TAXON 61 (2) • April 2012: 437–455 Funk & al. • Phylogeny and biogeography of

Phylogeny and biogeography of the Liabeae (Compositae subfamily )

Vicki A. Funk, Carol Kelloff & Raymund Chan

U.S. National Herbarium, Department of Botany, Smithsonian Institution MRC 166, P.O. Box 37012, Washington D.C. 20013, U.S.A. Author for correspondence: Vicki A. Funk, [email protected]

Abstract The tribe Liabeae (Compositae) contains ca. 175 distributed in 18 genera and its members occupy a variety of habitats in the Andes of South America as well as in Mexico, , and the West Indies. The tribe is recognizable by a combination of morphological characters. DNA sequence data from the nuclear ribosomal ITS region and three chloroplast regions (trnL-F, 3′ end of ndhF, matK; a total of more than 5 kb of sequence data) were used to infer a phylogeny. The data were analyzed using maximum parsimony, maximum likelihood, and Bayesian inference. The results support the of the tribe and show a consistent placement for all genera except . Four well-supported clades are recovered in the remainder of the tribe, all recognized as subtribes. Liabineae (Ferreyranthus, Dillandia, , , ) are the sister group of the rest of the tribe. Sinclairineae (Sinclairiopsis and with segregates Liabellum and Megaliabum) are the sister group of Munnoziinae (Chrysactinium nested inside s.l.) plus Paranepheliinae (Stephenbeckia, ­ , Pseudonoseris, , Chionopappus, , ). Cacosmia is placed as the sister group of either all the rest of the tribe or of subtribe Liabineae; morphologically its characters are either autapomorphic or plesiomorphic. Bishopanthus could not be confidently placed in any of the subtribes; it is only known from scraps of the type and a molecular study is not possible. The phylogeny slightly alters previous assumptions about the biogeography and it seems that Liabeae originated in the central and northern Andes and spread north and south with several independent introductions into Mexico and Central America and one into the Caribbean. With the exception of the Liabeae (Andes) and Moquineae (Brazil), all of the tribes in the subfamily Cichorioideae are either restricted to or have their basal grade in Africa.

Keywords ; Caribbean; molecular systematics; North America; South America

Supplementary Material Figure S1 is available in the Electronic Supplement to the online version of this article (http://www .ingentaconnect.com/content/iapt/tax).

Introduction Liabeae were subject of a recent review (Dillon & al., 2009) that included an overview of its taxonomic history which need Liabeae are concentrated in the northern and central Andes not be repeated here. However, it is useful to note that all re- although they extend north to central Mexico and the Carib- cent phylogenetic studies have supported the monophyly of bean, and as far south as the mountains of northern . the tribe and its placement in subfamily Cichorioideae near The tribe contains ca. 175 species arranged in 18 genera and and Arctotidieae and more distantly as such it represents one of the smaller tribes in the . Its (Funk & Chan, 2009). The paper by Dillon & al. (2009) also component taxa have been variously placed in several tribes, provided a detailed discussion on the structure of the pollen most frequently Vernonieae or . Despite work by and chromosome numbers in relation to the preliminary Rydberg (1927), who established the tribe, and subsequent work that they had at the time. Although the present study alters the by Blake (1935), Cabrera (1954), and Sandwith (1956), the tribe tree, it does not affect the conclusions by Dillon & al. (2009) Liabeae was not adopted until Robinson and co-workers pub- that Liabeae completely lack lophate pollen and that the most lished a series of papers bringing the genera together (Robin- likely base chromosome number is x = 9. son & Brettell, 1973, 1974; see Robinson, 1983a for additional The majority of taxa in Liabeae often are confined to a references). small geographic area. Members of the tribe occupy sites in Table 1 documents the center of generic diversity in the forest communities (50–4750 m) or high-elevation areas such Liabeae; it lies in Peru where 13 of the 18 genera are found, fol- as subpáramo, páramo, jalca, and puna (> 3000 m). More rarely lowed by (9 genera), Colombia (8), Bolivia (7), Costa they are found in seasonally dry scrub, desert habitats, or dis- Rica and Panama (5 each), Venezuela and Argentina (4), Mex- turbed areas. ico (3), , El Salvador, Honduras, and (2), Since the Dillon & al. (2009) paper, a new has been and the Caribbean (1). Recently two collections were identified added, Stephanbeckia (Robinson & Funk, 2011), based on a from Acre (Brazil) that extend the range of the tribe into that recent collection from Bolivia by Stephan Beck for whom the country: Liabum acuminatum Rusby (Prance & al. 7310, US) genus is named. It seems that every treatment of the tribe is and L. amplexicaule Poepp. & Endl. (Daly & al. 9631, US). fated to be followed by a new genus (Robinson, 1983a was

437 Funk & al. • Phylogeny and biogeography of Liabeae TAXON 61 (2) • April 2012: 437–455

followed by Bishopanthus, Robinson, 1983b; Funk & al., 1996 Materials and Methods by Dillandia, Funk & Robinson, 2001; Funk & al., 2007b by Sampera, Funk & Robinson, 2009; and Dillon & al., 2009 by DNA amplification and sequencing. — DNA extractions Stephanbeckia, Robinson & Funk, 2011). This may happen were performed using DNeasy Mini Kits (Qiagen, Ger- again as the new monotypic genus Inkaliabum D.G. Gut. has mantown, Maryland, U.S.A.) following the manufacturer’s recently been published (Gutiérrez, 2010) but any decision on its instructions, but with an extended incubation period (up to acceptance has been deferred until specimens can be examined. 40 minutes) for herbarium material. For some samples, besides The purpose of this study is to produce a molecular phy- the usual preference for young leaflets, removing debris such as logeny of the Liabeae and to investigate how such a phylogeny epidermal hairs from the plant tissue under a dissecting scope affects our ideas about the origin and evolution of this tribe. proved essential to avoid amplifying contaminating fungal A preliminary version of this tree was used in Dillon & al. DNA. The extracted DNA was further purified using the Ultra (2009) by permission of the authors of this study. However, Clean 15 DNA purification kit (MO BIO Laboratories, Carlsbad, since that paper was published 17 additional taxa have been California, U.S.A.) when the initial PCR amplification was un- added to the ingroup as well as 2.8 kb of matK sequence data successful. The Ultra Clean protocol for large DNA fragments for each taxon. was followed to avoid shearing the extracted genomic DNA.

Table 1. Genera of Liabeae (with abbreviations in parentheses) and their distributions.

Latitude Genus Abbr. Species Distribution range

Bishopanthus H. Rob. 1 Peru, type collection only 3°S–7°S

Cacosmia Kunth Cac 3 Ecuador, Peru 0.0°–10.5°S

Chionopappus Benth. Chi 1 Peru 7°S–13°S

Chrysactinium Wedd. Cry 8 Ecuador, Peru 0.5°N–11°S

Dillandia V.A. Funk & H. Rob. Dil 3 Colombia, Peru 1.37°N–5.7°S

Erato DC. Era 5 Costa Rica, Panama, Venezuela, Colombia, Ecuador, Peru, Bolivia 11°N–18.5°S

Ferreyranthus H. Rob. & Brettell Fer 8 Ecuador, Peru 1°S–15°S

Liabum Adans. Lia 37 Mexico, Caribbean, Guatemala, El Salvador, Honduras, Nicaragua, 21.5°N–23°S Costa Rica, Panama, Venezuela, Colombia, Ecuador, Peru, Bolivia, Brazil, Argentina

Microliabum Cabrera Mic 5 Bolivia, Argentina 19°S–33°S

Munnozia Ruiz & Pav. Mun 46 Costa Rica, Panama, Venezuela, Colombia, Ecuador, Peru, Bolivia, 11°N–23.5°S Argentina

Oligactis Cass. Oli 7 Ecuador, Colombia, Venezuela, Costa Rica, Panama 10°N–4°S

Paranephelius Poepp. Par 7 Peru, Bolivia, Argentina 3°S–22.5°S

Philoglossa DC. Phi 5 Colombia, Ecuador, Peru, Bolivia 0.5°N–17°S

Pseudonoseris H. Rob. & Brettell Psu 3 Peru 3°S–17°N

Sampera V.A. Funk & H. Rob. Sam 8 Colombia, Ecuador, Peru 4°N–6°S

Sinclairiopsis Rydb. Sio 2 Mexico 17°–18°N

Sinclairia Hook. & Arn., Liabellum Sin, Lib, 25 Mexico, Guatemala, El Salvador, Honduras, Nicaragua, Costa Rica, 23°N–4°N Rydb., Megaliabum Rydb. Meg Panama, Colombia

Stephanbeckia H. Rob. & V.A. Ste 1 Bolivia, type collection only 21°S–22°S Funk

438 TAXON 61 (2) • April 2012: 437–455 Funk & al. • Phylogeny and biogeography of Liabeae

All primer sequences and their sources are shown in DNA) for each subsequent reaction cycle. After a total of 40 Table 2. Primers ITS5A and ITS4 were used to amplify and reaction cycles, an additional 7-min extension at 72°C was al- sequence the complete ITS region. For a few samples, it was lowed for completion of unfinished DNA strands. The anneal- necessary to substitute primer ITS5A with ITS5HP and use a ing temperature was raised when necessary to obtain a single higher annealing temperature (up to 58°C) to avoid amplifying fragment product. All PCR products were quantified by agarose fungal ITS sequences. gel electrophoresis with comparison of an aliquot of products Primers trnL-C and trnL-F were used to amplify and se- with a known quantity of a 100-bp DNA ladder (GeneChoice, quence the plastid trnL-F intron and intergenic spacer region. Frederick, Maryland, U.S.A.) visualized with ethidium bromide. The 3′ end of the plastid ndhF gene was amplified and se- The remainder was stored at 4°C until utilized. quenced using primers ndhF1603 and ndhF+607. The entire PCR products used for sequencing were enzymatically pu- matK gene and part of the upstream and downstream flanking rified for sequencing using ExoSAP-IT (USB now Affymetrix, trnK introns were amplified and sequenced in two fragments Cleveland, Ohio, U.S.A.). This procedure involved mixing 10 μl using primer pairs trnK3914F/matK1541R and matK1240F/ of the PCR product with 1 μl of ExoSAP-IT and then incubating trnK2R. When sequencing each of these regions, an additional the mixture first at 37°C for 30 min to degrade excess prim- primer was used (i.e., matK1240R and matK1541F, respec- ers and dNTPs, and then raising the temperature to 80°C for tively) to ensure complete coverage with high-quality reads. 15 min, to denature the ExoSAP-IT enzymes. This method of For the PCR amplification reactions, each 25 μl PCR reac- purification without loss of PCR products (no filtration, pre- tion cocktail contained 12.9 μl of sterile water, 2.5 μl of 10× cipitation, or washes are necessary) is especially important for PCR reaction buffer (Bioline, Taunton, Massachusetts, U.S.A.), DNA extracted from herbarium vouchers, which is sometimes 2 μl of 20 mM dNTPs (Bioline) in an equimolar ratio, 2 μl only weakly amplified and yields barely sufficient PCR product of 25 mM magnesium chloride (2.5 μl for chloroplast DNA), for sequencing. 0.5 μl of 10 mg/ml Bovine Serum Albumin (Sigma-Aldrich, St. The cycle sequencing reactions were done using 96-well Louis, Missouri, U.S.A.), 1 μl of a 10 μM concentration of the microplates in the DNA Engine Tetrad thermal cycler. Each forward primer, 1 μl of a 10 μM concentration of the reverse 10 μl cycle sequencing reaction cocktail contained 50–150 ng of primer, 0.1 μl of Taq DNA polymerase enzyme (5 units/μl from the purified PCR product in 5 μl of sterile water, 2 μl of a 1 μM Bioline), and 2.5 μl of sample DNA. The amount of template concentration of the sequencing primer, 0.6 μl of a 5× reaction DNA was adjusted when necessary to generate sufficient PCR buffer (400 mM Tris-HCl, 10 mM magnesium chloride at pH products for DNA sequencing. 9.0), and 1 μl of the reagent pre-mix from the BigDye (Version The amplification reactions were done using thin-walled 3.1) dye terminator cycle sequencing pre-mix kit (Applied Bio- 0.2 ml PCR reaction tubes in a DNA Engine Tetrad thermal systems, Foster City, California, U.S.A.). The cycle sequencing cycler from MJ Research (now Bio-Rad, Hercules, California, program consisted of an initial preheating at 96°C for 30 s. U.S.A.). The PCR program consisted of an initial preheating Then, the first reaction cycle proceeded as follows: 10 s at 92°C at 94°C for 2 min. Then, the first reaction cycle proceeded as to denature the template DNA, followed by 5 s at 55°C to al- follows: 1 min at 94°C to denature the template DNA, followed low primer annealing and 4 min at 60°C for primer extension. by 1 min at 48°C (54ºC for trnL-F; up to 58°C for matK) to al- Unincorporated dye terminators were removed by Sephadex low primer annealing, and 2 min at 72°C for primer extension. (GE Healthcare, Piscataway, New Jersey, U.S.A.) gel filtration Primer extension time was increased by 4 s (7 s for chloroplast using MultiScreen plates (Millipore, Billerica, Massachusetts,

Table 2. Primer sequences and their sources. Name Sequence (5′ to 3′) Reference ITS5A GGA AGG AGA AGT CGT AAC AAG G Downie & Katz-Downie, 1996 ITS5HP GGA AGG AGA AGT CGT AAC AAG G Hershkovitz & Zimmer, 1996 ITS4 TCC TCC GCT TAT TGA TAT GC White & al., 1990 trnL-C CGA AAT CGG TAG ACG CTA CG Taberlet & al., 1991 trnL-F ATT TGA ACT GGT GAC ACG AG Taberlet & al., 1991 ndhF1603 CCT YAT GAA TCG GAC AAT ACT ATG C Jansen, 1992 ndhF+607 ACC AAG TTC AAT GYT AGC GAG ATT AGT C Jansen, 1992 trnK3914F GGG GTT GCT AAC TCA ACG G Johnson & Soltis, 1994 trnK2R AAC TAG TCG GAT GGA GTA G Steele & Vilgalys, 1994 matK1240F ACC TTA CCC AGC TCA TCT G Bayer & al., 2002 matK1240R CAG ATG AGC TGG GTA AGG T Bayer & al., 2002 matK1541F CGA TCA ACA TCT TCT GGA GC Bayer & al., 2002 matK1541R GCT CCA GAA GAT GTT GAT CG Bayer & al., 2002

439 Funk & al. • Phylogeny and biogeography of Liabeae TAXON 61 (2) • April 2012: 437–455

U.S.A.). The purified cycle sequencing products were then re- after consultating with Dr. Harold Robinson, the new combina- solved by capillary electrophoresis using a Hitachi 3730xl DNA tion was added to the Nomenclature section of this paper. For Analyzer (Applied Biosystems). Sequences from both strands the analyses shown in Figs. 3 and 4, only taxa with all sequence (six for matK) of each PCR product were assembled, examined, data from all four regions were used (as in Fig. 1). No outgroup compared, and corrected using Sequence Navigator software was included in the unrooted tree (Fig. 3) and the root used for (Applied Biosystems). the bootstrap and Bayesian analyses was further reduced to All of the sequences were aligned visually. Insertions and only populifolius (Lam.) Cass. and Pseudostifftia deletions were examined and tracked during the analyses but kingii H. Rob. (Fig. 4). ultimately gaps were not coded separately (see discussion of Phylogenetic analysis. — The nuclear (ITS) and chlo- Fig. 2). For all of the analyses, the names of the terminal taxa roplast data (trnL-trnF, ndhF, matK) were analyzed using consist of a three-letter abbreviation for the genus, the spe- Maximum parsimony (MP), Maximum likelihood (ML), and cies epithet, and the extraction number that serves as a unique Bayesian inference (BI). Congruence between datasets under identifier across all our papers. The identification of the three- parsimony was assessed with the incongruence length differ- letter abbreviations for the genera is listed in Table 1. The ITS ence (ILD) test (Farris & al., 1994) using a heuristic search and sequences produced for the Kim & al. (2003) paper were not the tree-bisection-and-reconnection (TBR) branch-swapping used in this study; instead new sequences were generated. option in PAUP* v.4.0b10 (Swofford, 2002). All characters were Selection of outgroup taxa. — The selection of outgroup unweighted and treated as unordered. Consistency indices (CIs) taxa was addressed by the addition and subtraction of various and rescaled consistency indices (RCs; Farris, 1989) were cal- clades. The selection of the clades to be examined was based on culated to evaluate the amount of homoplasy in the data. the most recent overall analyses of the family (Panero & Funk, The initial analyses were run in maximum parsimony and 2008; Funk & al., 2009). The process began with an analysis the maximum number of was set at 10,000. This limit was that used 150 outgroup taxa from the basal grade of the family, exceeded only when a large number of outgroups was used subfamily , and the other tribes of subfamily Cicho- (ca. 150). Once the number of outgroups was reduced to ca. rioideae; Gerbera L. from was selected as the root for 60 (see Selection of uutgroup taxa) the number of trees (when the trees. This group of outgroup taxa included representatives all the data were included) was reduced to under 8000, and of all clades in the family except for those in the strongly nested when the number of outgroups was reduced even further there subfamily Asteroideae and all ingroup taxa with sequence data were around 3000 equally parsimonious trees. Each of the four from all four regions (63 accessions). This analysis showed that markers was analyzed separately and in all pairwise and 3-way the sequences of the furthest removed outgroup clades were combinations. Topologies obtained from separate analyses of strongly divergent from those of the Liabeae and, therefore, the three chloroplast markers were mutually congruent. The posed alignment difficulties too great to justify their inclusion. ITS topology was largely congruent with that of combined The elimination of the basal grade and the use of Carduoideae as chloroplast data. Topological conflict was restricted to highly the root provided a stable ingroup topology but resulted in thou- nested clades where the terminal taxa had short branches sands of trees based on the rearrangement of the outgroup taxa. (within Liabum and Sinclairia). In order to find the minimum number of outgroup taxa that Maximum parsimony analyses were conducted using could be used and still maintain the same basic topology, taxa PAUP* v.4.0b10, with heuristic searches using 1000 stepwise were systematically removed. The outgroup taxa selected in- random addition sequences replicates, holding 10 trees at each cluded the first few branches of tribe Vernonieae (basal grade; step, with tree-bisection-reconnection (TBR) branch-swapping 15 species), all species of Moquineae (2) and Eremothamneae and MULTREES in effect (Felsenstein, 1985). Bootstrap analy- (2), and representatives of both subtribes of Arctotideae (11). ses were used for each analysis to assess character support. Moquineae were nested one branch up from the base of Vernon- We determined nucleotide substitution models for each of ieae, just above the tropical African/Madagascaran genus Dis- the genes and gene regions using jMODELTEST v.0.1.1 (Guindon tephanus Cass. (Vernonieae). It was clear from the phylogeny & Gascuel, 2003; Posada, 2008) using the Akaike information produced (Fig. 1) that except for Distephanus and Moquineae, criterion (Akaike, 1974). A GTR + I + Г model was selected for the branches of Vernonieae were much longer than those of all three chloroplast markers and a SYM + Г model was selected Liabeae. Ultimately, most of the outgroup taxa were removed for the ITS region. Because each of the chloroplast markers had from the analysis leaving only Distephanus and Moquineae. the same model selected by Modeltest and produced mutually This method of progressive elimination of potential outgroup congruent results in the parsimony analyses, the three datasets taxa resulted in a significant reduction in the number of trees were combined into one partition in the analyses described be- and did not alter the basic arrangement of the ingroup taxa. low. The ITS data were modeled in a separate partition. During the analysis that resulted in Fig. 2, all of the ingroup We also employed maximum likelihood and Bayesian ap- taxa that were missing only matK sequence data were added to proaches for analyzing the concatenated data matrix. Maximum the analysis increasing the pool from 63 to 81 samples. Also, likelihood optimization of the partitioned matrix was imple- four additional Distephanus species that were missing matK mented in GARLI v.0.97 (Zwickl, 2006) using 100 independent sequence data were added resulting in eight outgroup taxa. search replicates. Starting parameter values were set according During this study we realized that one species, mada- to the results of the model selection tests, and allowed to be fur- gascariensis Less., had never been transfered to Distephanus; ther optimized within each replicate. We enforced an automated

440 TAXON 61 (2) • April 2012: 437–455 Funk & al. • Phylogeny and biogeography of Liabeae

stopping criterion for each replicate when a stable −lnL score parsimonious trees were created by rearrangements of terminal was attained for 2 × 104 generations. Maximum likelihood boot- taxa within Liabum or Sinclairia. Clades that lose resolution in strap support values for the concatenated matrix were estimated the strict consensus tree are indicated on the figure with an as- using 1000 pseudoreplicates, each including three independent terisk (*). The tree values are as follows: PI = 611; L = 1887; CI = search replicates, but otherwise following the same search strat- 0.596; RC = 0.872. The strict consensus tree is shown in Fig. S1. egy as described above. For convenience, we refer to clades with An unrooted parsimony tree is shown in Fig. 3, it was ≥ 85% BP as being strongly supported, 84%–70% as moderately generated using only taxa that had sequence data from all four supported, and 69%–50% as poorly supported. markers (as in Fig. 1) but no outgroups were used. Note the Bayesian MCMC analyses (Yang & Rannala, 1997) were distinctness of the subtribes, the distance of Cacosmia from implemented in MrBayes v.3.1.2 (Huelsenbeck & Ronquist, all other taxa, and the taxa on long and short branches. The 2001; Ronquist & Huelsenbeck 2003; Altekar & al., 2004), label “root 1” indicates the position selected to root the tree in with the markers partitioned and modeled as described above. the parsimony analyses (see the strict consensus tree; Fig. S1) Within each partition, default priors were used for the rate ma- and it causes Cacosmia to be the sister clade to the rest of the trix, branch lengths, gamma shape parameter (Yang, 1993), and tribe. The position labeled “root 2” was selected by GARLI the proportion of invariant sites (where appropriate; Reeves, and MRBAYES (Fig. 4) and it positions Cacosmia as the sister 1992). A flat Dirichlet distribution was used for the base fre- group of clade B, the Liabinae. The “root 2” position was also quency parameters. An uninformative prior was used for the supported by a 9 bp deletion (see discussion below and Fig. 2). tree topology. The final analyses were conducted with four con- The values for the unrooted trees are as follows: number of trees current runs of eight incrementally heated chains for 2.0 × 107 = 4680; PI = 498; L = 1373; CI = 0.650; RC = 0.884. generations that were sampled every 100 generations, with An examination of the aligned sequences in the nuclear the heating parameter set to 0.15 to promote adequate mixing and chloroplast datasets revealed that there were a number between the chains. The program AWTY (Wilgenbusch & al., of indels that are informative (i.e., several indels that involve 2004; Nylander & al., 2008) was used to diagnose topological multiple genera or clades; Fig. 2). Most of the larger indels convergence, with emphasis placed on the results of the Cumu- occur in the trnL-F and matK datasets but the ITS dataset lative, Split, and Compare diagnostics. These results indicated does have an 18 bp deletion that is restricted to the ingroup a burn-in period of 5.0 × 106 generations, which was much more (based on the analysis done for Fig. 1 with many outgroup taxa) conservative than the approximately 2.0 × 105 generations sug- and a 12 bp deletion shared by Pseudonoseris and Paraneph- gested by the plot of the log likelihood values viewed in Tracer elius. The trnL-F dataset has a 9 bp deletion that defines clade v.1.5 (Rambaut & Drummond, 2003–2007). The average stan- A + clade B (Cacosmia and Liabinae; this supports “root 2” in dard deviation of split frequencies at 5.0 × 106 generations was Fig. 3); a 21 bp deletion shared by Sinclairia, Megaliabum, and 4.9 05 × 10 −3. Trees from the post burn-in period of each analysis Liabellum, but not found in Sinclairiopsis klattii (B.L. Rob. were pooled together (6.0 × 105 trees in total) to calculate Bayes- & Greenm.) Rydb. or Sinclairia ismaelis Panero & Villaseñor; ian posterior probabilities (PP). a 7 bp insertion for the yellow-rayed Munnozia (Fig. 2, clade D3); a 49 bp deletion for Pseudonoseris and Paranephelius; and a 18 bp insertion for Sinclairiopsis klattii and Sinclairia Results ismaelis. The matK dataset had fewer indels considering its length (2.8 kb): Stephanbeckia and Microliabum share a 5 bp Phylogenetic analysis. — One of the 7488 equally par- insertion; Liabinae (minus the first branch: Ferreyranthus) simonious trees of 69 Liabeae samples and 32 outgroup taxa have a 9 bp deletion; Philoglossa and Erato have three 6–7 bp is shown in Fig. 1. This particular analysis included only taxa deletions and one insertion; Munnoziinae members have a 6 bp that had sequence data from all four regions (see Appendix; insertion; Sinclairinae (minus Sinclairia ismaelis; no matK parsimony informative characters, PI = 881; length, L = 3794; sequence data for Sinclairiopsis klattii) have a 9 bp deletion; consistency index, CI = 0.483; rescaled consistency index, RC = Paranepheliinae plus Munnoziinae clades (Fig. 2, clades D–E) 0.812). The two clades indicated with an asterisk (*) (Sinclairia, share a 10 bp deletion; and finally there is a 30 bp duplication Liabum) lose resolution in the strict consensus tree (Fig. S1 in found in Liabinae but missing and presumed lost in the Carib- the Electronic Supplement). The outgroups used were Arcto- bean Liabum species. All of these groups are found in Figs. 2 tideae subtribe Arctotidinae, Arctotideae subtribe Gorteriinae, and/or 4 so they were not coded separately as characters. the small tribes Eremothamneae and Moquineae, and Vernon- ieae; the latter is the sister group of Liabeae. Vernonieae have longer branches than the other outgroups. The tribe Cichorieae, Discussion the sister group of all taxa shown in Fig. 1 (not shown), have branches similar in length to Vernonieae. Phylogeny. — It is clear from Figs. 1–4 that four clades Figure 2 shows one of the 5615 equally parsimonious phy- (A–D) are well-supported (93–100% bootstrap support and 1.0 logenetic trees produced by using fewer outgroups and all in- Bayesian posterior probability). However, although clade E was group taxa (including those that were missing matK (8 outgroup present in all of the equally parsimonious cladograms (Fig. S1) taxa; 81 samples of Liabeae). The outgroups were Moquin- it dissolved into three clades in the maximum likelihood and eae plus Distephanus (Vernonieae). Nearly all of the equally Bayesian trees (Fig. 4). Clade E has never been proposed as

441 Funk & al. • Phylogeny and biogeography of Liabeae TAXON 61 (2) • April 2012: 437–455

Hap nervosa 139 Hap ruppallii 76 Hap scaposa 77 Arh calendula 03 Arh sp 95 Arctotideae subtribe Arctotidinae Ari bellidifolia 08 Ari cuprea 10 Ari fastulosa 14 Ari hirsuta 271 Hop spinosum 297 Ere marlothianus 295 Eremothamnieae Ber spinosissima 28 Did spinosa 49 Arctotideae subtribe Gorteriinae Did carnosa 50 Dis populifolius K12 Distephanus Pse kingii 150 Moq racemosa 354 Moquineae Lin melleri K38 Bac adoensis K10

Hes arbuscula Hau2 Vernonieae Gym amygdalinum K11 Gym mesipifolium K112 Bac lasiopus K18 Erl misera 292 Pol poskeana 291 Cen pauciflorus K64 Led jonesii K40 Str arboreum 228 Cri sodiroi K47 Cep punctatum CJ Les warmingianus 293 Pro argentea 321 A Cac rugosa 174 Cac hieronymus 197 Cac harlingii 238 Fer verbasifolia 177 Fer rugosus 176 B Fer gentrii 259 Oli volubilis 121 Dil pefoliata 199 Sam cuatrecasasii 182 Sam coriacea 214 Lia grandiflorum 239 Lia solidagineum 178 Lia wurdackii 179 Lia igniarium 206 Lia floribundum 269 Lia bourgeaui 147 Lia asclepiadeum 257 Lia kingii 270 * Lia umbellatum 207 Lia fromcuba 461 Lia poiteaui 246 Lia baharonense 204 Lia subacaule 283 Lia selleanum 284 Sin ismaelis 281 C Sin caducifolia 193 Sin glabra 218 Sin sublobata 250 Meg andrieuxii 187 Sin polyantha 148 Sin deamii 194 Sin discolor 195 * Sin moorei 219 Meg pringlei 188 Lib cervinum 339 Sin liebmannii 280 Liabeae Sin simile 337 Lib angustissimum 185 Lib palmerii 186 Mun foliosa 249 Mun jussieui 180 Mun campii 210 D Mun lyrata 212 Chr acuale 175 Chr hieracioides 253 Chr hieracioides 254 Mun sagastiguii 181 Mun gigantea K39 Mun hastifolia 190 Mun maronii 191 Mun senecionidis 189 Mun wilberii 213 Mun wilberii 221 Chi benthamii 286 Chi benthamii 441 Phi mimuloides 120 Phi mimuloides 183 Phi purpureodisca 216 Era sodiroi 202 Era polymonoides 201 Era costaricinsis 211 Ste plumosa 426 E Mic mulgediifolium 209 Mic polymnioides 173 Mic polymnioides 235 Psu discolor 184 Par asperifolis 215 Psu szyszylowiczii 217 10 changes Par ovatus 192

Fig. 1. One of the 7488 equally parsimonious trees of 69 Liabeae and 32 outgroup samples that incorporated DNA sequence data from the ITS, trnL-F, 3′ end of ndhF, and matK regions (PI = 881; L = 3794; CI = 0.483; RC = 0.812). The outgroups used were Arctotideae subtribe Arctotidi- nae, Arctotideae subtribe Gorteriinae, and tribes Eremothamneae, Moquineae, and Vernonieae; the latter is the sister group of Liabeae. Note the long branches in Vernonieae as compared to the other tribes. This analysis included only taxa that had sequence data from all four regions mentioned above (see Appendix). The full taxon names can be found in Table 1 and the Appendix. An asterisk (*) indicates a clade that has some nodes within it that collapse in the strict consensus tree.

442 TAXON 61 (2) • April 2012: 437–455 Funk & al. • Phylogeny and biogeography of Liabeae Outgroups Moq racemosa 355 Pse kingii 150 Pse kingii 354 Dis barus 500 Dis ambongensis 502 Dis populifolius K12 Dis madagascariensis V415 ++9 bp D Dis garnierianus 501 A Cac rugosa 174

— Cac hieronymus 197 Cac harlingii 238 Fer verbasifolia 177 Fer rugosus 176 +30 bp dup Fer gentrii 259 Oli volubilis 121 — Oli sessiliflora 261

Dil pefoliata 199 Liabinae B — Sam cuatrecasasii 182 9 bp D Sam coriacea 214 Lia grandiflorum 239 Lia solidagineum 178 Lia wurdackii 179 — Lia igniarium 206 18 bp D Lia floribundum 269 Lia bourgeaui 147 Lia asclepiadeum 257 Lia kingii 270 * Lia acuminatum 203 Lia vargasii 266 Lia umbellatum 207 Lia fromcuba 461 — Lia poiteaui 246 +30 bp D Lia baharonense 204 Lia subacaule 283 18 bp I Lia selleanum 284 1 Sin ismaelis 281 — Sio klattii 240 C Sin caducifolia 193 Sin glabra 218 Sinclairiinae Sin sublobata 250 2 Meg andrieuxii 187 — Sin vagans 287 21 bp D Sin polyantha 148 9 bp D Sin deamii 194 Sin discolor 195 Sin moorei 219 Meg pringlei 188 * Lib angustissimum 185 Lib palmerii 186 Lib cervinum 339 Sin liebmannii 280 Sin simile 337 1 5 bp I Ste plumosa 426

— Mic mulgediifolium 209 Mic polymnioides 173 Paranepheliinae Mic polymnioides 235 Psu discolor 184

2 — Par ovatus 192 12 bp D Psu szyszylowiczii 217 E Par asperifolis 215 49 bp D Chi benthamii 208 3 Chi benthamii 286 Chi benthamii 441 Era sodiroi 202 Era polymonoides 201 4 Era costaricinsis 211 — Phi mimuloides 120

— 6&7 bp D Phi peruviana 232 10 bp D 6 bp I Phi mimuloides 183 Phi purpureodisca 216 Mun foliosa 249 1 Mun foliosa 520 Mun jussieui 180 Mun campii 210 D Munnoziinae Mun lyrata 212 — 6 bp I 2 Chr acuale 175 Chr hieracioides 253 Chr hieracioides 223 Chr hieracioides 254 Mun sagastiguii 181 Mun fosbergii 119 3 Mun senecionidis 189 — Mun wilberii 213 7 bp I Mun wilberii 221 4 Mun pinnatipartita 256 Mun nivea 258 Mun gigantea K39 10 changes Mun hastifolia 190 Mun maronii 191

Fig. 2. One of the 5615 equally parsimonious phylogenetic trees resulting from a parsimony analysis using a reduced number of outgroups (8) and adding all taxa of Liabeae (81 samples), including those that did not have matK sequence data (PI = 611; L = 1887; CI = 0.596; RC = 0.872). The outgroup used was Moquineae plus Distephanus (Vernonieae). Nearly all of the equally parsimonious trees were created by rearrangements of terminal taxa within Liabum, Sinclairia, or the Pseudonoseris-Paranephelius clade. Insertions and deletions are indicated. The full taxon names can be found in Table 1 and the Appendix. An asterisk (*) indicates a clade that has some nodes within it that collapse in the strict consensus tree; bp = base pair; D = deletion; dup = duplicaton; I = insertion; + = this duplication is lost in the highly nested Caribbean Liabum clade; ++ this deletion is also found in calde B.

443 Funk & al. • Phylogeny and biogeography of Liabeae TAXON 61 (2) • April 2012: 437–455

a monophyletic group in any of the morphological studies. In Cichorioideae are the yellow radiate heads, 2 rows of pappus fact, if one examines the phylogenies based on morphology as with the inner row consisting of bristles, and achenes with 5–10 proposed by Robinson (1983a) and Funk & al. (1996), clades ribs. The outer row of the pappus (when present) can be either A–D were all recognized, and only clade E is novel. Within scales or bristles in Liabeae and Vernonieae but in Distepha- clade E there are four well-supported clades (Fig. 4, clades nus and Moquineae the outer row consists of bristles, so that E1–4) all of which have been recognized as distinct groups or character was assumed to be plesiomorphic. taxa. The insertions and deletions are shown on Fig. 2 and show The monophyly of Liabeae is very strongly supported by additional support for many of the branches but not for clade E. its morphology. The genera have a series of characters that Morphology. — Figure 5 shows two generic level clado- make it relatively easy to place them in the tribe; however, as grams that provide a framework for examining the basic mor- in most tribes, there are taxa with exceptions to some of the phological characters of the tribe. Figure 5A displays characters tribal characters. The tribe is diagnosed by the following char- related to latex, leaves, corollas, and style branches; Fig. 5B acters: frequent occurrence of white latex (Fig. 6A; apparently displays pappus and achene characters. The basic structure of absent in clade B); opposite leaves (Fig. 6A; Chrysactinium, the tree is based on the results of the parsimony analysis where Paranephelius, Pseudonoseris possess leaves congested on Cacosmia is the sister group of the rest of the tribe (Figs. 1–2). short stems so they appear to be whorled); strongly tri-nervate As mentioned above, Cacosmia is the sister group of clade B leaf venation (Fig. 6A; pinnate in most genera in clade B and (Liabinae) in the maximum likelihood and Bayesian trees (see in Paranephelius and Pseudonoseris; palmate in Erato); blades arrow). However, the characters of Cacosmia are either poten- white-tomentose beneath (Fig. 6A; strigose or pilose in Erato; tial apomorphies for the tribe (latex, opposite and tri-nervate glabrous in Philoglossa); yellow fertile ray florets and yellow leaves, white tomentum) or are apomorphic for the genus (short hermaphroditic disk florets (Figs. 6 and 7; all florets reddish in style branches, no pappus, 4–5 angled achenes with no ribs). one species of Pseudonoseris; ray florets reddish or purplish in In fact, Cacosmia seems to have no synapomorphic characters Chionopappus and one species of Philoglossa; rays white in at that it shares with other close members of the tribe so it can least four species of Munnozia (Fig. 7G); heads discoid in a few be positioned as the sister group to the rest of the tribe or the Sinclairia (Figs. 6F, 7A); achenes that are usually prismatic or sister group of clade B without having much affect on our in- subterete with 5–10 ribs (4-ribbed in Erato, compressed with terpretations of character evolution. The arrows in Fig. 5A–B 2 ribs in Philoglossa and Stephenbeckia, and long-fusiform indicate the alternate placement of Cacosmia. Characters that in Paranephelius); echinate pollen with unevenly distributed are believed to be plesiomorphic in the context of subfamily spines (evenly spaced spines are found in Chrysactinium, Mun- nozia, Paranephelius, and Pseudonoseris), and without obvi- ously lophate or psilate variations. Liabum The pappus is more variable than the other characters dis- cussed above (Fig. 5B) but nine genera have a biseriate pappus: Sampera Dillandia outer scales and inner scabrous bristles, two rows of bristles, Oligactis or in one case (some taxa in Microliabum), two rows of scales. Ferreyranthus Cacosmia Sometimes it is easy to distinguish scales from bristles but in A Sinclairiopsis other taxa it can be difficult to distinguish a slightly flattened Stephanbeckia root 2 bristle from a narrow scale. Conversely, seven genera have only one row of bristles, two of which are plumose. Finally, two B 1 Sinclairia & seg. unrelated taxa have no pappus (Cacosmia, Philoglossa). Four root 1 1 C of the seven genera in clade E do not have a biseriate pappus Microliabum 2 and the positions of these genera in the phylogeny make it a possibility that the loss of the outer row is an apomorphy in Chi- E D Psudonoseris 2 2 onopappus plus the well-recognized Erato-Philoglossa clade. The tribe also possesses many of the plesiomorphic char- 4 1 Munnozia Paranephelius Munnozia 3 -Yellow acters often found in non-Asteroideae taxa: disc florets with -White narrow limbs and deeply lobed corollas; a single, wide, band 3 Erato of receptive tissue on the style branches, and hairs on the style. The characters of Liabeae contrast with those of the sister Chionopappus group, Vernonieae, whose members have no latex, alternate Philoglossa leaves, pinnate venation, no white tomentum, no ray florets, Chrsactinium blue or purplish corollas, and usually some form of lophate 10 changes pollen (lophate, sublophate, or echinolophate). The two tribes share a biseriate pappus of bristles and long style branches. Fig. 3. An unrooted parsimony phylogeny using taxa from Fig. 1 but without outgroups (no. of trees = 4680; PI = 498; L = 1373; CI = 0.650; Interestingly, the sister group to the remainder of Vernonieae is RC = 0.884]. Note the distinctness of the subtribes (see Fig. 2), the the mostly African genus Distephanus which has yellow flow- distance of Cacosmia from all other taxa, the two positions of the ers and tri-nervate leaf venation like most of Liabeae. The next root, and the taxa on long and short branches. branch above Distephanus is tribe Moquineae whose members

444 TAXON 61 (2) • April 2012: 437–455 Funk & al. • Phylogeny and biogeography of Liabeae

Dis populifolius K12 Pse kingii 354 Outgroups 1.00 Cac rugosa 174 A 1.00 100 Cac hieronymus 197 76 Cac harlingii 238 1.00 1.00 Fer verbasifolia 177 82 .91 Fer rugosus 176 100 1.00 72 Fer gentrii 259 B Oli volubilis 121 94 Dil pefoliata 199 1.00 1.00 Sam cuatrecasasii 182 99 100 Sam coriacea 214 Liabinae 1.00 .99 Lia bourgeaui 147 1.00 88 65 Lia asclepiadeum 257 94 Lia kingii 270 1.00 Lia grandiflorum 239 1.00 Lia solidagineum 178 100 66 1.00 Lia wurdackii 179 63 1.00 Lia igniarium 206 78 Lia floribundum 269 1.00 Lia umbellatum 207 Lia ‘from cuba’ 461 1.00 100 .77 Lia poiteaui 246 100 52 1.00 Lia baharonense 204 100 .98 Lia subacaule 283 1 63 Lia selleanum 284 Sin ismaelis 281 1.00 C Sin sublobata 250 100 21.00 Sin caducifolia 193 Sin glabra 218 Sinclairiinae 100 Meg andrieuxii 187 1.00 Sin polyantha 148 .96 100 Sin deamii 194 .96 64 Sin discolor 195 Sin moorei 219 58 .97 88 Meg pringlei 188 1.00 58 Sin liebmannii 280 1.00 95 Sin simile 337 76 .62 Lib cervinum 339 1.00 - .69 Lib angustissimum 185 91 54 Lib palmerii 186 1.00 1 Ste plumosa 426 Mic mulgediifolium 209 100 1.00

1.00 Paranepheliinae 100 Mic polymnioides 173 99 Mic polymnioides 235 1.00 2 Psu discolor 184 .99 Psu szyszylowiczii 217 E 100 Par ovatus 192 80 Par asperifolis 215 3 1.00 Chi benthamii 286 1.00 100 Chi benthamii 441 1.00 Phi mimuloides 120 100 1.00 93 1.00 Phi mimuloides 183 1.00 100 100 Phi purpureodisca 216 4 100 1.00 Era sodiroi 202 1.00 100 Era polymonoides 201 Africa 99 Era costaricinsis 211 General Africa 1.00 1 Mun foliosa 249 99 1.00 Mun jussieui 180 Mun campii 210 Munnoziinae South America 1.00 2 100 100 Mun lyrata 212 North & Central Andes 1.00 1.00 Chr acuale 175 .95 Chr hieracioides 253 Southern Andes, D 92 100 southern South America 3 .99 60 Chr hieracioides 254 Mun sagastiguii 181 North America 84 1.00 1.00 Mun gigantea K39 Mexico 1.00 Mun hastifolia 190 100 1.00 100 4 89 Mun maronii 191 Central America 100 1.00 Mun senecionidis 189 1.00 Mun wilberii 213 Caribbean 85 100 Mun wilberii 221

Fig. 4. The phylogeny of Liabeae produced by GARLI and MRBAYES using sequence data from ITS, trnL-F, ndhF, and matK datasets. Bootstrap values are below and Bayesian posterior probabilities are above each branch. Note the collapse of clade E, Paranepheliinae. In the MRBAYES tree in the C2 clade, the two-taxon clade containing “Sin morei 219” and the five-taxon clade containing “Sin liebmannii 280” were sister taxa with a posterior probability of 0.97; this grouping was also found in the parsimony strict consensus tree (Fig. S1 in the Electronic Supplement) but not in the GARLI tree. The colors follow those used in Funk & al. (2009) with one exception; the Caribbean has a yellow-green color in this figure. The full taxon names can be found in Table 1 and the Appendix.

445 Funk & al. • Phylogeny and biogeography of Liabeae TAXON 61 (2) • April 2012: 437–455

have a biseriate pappus of bristles but also have pinnate leaf ve- remained closer to one area? We cannot answer these questions nation and purple flowers as found in Vernonieae, so it is only with any certainty but these are topics for of future research. Distephanthus that shares these two characters with Liabeae. The genus Cacosmia (Fig. 6B). — Clade A, Cacosmia, In Vernonieae and Cichorieae the branches supporting consists of three species that have been collected in northern the genera and the groups of genera are often much longer Peru and southern Ecuador. This genus is found in two differ- than those in the Liabeae, Arctotidieae-Arctotidinae, and the ent positions in the phylogeny depending on the data analysis Arctotidieae-Gorteriinae clades (Fig. 1). The presence of these performed. The parsimony analyses place Cacosmia as the long branches poses an interesting question: Does this mean sister group to the remainder of the tribe. The maximum like- that character evolution is taking place faster in Vernonieae lihood and Bayesian analyses place it as sister group to clade and Cichorieae than in the other tribes? Is it because Vernon- B. Ferreyranthus is the sister group to the rest of clade B (Figs. ieae and Cichorieae have continually moved around expand- 1–4, Fig. S1) and while this genus shares some characters with ing their range (mostly dry tropical areas with more recent Cacosmia, such as an upright woody habit (also in Sinclairia), expansion into northern and northern temperate environments, bullate leaf surfaces (also in some members of Paranephelius respectively) while Arctotideae (mostly southern Africa) and and Pseudonoseris), and the presence of nodal sheaths, mor- Liabeae (Neotropics, mostly Andean) have, as far as we know, phologically it does not have much in common with either the

Fig. 5. Major characters used A B C D E within the tribe and to circum- scribe the tribe are mapped is ia on a cladogram derived from ibbean thus tinium a iopsis ia ** a n zia 1 zia 2 Fig. S1 (Electronic Supplement). zia 3 tis sa c oliabum o ey r A, osmia Latex, leaves, tomentum, y t anephelius r a ephenbec k ic r egaliabum unn o unn o unn o a c ampe r t hiloglossa seudonose r a r heads, and flowers; B, pappus e r C F Sinclai r Sinclai r Liabellum M M M Ch r Oliga c Dillandia S Liabum * Liabum -C a r P P P S M M Chionopappus E r and achenes. — The arrow indicates the alternative place- tri-nervate palmate red ment for Cacosmia; * = poorly red glabrous pinnate

resolved and includes Liabum- Outgroups Caribbean clade; ** = poorly 9 of 19 species 1 of 2 species are radiate is radiate white resolved and includes Liabellum and Megaliabum.

discoid short style branches

no latex pinnate

short style branches

latex, opposite leaves, tri-nervate leaves, dense white tomentum, A radiate heads, yellow corollas, long style branches

A B C D E is ia ibbean a r thus tinium C a iopsis ia ** * a n zia 3 zia 1 zia 2 r r tis sa c oliabum o ey r osmia y t anephelius r a ephenbec k ic r egaliabum unn o unn o unn o a c ampe r t hiloglossa seudonose r a r e r M Chionopappus E r P P S M Sinclai Sinclai Liabellum M M M Ch r P F Oliga c Dillandia S Liabum Liabum - C 2-sided plumose bristles lost lost 4-sided no pappus 2-sided bristles lost plumose Outgroups

lost lost

scales or bristles

scales or bristles scales or bristles

no pappus 4-5 angled, no ribs pappus outer row = scales B 2 rows of pappus, pappus inner row = bristles, achenes with 5–10 ribs

446 TAXON 61 (2) • April 2012: 437–455 Funk & al. • Phylogeny and biogeography of Liabeae

Fig. 6. Images of Liabeae. A, Sinclairia showing the opposite leaves with tri-nervate venation, note the white tomentum on undersurface and the latex where the stem was cut; B, Cacosmia with bullate leaf surfaces; C, Ferreyranthus is a large or a small tree with many heads; D, Oligactis has a very non-showy inflorescence and is often vine-like, note the white tomentum; E, Liabum is widespread with a clade confined to the Caribbean, these two images are from Caribbean members; F, Liabellum (now placed in Sinclairia) is a small herb on forest floors and is without ray flowers. — Photos by V. Funk except B and C “inset” by M. Dillon.

447 Funk & al. • Phylogeny and biogeography of Liabeae TAXON 61 (2) • April 2012: 437–455

Fig. 7. Images of the Liabeae. A, Megaliabum pringlei (B.L. Rob. & Greenm.) Rydb. (now placed in Sinclairia) has thick green stems that ema- nate from a woody base (upper insert) and large heads without ray florets (lower inset); B, Paranephelius has a basal rosette with sessile heads; C, Philoglossa has dark anther thecae; D, Microliabum; E, Munnozia also has dark anther thecae; F, Chrysactinium is a small herb and some spe- cies appear to have a basal rosette; G, Munnozia campii H. Rob.: one of the white flowered species. — Photos by V. Funk.

448 TAXON 61 (2) • April 2012: 437–455 Funk & al. • Phylogeny and biogeography of Liabeae

taxa in clade B or in the rest of the tribe, except of course Sinclairia, Megaliabum, and Liabellum form the core Sin- for having several of the defining characters of the tribe (see clairia complex which is the sister group of Sinclairiopsis and Fig. 5). All three Cacosmia species have disc flowers with contains ca. 20 species, principally confined to open forests in short style branches, tri-nervate leaf venation, glabrous achenes Mexico and Central America (150–2500 m). A few of the Mexi- without distinct ribs, often angled in shape, no pappus, cylindri- can species are found in more arid environments associated cally shaped heads with an involucre of 5–6 series with bracts with tropical scrub and deciduous forests. Turner (1989, 2007) that are ovate to elliptical with an obtuse or rounded apex, a treated Sinclairiopsis, Megaliabum, and Liabellum as part of glabrous receptacle and latex. Sinclairia, while Robinson (1983a) recognized Liabellum as Ferreyranthus has elongate style branches in the disc separate. Robinson (1983a) based his recognition of Liabellum florets, pinnate leaf venation, setiferous and glanduliferous on the fact that it is an annual with a swollen tuberous root. achenes, ~10 ribs on the prismatic achenes, a pappus with an However, recent field work (Funk) has shown that two species inner row of bristles and outer row of short setae, heads that of Megaliabum and several species of Sinclairia also have a are broadly campanulate with strongly graduated involucral tuberous root, however, species that form trees have tuber- bracts that are ovate to lanceolate with an acute or acuminate ous roots only when they are young. While the three species apex, a receptacle with short chaff, and no latex reported. It of Liabellum sampled are grouped together on the phylogeny seems best at this time to leave Cacosmia as the sister group to (Figs. 1–3), they are nested within Sinclairia. the remainder of the tribe and not to place it in its own subtribe Two species of Megaliabum were sampled: one, M. prin- until more data are available (Fig. 5). glei (B.L. Rob. & Greenm.) Rydb., was the sister group of Sin- Subtribe Liabinae (Fig. 6C–E). — Clade B consists of Dil- clairia moorei (H. Rob. & Brettell) H. Rob. & Brettell. A close landia, Ferreyranthus, Liabum, Oligactis, and Sampera, which examination of the herbarium material of S. moorei showed it make up ca. 35% of the species diversity in the tribe; Liabum to resemble M. pringlei in head size and potentially in growth has the widest distribution in the tribe occurring from Mexico form as well (thick herbaceous stems from a woody caudex). to northwestern Argentina and reaching into the Caribbean. These two taxa were distinct from Megaliabum andrieuxii The species range in habit from herbs to small trees. The mem- (DC.) Rydb., which has large heads but has ray florets and is bers of this clade have long style branches, pinnate leaf venation a shrub. Field observations (Funk) support the unique growth (Fig. 6D), lack latex, and most members have a single row of form of M. pringlei, however, all three species are nested pappus bristles ca. 4–5 mm long. However, Ferreyranthus has within Sinclairia and so it seems best at this time to treat all an outer row of 0.1–1.0 mm long broad-tipped flattened scales, of them part of this larger genus. The species of Megaliabum Sampera has an outer row of ca. 1 mm long acuminate scales, were placed into Sinclairia by Robinson & Brettell (1974) and and Oligactis has an outer row of much shorter bristles (Figs. 5). Liabellum was sunk into Sinclairia by Turner (1989); there Subtribe Sinclairiinae (Figs. 6A, F, 7A). — Clade C, consist- are, therefore, no nomenclature changes that need to be made. ing of Liabellum, Megaliabum, Sinclairia, and Sinclairiopsis, Subtribe Munnoziinae (Figs. 7E–G). — Clade D consists has ca. 15% of the species diversity of the tribe and is essentially only of Chrysactinium and Munnozia but it contains ca. 31% of a Northern Hemisphere group which has experienced consider- all species diversity in the tribe; all but two species are found in able radiation. The four genera, sometimes treated as synony- the Andes. They are found in a wide variety of habitats. Mun- mous, are all found in Mexico and Central America with the nozia ranges from Costa Rica and Panama, where two species exception of the widespread Sinclairia polyantha (Klatt) Rydb. have been recorded, to the Andes of Bolivia and Argentina, which is found from southern Mexico to Colombia (Table 1). All while Chrysactinium is confined to Ecuador and Peru. Mem- four genera have latex, long style branches, and tri-nervate vena- bers of clade E have short style branches, tri-nervate leaf vena- tion in the leaves. Their habit varies from herbs and scandent tion, and latex (not reported in Chrysactinium). Chrysactinium to trees. The pappus morphology is consistently two rows is a small herb while most species of Munnozia are scandent with the inner being bristles and the outer being shorter scales shrubs although a few approach Chrysactinum in habit. except in Sinclairiopsis which has only one row of bristles. All of the phylogenies (Figs. 1–4, Fig. S1) show Chrys- Sinclairiopsis was described as a monotypic Mexican ge- actinium to be nested inside of the much larger Munnozia. nus (Rydberg, 1927). As mentioned above, the DNA sequences However, the 13 Munnozia species sampled (out of ~40 spe- obtained from the involucral bracts of S. klattii (ITS, trnL-F) cies) fell into three distinct groups. The sister group to the matched almost exactly the data obtained from the leaf ma- remainder of the clade, “Munnozia 1” (Fig. 4, clade D1) has terial of the recently described Sinclairia ismaelis. In their three species all of which have white ray florets. An exami- paper, Panero & Villaseñor (1996) mentioned that S. ismaelis nation of the lectotype of the genus (Munnozia lanceolata is similar to Sinclairia (Sinclairiopsis) klattii differing in its Ruiz & Pavón) shows that morphologically it belongs in this “open paniculiform capitulescence, larger heads, and reproduc- group, and the herbarium label states that it has white ray tive phenology”. Based on the many molecular characters that florets. “Munnozia 3” contains the bulk of the genus. It is separate the two from the rest of this clade (Fig. 2) and their interesting to note that M. subg. Kastnera (Sch.Bip.) H. Rob. likewise distinct morphology: few involucral bracts, deeply al- & Brettell, represented in the analysis by M. nivea (Hieron.) veolate receptacle with erose edges and discoid heads, it seems H. Rob. & Brettell and M. pinnatipartita (Hieron.) H. Rob. best (Funk, 1985) to resurrect Rydberg’s genus Sinclairiopsis & Brettell, is nested within “Munnozia 3”. The length of the and to move Sinclairia ismaelis into it (see Nomenclature). branches indicates that they form a distinct clade, and this

449 Funk & al. • Phylogeny and biogeography of Liabeae TAXON 61 (2) • April 2012: 437–455

supports Robinson’s (1983a) decision to keep them in Mun- Since the outer row of bristles is quite variable in its presence nozia rather than separate them out. it seems best at this time to propose that it belongs somewhere “Munnozia 2” (Fig. 4, clade D2) contains only M. lyrata near Liabum, but it is not a perfect fit. (A. Gray) H. Rob. & Brettell and has no characters to distin- Biogeography. — In Fig. 4 and Fig. S1 various colors are guish it from “Munnozia 3” at this time. Before any nomencla- used to represent the global distribution of the species of Lia- tural changes can be recommended (e.g., sinking Chrysacti- beae. The phylogenies shown are the tree from GARLI and nium into Munnozia or describe “Munnozia 3” as a new genus), MRBAYES (Fig. 4) and the strict consensus tree (Fig. S1), the more species from Munnozia need to be sampled. Should the latter is based on the analysis used to generate the tree in Fig. 2. three groups of Munnozia prove to have sufficient morphologi- For the purpose of examining the distribution and phylogeny cal differences, it might be prudent to describe them as separate of Liabeae, the Andes can be divided into four informal and genera in order to maintain Chrysactinium, which strongly somewhat overlapping areas (Funk & al., 1995). There are two contrasts morphologically with Munnozia. northern areas: the Northeastern area, which begins in Venezu- Subtribe Paranepheliineae (Figs. 7B–D). — Clade E ela and continues into central Ecuador, and the Northwestern contains Microliabum, Stephanbeckia, Pseudonoseris, Para- area extending from northwestern Colombia to northern Peru. nephelius, Chionopappus, Philoglossa, and Erato and ca. 15.5% The Central Andean area stretches from northern to south- of the species diversity of the tribe. With the exception of one central Peru while a Southern area extends from southern Peru species of Erato in Costa Rica, members of clade E are en- into Bolivia, northern Argentina and adjacent Chile. Liabeae tirely South American in distribution. The most southern genus, have not been found in the temperate Andes. About half the Micro­liabum, is found from southern Bolivia to northwestern species and most of the genera in the tribe (Table 1) are found Argentina (Funk & Brooks, 1990). Erato is the northernmost in the Northwestern and Central areas. genus, found mostly in Ecuador with one endemic species in The Southern area is the oldest, its major uplift was in Costa Rica, and a few others that reach as far south as Bolivia. the Oligocene ca. 30 million years (Ma) ago in northern Chile Although Erato and Philoglossa were suggested as closely re- and southern Peru (James, 1973; Jordan & al., 1983; Windley, lated by Robinson (1983a), the addition of the other genera to 1984; Taylor, 1991). Gregory-Wodzicki (2011) estimated that the this clade has never been proposed. In fact Robinson thought Western Cordillera of this area of the Andes (referred to as the that Erato and Philoglossa were most closely related to Mun- Central Andean Plateau) was at no more than 50% of its current nozia and Chrysactinium (clade E) because of their dark anther elevation 25 Ma ago and the Eastern Cordillera and Altiplano thecae. The members of this subtribe have short style branches, reached no more than 50% of their modern elevation 10 Ma ago. tri-nervate leaf venation, latex (except for Chionopappus and The Central zone (central and northern Peru) is younger with Stephenbeckia), and are herbs or shrubs. The pappus morphology the uplift occurring 10–20 Ma ago (Garver & al., 2005). The is variable, ranging from absent to plumose to two rows of scales; northwestern Andes experienced their primary uplift in the last only one genus, Pseudonoseris, has the more usual arrangement 5 Ma (Hammen, 1974; Gentry, 1982) and the páramo-puna area of two rows consisting of inner bristles and outer, shorter scales. (the Northeastern zone) is considered the most recent, appear- Until recently, no documented evidence of interspecific ing during the Quaternary (2.0–0.1 Ma ago, Vuilleumier, 1969; or intergeneric hybridization had been reported for Liabeae, Simpson, 1975). For lower elevations Gregory-Wodzicki (2011) and it was thought that this was unlikely because the genera estimated that the Eastern Cordillera of the Colombian Andes were either chromosomally, geographically, and/or elevation- was at no more than 40% of its modern elevation by 4 Ma ago. ally separated. However, there is now evidence of interspecific The Andean Cordillera was and continues to be uplifted by the and intergeneric hybridization in northern Peru between spe- Nazca plate colliding with the South American plate along the cies in Paranephelius and Psuedonoseris (Soejima & al., 2008). Peru-Chile trench (James, 1973; Jordan & al., 1983). Bishopanthus solisceps H. Rob., . — Bishop- The pattern of distribution shown in Fig. 4 and Fig. S1 sup- anthus is a monotypic genus for which very little morphologi- ports the hypothesis that extant members of Liabeae have ori- cal information is known, let alone any chromosome counts gins located in the central and northern areas of the Andes, or molecular data, because all specimens of it were destroyed most likely in northern Peru (Fig. 4 and Fig. S1, clades A, B, except for a few fragments preserved in alcohol (Dillon & al., D, E). A number of genera from clade B are confined to the 2009). There are field notes and one photo taken by Earl Bishop, northern Andes or have most of their diversity in that area but for whom the genus is named (see Dillon & al., 2009). Bishop­ these are nested in clade B (Liabum, Sampera, Oligactis). Also, anthus is only known from the type collection in Peru; several nested high in the cladogram in clades E1 and E2 (Fig. S1) are expeditions to the type locality have failed to re-collect it (Dil- most of the taxa found in the southern Andes and this supports lon, pers. comm.). The shrubby habit, lack of latex, bullate the conclusion that the movement of Liabeae into the southern leaves, and long style branches place it close to members of the Andes was more recent than Liabeae of the central Andes de- basal grade of Liabinae (Ferreyanthus, Oligactis, Dillandia; spite the fact that the Andes are geologically older there. Fig. 5, clade B). In fact, it would fit reasonably well as a relative There is one subtribe (Sinclairiinae; Fig. 4 and Fig. S1, of Dillandia because both have a pappus with only one row of clade C) with four genera whose members are found in Mexico bristles. However, Bishopanthus has tri-nervate leaf venation and Central America, with one species also occurring in Co- and in Liabinae all the genera have pinnate leaf venation except lombia. There are at least two different opinions regarding Liabum which has tri-nervate venation but two rows of bristles. the geological history of Central America. Coates & Obando

450 TAXON 61 (2) • April 2012: 437–455 Funk & al. • Phylogeny and biogeography of Liabeae

(1996) and Coates & al. (2004) suggested that the isthmus was Mexico + Caribbean a series of volcanic islands ca. 15 Ma ago, while Kirby & al. Central 5–11 (2008) proposed that a peninsula existed as early as 19 Ma America ago, rather than the islands. However, both papers agree that 1 present day Central America was joined to South America ca. radiation A. Liabum 3 Ma ago. Prior to that time there was a deep marine channel. speciation If members of the Liabeae traveled by land in either direction it Andes must have been after 3 Ma ago. There is, of course, the “Proto- ~30

Greater Antilles” formed by the movement of the Caribbean Costa Rica Costa Rica + Panama Plate eastward which provided a single magmatic arc during the 1 Paleocene-Eocene (Draper & Barros, 1994) until the Oligocene 1 (Iturralde-Vinet, 1994) or early Miocene (Pindell & Barrett, B. Munnozia 1990) but these dates are all too old to be of significance here. range speciation extension The repeated movement of Liabeae from the Andes to North and Central America and the Caribbean seems to fall Andes ~39 into two types of hypotheses. The first is that it happened rather Costa Rica recently possibly by land during the last 3 Ma; these dispersal + Panama events resulted in little or no speciation (Munnozia, Oligactis, 1 Erato, Liabum bourgeaui Hieron.; Fig. 8A–D). The other appar- C. Oligactis ently happened long enough ago to result in a significant num- speciation ber of speciation events (Sinclairiinae and Caribbean Liabum; Fig. 8A, E). These were, most likely, the result of long distance Andes 6 dispersal from the northern Andes to the Caribbean (Fig. 8A) Costa Rica and from the central Andes to Mexico (Fig. 8E). 1 Supporting the first hypothesis is the fact that each one of D. Erato the Liabeae species in Mesoamerica (in Fig. 8A–D) is closely speciation related to one from the northern Andes, principally found in Co- lombia: Liabum bourgeaui is related to L. asclepiadeum Sch.- Andes Bip. (Colombia and Venezuela), Erato costaricensis V.A. Funk southern 4 & E. Moran is related to E. vulcanica (Klatt) H. Rob. (Colom- Mexico + Guatemala bia, Venezuela, Ecuador), and Munnozia wilberii H. Rob. is 26 range related to M. senecionidis Benth. (Costa Rica to Bolivia) or extension M. fosbergii H. Rob. (Colombia). Molecular data are not yet speciation and/or available for Oligactis valeri (Standl.) H. Rob. & Brettell but radiation all other species of this genus are from Colombia and Venezu- ela. The fact that most closely related taxa are in the northern Central Central Andes, mostly Colombia, supports the hypothesis that these America America 3 1 are recent events probably in the last three million years since radiation the connection of Panama with Colombia. range Although the distribution of Liabum is wide, most of the extension species are in Colombia and Ecuador as are those of its close E. Sinclairia & relatives, Oligactis and Sampera. The radiation of Liabum Sinclairiopsis Colombia into the Caribbean (Cuba, Jamaica, Hispaniola) has resulted Andes 1 in 5–11 species, depending on the species concept used. For Ancestor instance, Turner (1996) has proposed that all seven described taxa from Hispanola be sunk under L. poiteaui (Cass.) Urb. The Caribbean clade in Liabum is well-supported and highly nested Fig. 8. Dispersal figures: schematic illustration of the possible history in the genus (Figs. 2, 4, 8) but the rest of the species in the genus of five genera with distribution patterns that include areas outside the are mostly on short, unresolved branches (Fig. 2) because of few Andes. The arrows indicate dispersal or other movement of taxa. The differences in their molecular data. Likewise, the characters patterns found in Oligactis (C), Munnozia (B), Erato (D), and one spe- that separate them are sometimes difficult to discern because cies of Liabum (L. bourgeaui Hieron.) (A) show only range extensions of variability. The six “species” of Liabum in the Caribbean that or one new separate species and most likely reflect a recent movement from the Andes to Costa Rica and Panama, possibly 2 Ma when South were sampled for this study included representatives from all America was connected to Central America via the isthmus. In con- three islands and four countries. The Caribbean Liabum clade trast, the patterns in Liabum (A) with 5–11 species in the Great Antil- most likely represents a single introduction from the Andes les and Sinclairia (E) with 25 species in Mexico and Central America before the formation of the isthmus but after the magmatic arc reflect older connections. Numbers indicate the number of species in broke up and formed the Greater Antilles. that geographic area.

451 Funk & al. • Phylogeny and biogeography of Liabeae TAXON 61 (2) • April 2012: 437–455

All of the subtribes except Paranepheliinae (clade E) are Universidad Mayor de San Andrés, Bolivia; MEXU–Herbario Nacional very well supported (Fig. 4, Fig. S1), but of these four subtribes, at Universidad Nacional Autónoma de México; QCA–Herbario, Pontifi- the Sinclairiinae (clade C) has the most characters supporting cia Universidad Católica del Ecuador, Quito, Ecuador; USM–Herbario, it (Fig. 2). The Sinclairiinae is the only clade with an extensive Museo de Historia Natural, Universidad Nacional Mayor de San Marcos, radiation outside the Andes (ca. 26–30 species) and its place- Lima, Peru. Sadly, Raymundo Ramirez Delgadillo (ENSJ) who ac- ment as sister taxon of clades D + E makes it seem likely that companied Funk and A. Delgado S. (MEXU) into the field in Mexico this was the result of an older single colonizing event, possibly in 2005 died suddenly in 2010. Without his help we would never have a dispersal event from the central Andes. found so many of the we were looking for. We are grateful that Cody & al. (2010) examined the Great American Biotic fieldwork was made possible by a variety of grants from the Smithson- Interchange and concluded that plants had a greater propensity ian Institution (Scholarly Studies, Restricted Endowment, Lowland for dispersal over the isthmus before its closure as compared Tropical Ecosystem, and NMNH Research Opportunity Fund and its with animals. Cody & al.’s (2010) study also supported the idea successors). The cost of the lab work was defrayed by funds from the that plants made it from South America to Mexico, northern LAB at the NMNH. We thank I. Ventosa (HAC) and M. Dillon (F) who Central America and the Caribbean both before and after the kindly sent samples of leaf material from Cuba and Peru, respectively. closure of the isthmus. Our results, with older dispersal events We are grateful for the DNA for that was supplied and younger vicariant events support the idea that dispersal by the Hawaiian Plant DNA Library (Morden & al., 1996; Randell events occurred prior to the closure of the isthmus. & Morden, 1999; http://www.botany.hawaii.edu/faculty/morden/HPDL. htm); S.C. Harbin collected the Hesperomannia sample, extracted the DNA, and allowed us to use it before her project was finished. We also Nomenclature thank R. Capers (CONN), M. Dillon (F), N. Hind (K), L. Katinas (LP), S. Keeley (HAW), S. Knap (BM), G. McPherson (MO), P.B. Phillipson Sinclairiopsis is resurrected and the following name (MO), R. Rabeler (MICH), N. Roque (SPF), G. Schatz (MO), J. Soloman changes are recognized here: (MO), and T. Wendt (TEX) who helped track down vouchers and types.

Sinclairiopsis Rydb. in N. Amer. Fl. 34(4): 292–293. 1927 – Type: Sinclairiopsis klattii (B.L. Rob. & Greenm.) Rydb. Literature Cited in N. Amer. Fl. 34(4): 293. 1927 ≡ Liabum klattii B.L. Rob. & Greenm. in Amer. J. Sci., ser. 3, 50: 156. 1895 – Type: Akaike, H. 1974. A new look at the statistical model identification. Mexico. , Monte Alban, 8 Oct 1894, C.G. Pringle IEEE Trans. Automatic Control 19: 716–723. 6059 (holo­type: GH; isotypes: NY(1), US(3)!) Altekar, G., Dwarkadas, S., Huelsenbeck, J.P. & Ronquist, F. 2004. Parallel Metropolis coupled Markov chain Monte Carlo for Bayes- ian phylogenetic inference. Bioinformatics 20: 407–415. Sinclairiopsis ismaelis (Panero & Villaseñor) V.A. Funk, Bayer, R.J., Greber, D.G. & Bagnall, N.H. 2002. Phylogeny of Aus- comb. nov. ≡ Sinclairia ismaelis Panero & Villaseñor tralian Gnaphalieae (Asteraceae) based on chloroplast and nuclear in Brittonia 48(1): 85. 1996 – Type specimen: Mexico. sequences, the trnL intron, trnL/trnF intergenic spacer, matK and Oaxaca, Pto. Angel, 30 Sep 1993, Panero, Calzada & Sa- ETS. Syst. Bot. 27: 801–814. linas 3572 (holotype: MEXU; isotypes: MSC, TEX, US!) Blake, S.F. 1935. The genus Chionopappus of Bentham (Asteraceae). J. Washington Acad. Sci. 25: 488–493. Cabrera, A.L. 1954. Compuestas sudamericanas nuevas o críticas, II. Distephanus madagascariensis (Less.) H. Rob. & V.A. Funk, Notas Mus. La Plata 17: 71–80. comb. nov. ≡ Vernonia madagascariensis Less. in Linnaea Coates, A.G. & Obando, J.A. 1996. The geologic evolution of the 6: 644–645. 1831 – Type: Madagascar. Near Marou- Central American Isthmus. Pp. 21–56 in: Jackson, J.B.C., Budd, voai, Bojer s.n. (holotype: B, probably destroyed). A.F. & Coates, A.G. (eds.), Evolution and environment in tropical Lanjouw & Stafleu (1954) say the following: “Wenceslas America. Chicago: University of Chicago Press. Coates, A.G., Collins, L.S., Aubry, M.-P. & Berggren, W.A. 2004. (Wenzel) Bojer, Madagascar (1822–23, 1835): B, BM, C, G, The geology of the Darien, Panama, and the late Miocene-Pliocene G-DC, P, W(orig)”. Further study is necessary before a lecto- collision of the Panama Arc with northwestern South America. type or neotype can be designated. Bull. Geol. Soc. Amer. 116: 1327–1344. Cody, S., Richardson, J.E., Rull, V., Ellis, C. & Pennington, R.T. 2010. The Great American Biotic Interchange revisited. Ecography Acknowledgements 33: 326–332. Dillon, M.O., Funk, V.A., Robinson, H. & Chan, R. 2009. Liabeae. Pp. 417–437 in: Funk, V.A., Susanna, A., Stuessy, T.F. & Bayer, Funk wishes to thank her colleagues from a number of herbaria R.J. (eds.), Systematics, evolution, and biogeography of Composi- throughout for their help over the last twenty years in the tae. Vienna: International Association for Plant . field and in processing collected specimens (in alphabetical order based Downie, S.R. & Katz-Downie, D.S. 1996. A molecular phylogeny on herbarium abbreviation): COL–Herbario Nacional Colombiano, Uni- of Apiaceae subfamily Apioideae: Evidence from nuclear ribo- somal DNA internal transcribed spacer sequences. Amer. J. Bot. versidad Nacional de Colombia, Bogotá; CR–Museo Nacional de Costa 83: 234–251. Rica, San José, Costa Rica; INB–Instituto Nacional de Biodiversidad, Draper, G. & Barros, J.A. 1994. Cuba. Pp. 65–86 in: Donovan, S.K. & Santo Domingo, Costa Rica; LP–Herbario, División Plantas Vasculares, Jackson, T.A. (eds.), Caribbean geology: An introduction. Kings- Museo de La Plata, Argentina; LPB–Herbario Nacional de Bolivia, ton: University of the West Indies Publishers’ Association.

452 TAXON 61 (2) • April 2012: 437–455 Funk & al. • Phylogeny and biogeography of Liabeae

Farris, J.S. 1989. The retention index and the rescaled consistency Gregory-Wodzicki, K. 2011. Uplift history of the Central and Northern index. Cladistics 5: 417–419. Andes: A review. Bull. Geol. Soc. Amer. 112: 1091–1105. Farris, J.S., Källersjö, M., Kluge, A.G. & Bult, C. 1994. Testing Guindon, S. & Gascuel, O. 2003. A simple, fast, and accurate algorithm significance of incongruence. Cladistics 10: 315–319. to estimate large phylogenies by maximum likelihood. Syst. Biol. Felsenstein, J. 1985. Confidence limits on phylogenies: An approach 52: 696–704. using the bootstrap. Evolution 39: 783–791. Gutiérrez, D.G. 2010. Inkaliabum, a new Andean genus of Liabeae Funk, V.A. 1985. Cladistics and generic concepts in the Compositae. (Asteraceae) from Peru. Bol. Soc. Argent. Bot. 45: 363–372. Taxon 34: 72–80. Hammen, T. van der. 1974. The Pleistocene changes of vegetation and Funk, V.A. & Brooks, D.R. 1990. Phylogenetic systematics as the climate in tropical South America. J. Biogeogr. 1:3–26. basis of comparative biology. Smithsonian Contr. Bot. 73: 1–45. Hershkovitz, M.A. & Zimmer, E.A. 1996. Conservation patterns in Funk, V.A. & Chan, R. 2008. Phylogeny of the Spiny African Daisies angiosperm rDNA ITS2 sequences. Nucl. Acids Res. 24: 2857–2867. (Compositae, tribe Arctotideae, subtribe Gorteriinae) based on trnL- Huelsenbeck, J.P. & Ronquist, F. 2001. MRBAYES: Bayesian infer- F, ndhF, and ITS sequence data. Molec. Phylogenet. Evol. 48: 47–60. ence of phylogeny. Bioinformatics 17: 754–755. Funk, V.A. & Chan, R. 2009. Cichoroideae. Pp. 335–342 in: Funk, Iturralde-Vinet, M.A. 1994. Cuban geology: A new plate tectonic V.A., Susanna, A., Stuessy, T.F. & Bayer, R.J. (eds.) Systematics, synthesis. J. Petrol. Geol. 17: 39–70. evolution, and biogeography of Compositae. Vienna: International James, D.E. 1973. The evolution of the Andes. Sci. Amer. 229: 60–69. Association for Plant Taxonomy. Jansen, R.K. 1992. Current research. Pl. Molec. Evol. Newslett. 2: 13–14. Funk, V.A. & Robinson, H. 2001. A bully new genus from the Andes Johnson, L.A. & Soltis, D.E. 1994. matK DNA sequences and phyloge- (Compositae: Liabeae). Syst. Bot. 26: 216–225. netic reconstruction in Saxifragaceae s.str. Syst. Bot. 19: 143–156. Funk, V.A. & Robinson, H. 2009. Sampera, a new genus of Liabeae Jordan, T.E., Isacks, B.L., Allmendiger, R.W., Frewer, J.A., Ramos, (Compositae or Asteraceae) from the northern Andes. Proc. Biol. V.A. & Ando, C.J. 1983. Andean tectonics related to geometry of Soc. Washington 122: 155–161. subducted Nazca Plate. Bull. Geol. Soc. Amer. 94: 341–361. Funk, V.A., Anderberg, A.A., Baldwin, B.G., Bayer, R.J., Boni- Keeley, S.C., Forsman, A.H. & Chan, R. 2007. A phylogeny of the facino, J.M., Breitwieser, I., Brouillet, L., Carbajal, R., Chan, “evil tribe” (Vernonieae: Compositae) reveals Old/New World long R., Coutinho, A.X.P., Crawford, D.J., Crisci, J.V., Dillon, M.O., distance dispersal: Support from separate and combined congruent Freire, S.E., Galbany-Casals, M., Garcia-Jacas, N., Gemein- datasets (trnL-F, ndhF, ITS). Molec. Phylogenet. Evol. 44: 89–103. holzer, B., Gruenstaeudl, M., Hansen, H.V., Himmelreich, S., Kim, H.-G., Funk, V.A., Vlasek, A. & Zimmer, E.A. 2003. A phylog- Kadereit, J.W., Källersjö, M., Karaman-Castro. V., Karis, P.O., eny of the Munnoziinae (Compositae, Liabeae): Circumscription Katinas, L., Keeley, S.C., Kilian, N., Kimball, R.T., Lowrey, of Munnozia and a new placement of M. perfoliata. Pl. Syst. Evol. T.K., Lundberg, J., McKenzie, R.J., Mort, M.E., Nordenstam, 239: 171–186. B., Oberprieler, C., Ortiz, S., Pelser, P.B., Randle, C.P., Rob- Kirby, M.X., Jones, S.J. & MacFadden, B.J. 2008. Lower Miocene inson, H., Roque, N., Sancho, G., Semple, J.C., Serrano, M., stratigraphy along the Panama Canal and its bearing on the Cen- Stuessy, T.F., Susanna, A., Tadesse, M., Unwin, M., Urbatsch, tral American Peninsula. PLoS ONE 3: e2791, doi:10.1371/journal. L., Urtubey, E., Vallès, J., Vogt, R., Wagstaff, S. , Ward, J.M. pone.0002791 & Watson, L.E. 2009. Compositae metatrees: The next generation. Lanjouw, J. & Stafleu, F.A. 1954. Index herbariorum, pt. 2(1): Col- Pp. 747–777 in: Funk, V.A., Susanna, A., Stuessy, T.F. & Bayer, R. lectors A–D. Regnum Vegetabile 2. Utrecht: International Bureau (eds.), Systematics, evolution, and biogeography of Compositae. for Plant Taxonomy and Nomenclature. Vienna: IAPT. Morden, C.W., Caraway, V.C. & Motley, T.J. 1996. Development of a Funk, V.A., Chan, R. & Holland, A. 2007a. (Compositae: DNA library for native Hawaiian plants. Pacific Sci. 50: 324–335. Arctotideae, Arctotidinae): An endemic Australian genus embed- Nylander, J.A., Wilgenbusch, J.C., Warren, D.L. & Swofford, D.L. ded in a southern African clade. Bot. J. Linn. Soc. 153: 1–8. 2008. AWTY (Are We There Yet?): A system for graphical explo- Funk, V.A., Chan, R. & Keeley, S. 2004. Insights into the evolution ration of MCMC convergence in Bayesian . Bioin- of the tribe Arctoteae (Compositae) using trnL, ndhF, and ITS. formatics 24: 581–583. Taxon 53: 637–655. Panero, J.L. & Funk, V.A. 2008. The value of sampling anomalous Funk, V.A., Robinson, H. & Dillon, M.O. 1996. Liabeae: Taxon- taxa in phylogenetic studies: Major clades of the Asteraceae re- omy, phylogeny and biogeography. Pp. 545–567 in: Hind, D.J.N., vealed. Molec. Phylogenet. Evol. 47: 757–782. Beentje, H.J. & Caligari, P.D.S (eds.), Compositae: Systematics. Panero, J.L. & Villaseñor, J.L. 1996. Novelties in Asteraceae from Proceedings of the International Compositae Conference, Kew, southern Mexico. Brittonia 48: 79–90. 1994, vol. 1. Kew: Royal Botanic Gardens. Pindell, J. & Barrett, S.F. 1990. Geological evolution of the Caribbean: Funk, V.A., Robinson, H. & Dillon, M.O. 2007b. Liabeae (Astera- A plate tectonic perspective. Pp 405–432 in: Dengo, G. & Chase, ceae). Pp. 175–180 in: Kadereit, J.W. & Jeffrey, C. (eds.), The J.E. (eds.), The geology of North America, vol. H, The Caribbean families and genera of vascular plants, vol. 8, Flowering plants: Region. Boulder: Geological Society of America. ; . Berlin: Springer. Posada, D. 2008. jModelTest: Phylogenetic model averaging. Molec. Funk, V.A., Robinson, H., McKee, G.S. & Pruski, J.F. 1995. Neo- Biol. Evol. 25: 1253–1256. tropical montane Compositae with an emphasis on the Andes. Pp. Rambaut, A. & Drummond, A.J. 2003–2007. Tracer, version 1.5: MCMC 451–471 in: Churchill, S.P., Balslev, H., Forero, E. & Luteyn, J.L. trace analysis package. http://tree.bio.ed.ac.uk/software/tracer. (eds.), Biodiversity and conservation of neotropical montane for- Randell, R.A. & Morden, C.W. 1999. Hawaiian plant DNA library II: En- ests. New York: New York Botanical Garden. demic, indigenous, and introduced species. Pacific Sci. 53: 401–417. Garver, J.I., Montario, M., Perry, S.E., Reiners, P.W. & Ramage, Reeves, J.H. 1992. Heterogeneity in the substitution process of amino J.R. 2005. Uplift and exhumation of the northern Peruvian Andes. acid sites of proteins coded for by mitochondrial DNA. J. Molec. Pp. 305–307 in: Sempéré, T. (ed.) 6th International Symposium on Evol. 35, 17–31. Andean Geodynamics: Extended abstracts. IRD Éditions, Univ. de Robinson, H. 1983a. A generic review of the tribe Liabeae (Asteraceae). Barcelona, Instituto Geológico y Minero de España. Smithsonian Contr. Bot. 54: 1–69. Gentry, A.H. 1982. Neotropical floristic diversity: Phytogeographical Robinson, H. 1983b. Studies in the Liabeae (Asteraceae). XVI. New connections between Central and South America, Pleistocene cli- Taxa from Peru. Phytologia 54: 62–65. matic fluctuations, or an accident of the Andean orogeny? Ann. Robinson, H. & Brettell, R.D. 1973. Tribal revisions in the Asteraceae Missouri Bot. Gard. 69: 557–593. III: A new tribe, Liabeae. Phytologia 25: 404–407.

453 Funk & al. • Phylogeny and biogeography of Liabeae TAXON 61 (2) • April 2012: 437–455

Robinson, H. & Brettell, R.D. 1974. Studies in the Liabeae (Astera- Turner, B.L. 1989. Revisionary treatment of the genus Sinclairia, in- ceae), II: Preliminary survey of the genera. Phytologia 28: 43–63. cluding Liabellum (Asteraceae, Liabeae). Phytologia 67: 168–206. Robinson, H. & Funk, V.A. 2011. Stephanbeckia plumosa (Liabeae: Turner, B.L. 1996. The genus Liabum (Asteraceae: Liabeae) in the Compositae): A new genus and species from southern Bolivia. Dominican Republic and Haiti. Phytologia 80: 115–117. Brittonia 63: 75–82. Turner, B.L. 2007. The comps of Mexico: A systematic account of the Ronquist, F. & Huelsenbeck, J.P. 2003. MRBAYES 3: Bayesian phylo- family Asteraceae, vol. 8, Liabeae and Vernonieae. Phytologia genetic inference under mixed models. Bioinformatics 19: 1572–1574. Mem. 12: 1–144. Rydberg, A. 1927. (Carduales) Carduaceae, Liabeae, Neurolaeneae, Vuilleumier, F. 1969. Pleistocene speciation in birds living in the high Senecioneae (pars). North Amer. Fl. 34: 289–360. Andes. Nature 223: 1179–1180. Sandwith, N.Y. 1956. Contributions to the Flora of Tropical America, White, T.J., Bruns, T., Lee, S. & Taylor, J. 1990. Amplification and di- LXI: Notes on Philoglossa. Kew Bull. 11: 289–293. rect sequencing of fungal ribosomal RNA genes for phylogenetics. Simpson, B.B. 1975. Pleistocene changes in the flora of the high tropical Pp. 315–322 in: Innis, M.A., Gelfand, D.H., Sninsky, J.J. & White, Andes. Paleobiology 1: 273–294. T.J. (eds.), PCR protocols: A guide to methods and applications. Soejima, D.D., Wen, J., Zapata, M. & Dillon, M.O. 2008. Phylogeny San Diego: Academic Press. and putative hybridization in the subtribe Paranepheliinae (Lia- Wilgenbusch, J.C., Warren, D.L. & Swofford, D.L. 2004. AWTY: A beae, Asteraceae), implications for classification, biogeography, system for graphical exploration of MCMC convergence in Bayes- and Andean orogeny. J. Syst. Evol. 46: 375–390. ian phylogenetic inference. http://ceb.csit.fsu.edu/awty. Steele, K.P. & Vilgalys, R. 1994. Phylogenetic analyses of Polemonia- Windley, B.F. 1984. The evolving continents, 2nd ed. Chichester, New ceae using nucleotide sequences of the plastid gene matK. Syst. York, Brisbane, Toronto, Singapore: John Wiley. Bot. 19: 126–142. Yang, Z., 1993. Maximum-likelihood estimation of phylogeny from Swofford, D.L. 2011. PAUP*: Phylogenetic analysis using parsimony DNA sequences when substitution rates differ over sites. Molec. (*and other methods), version 4.0a114. Sunderland: Sinauer. Biol. Evol. 10: 1396–1401. Taberlet, P., Gielly, L., Pautou, G. & Bouvet, J. 1991. Universal prim- Yang, Z. & Rannala, B. 1997. Bayesian phylogenetic inference using ers for amplification of three non-coding regions of chloroplast DNA sequences: A Markov chain Monte Carlo method. Molec. DNA. Pl. Molec. Biol. 17: 1105–1109. Biol. Evol. 14: 717–724. Taylor, D.W. 1991. Paleobiogeographic relationships of Andean angio- Zwickl, D.J. 2006. Genetic algorithm approaches for the phylogenetic sperms of Cretaceous to Pliocene age. Palaeogeogr. Palaeoclima- analysis of large biological sequence datasets under the maximum tol. Palaeoecol. 88: 69–84. likelihood criterion. Ph.D. Dissertation, University of Texas, Austin.

Appendix. Voucher Table: Each sentence is a separate voucher and follows the same format: Tube number, name, country, 1st political division, date, col- lector & number, herbarium, ITS, trnL-F, ndhf, matK. A dash (–) indicates a missing sequence. In order to check the identifications an attempt was made to find every voucher from previously published sequences (“?” next to the herbarium designation: the specimen has not been located; “none*”: endangered species that has been collected before, no specimen taken). Three samples were taken from plants grown from seeds harvested from the living collection at Kew in the early days of collecting molecular data; these are indicated by the long three-part number in the “collection number” field. Unfortunately, these three vouchers could not be found at CONN or K; they are listed as “none”. Previously published sequences are indicated by a superscript: 1 = Funk & al., 2004; 2 = Funk & al., 2007a; 3 = Funk & Chan, 2008; 4 = Keeley & al., 2007. Ingroup Liabeae: 238, Cacosmia harlingii B. Nord., Ecuador: Loja, 21 Feb 1993, Harling & Stahl 26577, US, JN837120, JN837210, JN837300, JN837409. 197, C. hieronymus H. Rob., Ecuador: Azuay, 24 Oct 1997, Lewis 3645, US, JN837119, JN837209, JN837299, JN837408. 174, C. rugosa Kunth in HBK, Peru: Amazonas, 1 Jul 1997, Sánchez V. & Dillon 9083, F, MO, JN837118, JN837208, JN837298, JN837407. 441, Chionopappus benthamii S.F. Blake, Peru: Ancash, 3 Nov 2007, Leiva & al. 4169, US, JN837177, JN837267, JN837357, JN837461. 286, Ch. benthamii S.F. Blake, Peru: Cajamarca, 14 Jul 1991, Sagastegui & al. 14448, F, JN837176, JN837266, JN837356, JN837460. 208, Ch. benthamii S.F. Blake, Peru: La Libertad, 11 May 2004, Sagastegui & al. 17543, F, JN837175, JN837265, JN837355, JN837459. 175, Chrysactinium acuale (Kunth in HBK) Wedd., Peru: Amazonas, 12 Nov 2000, Sánchez V., Dillon & Zapata 10327, F, JN837158, JN837248, JN837338, JN837442. 254, Ch. hieracioides (Kunth in HBK) H. Rob. & Brettell, Peru: Cajamarca, 1 Apr 1987, Becker & al. 1674, US, JN837160, JN837250, JN837340, JN837444. 223, Ch. hieracioides (Kunth in HBK) H. Rob. & Brettell, Peru: Cajamarca, 1 Mar 1988, Panero 1130, US, JN837159, JN837249, JN837339, JN837443. 199, Dillandia perfoliata (Blake) H. Rob. & Brettell, Ecuador: Carchi, 20 Jul 1992, Panero & Clark 3038, US, JN837125, JN837215, JN837305, JN837414. 211, Erato costaricensis E. Moran & V.A. Funk, Costa Rica: San José, 18 Aug 1994, Kress (Funk & Zermoglio) 4814, US, JN837166, JN837256, JN837346, JN837450. 201, E. polymnioides DC., Ecuador: Morona-Santiago, 26 Oct 1995, Funk & Torracchi 11455, US, JN837164, JN837254, JN837344, JN837448. 202, E. sodiroi (Hieron) H. Rob., Ecuador: Chimborozo, 5 Jul 1992, Panero 2930, US, JN837165, JN837255, JN837345, JN837449. 259, Ferreyranthus gentrii H. Rob., Peru: Amazonas, 19 Jul 1995, Sánchez V. 8137, MO, US, JN837123, JN837213, JN837303, JN837412. 176, F. rugosus (Ferreyra) H. Rob. & Brettell, Peru: La Lib- ertad, 18 May 2004, Sagastegui & al. 17569, F, JN837121, JN837211, JN837301, JN837410. 177, Ferreyranthus verbasifolia (Kunth) H. Rob. & Brettell, Peru: Cajamarca, 11 Jan 1990, Dillon 6099, F, JN837122, JN837212, JN837302, JN837411. 185, Liabellum angustissimum (A. Gray) Rydburg, Mexico: , 23 Sep 2005, Funk, Delgado & Ramírez 12605, US, JN837189, JN837279, JN837369, JN837472. 339, L. cervinum Rydb., Mexico: , 30 Aug 1957, McVaugh 16583, US, JN837188, JN837278, JN837368, JN837471. 186, L. palmerii (A. Gray) Rydburg, Mexico: Jalisco, 23 Sep 2005, Funk, Delgado & Ramírez 12603, US, JN837190, JN837280, JN837370, JN837473. 203, Liabum acuminatum Rusby, Bolivia: Santa Cruz, 17 Oct 1999, Nee 50174, US, JN837131, JN837221, JN837311, –. 257, L. asclepiadeum Sch. Bip., Colombia: Putumayo, 2 Jan 1963, Bristol 441, US, JN837134, JN837224, JN837314, JN837422. 204, L. barahonense Urban, Dominican Republic: Barahona, 21 Jul 1996, Funk & Zermoglio 11464, US, JN837139, JN837229, JN837319, JN837426. 147, L. bourgeaui Hieron, Costa Rica: San Isidro, 17 Jun 2002, Redden 999, US, JN837128, JN837218, JN837308, JN837417. 461, Liabum sp., Cuba: Guantánamo, 15 Mar 2009, Ventosa s.n., HAC, JN837143, JN837233, JN837323, JN837430. 269, L. floribundum Less., Ecuador: Loja, 23 Jul 1996, Lewis 2460, US, JN837136, JN837226, JN837316, JN837423. 239, L. grandiflorum Less., Ecuador: Loja, 1 Jun 2003, Croat & Menke 89956, MO, JN837133, JN837223, JN837313, JN837421. 206, L. igniarium (Kunth in HBK) Less, Ecuador: Pi- chincha, 17 Jul 1992, Panero & Clark 3012, US, JN837132, JN837222, JN837312, JN837420. 270, L. kingii H. Rob., Ecuador: Pichincha, 27 Oct 1989, Borchsenius 91419, US, JN837137, JN837227, JN837317, JN837424. 246, L. poiteaui Urban, Dominican Republic: Barahona, 26 Jun 2006, Pruski & Ortiz 4061, US, JN837138, JN837228, JN837318, JN837425. 284, L. selleanum Urban, Dominican Republic: Elías Piña, 24 Jun 2003, Acevedo 13366, US, JN837142, JN837232, JN837322, JN837429. 178, L. solidagineum DC., Peru: Amazonas, 1 Jul 1997, Sánchez V. & Dillon 9085, F, JN837129, JN837219, JN837309, JN837418. 283, L. subacaule Rydb., Dominican Republic: Santiago, 1 Jun 2004, Acevedo 14178, US, JN837141, JN837231, JN837321, JN837428. 207, L. umbellatum Sch. Bip., Jamaica: St. Andrew, 9 Jul 1996, Funk & Zermoglio 11462, US, JN837140, JN837230, JN837320, JN837427. 266, L. vargasii H. Rob., Peru: Cuzco, 15 Sep 2002, Galiano & al. 4418, MO, JN837135, JN837225, JN837315, –. 179, L. wurdackii Ferreyra, Peru: Amazonas, 7 Jul 1997, Sánchez V. & Dillon 9094, F, MO, JN837130, JN837220, JN837310, JN837419. 187, Megaliabum andrieuxii (DC) Rydburg, Mexico: , 27 Sep 2005, Funk & Montero-Castro 12617, US, MEXU, JN837191, JN837281, JN837371, JN837474. 188, M. pringlei (R&G) Rydburg, Mexico: Jalisco, 23 Sep 2005, Funk, Delgado & Ramírez 12606, US, MEXU, JN837192, JN837282, JN837372, JN837475. 209, Microliabum mulgediifolium (Muschler) H. Rob., Bolivia: Tarija, 23 Mar 1986, Ehrich 188, US, JN837170, JN837260, JN837350,

454 TAXON 61 (2) • April 2012: 437–455 Funk & al. • Phylogeny and biogeography of Liabeae

Appendix. Continued JN837454. 173, M. polymnioides (R.E. Fr.) H. Rob., Argentina: Salta, 4 Jan 1998, Funk & Rankin 12086, US, LP, JN837168, JN837258, JN837348, JN837452. 235, M. polymnioides (R.E. Fr.) H. Rob., Argentina: Salta, 14 Mar 1996, Hawkes & Rahn 3852, MO, JN837169, JN837259, JN837349, JN837453. 210, Munnozia campii H. Rob., Ecuador: Morona-Santiago, 26 Oct 1995, Funk & Torracchi 11456, US, JN837152, JN837242, JN837332, JN837438. 249, M. foliosa Rusby, Bolivia: La Paz, 29 Jun 1988, Lewis 88161, US, JN837144, JN837234, JN837324, JN837431. 520, M. foliosa Rusby, Bolivia: La Paz, 29 Feb 2002, Michel 3030, US, JN837145, JN837235, JN837325, –. 119, M. fosbergii H. Rob., Colombia: Boyaca, 21 Aug 1997, Funk & Mendoza 12039, US, JN837146, JN837236, JN837326, –. K39, M. gigantia (Rusby) Rusby, Peru: Madre de Dios (cult.), 27 Nov 1985, Dillon 9200, F, AY5046971, AY5047391, AY5047791, JN837432. 190, M. hastifolia (Poepp. & Endl.) H. Rob. & Brettell, Colombia: Cundinamarca, 27 Aug 1997, Funk & Mendez 12050, US, JN837150, JN837240, JN837330, JN837436. 180, M. jussieui (Cass.) H. Rob. & Brettell, Peru: Piura, 19 Oct 2001, Sagastegui & al. 16768, F, HAO, JN837147, JN837237, JN837327, JN837433. 212, M. lyrata (A. Gray) H. Rob. & Brettell, Peru: Cajamarca, 10 Mar 1988, Panero 1201, US, JN837157, JN837247, JN837337, JN837441. 191, M. maronii (André) H. Rob., Argentina: Salta, 4 Jan 1998, Funk & Rankin 12087, US, LP, JN837151, JN837241, JN837331, JN837437. 258, M. nivea (Hieron.) H. Rob. & Brettell, Ecuador: Tungurahua, 3 Mar 1989, Buitron 464, US, JN837156, JN837246, JN837336, –. 256, M. pinnatipartita (Hieron.) H. Rob. & Brettell, Ecuador: Morona-Santiago, 26 Jun 2003, Clark & Kat- zenstein 8388, US, JN837155, JN837245, JN837335, –. 181, M. sagasteguii H. Rob., Peru: Cajamarca, 12 May 2004, Sagastegui & al. 17562, F, JN837148, JN837238, JN837328, JN837434. 189, M. senecionidis Benth., Colombia: Boyaca, 21 Aug 1997, Funk & Mendoza 12028, US, LP, JN837149, JN837239, JN837329, JN837435. 221, M. wilburii H. Rob., Costa Rica: Alajuela, 25 Feb 1992, Almeda & Daniel 7068, US, JN837154, JN837244, JN837334, JN837440. 213, M. wilburii H. Rob., Costa Rica: Puntarenas, 14 Aug 1994, Kress (Funk & Zermoglio) 4802, US, JN837153, JN837243, JN837333, JN837439. 261, Oligactis sessiliflora (Kunth) H. Rob & Brettell, Colombia: Boyaca, 21 Aug 1997, Funk & Mendoza 12031, US, JN837124, JN837214, JN837304, –. 121, O. volubilis (Kunth) Cass., Colombia: Cundina- marca, 26 Aug 1997, Funk & Méndez 12042, US, AY5046981, AY5047801, AY5047401, JN837413. 215, Paranephelius asperifolius (Muschl.) H. Rob. & Brettell, Argentina: Salta, 6 Jan 1998, Funk & Rankin 12088, US, LP, JN837174, JN837264, JN837354, JN837458. 192, P. ovatus Wedd., Bolivia: La Paz, 27 Apr 1995, Funk 11393, US, LPB, JN837173, JN837263, JN837353, JN837457. 120, Philoglossa mimuloides (Hieron.) H. Rob. & Cuatr., Ecuador: Azuay, 25 Oct 1995, Funk & Tor- racchi 11453, US, AY5046991, AY5047811, AY5047411, JN837445. 183, Ph. mimuloides (Hieron.) H. Rob. & Cuatr., Peru: La Libertad, 11 May 2004, Sagastegui & al. 17539, F, JN837161, JN837251, JN837341, JN837446. 232, Ph. peruviana DC., Peru: Lima, 14 Sep 1986, Knapp 8311, US, JN837163, JN837253, JN837343, –. 216, Ph. purpureodisca H. Rob., Peru: Trujillo, 6 Oct 1996, Leiva & Suárez 1889, US, JN837162, JN837252, JN837342, JN837447. 184, Pseudonoseris discolor (Muschl.) H. Rob. & Brettell, Peru: Puno, 28 May 2005, Quipuscoa & Caceres 3338, F, JN837171, JN837261, JN837351, JN837455. 217, P. szyszylowiczii (Hieron.) H. Rob & Brettell, Peru: Amazonas, 22 May 1962, Wurdack 467, US, JN837172, JN837262, JN837352, JN837456. 214, Sampera coriacea (Hieron.) V.A. Funk & H. Rob., Ecuador: Loja, 2 Jun 1984, Ollgaard 74615, US, JN837127, JN837217, JN837307, JN837416. 182, S. cuatrecasasii (M.O. Dillon & Sagast.) V.A. Funk & H. Rob., Peru: Piura, 20 Oct 2001, Sagastegui & al. 16823, F, JN837126, JN837216, JN837306, JN837415. 193, Sinclairia caducifolia Rydb., Mexico: Chiapas, 27 Sep 2005, Funk & Montero-Castro 12614, US, JN837179, JN837269, JN837359, JN837463. 194, S. deamii Rydb., Mexico: Chiapas, 27 Sep 2005, Funk & Montero- Castro 12616, US, JN837180, JN837270, JN837360, JN837464. 195, S. discolor Hook. & Arn., Mexico: Chiapas, 29 Sep 2005, Funk & Montero-Castro 12624, US, JN837181, JN837271, JN837361, JN837465. 218, S. glabra var. hypoleuca (Greenm.) B.L. Turner, Mexico: Chiapas, 27 Sep 2005, Funk & Montero-Castro 12618, US, JN837182, JN837272, JN837362, JN837466. 281, S. ismaelis Panero & Villaseñor, Mexico: Oaxaca, 30 Sep 1993, Panero 3572, MEXU, US, JN837193, JN837283, JN837373, JN837476. 280, S. liebmannii Sch. Bip., Mexico: Colima, 10 Dec 1959, McVaugh & Koelz 1642, MICH, US, JN837185, JN837275, JN837365, JN837469. 219, S. moorei (H. Rob. & Brettell) H. Rob & Brettell, Mexico: Jalisco, 23 Sep 2005, Funk, Delgado & Ramírez 12607, US, JN837183, JN837273, JN837363, JN837467. 148, S. polyantha Rydb., Costa Rica: Alajuela, 30 Apr 1987, Funk & al. 10106, US, JN837178, JN837268, JN837358, JN837462. 250, S. sublobata Rydb., El Salvador: Usulatán, 29 Jan 1999, Williams 15, US, JN837184, JN837274, JN837364, JN837468. 287, S. vagans (S.F. Blake) H. Rob. & Brettell, Guatemala: Chimaltenango, 27 Nov 1993, Castillo & al. 2073, F, JN837186, JN837276, JN837366, –. 337, S. similis (McVaugh) H. Rob. & Brettell, Mexico: Jalisco, 24 Sep 2005, Funk, Delgado & Ramírez 12609, US, JN837187, JN837277, JN837367, JN837470. 240, Sinclairiopsis klattii Rydb., Mexico: Oaxaca, 30 Oct 1974, Breedlove 39185, MO, US, JN837194, JN837284, JN837374, –. 426, Stephenbeckii plumosa H. Rob. & V.A. Funk, Bolivia: Tarija, 17 Apr 2000, Beck 27047, LPB, US, JN837167, JN837257, JN837347, JN837451. — Outgroups: Arctoteae: 3, calendula (L.) Levyns, South Africa: NW Cape, 21 Aug 1999, Trinder-Smith 143, US, DQ8896292, DQ8896452, DQ8896612, JN837378. 95, Arctotheca sp., South Africa: E Camp, 8 Sep 2000, Funk, Cowling, MacKinnon & Clark 12266, US, AY5047031, AY5047851, AY5047451, JN837379. 8, Arctotis bellidifolia Berg., South Africa: W Cape, 13 Sep 2000, Koekemoer & Funk 1926, PRE, AY5047041, AY5047861, AY5047461, JN837380. 10, A. cuprea Jacq., South Africa: W Cape, 14 Sep 2000, Koekemoer & Funk 1939, PRE, DQ8896312, DQ8896472, DQ8896632, JN837381. 14, A. fastuosa Jacq., South Africa: NW Cape, 25 Aug 1999, Trinder-Smith 238, US, AY5047051, AY5047871, AY5047471, JN837382. 271, A. hirsuta Beauv., South Africa: N Cape, 22 Aug 2004, Funk & Koekemoer 12491, US, JN837105, JN837195, JN837286, JN837383. 28, Berkheya spinosissima Willd., South Africa: N Cape, 30 Aug 1999, Trinder-Smith 346, US, EU5272003, EU5272503, EU5273003, JN837386. 50, Didelta carnosa Ait., South Africa: W Cape, 2 Jan 2000, Koekemoer & Funk 1943, PRE, AY5047161, AY5047981, AY5047581, JN837385. 49, D. spinosa (L. f.) Ait., South Africa: W Cape, 14 Sep 2000, Koekemoer & Funk 1936, PRE, EU5272213, EU5272713, EU5273183, JN837384. 139, nervosa Beauv., South Africa: Lesotho, 14 Jan 2003, Funk & Koekemoer 12417, US, PRE, DQ8896432, DQ8896592, DQ8896742, JN837375. 76, H. ruppallii (Sch. Bip.) A. Rich., Kenya: Meru District, 16 Aug 1985, Robertson & al. 3960, MO, DQ8896402, DQ8896562, JN837285, JN837376. 77, H. scaposa Harv., South Africa: Lesotho, 3 Feb 2000, Trinder-Smith 191, US, AY5047081, AY5047901, AY5047501, JN837377. Eremothamneae: 295, Eremothamnus marlothianus O. Hoffm., Namibia: Karas, 11 Aug 2007, Funk & Koekemoer 12684, US, JN837107, JN837197, JN837288, JN837388. 297, Hoplophyllum spinosum DC., South Africa: N Cape, 6 Aug 2007, Funk & Koekemoer 12650, US, JN837106, JN837196, JN837287, JN837387. Moquinieae: 355, racemosa DC., Brazil: Bahia, 9 Nov 2007, Roque 1691, ALCB, JN837117, JN837207, –, JN837406. 354, Pseudostifftia kingii H. Rob., Brazil: Bahia, 1 Feb 2008, Roque 1754, ALCB, JN837116, JN837206, JN837297, JN837405. Ver- nonieae: K10, Baccharoides adoensis (Sch.Bip ex Walp.) H. Rob., Nigeria (cult.), 1968, Kew living collection, now dead (donor: Ibadan Univ.) 453-68-45301, none, EF1557454, EF1556574, EF1558334, JN837399. K18, B. lasiopus (O. Hoffm.) H. Rob., Africa (cult.), n.d., Keeley s.n., accession #98542, CONN, EF1557964, EF1557084, EF1558844, JN837400. K64, Centrapalus pauciflorus (Willd.) H. Rob., Ethiopia: Hararge, 9 Dec 1964, Perdue 6333, K, USDA, EF1557514, EF1556634, EF1558394, JN837402. CJ, punctatum Cass., Brazil (cult.), 24 Jun 1905, Panero s.n., TEX, EF1557544, EF1556664, EF1558424, JN837403. K47, Critoniopsis sodiroi (Hieron. Ex Sodiro) H. Rob., Ecuador: Pichincha, 1986, Keeley s.n., US, EF1557604, EF1556724, EF1558484, JN837401. 502, Distephanus ambongensis (Humbert) H. Rob., Madagascar: Antsiranana, 23 Oct 2009, van Ee & al. 1065, US, JN837115, JN837205, JN837296, –. 500, D. barus (Humbert) H. Rob., Madagascar: Antananarivo, 20 Oct 2009, van Ee & al. 1015, US, JN837113, JN837203, JN837294, –. 501, D. garnierianus (Klatt) H. Rob. & B. Kahn, Madagascar: Antananarivo, 20 Oct 2009, van Ee & al. 1016, US, JN837114, JN837204, JN837295, –. V415, D. madagascariensis (Less.) H. Rob. & V.A. Funk, Madagascar: Mahajanga, 20 Jun 1987, Phillipson 1905, MO, EF1557614, EF1556734, EF1558494, –. K12, D. populifolius (Lam.) Cass., Mascarene Islands (cult.), 1985, Kew living collection (donor: Strahm) 282-85-03275, none, EF1557624, EF1556744, EF1558504, JN837404. 292, misera (Oliv. & Hiern) S. Moore, Namibia: Kavango, 21 Aug 2007, Funk & Koekemoer 12708, US, JN837109, JN837199, JN837290, JN837391. K11, amygdalinum Sch.Bip. Ex Walp., West Africa (cult.), 1986, Kew living collection, now dead (doner Badra) 318-86-2802, none, AY5046951, AY5047371, AY5047771, JN837394. K112, G. mesipifolium (Less.) H. Rob., Africa (cult.), not available, Jansen 995, MICH?, EF1557754, EF1556874, EF1558634, JN837395. Hau2, Hillebr., Hawaii: Oahu, Makaha Valley, not available, Ching-Harbin P2, accession #2753, none*, JN837112, JN837202, JN837293, JN837397. K40, Lepidonia jonesii (B.L. Turner) H. Rob.& V.A. Funk, Mexico: Oaxaca, 7 Jan 1989, Todzia & al. 2835, TEX, EF1557884, EF1557004, EF1558764, JN837389. 293, Lessingiant- hus warmingianus (Baker) H. Rob., Brazil: Minas Gerais, 29 Sep 1998, Roque & Funk 485, US, JN837110, JN837200, JN837291, JN837392. K38, Linzia melleri (Oliv. & Hiern.) H. Rob., Burundi: Bururi, 1981, Reekmans 10202, MO?, EF1557924, EF1557044, EF1558804, JN837398. 291, Polydora poskeana (Vatke & Hildebr.) H. Rob., Botswana: Central District, 24 Aug 2007, Funk & Koekemoer 12713, US, JN837108, JN837198, JN837289, JN837390. 321, Protiopsis argentea Mart. & Zucc. ex DC., Brazil: Minas Gerais, 30 Sep 1998, Roque & Funk 496, US, SPF, JN837111, JN837201, JN837292, JN837393. 228, arboreum (Buch.- Ham.) H. Rob., Singapore: Bukit Timah Nature Reserve, 20 Apr 2003, Lum & Chan s.n., US, EF1557744, EF1556864, EF1558624, JN837396.

455