ndash South Africa Cape Peninsula Esterhuysen 22946 (BM) Ch_namaquens GU935482 GU935491 GU935529 GU935560 GU935584 Cheilanthes parviloba (Sw) Sw South Africa Western Cape Prince Albert Roux 4163 (NBG) Ch_parvilo_1 GU935475 GU935517 GU935534 GU935571 GU935591 Cheilanthes pentagona Schelpe amp NC Anthony ndash Schelpe 4820 (BM) Ch_pentagona ndash ndash GU935551 ndash ndash Cheilanthes pulchella Bory ex Willd Spain Tenerife s coll sn (HBG) Ch_pulchella ndash GU935503 ndash ndash ndash Cheilanthes quadripinnata (Forssk) Kuhn South Africa Transvaal Schelpe 5926 (BM) Ch_quadripin GU935484 GU935496 GU935550 GU935566 ndash Cheilanthes rawsonii (Pappe) Mett ex Kuhn South Africa Richtersveld BIOTA 127477 (HBG) Ch_rawsonii GU935489 GU935520 GU935552 GU935577 GU935615 Cheilanthes robusta (Kunze) RM Tryon South Africa Namaqualand Vandersterrberg BIOTA 127478 (HBG) Ch_robusta_1 GU935466 GU935507 GU935535 GU935573 GU935609 South Africa Namaqualand Paulshoek BIOTA sn (HBG) Ch_robusta_2 ndash ndash ndash ndash GU935610 Cheilanthes venusta Feacutee Brazil Minas Gerais JC Yesliyurt amp J Prado 549 (BM) Ch_venusta ndash JN122014 ndash ndash JN122019 Cheilanthes viridis (Forssk) Sw South Africa Natal Scottburgh Schelpe 2525 (BM) Ch_viridis_2 GU935485 GU935494 ndash GU935567 GU935601 ndash Curle amp Schelpe 56 (BM) Ch_viridis_3 GU935486 ndash GU935547 ndash ndash Zimbabwe Great Zimbabwe J Liedtke sn (HBG) Ch_cf_involu ndash ndash ndash ndash GU935603 Cheilanthes viridis var glauca (Sim) Schelpe amp NC Anthony South Africa Natal Schelpe 2959 (BM) Ch_virid_g_1 GU935487 GU935495 GU935548 GU935568 GU935602 Doryopteris lomariacea (Kunze) Kl Brazil Satildeo Paulo JC Yesliyurt amp J Prado 547 (BM) D_lomariacea ndash ndash ndash ndash JN122020 Doryopteris lorentzii (Hieron) Diels Brazil Rio Grande do Sul JC Yesliyurt amp RN Cislinski 527 (BM) D_lorentzii ndash JN122015 ndash ndash ndash Doryopteris pedata (L) Feacutee Dominican Republic HA Allard 14835 (US) D_pedata ndash ndash ndash ndash JN122021 Doryopteris pedatoides (Desv) Kuhn amp Decken Madagascar G amp U Benl 6 (US) D_pedatoides ndash JN122016 ndash ndash JN122022 Doryopteris pilosa (Poir) Kuhn Mauritius M Gibby sn RBGE D_pilosa ndash JN122017 ndash ndash JN122023 Doryopteris sagittifolia (Raddi) J Sm Brazil Paranaacute JC Yesliyurt amp J Prado 510 (BM) D_sagittifol ndash ndash ndash ndash JN122024 Pellaea calomelanos (Sw) Link Namibia Fyndraai BIOTA sn (HBG) P_calomela_1 GU935483 GU935497 GU935549 GU935565 GU935600 Pellaea dura (Willd) Hook Zambesi Escarpment DS Mitchell 559 (BM) P_dura_1 GU935458 GU935490 GU935523 GU935556 GU935579 Pellaea pectiniformis Baker South Africa Transvaal Esterhuysen 21468 (BM) P_pectinifor ndash ndash ndash GU935557 ndash Pellaea pinnata (Kaulf) Prantl Brazil Rio de Janeiro JC Yesliyurt amp J Prado 550 (BM) P_pinnata_1 ndash ndash ndash ndash JN122025 Pellaea pteroides (L) Prantl South Africa Western Cape Paarl W Eiserhardt WE-062a (HBG) P_pteroide_1 GU935479 GU935502 GU935545 GU935564 GU935599 South Africa Cape Peninsula Salter 9632 (BM) P_pteroide_2 ndash ndash ndash ndash GU935598 Pellaea rufa ARTryon South Africa Western Cape Laingsburg Roux 4220 (NBG) P_rufa GU935488 GU935521 GU935553 GU935576 GU935616

Appendix Continued

1270


Richardson amp al 2001) is increasingly challenged by molecular age estimation studies (Verboom amp al 2009b Schnitzler amp al 2011) Many cape clades are too old for this explanation some dating back to the early Cenozoic (Verboom amp al 2009b) There is evidence that it applies to radiations in the SK and lowlands of the CFR (eg Klak amp al 2004) whereas montane lineages are older indicating that the Cape Fold Mountains have served as a refuge for Fynbos vegetation and associated taxa (Verboom amp al 2009b Schnitzler amp al 2011)

Several factors have likely acted in concert to cause the exceptional diversity of the GCFR The origin of this biodiver-sity hotspot may thus be best addressed by reconstructing the history of a diverse range of lineages with substantial differ-ences in their biology Many angiosperm Cape Clades are well studied whereas comparable studies are lacking for species-rich lineages of non-angiosperms such as ferns liverworts and mosses Understanding the origin of these lineages will help elucidate how the mechanisms underlying the diversification of the Cape Clades depend on particular life history aspects such as dispersal ability and reproductive biology

The Cape region also harbours a high diversity of ferns (Burrows 1990 Jacobsen 1983 Roux 2001 Schelpe amp Anthony 1986) but relatively little attention has been given to the diversification of ferns in this or other biodiversity hot-spots Studies on fern diversification found remarkable simi-larities to angiosperm diversification eg the colonization and radiation of the Diellia clade (Aspleniaceae) in Hawailsquoi (Schneider amp al 2005) the rapid radiation of tree ferns in the Madagascan biodiversity hotspot (Janssen amp al 2008 Sch-neider amp al 2010a) and the diversification of derived ferns in New Zealand (Perrie amp Brownsey 2007) In these examples the

high dispersal capacity of some ferns plays a role but patterns of local diversification were also found (Wolf amp al 2001 Schnei-der amp al 2005 Janssen amp al 2007 2008 Perrie amp Brownsey 2007) The majority of extant fern species belong to lineages that diversified in the shadow of angiosperms (Schneider amp al 2004 Schuettpelz amp Pryer 2009) and spatial diversity patterns are explained by the same environmental variables in ferns and angiosperms although humidity has a stronger impact on ferns (Kreft amp al 2010) However the two groups differ strongly in size with only about 9000 species of ferns (Smith amp al 2006) but at least 220000 angiosperm species (Scotland amp Wortley 2003) The importance of local radiation events needs to be seen in relation to the total diversity of the group for example a local radiation resulting in 10 fern species contributes about 01 to the global fern diversity and corresponds roughly to an angiosperm radiation generating about 220 new species The lower number of fern species coincides with the lower number of detected local fern radiations (Schneider amp al 2010b)

Cheilanthoid ferns (Pteridaceae subfam Cheilanthoideae) form a group of 278 or more species that generally have a prefer-ence for xeric and semi-xeric seasonal climates (Tryon amp al 1990 Schuettpelz amp al 2007) This is a less common adaption since ferns are most diverse in regions with abundant rainfall and equable temperatures (Barrington 1993) Cheilanthoids are an important element of the southern African fern flora (Fig 1) and show a higher degree of endemism than other ferns within the GCFR including many narrow-ranged endemics (Jacobsen 1983 Anthony 1984 Schelpe amp Anthony 1986 Burrows 1990) Based on this Anthony (1984) hypothesised that the south-western tip of Africa has been a centre of chei-lanthoid speciation resulting in the high number of endemics in the GCFR similar to the high endemism levels of several angiosperm groups in the Cape flora In situ diversification as such does not appear to be unusual as globally six major centres of cheilanthoid diversity have been described (Tryon amp Tryon 1973) one of which is South and East Africa together with Madagascar (henceforth Afro-Madagascar) These centres were proposed to be ldquomajor centres of autochthonous evolutionrdquo (Tryon amp Tryon 1973) which has since been corroborated with phylogenetic evidence (Gastony amp Rollo 1995 1998 Kirkpat-rick 2007 Prado amp al 2007 Rothfels amp al 2008 Windham amp al 2009) These studies support the scenario of a limited impact of long-distance dispersal on the diversification history of cheilanthoids Evidence for frequent long-distance dispersal was found in other fern groups such as the Afro-Madagascan polygrammoid ferns (Janssen amp al 2007) but not in the Mala-gasy scaly tree ferns (Janssen amp al 2008) and African mar-sileaceous ferns (Nagalingum amp al 2007) Previous studies on cheilanthoids included too few Afro-Madagascan species to test the phylogenetic coherence of this particular region or explore the relationships of the GCFR-endemics Here we aim to explore evidence for one or several in situ radiations of these xeric ferns in the GCFR We build a phylogeny from five plas-tid DNA loci that have previously been used with success to reconstruct relationships among cheilanthoid ferns (Gastony amp Rollo 1995 1998 Kirkpartrick 2007 Rothfels amp al 2008) We employ ancestral range reconstruction methods and explore

Fig 1 Distribution and species richness of cheilanthoid ferns in south-ern Africa based on distribution data provided in Burrows (1990) Cir-cle size scales with the number of taxa found in a 05 times 05deg grid cell centred on the circle midpoint Maximum circle size equals 16 spe-cies Circles with solid fill represent roughly the extent of the Greater Cape Floristic Region

0 1000km

1271












1272








results




1273









1274





002

Pellaea brachyptera


Pellaea breweri

Cheilanthes aurea




Notholaena rosei


Notholaena candida






Notholaena aliena




Pellaea ternifolia

Pellaea rufa


Pellaea truncata

Pellaea notabilis





Notholaena bryopoda

Pellaea cordifolia


Notholaena grayi

Pellaea ovata


Bommeria hispida

Pellaea pringlei



Pellaea wrightiana


Cheilanthes lanosa





Pellaea mucronata



Notholaena lemmonii


Astrolepis sinuata

Pellaea sagittata

Notholaena greggii

Notholaena neglecta


Notholaena affinis





Pellaea glabella








Argyrochosma nivea





Notholaena rigida

Pellaea bridgesii

Pellaea intermedia

Argyrochosma incana

B

P

S

N

M

Fig 2B

1275



Doryopteris pilosa





Cheilanthes decora


Pellaea patula

Pellaea dura

Pellaea doniana





Sinopteris albfusca




Pellaea calomelanos

Doryopteris pedata






Cheilanthes kunzei

Doryopteris collina

Adiantopsis radiata



Hemionitis rufa



Pellaea paupercula

Cheilanthes distans

Doryopteris nobilis






Pellaea pteroides


Cheilanthes dinteri



Cheilanthes robusta








Pellaea maxima

Cheilanthes induta




Cheilanthes leachii

Pellaea nitidula

Pellaea longipilosa

Cheilanthes hirta



Cheilanthes kuhnii


Cheilanthes viridis


Pellaea nitidula


Pellaea boivinii


Hemionitis palmata




Cheilanthes hastata



Aspidotis densa

Ormopteris riedelii

Pellaea cf dura


Cheilanthes venusta




Hemionitis levyi





D

C

A

1

1b

1a

2

2a

3

3a

3b


002

1c

HFig 2A

1276





ee

00100200300

[1147]

[881762]

[78171]

[175265]

[119194]

[109204]

[41123]

[209296]

[126200]

[187278][99204]

[73147]

[48115]

[44132]

[84187]

[95168][86159]

[130270]

[114207]

[85156]

[1866]

[157247]

[208294]

[64163]

[155234]

[130214]

[172254]

[37104]

[67134]

[154244]

[841905]

[55143]

[48114]

[179265]

[2266]

[49143]


[215305]

[233332]

[213302]

[254363]

baab

cabce

deff

f

bf

bb

bb

fef

efe

efe

efe

ff

fb

aa

babc

abca

cabc

Asian clade (A)

aa


cefe


1

3

2

1277










dIscussIon




100200

Ch parviloba

Ch contracta

Ch depauperata

Ch hirta

Ch eckloniana

Ch marlothii

Ch dinteri

Ch deltoidea

Ch capensis

Ch hastata

Ch kunzei

Ch robusta

CFRNE

CFR

CFR

CFRNE

CFRNE

N

N

SK

SKCFR

SKCFR

SK

SK

5466

28

69

88

84

9399

93

00

87 55

CFRCFRN

NSK

N

SK

(84)

1278







1279






1280




acknowledgeMents
























lIterature cIted

1281

















































1282

































1283



Appendix Continued

1271












1272








results




1273









1274





002

Pellaea brachyptera


Pellaea breweri

Cheilanthes aurea




Notholaena rosei


Notholaena candida






Notholaena aliena




Pellaea ternifolia

Pellaea rufa


Pellaea truncata

Pellaea notabilis





Notholaena bryopoda

Pellaea cordifolia


Notholaena grayi

Pellaea ovata


Bommeria hispida

Pellaea pringlei



Pellaea wrightiana


Cheilanthes lanosa





Pellaea mucronata



Notholaena lemmonii


Astrolepis sinuata

Pellaea sagittata

Notholaena greggii

Notholaena neglecta


Notholaena affinis





Pellaea glabella








Argyrochosma nivea





Notholaena rigida

Pellaea bridgesii

Pellaea intermedia

Argyrochosma incana

B

P

S

N

M

Fig 2B

1275



Doryopteris pilosa





Cheilanthes decora


Pellaea patula

Pellaea dura

Pellaea doniana





Sinopteris albfusca




Pellaea calomelanos

Doryopteris pedata






Cheilanthes kunzei

Doryopteris collina

Adiantopsis radiata



Hemionitis rufa



Pellaea paupercula

Cheilanthes distans

Doryopteris nobilis






Pellaea pteroides


Cheilanthes dinteri



Cheilanthes robusta








Pellaea maxima

Cheilanthes induta




Cheilanthes leachii

Pellaea nitidula

Pellaea longipilosa

Cheilanthes hirta



Cheilanthes kuhnii


Cheilanthes viridis


Pellaea nitidula


Pellaea boivinii


Hemionitis palmata




Cheilanthes hastata



Aspidotis densa

Ormopteris riedelii

Pellaea cf dura


Cheilanthes venusta




Hemionitis levyi





D

C

A

1

1b

1a

2

2a

3

3a

3b


002

1c

HFig 2A

1276





ee

00100200300

[1147]

[881762]

[78171]

[175265]

[119194]

[109204]

[41123]

[209296]

[126200]

[187278][99204]

[73147]

[48115]

[44132]

[84187]

[95168][86159]

[130270]

[114207]

[85156]

[1866]

[157247]

[208294]

[64163]

[155234]

[130214]

[172254]

[37104]

[67134]

[154244]

[841905]

[55143]

[48114]

[179265]

[2266]

[49143]


[215305]

[233332]

[213302]

[254363]

baab

cabce

deff

f

bf

bb

bb

fef

efe

efe

efe

ff

fb

aa

babc

abca

cabc

Asian clade (A)

aa


cefe


1

3

2

1277










dIscussIon




100200

Ch parviloba

Ch contracta

Ch depauperata

Ch hirta

Ch eckloniana

Ch marlothii

Ch dinteri

Ch deltoidea

Ch capensis

Ch hastata

Ch kunzei

Ch robusta

CFRNE

CFR

CFR

CFRNE

CFRNE

N

N

SK

SKCFR

SKCFR

SK

SK

5466

28

69

88

84

9399

93

00

87 55

CFRCFRN

NSK

N

SK

(84)

1278







1279






1280




acknowledgeMents
























lIterature cIted

1281

















































1282

































1283



Appendix Continued

1272








results




1273









1274





002

Pellaea brachyptera


Pellaea breweri

Cheilanthes aurea




Notholaena rosei


Notholaena candida






Notholaena aliena




Pellaea ternifolia

Pellaea rufa


Pellaea truncata

Pellaea notabilis





Notholaena bryopoda

Pellaea cordifolia


Notholaena grayi

Pellaea ovata


Bommeria hispida

Pellaea pringlei



Pellaea wrightiana


Cheilanthes lanosa





Pellaea mucronata



Notholaena lemmonii


Astrolepis sinuata

Pellaea sagittata

Notholaena greggii

Notholaena neglecta


Notholaena affinis





Pellaea glabella








Argyrochosma nivea





Notholaena rigida

Pellaea bridgesii

Pellaea intermedia

Argyrochosma incana

B

P

S

N

M

Fig 2B

1275



Doryopteris pilosa





Cheilanthes decora


Pellaea patula

Pellaea dura

Pellaea doniana





Sinopteris albfusca




Pellaea calomelanos

Doryopteris pedata






Cheilanthes kunzei

Doryopteris collina

Adiantopsis radiata



Hemionitis rufa



Pellaea paupercula

Cheilanthes distans

Doryopteris nobilis






Pellaea pteroides


Cheilanthes dinteri



Cheilanthes robusta








Pellaea maxima

Cheilanthes induta




Cheilanthes leachii

Pellaea nitidula

Pellaea longipilosa

Cheilanthes hirta



Cheilanthes kuhnii


Cheilanthes viridis


Pellaea nitidula


Pellaea boivinii


Hemionitis palmata




Cheilanthes hastata



Aspidotis densa

Ormopteris riedelii

Pellaea cf dura


Cheilanthes venusta




Hemionitis levyi





D

C

A

1

1b

1a

2

2a

3

3a

3b


002

1c

HFig 2A

1276





ee

00100200300

[1147]

[881762]

[78171]

[175265]

[119194]

[109204]

[41123]

[209296]

[126200]

[187278][99204]

[73147]

[48115]

[44132]

[84187]

[95168][86159]

[130270]

[114207]

[85156]

[1866]

[157247]

[208294]

[64163]

[155234]

[130214]

[172254]

[37104]

[67134]

[154244]

[841905]

[55143]

[48114]

[179265]

[2266]

[49143]


[215305]

[233332]

[213302]

[254363]

baab

cabce

deff

f

bf

bb

bb

fef

efe

efe

efe

ff

fb

aa

babc

abca

cabc

Asian clade (A)

aa


cefe


1

3

2

1277










dIscussIon




100200

Ch parviloba

Ch contracta

Ch depauperata

Ch hirta

Ch eckloniana

Ch marlothii

Ch dinteri

Ch deltoidea

Ch capensis

Ch hastata

Ch kunzei

Ch robusta

CFRNE

CFR

CFR

CFRNE

CFRNE

N

N

SK

SKCFR

SKCFR

SK

SK

5466

28

69

88

84

9399

93

00

87 55

CFRCFRN

NSK

N

SK

(84)

1278







1279






1280




acknowledgeMents
























lIterature cIted

1281

















































1282

































1283



Appendix Continued

1273









1274





002

Pellaea brachyptera


Pellaea breweri

Cheilanthes aurea




Notholaena rosei


Notholaena candida






Notholaena aliena




Pellaea ternifolia

Pellaea rufa


Pellaea truncata

Pellaea notabilis





Notholaena bryopoda

Pellaea cordifolia


Notholaena grayi

Pellaea ovata


Bommeria hispida

Pellaea pringlei



Pellaea wrightiana


Cheilanthes lanosa





Pellaea mucronata



Notholaena lemmonii


Astrolepis sinuata

Pellaea sagittata

Notholaena greggii

Notholaena neglecta


Notholaena affinis





Pellaea glabella








Argyrochosma nivea





Notholaena rigida

Pellaea bridgesii

Pellaea intermedia

Argyrochosma incana

B

P

S

N

M

Fig 2B

1275



Doryopteris pilosa





Cheilanthes decora


Pellaea patula

Pellaea dura

Pellaea doniana





Sinopteris albfusca




Pellaea calomelanos

Doryopteris pedata






Cheilanthes kunzei

Doryopteris collina

Adiantopsis radiata



Hemionitis rufa



Pellaea paupercula

Cheilanthes distans

Doryopteris nobilis






Pellaea pteroides


Cheilanthes dinteri



Cheilanthes robusta








Pellaea maxima

Cheilanthes induta




Cheilanthes leachii

Pellaea nitidula

Pellaea longipilosa

Cheilanthes hirta



Cheilanthes kuhnii


Cheilanthes viridis


Pellaea nitidula


Pellaea boivinii


Hemionitis palmata




Cheilanthes hastata



Aspidotis densa

Ormopteris riedelii

Pellaea cf dura


Cheilanthes venusta




Hemionitis levyi





D

C

A

1

1b

1a

2

2a

3

3a

3b


002

1c

HFig 2A

1276





ee

00100200300

[1147]

[881762]

[78171]

[175265]

[119194]

[109204]

[41123]

[209296]

[126200]

[187278][99204]

[73147]

[48115]

[44132]

[84187]

[95168][86159]

[130270]

[114207]

[85156]

[1866]

[157247]

[208294]

[64163]

[155234]

[130214]

[172254]

[37104]

[67134]

[154244]

[841905]

[55143]

[48114]

[179265]

[2266]

[49143]


[215305]

[233332]

[213302]

[254363]

baab

cabce

deff

f

bf

bb

bb

fef

efe

efe

efe

ff

fb

aa

babc

abca

cabc

Asian clade (A)

aa


cefe


1

3

2

1277










dIscussIon




100200

Ch parviloba

Ch contracta

Ch depauperata

Ch hirta

Ch eckloniana

Ch marlothii

Ch dinteri

Ch deltoidea

Ch capensis

Ch hastata

Ch kunzei

Ch robusta

CFRNE

CFR

CFR

CFRNE

CFRNE

N

N

SK

SKCFR

SKCFR

SK

SK

5466

28

69

88

84

9399

93

00

87 55

CFRCFRN

NSK

N

SK

(84)

1278







1279






1280




acknowledgeMents
























lIterature cIted

1281

















































1282

































1283



Appendix Continued

1274





002

Pellaea brachyptera


Pellaea breweri

Cheilanthes aurea




Notholaena rosei


Notholaena candida






Notholaena aliena




Pellaea ternifolia

Pellaea rufa


Pellaea truncata

Pellaea notabilis





Notholaena bryopoda

Pellaea cordifolia


Notholaena grayi

Pellaea ovata


Bommeria hispida

Pellaea pringlei



Pellaea wrightiana


Cheilanthes lanosa





Pellaea mucronata



Notholaena lemmonii


Astrolepis sinuata

Pellaea sagittata

Notholaena greggii

Notholaena neglecta


Notholaena affinis





Pellaea glabella








Argyrochosma nivea





Notholaena rigida

Pellaea bridgesii

Pellaea intermedia

Argyrochosma incana

B

P

S

N

M

Fig 2B

1275



Doryopteris pilosa





Cheilanthes decora


Pellaea patula

Pellaea dura

Pellaea doniana





Sinopteris albfusca




Pellaea calomelanos

Doryopteris pedata






Cheilanthes kunzei

Doryopteris collina

Adiantopsis radiata



Hemionitis rufa



Pellaea paupercula

Cheilanthes distans

Doryopteris nobilis






Pellaea pteroides


Cheilanthes dinteri



Cheilanthes robusta








Pellaea maxima

Cheilanthes induta




Cheilanthes leachii

Pellaea nitidula

Pellaea longipilosa

Cheilanthes hirta



Cheilanthes kuhnii


Cheilanthes viridis


Pellaea nitidula


Pellaea boivinii


Hemionitis palmata




Cheilanthes hastata



Aspidotis densa

Ormopteris riedelii

Pellaea cf dura


Cheilanthes venusta




Hemionitis levyi





D

C

A

1

1b

1a

2

2a

3

3a

3b


002

1c

HFig 2A

1276





ee

00100200300

[1147]

[881762]

[78171]

[175265]

[119194]

[109204]

[41123]

[209296]

[126200]

[187278][99204]

[73147]

[48115]

[44132]

[84187]

[95168][86159]

[130270]

[114207]

[85156]

[1866]

[157247]

[208294]

[64163]

[155234]

[130214]

[172254]

[37104]

[67134]

[154244]

[841905]

[55143]

[48114]

[179265]

[2266]

[49143]


[215305]

[233332]

[213302]

[254363]

baab

cabce

deff

f

bf

bb

bb

fef

efe

efe

efe

ff

fb

aa

babc

abca

cabc

Asian clade (A)

aa


cefe


1

3

2

1277










dIscussIon




100200

Ch parviloba

Ch contracta

Ch depauperata

Ch hirta

Ch eckloniana

Ch marlothii

Ch dinteri

Ch deltoidea

Ch capensis

Ch hastata

Ch kunzei

Ch robusta

CFRNE

CFR

CFR

CFRNE

CFRNE

N

N

SK

SKCFR

SKCFR

SK

SK

5466

28

69

88

84

9399

93

00

87 55

CFRCFRN

NSK

N

SK

(84)

1278







1279






1280




acknowledgeMents
























lIterature cIted

1281

















































1282

































1283



Appendix Continued

1275



Doryopteris pilosa





Cheilanthes decora


Pellaea patula

Pellaea dura

Pellaea doniana





Sinopteris albfusca




Pellaea calomelanos

Doryopteris pedata






Cheilanthes kunzei

Doryopteris collina

Adiantopsis radiata



Hemionitis rufa



Pellaea paupercula

Cheilanthes distans

Doryopteris nobilis






Pellaea pteroides


Cheilanthes dinteri



Cheilanthes robusta








Pellaea maxima

Cheilanthes induta




Cheilanthes leachii

Pellaea nitidula

Pellaea longipilosa

Cheilanthes hirta



Cheilanthes kuhnii


Cheilanthes viridis


Pellaea nitidula


Pellaea boivinii


Hemionitis palmata




Cheilanthes hastata



Aspidotis densa

Ormopteris riedelii

Pellaea cf dura


Cheilanthes venusta




Hemionitis levyi





D

C

A

1

1b

1a

2

2a

3

3a

3b


002

1c

HFig 2A

1276





ee

00100200300

[1147]

[881762]

[78171]

[175265]

[119194]

[109204]

[41123]

[209296]

[126200]

[187278][99204]

[73147]

[48115]

[44132]

[84187]

[95168][86159]

[130270]

[114207]

[85156]

[1866]

[157247]

[208294]

[64163]

[155234]

[130214]

[172254]

[37104]

[67134]

[154244]

[841905]

[55143]

[48114]

[179265]

[2266]

[49143]


[215305]

[233332]

[213302]

[254363]

baab

cabce

deff

f

bf

bb

bb

fef

efe

efe

efe

ff

fb

aa

babc

abca

cabc

Asian clade (A)

aa


cefe


1

3

2

1277










dIscussIon




100200

Ch parviloba

Ch contracta

Ch depauperata

Ch hirta

Ch eckloniana

Ch marlothii

Ch dinteri

Ch deltoidea

Ch capensis

Ch hastata

Ch kunzei

Ch robusta

CFRNE

CFR

CFR

CFRNE

CFRNE

N

N

SK

SKCFR

SKCFR

SK

SK

5466

28

69

88

84

9399

93

00

87 55

CFRCFRN

NSK

N

SK

(84)

1278







1279






1280




acknowledgeMents
























lIterature cIted

1281

















































1282

































1283



Appendix Continued

1276





ee

00100200300

[1147]

[881762]

[78171]

[175265]

[119194]

[109204]

[41123]

[209296]

[126200]

[187278][99204]

[73147]

[48115]

[44132]

[84187]

[95168][86159]

[130270]

[114207]

[85156]

[1866]

[157247]

[208294]

[64163]

[155234]

[130214]

[172254]

[37104]

[67134]

[154244]

[841905]

[55143]

[48114]

[179265]

[2266]

[49143]


[215305]

[233332]

[213302]

[254363]

baab

cabce

deff

f

bf

bb

bb

fef

efe

efe

efe

ff

fb

aa

babc

abca

cabc

Asian clade (A)

aa


cefe


1

3

2

1277










dIscussIon




100200

Ch parviloba

Ch contracta

Ch depauperata

Ch hirta

Ch eckloniana

Ch marlothii

Ch dinteri

Ch deltoidea

Ch capensis

Ch hastata

Ch kunzei

Ch robusta

CFRNE

CFR

CFR

CFRNE

CFRNE

N

N

SK

SKCFR

SKCFR

SK

SK

5466

28

69

88

84

9399

93

00

87 55

CFRCFRN

NSK

N

SK

(84)

1278







1279






1280




acknowledgeMents
























lIterature cIted

1281

















































1282

































1283



Appendix Continued

1277










dIscussIon




100200

Ch parviloba

Ch contracta

Ch depauperata

Ch hirta

Ch eckloniana

Ch marlothii

Ch dinteri

Ch deltoidea

Ch capensis

Ch hastata

Ch kunzei

Ch robusta

CFRNE

CFR

CFR

CFRNE

CFRNE

N

N

SK

SKCFR

SKCFR

SK

SK

5466

28

69

88

84

9399

93

00

87 55

CFRCFRN

NSK

N

SK

(84)

1278







1279






1280




acknowledgeMents
























lIterature cIted

1281

















































1282

































1283



Appendix Continued

1278







1279






1280




acknowledgeMents
























lIterature cIted

1281

















































1282

































1283



Appendix Continued

1279






1280




acknowledgeMents
























lIterature cIted

1281

















































1282

































1283



Appendix Continued

1280




acknowledgeMents
























lIterature cIted

1281

















































1282

































1283



Appendix Continued

1281

















































1282

































1283



Appendix Continued

1282

































1283



Appendix Continued

1283



Appendix Continued

evidence for radiations of cheilanthoid ferns in the greater cape floristic region

Documents