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Botanical Journal of the Linnean Society, 2013, 173, 387–406. With 2 figures

Phylogenetic relationships and new tribal delimitations in subfamily ()

KENT KAINULAINEN1,2*, SYLVAIN G. RAZAFIMANDIMBISON1,2 and BIRGITTA BREMER1,2

1Bergius Foundation, The Royal Swedish Academy of Sciences, SE-106 91 Stockholm, Sweden 2Department of Ecology, Environment and Sciences, Stockholm University, SE-106 91 Stockholm, Sweden

Received 4 April 2012; revised 2 October 2012; accepted for publication 31 January 2013

Subfamily Ixoroideae is one of three major lineages in Rubiaceae, with approximately 4000 . Previous molecular phylogenetic studies have indicated that many genera and tribes previously placed in other subfamilies are better considered as part of Ixoroideae. However, the internal resolution and clade support have generally been low, and several genera found to be nested in the subfamily do not appear to be associated with any described tribe. In order to resolve the phylogeny and assess the tribal delimitations in the expanded Ixoroideae, phylogenetic reconstructions were performed using Bayesian and parsimony analyses of six plastid DNA regions and a broad sampling of genera from all tribes of the subfamily. In the inferred phylogenetic hypotheses, the tribal relationships were mostly well supported, with Ixoroideae consisting of the and the alliances as sister groups and a grade comprising , Henriquezieae, Posoquerieae, Retiniphylleae, Sipaneeae and the Steenisia. A revised tribal classification, including the description of five new tribes, Airospermeae, Augusteae, Scyphiphoreae, Steenisieae and Trailliaedoxeae, is provided. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173, 387–406.

ADDITIONAL KEYWORDS: Airospermeae – Augusteae – molecular phylogenetics – Scyphiphoreae – Steenisieae – systematics – Trailliaedoxeae.

INTRODUCTION include Ixoroideae in a wide (see also Robbrecht & Manen, 2006). In a comprehensive clas- With >13 000 species (Davis et al., 2009), Rubiaceae is sification of the , Robbrecht (1988) restricted one of the largest and most diverse families of Ixoroideae to include genera with contorted corolla angiosperms. Well-known representatives include lobe aestivation and fleshy . In his system, L., L. and J.Ellis, all part of Ixoroideae comprised tribes Cremasporeae, Garde- Ixoroideae, a subfamily comprising about 4000 species nieae and Ixoreae sensu Bremekamp (the correct of pantropical and subtropical distributions. Ixoroi- name of which would have been Coffeeae; Darwin, deae, as recognized by Bremekamp (1952, 1966), 1976), i.e. Aulacocalyceae, Coffeeae, , included taxa characterized by secondary pollen pres- Octotropideae and Pavetteae. For an overview of the entation (‘ixoroid pollination mechanism’), a feature changes in the classification of Ixoroideae, the reader that Bremekamp considered to be of high taxonomic is referred to Andreasen & Bremer (2000). value because of its requirement for a combina- Results of molecular phylogenetic studies have since tion of characters for functionality. Included were then indicated many unknown relationships and also, tribes Acranthereae, Chiococceae, Coptosapelteae, consequently, possible improvements in the classifica- Cremasporeae, Gardenieae, Ixoreae and Vanguerieae. tion of Rubiaceae. This includes the expansion of Verdcourt (1958), however, considered pollen presen- Ixoroideae, as many additional tribes or segregates of tation to be of less importance, and preferred to tribes and a number of taxa of uncertain taxonomic placement have been shown to be associated with the *Corresponding author. E-mail: [email protected] subfamily. Ixora, the type of the subfamily, was shown

© 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173, 387–406 387 388 K. KAINULAINEN ET AL. to form a clade with Vanguerieae (Bremer et al., 1999), Kainulainen et al., 2009; Razafimandimbison et al., confirming the position of Vanguerieae in Ixoroideae, 2011). Subsequent tribal recognition of the lineages as suggested by Bremekamp (1952, 1966), and sup- represented by these genera has so far only been porting the exclusion of Ixora from Pavetteae. performed for Aleisanthia and Aleisanthiopsis and Andreasen & Bremer (2000) resurrected Ixoreae and (Aleisanthieae and Greeneeae, respectively; provided new circumscription for several other tribes Mouly et al., 2009a). in the subfamily. Many additional taxa have been In this study, we analyse data from six combined found to be associated with Ixoroideae sensu Robbrecht plastid DNA regions (matK, ndhF, rbcL, the rps16 (1988), including Aubl. (Bremer, Andreasen & intron, trnS-G and trnT-F) using Bayesian and par- Olsson, 1995), and Sabiceeae, (Bremer & simony methods of phylogenetic reconstruction to Thulin, 1998), Alberteae and C.F.Gaertn. resolve the tribal relationships of the expanded (Bremer et al., 1999), Condamineeae and many genera Ixoroideae and, in particular, the relationships of the formerly placed in or Rondeletieae (e.g. genera which, in previous studies, have not been Bremer et al., 1999; Rova et al., 2002; Delprete & associated with any previously described tribes. Fol- Cortés-B., 2004; Kainulainen et al., 2010). Crossop- lowing the inferred phylogenetic hypothesis, we also teryx Fenzl, part of Ixoroideae sensu Bremekamp provide a revised tribal classification of the subfamily. (included in Coptosapelteae), but considered part of Cinchoneae by Robbrecht (1988), has also been found to be nested in Ixoroideae (Razafimandimbison & MATERIAL AND METHODS Bremer, 2001), as have Mussendopsis (Razafimandim- bison & Bremer, 2001), Hekistocarpa Hook.f. (Dessein TAXON SAMPLING et al., 2001), Standl., Humb. This study includes genera representing all tribes & Bonpl. and Sipaneeae (Rova et al., 2002; Delprete & associated with Ixoroideae in previous molecular phy- Cortés-B., 2004), Boholia Merr. and Steenisia Bakh.f. logenetic studies. The ingroup included 110 species (Bremer & Eriksson, 2009), Airosperma K.Schum. & from 87 genera. An effort was made to sample prima- Lauterb. (Kainulainen et al., 2009), Merr. rily material representing the types of genera. Gelse- (Alejandro et al., 2010), Glionnetia Tirveng., Jackiop- mium sempervirens (L.) J.St.-Hil. (Gelsemiaceae), sis Ridsdale and Trailliaedoxa W.W.Sm. & Forrest Logania vaginalis (Labill.) F.Muell. (Loganiaceae), (Razafimandimbison et al., 2011). Although the gratissima (Wall.) Sweet, Colletoecema dew- expanded Ixoroideae is well supported by molecular evrei (De Wild.) E.M.A.Petit, Ophiorrhiza mungos L. data, internal resolution and clade support, especially and six species of Cinchonoideae were utilized as among the early divergent clades, have generally been outgroup. Because the focus of the study is primarily low. concerned with the phylogeny of the early divergent Ixoroideae, as widely delimited according to recent lineages of the subfamily, sampling was limited in the molecular phylogenetic studies, includes all tribes of large and problematic Gardenieae and associated Robbrecht’s (1988) narrow circumscription of the sub- tribes (Octotropideae and Pavetteae), the phylogeny family, whereas tribes Acranthereae, Chiococceae and of which will be investigated further in an upcoming Coptosapelteae are not part of Ixoroideae as sug- study (A. Mouly et al., Université de Franche-Comté, gested by Bremekamp (1966). Chiococceae has been Besançon, unpubl. data). Sampling was increased for shown to be part of Cinchonoideae (Bremer et al., Airosperma, and , all of which 1995), and Acranthera Arn. ex Meisn. and Cop- have widely disjunct distributions and in previous tosapelta Korth. appear to represent a separate, early molecular phylogenetic studies have been resolved in divergent clade in the family (Rydin et al., 2009). The Ixoroideae, but not in association with any previously expanded Ixoroideae is morphologically diverse and recognized tribes (Rova et al., 2002; Kainulainen can no longer be easily characterized morphologically. et al., 2009). We were able to obtain sequences Molecular phylogenetic studies have also shown that of Airosperma taxa from New Guinea and , includ- some tribes of the subfamily are not monophyletic (i.e. ing A. trichotomum (Gillespie) A.C.Sm., originally Gardenieae including Aulacocalyceae, and Pavetteae; described as Abramsia trichotoma Gillespie. The Andreasen & Bremer, 2000; Persson, 2000; Bremer & Augusta sample comprised specimens of the type Eriksson, 2009) and that several genera found to be A. longifolia (Spreng.) Rehder from , A. austro- nested in Ixoroideae do not appear to be associated caledonica (Brongn.) J.H.Kirkbr. from with any described tribe, i.e. Airosperma, Aleisanthia and A. rivalis (Benth.) J.H.Kirkbr. from Central Ridl., Aleisanthiopsis Tange, Augusta Pohl, Boholia, America, the last two species of which were previ- Glionnetia, Greenea Wight & Arn., Scyphiphora, ously recognized as Lindenia Benth. The Wendlandia Steenisia, Trailliaedoxa and Wendlandia Bartl. ex sample included W. arabica Deflers and W. ligus- DC. (Rova et al., 2002; Bremer & Eriksson, 2009; troides (Boiss. & Hohen.) Blakelock, the inclusion of

© 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173, 387–406 PHYLOGENY OF IXOROIDEAE 389 which in Wendlandia was considered to be doubtful in L. plastid genome (GenBank accession the last comprehensive revision of the genus by number, EF044213; Samson et al., 2007). An AT-rich Cowan (1932). We were unsuccessful in obtaining region in the trnS-G spacer, corresponding to posi- DNA sequences from any Steenisia spp. other than tions 9184–9203 of the C. arabica plastid genome, S. pleurocarpa (Airy Shaw) Bakh.f. An overview of the was excluded from the analyses because of difficulties sampling and voucher specimens is given in the in finding an unambiguous alignment. The combined Appendix, and a few species representing the differ- dataset consisted of 10 165 characters, 10 003 of ent tribes of Ixoroideae are shown in Figure 1. which were included in the analyses. Methods of phylogenetic reconstruction included maximum par- simony (MP) and Bayesian inference (BI; Yang & DNA EXTRACTION, AMPLIFICATION AND SEQUENCING Rannala, 1997), using the programs PAUP* v4.0b10 Three protein coding genes (matK, ndhF and rbcL) (Swofford, 2002) and MrBayes v3.1.2 (Huelsenbeck & and three noncoding regions [the rps16 intron, the Ronquist, 2001; Ronquist & Huelsenbeck, 2003), trnS-G spacer including the trnG intron (trnSGCU- respectively. Gaps were treated as missing data. MP trnGUUC-trnGUUC) and the trnT-F region (trnTUGU- analyses were performed using heuristic searches trnLUAA-trnLUAA-trnFGAA)] of the plastid DNA, with the bisection–reconnection (TBR) branch previously shown to be phylogenetically informative swapping algorithm, Multrees on, 1000 random in Rubiaceae (Andersson & Rova, 1999; Bremer et al., sequence addition replicates and a maximum of 10 1999; Razafimandimbison & Bremer, 2002; Rova saved per replicate. Clade support was esti- et al., 2002; Andersson & Antonelli, 2005; mated using 1000 bootstrap replicates, with three Kainulainen et al., 2009), were used in this study. random addition replicates per replicate. For the BI DNA was extracted from material using a Mini- analyses, the protein coding and noncoding data were Beadbeater 3110BX (BioSpec), following the protocol analysed as separate partitions with unlinked model of Doyle & Dickson (1987), and was subsequently parameter estimates (except topology). Substitution purified using the QIAquick® PCR purification kit models suggested as best fit to the data under the according to the manufacturer’s instructions corrected Akaike information criterion (AICc), as (Qiagen). PCR amplifications were performed using implemented in MrAIC v1.4.4 (Nylander, 2004; a standard PCR settings and the following primers: script dependent on the program PHYML v3.0; matK1198F and matK2053R (Andersson & Antonelli, Guindon & Gascuel, 2003), were used for each parti- 2005) for matK; 2F, 1000R, 720F, 1700R, 1320F and tion (the GTR + G + I model for both datasets). The 2280R (Rydin, Razafimandimbison & Bremer, 2008) analysis comprised two runs of four chains each and for ndhF; rbcL5’F, rbcL_Z895F, rbcL_Z895R and was monitored for 107 generations, with every 1000th rbcL3’R (Bremer et al., 2002) for rbcL; rsp16_2F generation being sampled. The chain heating param- (Bremer et al., 2002) and rpsR2 (Oxelman, Lidén & eter was 0.10. The initial 25% of the sampled trees Berglund, 1997) for the rps16 intron; and cF, eF, dR, were considered as burn-in and excluded from the fR (Taberlet et al., 1991), A1F, IR (Bremer et al., consensus. The effective sample sizes (ESSs) of the 2002), 820R, 940F, 1250F, 1880F and 2670R (Rydin model parameters were checked using the program et al., 2008) for trnT-F. The trnS-G region was ampli- Tracer v1.5 (Rambaut & Drummond, 2007), in order fied using the primers and protocols of Shaw et al. to ensure an ESS > 200, as recommended by (2005). Sequences were assembled using the Staden Drummond et al. (2007) for the adequate representa- package v1.5.3 (Staden, 1996). Sequences new to this tion of the posterior probability (PP). study (324) were deposited in GenBank (Appendix). Additional sequences (361) were obtained from RESULTS GenBank (for references, see Appendix). The combined dataset contained 10 003 DNA charac- ters, 2410 of which were potentially phylogenetically DATA ANALYSES informative in the MP analysis. Analyses of the coding The sequences were preliminarily aligned using and noncoding datasets separately (data not shown) Clustal W (default settings; Thompson, Higgins & showed no incongruences with bootstrap support (BS) Gibson, 1994), as implemented in BioEdit (Hall, values Ն70% or PPs Ն 0.95, with the exception of the 1999), and then adjusted manually. Sequence inver- position of L., which formed a sister group to sions were dealt with by separating the alternative Salisb. (PP, 0.99; BS, 73%) in the coding sequence versions in the alignment. Inversions in the dataset, but was resolved as sister group to a clade included sequences occurred in regions corresponding comprising Fagerl. and to positions 5417–5430 (rps16), 9082–9113, 9164– Aubl. in the noncoding dataset (PP, 0.95; BS, 84%). The 9169, 9330–9335 and 9714–9723 (trnS-G)ofthe inferred phylogenetic hypotheses of the MP and BI

© 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173, 387–406 390 K. KAINULAINEN ET AL.

ABC D

E F G

GH I J

KLJ M N

Figure 1. Selected taxa of Ixoroideae: A, longiflora;B,Schizomussaenda henryi;C, pervillei;D, Sabicea diversifolia;E,Steenisia pleurocarpa;F,Scyphiphora hydrophylacea;G,Glionnetia sericea;H,Ixora borboniae;I, Augusta rivalis;J,Wendlandia tinctoria;K,Bertiera rufa;L, sp.; M, axillare;N,Rosenbergiodendron densiflorum (image credits: A–D, G, H, K–N, by Kent Kainulainen; E, F, J, © Christian Puff, Faculty Centre of Biodiversity, University of Vienna; used with permission; I, by Indiana Coronado, Tropicos, botanical information system at the Missouri Botanical Garden – http://www.tropicos.org, © MBG, licensed under a Creative Commons by-nc-nd 3.0 licence).

© 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173, 387–406 PHYLOGENY OF IXOROIDEAE 391 analyses were overall mostly congruent, well resolved six combined plastid regions. The phylogenetic and strongly supported. Figure 2 shows the majority hypotheses indicate that Ixoroideae can be con- rule consensus tree from the BI analyses of the com- sidered as comprising the Coffeeae alliance, the Van- bined data, including the PPs of the clades and the guerieae alliance and a grade consisting of Retini- clade BS values of the MP analysis. phylleae, Steenisia, Mussaendeae with Sabiceeae The clades corresponding to the Coffeeae and Van- and a Condamineeae–Henriquezieae–Posoquerieae– guerieae alliances, respectively (cf. Razafiman- Sipaneeae clade (Fig. 2). Henriquezieae, Posoquerieae dimbison et al., 2011), were resolved as sister groups and Sipaneeae form a clade, as in the study by Rova (‘the core Ixoroideae’; PP, 1.00; BS, 100%). The Coffeeae et al. (2002). This clade forms a weakly supported alliance comprised the tribes Alberteae, Bertiereae, sister group to Condamineeae, and the Condami- Coffeeae, Gardenieae, Octotropideae and Pavetteae neeae–Henriquezieae–Posoquerieae–Sipaneeae clade and the genera Airosperma, Augusta, Boholia and is, in turn, sister group to the remainder of the Wendlandia. Gardenieae did not appear to be mono- subfamily. This is congruent with the results of Rydin phyletic, because Burchellia R.Br. was resolved as et al. (2009). However, in the phylogenetic hypotheses sister group to Octotropideae (PP, 1.00; BS, 64%), and of Kainulainen et al. (2009), Condamineeae appeared Mitriostigma Hochst. and DC. formed a closer to core Ixoroideae. Further research is needed sister group to Pavetteae, although with weak support in order to better understand the relationships of (PP, 0.80; BS, 54%). The Gardenieae–Pavetteae clade these early divergent lineages. As in previous studies was resolved as sister group to the Burchellia– (Rova et al., 2002; Bremer & Eriksson, 2009; Octotropideae clade (PP, 1.00; BS, 65%), and a clade Kainulainen et al., 2010), the resolution in Condami- comprising Bertiereae and Coffeeae (PP, 1.00; BS, neeae was low. 98%), in turn, formed a sister group to this clade (PP, In the phylogenetic tree presented, Mussaendeae is 1.00; BS, 100%). Strongly supported successive sister resolved as sister group to Sabiceeae with strong groups were Alberteae (PP, 1.00; BS, 85%), an support, in agreement with the results of Alejandro, Augusta–Wendlandia clade (PP, 1.00; BS, 100%) and Razafimandimbison & Liede-Schumann (2005). As, in an Airosperma–Boholia clade (PP, 1.00; BS 100%). their study, DC. forms a sister group to the In the Vanguerieae alliance, Aleisanthieae formed a remaining genera of the tribe, and L. sister group to Ixoreae (PP, 1.00; BS, 100%) and this forms a clade with Pseudomussaenda Wernham., clade was, in turn, sister to Greeneeae (PP, 1.00; BS, Schizomussaenda H.L.Li (Fig. 1B) from South-East 100%). Relationships between the Aleisanthieae– Asia is here further resolved as sister group to a clade Greeneeae–Ixoreae clade, Vanguerieae and Glionne- comprising the Malagasy–Mascarene genera Breme- tia were unresolved. Although strongly supported by ria Razafim. & Alejandro (Fig. 1C) and Landiopsis the BI analyses (PP, 1.00), the support for this tri- Capuron ex Bosser. In Sabiceeae, Bremek. chotomy was low in the MP analysis (< 50%). Trail- is resolved as sister group to Tamridaea Thulin & liaedoxa was resolved as sister to the clade formed by B.Bremer, as in previous studies by Bremer & Thulin this trichotomy. However, this clade also had low (1998) and Khan et al. (2008). These genera, together support (PP, 0.83; BS, 54%). Successive sister groups with Hekistocarpa, have been suggested as constitut- were Scyphiphora (PP, 1.00; BS, 87%), Jackiopsis (PP, ing a clade recognized at tribal level (Virectarieae s.l., 1.00; BS, 97%) and (PP, 1.00; BS, 91%). based on molecular data) by Dessein et al. (2001), and A grade of successive sister clades to core Ixoroideae the three genera also formed a well-supported clade comprised, in turn, Retiniphylleae (PP, 1.00; BS, in the study by Bremer & Eriksson (2009). These 100%), Steenisia (PP, 1.00; BS, 100%) and a clade relationships are not supported in our study, in which comprising the sister tribes Mussaendeae and Sabi- Hekistocarpa instead forms a poorly supported sister ceeae (PP, 0.98; BS, 54%). The Mussaendeae– group to Sabicea Aubl. However, previously, Hekisto- Sabiceeae clade was well supported (PP, 1.00; BS, carpa has also been resolved as sister group to the 96%). A Condamineeae–Henriquezieae–Posoquerieae– remaining Sabiceeae (Khan et al., 2008; based on Sipaneeae clade was resolved as the earliest diverging nrDNA data). Phylogenetic relationships in this tribe clade in relation to the rest of the subfamily. However, should be investigated further. The position of Retini- the support for this clade was low (PP, 0.79; BS, 72%). phyllum indicated in this study conforms with that Subfamily Ixoroideae received strong support (PP, first found in the study by Rova et al. (2002). 1.00; BS, 100%). Cortés-B., Delprete & Motley (2009) have subse- quently demonstrated the monophyly of the monoge- neric Retiniphylleae. DISCUSSION The South-East Asian genus Steenisia (Fig. 1E) was In this study, we infer the tribal relationships segregated from Neurocalyx Hook. (subfamily Rubio- in Ixoroideae by the phylogenetic reconstruction of ideae) by Bakhuizen van den Brink (1952). In a

© 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173, 387–406 392 K. KAINULAINEN ET AL.

Gelsemium sempervirens Logania vaginalis Luculia gratissima 0.61/- Colletoecema dewevrei 1.00/100 Ophiorrhiza mungos Cinchona pubescens Rondeletia odorata 1.00/100 1.00/100 speciosa Cubanola domingensis 0.54/- Cephalanthus occidentalis 1.00/100 Nauclea orientalis 1.00/100 nitida - HEN 1.00/ Molopanthera paniculata - POS 100 1.00/100 - POS 1.00/100 Posoqueria longiflora - POS 1.00/ Chalepophyllum guyanense - SIP 100 1.00/100 Dendrosipanea spigelioides - SIP 1.00/100 Neobertiera gracilis - SIP 1.00/100 Sipanea aff. biflora - SIP 0.79/72 1.00/100 Sipanea hispida - SIP 1.00/97 bracteata - CON flavidum - CON henryi - CON 1.00/100 Dioicodendron dioicum - CON corymbosa - CON Pogonopus speciousus - CON speciosa - CON 0.99/84 cordifolia - CON 0.82/- triflora - CON 0.97/56 thibaudioides - CON candidissimum - CON 1.00/100 hookeri - CON 1.00/100 Ixoroideae 1.00/100 stipulata - CON Virectaria multiflora - SAB 1.00/100 Tamridaea capsulifera - SAB 1.00/ Hekistocarpa minutiflora - SAB 100 0.86/- Sabicea diversifolia - SAB 1.00/100 Sabicea aspera - SAB 1.00/96 0.54/72 Sabicea africana - SAB - MUS Pseudomussaenda flava - MUS 1.00/100 1.00/ - MUS 100 1.00/100 Mussaenda arcuata - MUS 1.00/100 Schizomussaenda henryi - MUS STEENISIEAE 1.00/96 Bremeria landia - MUS 0.98/54 1.00/99 Landiopsis capuronii - MUS SCYPHIPHOREAE Steenisia pleurocarpa Retiniphyllum pilosum - RET TRAILLIAEDOXEAE Crossopteryx febrifuga - CRO Jackiopsis ornata - JAC 1.00/91 Scyphiphora hydrophylacea 1.00/97 Trailliaedoxa gracilis 1.00/100 Glionnetia sericea 1.00/87 Greenea oblonga - GRE Aleisanthia rupestris - ALE 0.83/54 1.00/ 1.00/100 Aleisanthiopsis distantiflora - ALE 1.00/100 100 Ixora novoguineensis - IXO Vanguerieae-alliance 1.00/100 Ixora margaretae - IXO 1.00/- 1.00/100 1.00/66 - IXO 0.90/- Ixora ferrea - IXO 0.87/- Ixora borboniae - IXO 1.00/92 Ixora trilocularis - IXO coromandelicum - VAN senegalensis - VAN 1.00/ 1.00/ tetraquerta - VAN 1.00/100 core Ixoroideae 100 1.00/98 100 madagascariensis - VAN 1.00/99 concrescens - VAN 1.00/100 obovata - VAN Afrocanthium lactescens - VAN 1.00/100 1.00/87 gueinzii - VAN deplanchei - VAN 1.00/88 horridum - VAN AIROSPERMEAE Coffeeae-alliance 0.98/74 hystrix - VAN Boholia nematostylis Airosperma sp. 1.00/100 1.00/88 Airosperma psychotrioides 0.93/59 Airosperma trichotomum 0.94/73 Airosperma vanuense

Augusta austrocaledonica AUGUSTEAE 1.00/100 Augusta longifolia 1.00/83 1.00/83 Augusta rivalis 1.00/100 Wendlandia ligustroides 1.00/100 Wendlandia formosana 1.00/100 Wendlandia tinctoria 1.00/91 Wendlandia arabica 0.80/72 Wendlandia paniculata 1.00/100 - ALB Bertiera guianensis - BER 1.00/100 Bertiera longithyrsa - BER 1.00/76 Bertiera aethiopica - BER Henriquezieae 1.00/98 Empogona coriacea - COF Posoquerieae 1.00/85 1.00/100 Coffea ebracteolata - COF Sipaneeae 1.00/100 Coffea sessiliflora - COF 1.00/100 Coffea arabica - COF Condamineeae Burchellia bubalina - GAR triflora - OCT 1.00/100 1.00/64 Paragenipa lancifolia - OCT Sabiceeae 1.00/100 0.65/50 Hypobathrum racemosum - OCT 1.00/100 buxifolia - OCT Mussaendeae 0.97/85 apodanthera - OCT Retiniphylleae Oxyanthus speciosus - GAR Trailliaedoxeae 1.00/65 1.00/100 Mitriostigma axillaris - GAR Aleisanthieae Pavetta abyssinica - PAV Ixoreae 0.80/54 1.00/ Pavetta indica - PAV 1.00/100 100 Tennantia sennii - PAV Vanguerieae 1.00/94 supra-axillare - PAV americana - GAR 1.00/76 Airospermeae 1.00/100 Gardenia hansemanni - GAR 1.00/100 rudis - GAR Augusteae 1.00/98 tentaculata - GAR capensis - GAR Bertiereae 1.00/95 1.00/99 jasminiflora - GAR Coffeeae balansaeanus - GAR Octotropideae pavettifolia - GAR 1.00/83 1.00/98 spinosa - GAR Pavetteae 0.65/- penduliflora - GAR 0.92/68 grevei - GAR Gardenieae 1.00/97 Euclinia longiflora - GAR - GAR 0.02 1.00/100 0.90/51 - GAR 1.00/100 Rosenbergiodendron densiflorum - GAR

Figure 2. The majority-rule consensus tree from the Bayesian inference analysis of the combined plastid data (ndhF, matK, rbcL, rps16 intron, trnS-G and trnT-F). Posterior probabilities and bootstrap support values from the parsimony analysis are indicated below the clades (a dash indicates bootstrap support of <50%). Phylogram of the tree with the highest marginal likelihood in inset. New tribal names presented in this article are highlighted. ALB, Alberteae; ALE, Aleisanthieae; BER, Bertiereae; COF, Coffeeae; CON, Condamineeae; CRO, Crossopterygeae; GAR, Gardenieae; GRE, Greeneeae; HEN, Henriquezieae; IXO, Ixoreae; JAC, Jackieae; MUS, Mussaendeae; OCT, Octotropideae; PAV, Pavetteae; POS, Posoquerieae; RET, Retiniphylleae; SAB, Sabiceeae; SIP, Sipaneeae; VAN, Vanguerieae.

© 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173, 387–406 PHYLOGENY OF IXOROIDEAE 393 revision of the genus, Bremer (1984) proposed that it ered as restricted to the New World; cf. Manns & should be transferred from Argostemmateae to Ron- Bremer, 2010). Despite the inclusion of more sequence deletieae (subfamily Cinchonoideae). However, in the data in this study, the phylogenetic position of Glion- molecular phylogenetic study of Bremer & Eriksson netia is still unresolved. (2009), Steenisia was found nested in Ixoroideae. Stee- As in the study by Razafimandimbison et al. (2011), nisia can be readily characterized by a suffrutescent successive sister groups to the Aleisanthieae– unicaulous habit, rotate corolla and anthers with Glionnetia–Greeneeae–Ixoreae clade are Trailliae- apical appendages fused to form a hollow cone around doxa, Scyphiphora, Jackieae and Crossopterygeae. the style (in a manner reminiscent of the flowers of The little-studied Trailliaedoxa has hitherto re- Solanum L.), characters which are all unusual (the mained without taxonomic placement in Rubiaceae. It latter unique) in Ixoroideae. A region of the style occurs in the highlands of southern , although it covered with hairs that probably catch released pollen has rarely been collected. Trailliaedoxa is an erect is usually found level with the pollen sacs (Bremer, shrublet with minute schizocarpic fruits with solitary 1984; Puff et al., 1995). However, as this region does pendulous ovules. The corolla aestivation is left- not appear to extend above the surrounding staminal contort. Whether or not secondary pollen presentation tube, pollen is not presented in a secondary manner. occurs is not known. Our results support the recog- Instead, the mode of pollination is most likely by buzz nition of a monogeneric tribe, Trailliaedoxeae (see pollination (Puff et al., 1995). Steenisia is not associ- Taxonomic synopsis section). ated with any previously described tribe in our analy- The plant Scyphiphora (Fig. 1F) occurs ses and, consequently, the recognition of the Steenisia in coastal areas of tropical Asia and the western clade at the tribal level is proposed in the Taxonomic Pacific (Puff & Rohrhofer, 1993). Scyphiphora has a synopsis section. complex taxonomic history (see Puff & Rohrhofer, 1993; Mouly et al., 2009a), as the highly modified fruits, in particular, which are adapted for sea dis- THE VANGUERIEAE ALLIANCE persal, have caused difficulties in assessing its sys- The Vanguerieae alliance comprises Aleisanthieae, tematic placement. The fruits are indehiscent and Crossopterygeae, Glionnetia, Greeneeae, Ixoreae, corky, with one ascending and one descending ovule Jackieae, Scyphiphora, Trailliaedoxa and Van- per locule (Puff & Rohrhofer, 1993). However, the guerieae. The clades resolved in Vanguerieae in this flowers have secondary pollen presentation and left- study are congruent with the results of Lantz & contort corolla aestivation, characteristics shared by Bremer (2004). However, the position of Psydrax most of the core Ixoroideae. Andreasen & Bremer Gaertn. is here further resolved in a clade consisting (2000) tentatively included the genus in Ixoreae. of Afrocanthium (Bridson) Lantz & B.Bremer, Keetia However, in the studies by Bremer et al. (1999), E.Phillips and a clade comprising Cyclophyllum Bremer & Eriksson (2009) and Cortés-B. et al. (2009), Hook.f., Peponidium (Baill.) Arènes and Pyrostria Scyphiphora was resolved as sister to the Ixoreae– Comm. ex Juss., representing the ‘dioecious group’ of Vanguerieae clade, a position congruent with this Razafimandimbison et al. (2009). In agreement with study, in which Scyphiphora is resolved as sister to the results of Rova et al. (2002) and Mouly et al. the Aleisanthieae–Glionnetia–Greeneeae–Ixoreae– (2009a), Ixoreae forms a sister group to Aleisanthieae, Trailliaedoxa–Vanguerieae clade. Consequently, we and these are, in turn, sister to Greeneeae. Aleisan- propose the recognition of tribe Scyphiphoreae (see thia and Greenea both have traditionally been consid- Taxonomic synopsis section). ered part of Rondeletieae (Hooker, 1873), a tribe characterized by contorted or imbricate corolla aesti- vation and capsular fruits. Molecular phylogenetic THE COFFEEAE ALLIANCE studies have indicated that this is an unnatural The Coffeeae alliance comprises Airosperma, Augusta, group, and Rondeletieae has since been much reduced Boholia, Wendlandia and tribes Alberteae, Coffeeae, (see Rova et al., 2002; Manns & Bremer, 2010; Rova, Gardenieae, Octotropideae and Pavetteae. As in pre- Delprete & Bremer, 2009). vious molecular studies (Andreasen & Bremer, 1996; Glionnetia (Fig. 1G), an endemic of the Seychelles, Bremer & Eriksson, 2009), Gardenieae is not mono- has also been suggested as part of Rondeletieae by its phyletic. Mitriostigma (Fig. 1M) and Oxyanthus form author (Tirvengadum, 1984). However, in a recent a poorly supported sister group to Pavetteae, and study by Razafimandimbison et al. (2011), Glionnetia Burchellia forms a sister group to Octotropideae. was found nested in the Vanguerieae alliance, Further research is needed to better understand although with an unresolved position relative to Van- the phylogenetic relationships in the Gardenieae– guerieae and the Aleisanthieae–Greeneeae–Ixoreae Octotropideae–Pavetteae complex. Successive sister clade (Rondeletieae s.s. can consequently be consid- groups to this complex are the Bertiereae–Coffeeae

© 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173, 387–406 394 K. KAINULAINEN ET AL. clade, Alberteae, the Augusta–Wendlandia clade and included species from the respective areas form sister the Airosperma–Boholia clade. The included repre- groups). One of the Fijian species, A. trichotomum, sentatives of Bertiera form a monophyletic group was originally described by Gillespie (1932) as sister to Coffeeae, a relationship also inferred in other Abramsia trichotoma. Smith (1945: 108) considered studies (e.g. Bremer et al., 1995; Persson, 2000; Davis the generic concepts as ‘essentially identical’ and sub- et al., 2007; see the latter for a discussion on the sumed Abramsia. This synonymization is supported delimitation and morphological differentiation of by the molecular data, because A. trichotomum is these tribes, which some authors have treated as one, nested in Airosperma, forming a sister group to i.e. Andreasen & Bremer, 2000; Robbrecht & Manen, A. vanuense S.P.Darwin (Fig. 2). Secondary pollen 2006). presentation is not present in Airosperma (Darwin, Augusta and Wendlandia have traditionally 1979) and at least some of the New Guinean species (Hooker, 1873) been considered as part of Rondele- of the genus appear to be functionally dioecious tieae. However, in the molecular phylogenetic study of (Darwin, 1980). The status of this character in Rova et al. (2002), they were resolved as a clade in Boholia is not known. Although having been Ixoroideae. Both genera have widely disjunct distri- described as right-contorted or imbricate, the corolla butions. In Augusta, A. longifolia occurs in north- aestivation is left-contorted in both genera (K. Kai- eastern and central Brazil (Delprete, 1997), A. rivalis nulainen, pers. observ.). The fruits are fleshy and (Fig. 1I) in Central America, A. austrocaledonica in contain pyrenes, and were interpreted by Robbrecht New Caledonia and A. vitiensis (Seem.) J.H.Kirkbr. in (1988) as adapted for dispersal by sea currents. Fiji. The last three species have previously been rec- However, animal dispersal also seems a possibility ognized as Lindenia, a name that was reduced to considering the vivid colour of the fruits (blue–purple, synonymy by Kirkbride (1997). This is supported by white or yellow; Darwin, 1980). In this study, as in our results, because A. rivalis and A. austrocal- that of Kainulainen et al. (2009), they form a clade edonica are paraphyletic with respect to A. longifolia separate from Alberteae; consequently, we propose (Fig. 2). Wendlandia (Fig. 1J) has a wide distribution that this clade should be recognized at the tribal level in East Asia and Australasia (centre of diversity in (see Taxonomic synopsis section). China), with one species, W. ligustroides, in south- eastern Turkey–northern Iraq and one species, W. arabica, in , and Yemen (see Puff, TAXONOMIC SYNOPSIS 1990). Cowan (1932), who provided the last compre- Following the results of the molecular phylogenetic hensive revision of the genus, considered the last two analyses, we present here a revised tribal classifica- species as exceptional, and their inclusion in Wend- tion of Ixoroideae, although we do not address the landia as doubtful. However, Wendlandia appears to tribal delimitation in the problematic Gardenieae be monophyletic in our phylogenetic analyses, in complex, i.e. Gardenieae, Octotropideae (including which W. ligustroides forms a sister group to the Cremasporeae) and Pavetteae, which will be dis- remaining included species, whereas W. arabica is cussed further by A. Mouly et al. (Université de nested in the East Asian clade. Secondary pollen Franche-Comté, Besançon; unpubl. data). Including presentation appears to be absent in Wendlandia (D. these three tribes, Ixoroideae comprises 24 tribes, five Zhang, South China Botanical Garden, Guangzhou, of which are newly and formally described below. Guangdong, China, pers. comm.). The status of this Although the molecular support for Ixoroideae is character in Augusta is not known. Both genera have strong, morphologically the members are diverse and left-contorted corolla aestivation. In the Coffeeae alli- not easily characterized; no potential morphological ance, the Augusta–Wendlandia clade is distinguished synapomorphies for the subfamily are known at this by capsular fruits. This clade is not associated with point. In particular, Condamineeae and Hen- any previously described tribe; consequently, we riquezieae contain morphologically aberrant genera propose that it should be recognized at tribal level (Rogers, 1984; Kainulainen et al., 2010), some of (see Taxonomic synopsis section). which, at times, have been classified outside Airosperma and Boholia were placed in Alberteae Rubiaceae [e.g. Kuhlm. in Dialyp- by their respective authors (Schumann & Lauterbach, etalanthaceae (Rizzini & Occhioni, 1949) and Hen- 1900; Merrill, 1926) based on the presence of solitary riquezia Spruce ex Benth. and Platycarpum Humb. & pendulous ovules and contorted corolla aestivation. Bonpl. in Henriqueziaceae (Bremekamp, 1957)]. Boholia occurs in the , but has also been collected on the island of Flores in Indonesia (Schmutz 2691; P; image seen). Airosperma has a SUBFAMILY IXOROIDEAE RAF. disjunct distribution, occurring in New Guinea and Trees, , monocaulous treelets, subshrubs or the Fiji islands (in the phylogenetic hypothesis, the rarely herbs. Raphides absent. Stipules entire, bifid

© 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173, 387–406 PHYLOGENY OF IXOROIDEAE 395 or rarely fimbricate; interpetiolar or rarely intrapeti- Inclusion based on Alejandro et al. (2010): Green- olar. usually actinomorphic, protandrous, iopsis, occurring in South-East Asia. often with secondary pollen presentation; rarely zygo- morphic, heterostylous, dioecious, monoecious or pro- Tribe Augusteae Kainul. & B.Bremer, trib. nov. togynous. Calyces mostly persistent, in several taxa Type: Augusta Pohl, Pl. Bras. Icon. Descr. 2: 1 with one or several lobes expanded and showy. (1828) Corolla aestivation contorted to left, rarely to right, Frutices vel arbusculae. Inflorescentiae cymae termi- imbricate, valvate or rarely open. Fruits fleshy and nales compositae aut raro corymbi. Flores 5-meri. indehiscent, dry and indehiscent or capsular, with Calyx persistens. Corolla tubulari-infundibuliformis, numerous, few or solitary ovules per locule. lobis in aestivatione sinistrorsum contortis. Ovarium biloculare, ovulis numerosis horizontaliter insertis. Tribe Airospermeae Kainul. & B.Bremer, trib. nov. Stylus exsertus. Fructus capsulares, dehiscentia sep- Type: Airosperma K.Schum. & Lauterb., Fl. ticida (Augusta) aut loculicida (Wendlandia). Semina Schutzgeb. Südsee: 565 (1900) minuta, complanata, testa reticulata. Suffrutices, frutices vel arbusculae unicaules. Flores Shrubs to small trees. Inflorescences terminal com- 5-meri in paniculis terminalibus. Calyx persistens. pound cymes or rarely corymbs. Flowers 5-merous. Corolla brevi- vel longi-tubularis, lobis in aestivatione Calyx persistent. Corolla tubular-funnel-shaped, with sinistrorsum contortis. Ovarium biloculare, ovulo uno left-contorted aestivation. Ovary bilocular, pendulo in quoque loculo. Stylus inclusus vel (longi-) with numerous ovules horizontally inserted. Style exsertus. Fructus carnosi, pyrenis duabus, in matu- exserted. Fruits capsular, with septicidal (Augusta)or ritate azureopurpurei, albidi vel lutescentes. loculicidal (Wendlandia) dehiscence. Seeds minute, Unicaulous subshrubs, shrubs or small trees. compressed, with reticulate testa. Flowers 5-merous in terminal panicles. Calyx persist- ent. Corolla short- to long-tubular, with left-contorted Included genera: Augusta, with four species occurring aestivation. Ovary bilocular, with one pendulous in , north-eastern and central Brazil, the Fiji ovule in each locule. Style included to (long-)exserted. islands and New Caledonia, and Wendlandia (c.80 Fruits fleshy, with two pyrenes, becoming blue– spp.), occurring in north-eastern , west Asia, purple, whitish or yellowish in maturity. South-East Asia and northern Australia.

Included genera: Airosperma with six species in Fiji Tribe Bertiereae Bridson and New Guinea, and the monotypic Boholia from the For a description, see Bridson & Verdcourt (2003). Philippines and the island of Flores, Indonesia. Bertiereae as currently defined is monogeneric.

Tribe Alberteae Hook.f. Distribution: Tropical to southern For descriptions, see Puff et al. (1984), Robbrecht Mexico, tropical Africa, and Mascarenes. (1988) and Kainulainen et al. (2009).

Included genus: Bertiera. Distribution: South-east Africa and Madagascar.

Tribe Coffeeae DC. Included genera (here investigated): Alberta. For a description, see Davis et al. (2007). Recent taxo- Included based on Kainulainen et al. (2009): Nema- nomic changes have been made by Tosh et al. (2009; tostylis Hook.f and Razafimandimbisonia Kainul. & resurrection of Empogona Hook.f.) and Davis et al. B.Bremer. (2011; inclusion of Psilanthus Hook.f. in Coffea).

Tribe Aleisanthieae Mouly, J.Florence & B.Bremer Distribution: Tropical Africa, Madagascar, Comores, For descriptions, see Mouly et al. (2009a) and Mascarenes, Seychelles and tropical Asia to Australia. Alejandro et al. (2010).

Included genera (here investigated): Coffea and Distribution: South-East Asia. Empogona. Inclusion based on Davis et al. (2007): Argocoffeop- Included genera (here investigated): Aleisanthia and sis Lebrun, Hook.f., Aleisanthiopsis. Pierre ex Robbr., DC.,

© 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173, 387–406 396 K. KAINULAINEN ET AL.

Dalzell, T.Durand, Sericanthe Robbr., Inclusion based on Rogers (1984) and Delprete & A.Rich. ex DC. and Xantonnea Pierre ex Cortés-B. (2004): Gleasonia Standl. and Platycarpum. Pit. Tribe Ixoreae A.Gray. Tribe Condamineeae Hook.f. For descriptions, see Mouly et al. (2009a, b). Ixoreae For a description, see Kainulainen et al. (2010). as currently defined is monogeneric.

Distribution: Neotropics, south-eastern USA (Pinck- Distribution: Neotropics, tropical Africa, Madagascar, neya Michx.), South-East Asia (Emmenopterys Oliv.) Comores, Mascarenes, Seychelles, tropical Asia to the and the Malesia-Pacific region ( Pacific. A.Gray, Mastixiodendron Melch. and Mussaendopsis Baill.). Included genus: Ixora.

Tribe Jackieae Korthals Included genera (here investigated): Bathysa C.Presl, For descriptions, see Ridsdale (1979) and Calycophyllum DC., Chimarrhis Jacq., Condaminea Razafimandimbison et al. (2011). Jackieae as cur- DC., Dioicodendron Steyerm., Emmenopterys, Ferdi- rently defined is monogeneric. nandusa Pohl, Hippotis Ruiz & Pav., Mastixioden- dron, Pinckneya, Pogonopus Klotzsch, Rustia Distribution: South-East Asia. Klotzsch and Simira Aubl. Inclusion based on Kainulainen et al. (2010): Schott, Hook.f., Spruce, Dia- Included genus: Jackiopsis. lypetalanthus, Dolichodelphys K.Schum. & K.Krause, Dolicholobium, Wedd., Holtonia Standl., Tribe Mussaendeae Hook.f. Macbrideina Standl., P.Browne, For descriptions, see Alejandro et al. (2005). Mussaendopsis, Parachimarrhis Ducke, Benth., Picardaea Urb., Schizocalyx Wedd., Sema- Distribution: Tropical Africa, Madagascar, Mas- phyllanthe L.Andersson, Schltdl., Tammsia carenes, South-East Asia to the Pacific. H.Karst., Klotzsch and Wittmackanthus Kuntze. Included genera (here investigated): Bremeria, Heinsia, Landiopsis, Mussaenda, Pseudomussaenda and Schizomussaenda. Tribe Crossopterygeae F.White ex Bridson Inclusion based on Tange (1994): Neomussaenda For a description, see Bridson & Verdcourt (2003). Tange. Crossopterygeae as currently defined is monogeneric. Tribe Posoquerieae Delprete Distribution: Tropical Africa. For a description, see Delprete (2009).

Included genus: Crossopteryx. Distribution: Neotropics.

Tribe Greeneeae Mouly, J.Florence & B.Bremer Included genera: Molopanthera Turcz. and Posoqueria For a description, see Mouly et al. (2009a). Aubl.

Distribution: South-East Asia. Tribe Retiniphylleae Hook.f. For a description, see Robbrecht (1988). Retiniphyl- leae as currently defined is monogeneric. Included genera (here investigated): Greenea. Inclusion based on Mouly et al. (2009a): Spathich- Distribution: Tropical South America. lamys R.Parker.

Included genus: Retiniphyllum. Tribe Henriquezieae Hook.f. For a description, see Rogers (1984). Tribe Sabiceeae Bremek. For a description, see Khan et al. (2008). Distribution: Tropical South America. Distribution: Neotropics, Tropical Africa, Madagascar, Included genera (here investigated): Henriquezia. São Tomé, Socotra, and Sri Lanka.

© 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173, 387–406 PHYLOGENY OF IXOROIDEAE 397

Included genera: Hekistocarpa, Sabicea, Tamridaea forming a tube surrounding the style. Fruits septi- and Virectaria. cidal capsules. Seeds minute, flattened, with a reticu- late, granulate testa. Tribe Scyphiphoreae Kainul. & B.Bremer, trib. nov. Type: Scyphiphora C.F.Gaertn., Suppl. Carp.: 91 Included genus: Steenisia, with five species in (1806) Sundaland (Borneo, Natuna Islands and Peninsular Frutices vel arbusculae. Flores 4(–5)-meri, in thyrsis ). axillaribus. Calyx tubularis, brevis, persistens, lobis indistinctis. Corolla tubularis, lobis in aestivatione Tribe Trailliaedoxeae Kainul. & B.Bremer, trib. nov. sinistrorsum contortis. Ovarium biloculare, ovulo uno Type: Trailliaedoxa W.W.Sm. & Forrest, Notes Roy. ascendenti et uno descendenti in quoque loculo. Stylus Bot. Gard. Edinburgh 10: 74 (1917) exsertus. Fructus sicci, indehiscentes, porcati, meso- Fruticuli erecti. Flores minuti, 5-meri, in cymis ter- carpio extenso. minalibus. Calyx persistens. Corolla infundibuli- Shrubs or small trees. Flowers 4(–5)-merous, formis, lobis in aestivatione sinistrorsum contortis. in axillary thyrses. Calyx tubular, short, persistent, Ovarium biloculare, ovulo uno pendulo in quoque with indistinct lobes. Corolla tubular, with left- loculo. Stylus exsertus. Fructus schizocarpi. contorted aestivation. Ovary bilocular, with one Erect shrublets. Flowers minute, 5-merous, in ter- ascending and one descending ovule in each locule. minal cymes. Calyx persistent. Corolla funnel-shaped, Style exserted. Fruits dry, indehiscent, ridged, with with left-contorted aestivation. Ovary bilocular, with extensive mesocarp. one pendulous ovule in each locule. Style exserted. Fruits schizocarpous. Included genus: The monotypic Scyphiphora, occur- ring in tropical Asia and the western Pacific. Included genus: The monotypic Trailliaedoxa, occur- ring in south-western China (Sichuan, Yunnan). Tribe Sipaneeae Bremek. Tribe Vanguerieae A.Rich. ex Dum. For a description, see Delprete & Cortés-B. (2004). For descriptions, see Robbrecht (1988) and Lantz & Bremer (2004). Distribution: Central America and tropical South America. Distribution: Southern Africa, tropical Africa, Mada- gascar, Comores, Mascarenes, Seychelles, (sub-) tropi- Included genera (here investigated): Chalepophyllum cal Asia to Australia and the Pacific. Hook.f., Dendrosipanea Ducke, Neobertiera Wernham and Sipanea Aubl. Included genera (here investigated): Afrocanthium, Inclusion based on Delprete & Cortés-B. (2004): Canthium Lam., Cyclophyllum, Fadogia Schweinf., Limnosipanea Hook.f., Maguireothamnus Steyerm., Keetia, Multidentia Gilli, Peponidium, Psydrax, Pyro- Neblinathamnus Steyerm., Pteridocalyx Wernham, stria, Rytigynia Blume and Vangueria Juss. Sipaneopsis Steyerm. and Steyermarkia Standl. Inclusion based on Robbrecht (1988), Lantz & Bremer (2004) and Razafimandimbison et al. (2009): Tribe Steenisieae Kainul. & B.Bremer, trib. nov. (Bridson) Razafim., Lantz & B.Bremer, Type: Steenisia Bakh.f., Webbia 8: 381 (1952) DC., Robyns, S.T.Rey- Suffrutices unicaules. Flores 4–5-meri, in thyrsis axil- nolds & R.J.F.Hend., Fadogiella Robyns, Hutchinsonia laribus. Calyx persistens, interdum uno lobo post Robyns, Perakanthus Robyns ex Ridl., anthesin expanso. Corolla rotata, lobis in aestivatione Robyns, Robyns, Robynsia Hutch., sinistrorsum contortis. Ovarium biloculare, ovulis Robyns ex Ridl., Vangueriella Verdc. and numerosis horizontaliter insertis. Stylus curvatus. Robyns. Antherae appendicibus apicalibus prolongatis, conna- tis tubumque stamineum stylum cingentem facienti- CONCLUSIONS bus. Fructus capsulares septicidales. Semina minuta, complanata, testa reticulata granulata. Here, we analysed the tribal relationships and delimi- Unicaulous subshrubs. Flowers 4–5-merous, in axil- tation in Ixoroideae using six plastid DNA regions and lary thyrses. Calyx persistent, sometimes with one Bayesian and parsimony methods of phylogenetic lobe expanding after anthesis. Corolla rotate, with reconstruction. Following the results of this study, left-contorted aestivation. Ovary bilocular, with Ixoroideae can be considered as being composed of two numerous ovules horizontally inserted. Style curved. well-supported lineages, the Coffeeae alliance and the Anthers with extended apical appendages, united and Vanguerieae alliance, and a grade consisting of the

© 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173, 387–406 398 K. KAINULAINEN ET AL. tribes Condamineeae, Henriquezieae, Mussaendeae, Andreasen K, Bremer B. 2000. Combined phylogenetic Posoquerieae, Retiniphylleae, Sabiceeae, Sipaneeae analysis in the Rubiaceae–Ixoroideae: morphology, nuclear and Steenisieae. The Vanguerieae alliance comprises and chloroplast DNA data. American Journal of Botany 87: Glionnetia (with unplaced tribal position) and the 1731–1748. tribes Aleisanthieae, Crossopterygeae, Greeneeae, Backlund M, Bremer B, Thulin M. 2007. Paraphyly of Ixoreae, Jackieae, Scyphiphoreae, Trailliaedoxeae and Paederieae, recognition of Putorieae and expansion of Vanguerieae. The Coffeeae alliance contains Airosper- Plocama (Rubiaceae–). Taxon 56: 315–328. meae, Alberteae, Augusteae, Bertiereae, Coffeeae, Backlund M, Oxelman B, Bremer B. 2000. Phylogenetic relationships within the based on ndhF and Gardenieae, Octotropideae and Pavetteae. Molecular rbcL sequences, with particular reference to the Loga- data furthermore support the transfer of Abramsia niaceae. American Journal of Botany 87: 1029–1043. to Airosperma and Lindenia to Augusta. Future Bakhuizen van den Brink RC Jr. 1952. ‘Steenisia’ a new studies should further investigate the phylogenetic genus of Malaysian Rubiaceae. Webbia 8: 381–382. positions of Glionnetia and Hekistocarpa and the rela- Bremekamp CEB. 1952. The African species of Oldenlandia tionships of the early divergent Condamineeae and L sensu Hiern et K. Schumann. Verhandelingen der Konin- Henriquezieae–Posoquerieae–Sipaneeae clades with klijke Nederlandsche Akademie van Wetenschappen, Afdeel- respect to the rest of the subfamily. ing Natuurkunde, Tweede sectie 18: 1–297. Bremekamp CEB. 1957. On the position of Platycarpum Humb. et Bonpl., Henriquezia Spruce and Gleasonia Standl. ACKNOWLEDGEMENTS Acta Botanica Neerlandica 6: 351–377. Bremekamp CEB. 1966. Remarks on the position, the We thank the curators of the herbaria AAU, BR, GB, delimitation and the subdivision of the Rubiaceae. Acta HUH, L, MO, S and UPS for providing loans and/or Botanica Neerlandica 15: 1–33. access to collections. We also thank Anbar Khodaban- Bremer B. 1984. The genus Steenisia (Rubiaceae) and its deh for laboratory assistance, Claes Persson for DNA taxonomic position. Nordic Journal of Botany 4: 333–345. samples, Christian Puff and the Missouri Botanical Bremer B, Andreasen K, Olsson D. 1995. Subfamilial and Garden for photographs and Dianxiang Zhang for tribal relationships in the Rubiaceae based on rbcL sequence sharing unpublished information on Wendlandia. Per data. Annals of the Missouri Botanical Garden 82: 383–397. Ola Karis and Philip Oswald checked the Latin diag- Bremer B, Bremer K, Heidari N, Erixon P, Olmstead noses. 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APPENDIX Overview of taxon sampling, including GenBank sequence accession numbers and voucher specimens for previously unpublished sequences (indicated with *). Italic sequences originate from a different specimen from that indicated

Species Voucher matK ndhF rbcL rps16 trnS-G trnT-L-F

Afrocanthium Luke & Luke 9045 HM119502* HM164346* HM164152* HM164189* HM164234* AJ6201271 lactescens (Hiern) (UPS) Lantz Airosperma Nyman 569 (UPS) – HM164347* – – – HM164302* psychotrioides K.Schum. & Lauterb. Airosperma sp. Sands 6819 (L) HM119503* HM164348* – – HM164235* HM164303* Airosperma Degener & – HM164349* – HM164190* – HM164304* trichotomum Ordonez 15542 (Gillespie) (S) A.C.Sm. Airosperma Smith 8214 (P) HM119504* AM9498452 – FM2047002 FM2047402 FM2071082 vanuense S.P.Darwin Alberta magna Tonkin 200 (UPS) HM119505* AJ2362823 Y187083 FM2047012 FM2047412 AJ6201181 E.Mey. Aleisanthia Tange 45171 HM119506* – HM164153* AF2429024 HM164236* HM164305* rupestris (Ridl.) (AAU) Ridl. Aleisanthiopsis Tange 46977 HM119507* HM16435028 HM164154* AF2429034 HM164237* HM164306* distantiflora (AAU) (Merr.) Tange Aoranthe Iverssen & Steiner HM119508* – Y118455 HM164191* HM164238* HM164307* penduliflora 86776 (UPS) (K.Schum.) Somers Atractocarpus Munzinger & HM119509* HM164351* HM164155* HM164192* HM164239* HM164308* balansaeanus McPherson 635 Guillaumin (MO) Augusta Mouly & Innocente HM119510* HM164352* EU8174126 HM164193* HM164240* EU8174546 austrocaledonica 237 (P) (Brongn.) J.H.Kirkbr. Augusta longifolia Kirkbride 5295 HM119511* HM164353* – – HM164241* HM164309* (Spreng.) Rehder (BR) Augusta rivalis McDowell 5017 HM119512* AM9498462 AM9498422 HM164194* FM2047522 FM2071182 (Benth.) (ETSU) J.H.Kirkbr. Aulacocalyx Schmidt et al. HM119513* HM164354* EU8174136 EF2056397 FM2047532 EU8174556 jasminiflora 1672 (MO) Hook.f. Bathysa stipulata Pirani & Bremer FJ9053378 FJ8719478 HM164156* FJ8846378 FJ9483028 FJ9483718 (Vell.) C.Presl 4901 (SPF) Bertiera aethiopica Dessein s.n. (BR) HM119514* HM164355* HM164157* HM164195* HM164242* HM164310* Hiern

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APPENDIX Continued

Species Voucher matK ndhF rbcL rps16 trnS-G trnT-L-F

Bertiera guianensis Andersson et al. HM119515* AM9498472 AJ2248459 AF20098310 FM2047542 FM2071192,33 Aubl. 2029 (GB) Bertiera longithyrsa Kårehed et al. 256 HM119516* HM164356* HM164158* HM164196* HM164243* HM164311* Baker (UPS) Boholia Bicknell 1561A HM119517* AM9498482 AM11721011 AM11728611 FM2047552 HM164312* nematostylis (SUNIV) Merr. Bremeria landia Andriambololonera HM119518* HM164357* – HM164197* HM164244* HM164313* (Poir.) Razafim. & 37 (MAU) Alejandro Burchellia Bremer 3129 HM119519* HM164358* Z6883312 HM164198* HM164245* HM164314* bubalina (UPS) (L.f.) Sims Calycophyllum Sanders 1805 FJ9053408 AJ2362853 X8362713 FM2047122 FM2047562 AJ84739814 candidissimum (FTG) (Vahl) DC. Canthium Andreasen 36 HM119520* HM164359* Z6885112 HM164199* HM164246* AJ84740114 coromandelicum (UPS) (Burm.f.) Alston Catunaregam Luke 8332A HM119522* – AM11721411 HM164200* HM164248* HM164315* spinosa (Thunb.) (UPS) Tirveng. Cephalanthus Forbes s.n. (S) AY53837715 AJ2362883 X8362913 AF00403316 – AJ34695517 occidentalis L. Chalepophyllum Sandwith 1346 HM119523* – HM164159* HM164201* HM164249* HM164316* guyanense Hook.f. (S) Chimarrhis hookeri Delprete & FJ9053448 AJ2362893 Y187123 FJ8846438 FJ9483098 FJ9483788 K.Schum. Verduga 6421 (UPS) Cinchona McDowell 4613 Z7019718 AJ23584319 X8363013 FM2047142 FM2047582 AJ34696317 pubescens Vahl. (DUKE) Coddia rudis Bremer 3764 HM119524* HM164361* AJ28669520 HM164202* HM164250* HM164317* (E.Mey. ex Harv.) (UPS) Verdc. Coffea arabica L. EF04421321 EF04421321 EF04421321 EF04421321 EF04421321 EF04421321 Coffea ebracteolata Andreasen 223 HM119560* HM164387* AJ28669220 HM164219* HM164283* HM164333* (Hiern) Brenan (UPS) Coffea sessiliflora Luke 9024 HM119525* HM164362* HM164160* HM164203* HM164251* HM164318* Bridson (UPS) Colletoecema Lisowski 47195 HM119526* EU14540922 EU14545722 AF12927223 HM164252* EU14553222 dewevrei (De (K) Wild.) E.M.A.Petit Condaminea Pennington & FJ9053478 FJ8719508 HM164161* FJ8846458 FJ9483128 FJ9483818 corymbosa (Ruiz Daza 17436 & Pav.) DC. (MOL) Coptosperma Bremer 3810 HM119527* HM164363* AJ28671120 HM164204* HM164253* HM164319* supra-axillare (UPS) (Hemsl.) Degreef Cremaspora triflora Andreasen 51 HM119528* AM9498502 Z6885612 FM2047162 FM2047592 FM2071212 ssp. confluens (UPS) (K.Schum.) Verdc. Crossopteryx Bremer 3097 FJ9053298 AM9498512 AM11722311 FM2047172 FM2047602 FM2071232 febrifuga (Afzel. (UPS) ex G.Don) Benth. Cubanola McDowell 4427 AY53838615 AM11734511 X8363213 FM2047182 FM2047612 FM2071242 domingensis (DUKE) (Britton) Aiello

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APPENDIX Continued

Species Voucher matK ndhF rbcL rps16 trnS-G trnT-L-F

Cyclophyllum Mouly & HM119529* – EU8174166 EF2056407 HM164254* EF2056317 deplanchei Innocente Hook.f. 228 (P) Dendrosipanea Prance et al. FJ9053248 FJ8719438 HM164162* FJ8846278 FJ9482918 FJ9483608 spigelioides 16199 (S) Ducke Dioicodendron Zarucchi & FJ9053498 FJ8719528 HM164163* FJ8846478 FJ9483148 FJ9483838 dioicum Echeverry 4780 (K.Schum. & (MO) Krause) Steyerm. Duperrea cult. Bergius HM119531* HM164365* AJ28670920 HM164205* HM164256* HM164320* pavettifolia (Kurz) Botanic Garden Pit. Emmenopterys Robbrecht s.n. FJ9053608 AJ2362943 Y187153 FM2047192 FM2047622 FM2071252 henryi Oliv. (UPS) Empogona coriacea Bremer et al. 4302 HM119574* HM164401* HM164184* HM164228* HM164295* HM164340* (Sond.) Tosh & (UPS) Robbr. Euclinia longiflora De Block 27 (BR) HM119532* HM164366* Z6883512 HM164206* HM164257* AJ84739914 Salisb. Fadogia tetraquetra Bremer & Bremer HM119533* HM164367* AM11722511 HM164207* HM164258* AJ6201391 K.Schum. & 3799 (UPS) Krause Ferdinandusa Malme 2442 (UPS) HM119534* EU14541222 AM11722611 FJ8846578 FJ9483278 EU14553422 speciosa Pohl Feretia apodanthera Luke 8385 (UPS) HM119535* HM164368* HM164164* HM164208* HM164259* HM164321* Delile Nat. Bot. Gard. HM119536* HM164369* AJ28670420 AF2448924 HM164260* EU14554022 Lam. Meise, no. 19940025-26 Gardenia Drozd & Molem HM119537* AM9498522 AJ31844624 AJ32007724 FM2047632 FM2071262 hansemannii 1998-11-13 K.Schum. Gelsemium Bremer 3026 AJ42932225 AJ01198426 L1439727 AJ43103325 – AJ43090825 sempervirens (L.) (UPS) J.St.-Hil. Genipa americana Kiehn, HBVsub HM119538* HM164370* Z6883912 HM164209* HM164261* HM164322* L. RR-420 (WU, UPS) Glionnetia sericea Beaver 17 (S) HM569720* HM53621728 HM53622328 HM53622928 HM569721* HM53623528 (Baker) Tirveng. Greenea oblonga Larsen & Larsen HM119539* HM164371 – EU8174396 HM164262* EU8174596 Craib 33451 (P) Guettarda speciosa Persson 141 (GB) AY53838915 FJ8719428 AY53848515 AF2429644 FJ9482898 FJ9483588,34 L. Heinsia crinita McPherson 16188A HM119540* HM164372* Y118495 HM164210* HM164263* HM164323* (Afzel.) G.Taylor (MO) Hekistocarpa Leuwenberg 6393 HM119541* HM164373* AF33236629 AF33236729 HM164264* HM164324* minutiflora (BR, K, WAG) Hook.f. Henriquezia nitida Liesner 8531 (MO) – HM164374* – HM164211* – HM164325* var. macrophylla (Ducke) Steyerm. Hippotis triflora Ståhl 2660 (GB) FJ9053658 FJ8719568 HM164165* FJ8846598 FJ9483298 FJ9483978 Ruiz & Pav. Hyperacanthus Razafimandimbison HM119542* HM164375* HM164166* HM164212* HM164265* HM164326* grevei Rakotonas. 547 (UPS) & A.P.Davis

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APPENDIX Continued

Species Voucher matK ndhF rbcL rps16 trnS-G trnT-L-F

Hypobathrum Ridsdale 81 HM119543* AM9498532 AJ28670520 AM11731811 HM164266* FM2071272 racemosum (Roxb.) Kurz Ixora borboniae Friedmann 3049 HM119552* HM164383* HM164173* FJ1506856 HM164275* FJ1506096 Mouly & (P) B.Bremer Ixora coccinea L. Bremer 2719 HM119544* HM164376* HM164167* EF2056417 FM2047642 EU8174646 (UPS) Ixora ferrea (Jacq.) Taylor 11693 (MO) HM119545* HM164377* EU8174226 EF2056427 HM164267* EU8174656 Benth. Ixora margaretae Mouly & Innocente HM119521* HM164360* EU8174156 EU8174366 HM164247* EU8174566 (N.Hallé) Mouly 222 (P) & B.Bremer Ixora Drozd & Molem HM119581* HM164406* AJ31845924 AJ32009024 HM164300* EU8174766 novoguineensis 1998-11-13 Mouly & B.Bremer Ixora trilocularis Lesouef 31 (TAN) HM119530* HM164364* EU8174176 EU8174376 HM164255* EU8174576 (Balf.f.) Mouly & B.Bremer Jackiopsis ornata K. S. Tan s.n. HM119546* HM53621328 HM53621928 HM53622528 HM164268* HM53623128 (Wall.) Ridsdale Kailarsenia Larsen 41627 HM119547* HM164378* HM164168* HM164213* HM164269* HM164327* tentaculata (AAU) (Hook.f.) Tirveng. Keetia gueinzii Bremer et al. 4349 HM119548* HM164379* HM164169* HM164214* HM164270* AJ6201431 (Sond.) Bridson (UPS) Landiopsis Gautier 4533 (MO) HM119549* HM164380* HM164170* HM164215* HM164271* HM164328* capuronii Capuron ex Bosser Logania vaginalis Bremer 3013 AJ42932425 AJ23583719 Z6882612 AJ43103525 HM164272* AJ43091025 (Labill.) F.Muell. (UPS) Luculia gratissima CT 870064 AJ42932525 AJ01198726 AM11724311 AJ43103625 FM2047652 AJ43091125 (Wall.) Sweet Mastixiodendron Rova & FJ9053698 FJ8719598 HM164171* AF2429794 FJ9483328 FJ9484018 flavidum (Seem.) Gustavsson 2429 A.C.Sm. (GB) Mitriostigma Bremer 2705 HM119550* HM164381* X8365013 HM164216* HM164273* HM164329* axillare Hochst. (UPS) Molopanthera Williams 6861 (S) – HM164382* HM164172* AF2429814 – FJ948361* paniculata Turcz. Multidentia Bidgood et al. 845 – – – – HM164274* AJ6201501 concrescens (K) (Bullock) Bridson & Verdc. Mussaenda arcuata McPherson 16213 HM119551* AJ2363013 Y118545 FM2047212 FM2047672 FM2071282 Poir. (MO) Mussaenda Gillis 10838 (FTG) FJ9053268 AJ1308363 X8365213 FJ8846288 FJ9482928 AJ6201161 erythrophylla Schumach. & Thonn. Nauclea orientalis Bremer 3001 AY53840715 EU14541022 X8365313 AJ32008024 HM164276* AJ34695817 (L.) L. (UPS) Neobertiera gracilis de Granville 2888 – – – AF2429844 – AF15267930 Wernh. (NY) Ophiorrhiza Bremer 3301 AY53840815 AJ1308383 X8365613 AF00406416 HM164277* DQ66215131 mungos L. (UPS)

© 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173, 387–406 PHYLOGENY OF IXOROIDEAE 405

APPENDIX Continued

Species Voucher matK ndhF rbcL rps16 trnS-G trnT-L-F

Oxyanthus speciosus Bremer 4348 (S) HM119554* HM164384* AM11725211 AM11733011 HM164278* AM11737511 DC. Paragenipa Persson 156 (GB) HM119555* HM164385* AJ28670720 AF00406616 HM164279* HM164330*35 lancifolia (Bojer ex Baker) Tirveng. & Robbr Pavetta abyssinica De Block 6 (BR) HM119556* AM9498542 Z6886312 FM2047262 FM2047722 FM2071332 Fresen. Pavetta indica L. Andreasen 202 HM119557* – – HM164217* HM164280* HM164331* (UPS) Peponidium Labat et al. 2236 – – – – HM164281* AJ6201531 horridum Arènes (K) Pinckneya bracteata Massey s.n. 15F FJ9053778 AJ1308393 X8366113 FJ8846688 FJ9483398 FJ9484088 (Bartram) Raf. Pogonopus speciosus Meier 2548 (GB) FJ9053788 FJ8719648 HM164174* FJ8846708 FJ9483418 FJ9484108 (Jacq.) K.Schum. Posoqueria latifolia Andreasen 90 FJ9053258 AM9498552 Z6885012 FM2047282 FM2047742 FM2071352 (Rudge) Roem. & (UPS) Schult. Posoqueria cult. Bergius HM119558* HM164386* HM164175* HM164218* HM164282* HM164332* longiflora Aubl. Botanic Garden Pseudomussaenda Nissen s.n. (UPS) HM119559* AM9498562 Y118555 FM2047292 FM2047752 FM2071362 flava Verdc. Psydrax obovata Bremer & Bremer HM119561* HM164388* HM164176* HM164220* HM164284* AJ6201611 (Klotzsch ex Eckl. 3762 (UPS) & Zeyh.) Bridson Pyrostria hystrix Bremer & Bremer HM119562* HM164389* AM11726211 AM11733811 HM164285* AJ6201681 (Bremek.) Bridson 3791 (UPS) Randia aculeata L. F.G.x.1–145 HM119563* HM164390* Z6883212 HM164221* HM164286* HM164334* Retiniphyllum Wurdack & – HM164392* AF33165432 AF00407616 FM2047762 FM2071372 pilosum (Spruce Adderley 43270 ex Benth.) Müll. (S) Arg. Rondeletia odorata Bremer & – AJ23584519 Y118575 AF2430104 FJ9482908 FJ9483598,36 Jacq. Andreasen 3504 (UPS) Rosenbergiodendron Jansen-Jacobs HM119566* HM164393* HM164177* HM164222* HM164287* HM164335* densiflorum (K. 3977 (GB) Schum.) Fagerl. Rothmannia Bremer et al. 4346 HM119567* HM164394* AM11726611 AM11734011 HM164288* AM11738411 capensis Thunb. (S) Rustia Delprete 6378 FJ9053858 AJ2363103 Y187163 FM2047312 FM2047772 FM2071382 thibaudioides (UPS) (H.Karst.) Delprete Rytigynia van den Berghen – – HM164178* HM164223* HM164289* AJ6201751 senegalensis 8746 (BR) Blume Sabicea africana (P. Wieringa 3591 – HM164397* HM164180* HM164225* HM164292* HM164337* Beauv.) Hepper (WAG) Sabicea aspera Andersson et al. AY53842015 EU14541622 AY53850815 AF00407916 FJ9482938 FJ9483628,37 Aubl. 1941 (GB) Sabicea diversifolia Bremer et al. HM119568* EU14541522 EU14545922 EU14549422 HM164290* AJ84739614 Pers. 4018-B18 (UPS) Schizomussaenda Puff 961116-1/1 HM119569* HM164395* AM11727311 HM164224* HM164291* AJ84738314 henryi (Hutch.) (WU) X.F.Deng & D.X.Zhang

© 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173, 387–406 406 K. KAINULAINEN ET AL.

APPENDIX Continued

Species Voucher matK ndhF rbcL rps16 trnS-G trnT-L-F

Scyphiphora Bremer et al. 99 FJ9053278 AJ2363113 Y187173 EU8174506 FM2047792 EU8174756 hydrophyllacea (S) C.F.Gaertn. Simira cordifolia Bremer 3361 FJ9053888 FJ8719708 HM164179* FJ8846778 FJ9483508 FJ9484198 (Hook.f.) Steyerm. (UPS) Sipanea aff. biflora Rova et al. 2005 AY53842115 EU14541322 AY53850915 AF00408516 FM2047802 FM2071412,38 (L.f.) Cham. & (GB) Schltdl. Sipanea hispida Irwin et al. 34756 – EU14541422 EU14545822 EU14549222 – HM164336* Benth. ex (UPS) Wernham Steenisia Puff BF FJ9053288 HM164396* AM11727911 FM2047352 FM2047812 FM2071422 pleurocarpa (Airy 990619-1/4 Shaw) Bakh.f. (WU) Tamridaea Thulin & Gifri – HM164398* Y118605 FM2047362 FM2047822 FM2071432 capsulifera (Balf. 8663 (UPS) f.) Thulin & B. Bremer Tennantia sennii Abdalla et al. HM119570* AM9498582 AM9498432 FM2047372 FM2047832 FM2071442 (Chiov.) Verdc. & 96/166 (P) Bridson Tocoyena pittieri Rova et al. 2369 HM119571* AM9498592 HM164181* FM2047382 FM2047842 FM2071452 (Standl.) Standl. (GB) Trailliaedoxa Boufford et al. HM119573* HM16440028 HM16418328 HM16422728 HM164294* HM16433928 gracilis W.W.Sm. 35041 (MO) & Forrest Vangueria Sanders 1798, HM119575* AJ1308403 X8367013 HM164229* FM2047852 AJ6201841 madagascariensis #62146 (FTG) J.F.Gmel. Wendlandia arabica Thulin et al. HM119576* HM164402* HM164185* HM164230* HM164296* HM164341* Deflers 10192 (UPS) Wendlandia Chung 1403 (S) HM119577* HM164403* HM164186* HM164231* HM164297* HM164342* formosana Cowan Wendlandia Rechinger 183 (S) HM119578* HM164404* HM164187* AF2430364 HM164298* HM164343*39 ligustroides (Boiss. & Hohen.) Blakelock Wendlandia Ridsdale 2179 HM119579* HM164405* HM164188* HM164232* HM164299* HM164344* paniculata (Roxb.) DC. Wendlandia Parker 3227 (S) HM119580* AM9498602 FM2076492 FM2047392 FM2047862 FM2071472 tinctoria (Roxb.) DC. Virectaria Adames 606 (UPS) HM119582* HM164407* Y118615 HM164233* HM164301* HM164345* multiflora (Sm.) Bremek.

Published sequences: 1Lantz & Bremer (2004); 2Kainulainen et al. (2009); 3Bremer et al. (1999); 4J. H. E. Rova (unpubl. data); 5Bremer & Thulin (1998); 6Mouly et al. (2009a); 7Mouly et al. (2007); 8Kainulainen et al. (2010); 9Andreasen, Baldwin & Bremer (1999); 10Persson (2000); 11Bremer & Eriksson (2009); 12Andreasen & Bremer (1996); 13Bremer et al. (1995); 14Alejandro et al. (2005); 15Andersson & Antonelli (2005); 16Andersson & Rova (1999); 17Razafimandimbison & Bremer (2002); 18Endress et al. (1996); 19Backlund, Oxelman & Bremer (2000); 20Andreasen & Bremer (2000); 21Samson et al. (2007); 22Rydin et al. (2008); 23Piesschaert et al. (2000); 24Novotny et al. (2002); 25Bremer et al. (2002); 26Oxelman, Backlund & Bremer (1999); 27Olmstead et al. (1993); 28 Razafimandimbison et al. (2011); 29Dessein et al. (2001); 30Rova et al. (2002); 31Backlund, Bremer & Thulin (2007); 32L. Andersson (unpubl. data). Combined sequences: 33with AF152670; 34with AF152725; 35with AF152672; 36with AF152741; 37with AY538475; 38with AF152675; 39with AF152661.

© 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173, 387–406