Reassessing the Taxonomic Status of Two Enigmatic Desmos Species

Journal of Systematics and Evolution 52 (1): 1–15 (2014) doi: 10.1111/jse.12064

Research Article Reassessing the taxonomic status of two enigmatic Desmos species (Annonaceae): Morphological and molecular phylogenetic support for a new genus, Wangia 1Xing GUO 1Jing WANG 1Bine XUE 1Daniel C. THOMAS† 1Yvonne C. F. SU‡ 2Yun‐Hong TAN 1Richard M. K. SAUNDERS* 1(School of Biological Sciences, The University of Hong Kong, Hong Kong, China) 2(Key Laboratory of Tropical Forest Ecology, Center for Integrative Conservation, Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Menglun 666303, Yunnan, China)

Abstract The systematic position of two enigmatic Annonaceae species from China, Desmos saccopetaloides (W. T. Wang) P. T. Li and Desmos yunnanensis (Hu) P. T. Li, has been controversial, with both species having been transferred between several different genera within subfamilies Annonoideae and Malmeoideae. Phylogenetic analyses of eight chloroplast regions (matK, ndhF, ndhF‐rpl32, psbA‐trnH, rbcL, rpl32‐trnL, trnL‐F, and ycf1; ca. 9.2 kb, 66 taxa) unambiguously placed D. saccopetaloides in a subclade of tribe Miliuseae, nested among the genera Monoon, Neo‐uvaria, Phaeanthus, Sageraea, and Stelechocarpus. This relationship was also supported by endosperm rumination patterns in the seed; other morphological characters furthermore indicated that D. saccopetaloides has closer affinities with Monoon, Neo‐uvaria, and Phaeanthus rather than either Sageraea or Stelechocarpus. Desmos saccopetaloides is distinguished from these genera by its leaf‐opposed inflorescences, sepaloid outer petals, saccate inner petals with basal glandular tissue, moniliform monocarps with uniseriate seeds, and rectangular disulculate pollen with two “cryptoapertures.” On the basis of the combined molecular phylogenetic and morphological data, we propose a new genus, Wangia, to accommodate D. saccopetaloides. The molecular phylogenetic analyses furthermore indicated that D. yunnanensis belongs to the genus Dasymaschalon: examination of the type collections revealed that it is conspecific with Dasymaschalon obtusipetalum, although the combination Dasymaschalon yunnanense has nomenclatural priority. Key words Dasymaschalon, molecular phylogeny, morphology, Phaeanthus, taxonomy.

Early studies of higher‐level taxonomic relation- several taxa remain obscure, including those of two ships within the pantropical flowering plant family enigmatic species from China, currently recognized as Annonaceae were based by necessity on morphological Desmos saccopetaloides (W. T. Wang) P. T. Li and characters, including pollen (e.g., Walker, 1971), Desmos yunnanensis (Hu) P.T. Li (Li et al., 2011). The flowers (e.g., Van Heusden, 1992), and fruits and seeds former species has been variously placed in Desmos (e.g., Van Setten & Koek‐Noorman, 1992). Reliance on Lour. (subfam. Annonoideae) and Phaeanthus Hook. f. these characters ultimately proved unsatisfactory due to & Thomson (subfam. Malmeoideae), and the latter in extensive morphological convergence (Saunders, Dasymaschalon (Hook. f. & Thomson) Dalla Torre 2010), and most recent higher‐level classifications of & Harms (subfam. Annonoideae), Desmos and the family have been based on DNA sequence data, Phaeanthus. particularly that of the chloroplast genome (e.g., The outer petals of D. saccopetaloides (ca. 4 mm Chatrou et al., 2012). Despite significant advances long) and D. yunnanensis (ca. 3 mm long) are much over the past decade, the phylogenetic affinities of shorter than the inner petals (27–35 mm and ca. 28 mm, respectively). This feature is characteristic of Phaean- thus, leading Hu (1940) and Wang (in Wu & Wang, Received: 18 July 2013 Accepted: 17 October 2013 † 1957) to describe these species as Phaeanthus Current address: Naturalis Biodiversity Centre, Section NHN, Leiden University, 2300 RA Leiden, The Netherlands. yunnanensis Hu and Phaeanthus saccopetaloides ‡ Current address: Duke‐NUS Graduate Medical School Singapore, W. T. Wang. Wang furthermore suggested an affinity Singapore 169857, Singapore. Author for correspondence. E‐mail: [email protected]. Tel.: 852‐ between the two species by comparing them in the 2299‐0608. Fax: 852‐2517‐6082. diagnosis of the latter species, in which he contrasted

© 2013 Institute of Botany, Chinese Academy of Sciences 2 Journal of Systematics and Evolution Vol. 52 No. 1 2014 the oblong to lanceolate inner petals and flat monocarps terized by flowers with greatly reduced sepaloid outer of P. saccopetaloides with the ovate inner petals and petals, and globose to ellipsoid monocarps, with one cylindrical monocarps of P. yunnanensis (Wu & Wang, (rarely two) seeds (Mols & Keßler, 2000). Li et al. 1957). (2011) recognized the doubtful affinities of the two Bân (1975) examined one of the type specimens of species, stating that their taxonomic placement “re- P. yunnanensis with fruits (C. W. Wang 79167, A) and quires further research.” The aims of the present study noted that the monocarps are moniliform (elongated, are therefore to: (i) clarify the phylogenetic relation- with constrictions between seeds), a fruit shape that had ships of the two species based on chloroplast DNA never previously been reported in Phaeanthus;he sequence data; (ii) assess the morphological support for accordingly transferred the species to Dasymaschalon, these relationships and to identify synapomorphies; and which is characterized by such fruits. Although Bân did (iii) validate consequential nomenclatural changes as not examine flowering material, he doubted whether necessary. there were truly six petals in the type specimen: he argued that Hu (1940) had erroneously interpreted an apical expansion of the pedicel as sepals (1 mm long in 1 Material and methods Hu’s description) and the sepals (3 mm long) as outer petals, and hence believed that the flowers only had 1.1 Taxon and DNA region sampling three petals (also characteristic of Dasymaschalon). The 66‐accession dataset included a paratype (T. T. Tsiang & Li (1979) subsequently dissected flowers Yu 16484, PE) and isotype (T. T. Yu 17336,A)of from another type specimen of P. yunnanensis (C. W. Phaeanthus (Desmos) saccopetaloides, the isotype Wang 79167, PE) and agreed with Hu’s (1940) (C. W. Wang 79167,A)ofPhaeanthus (Desmos) observation of six petals in two whorls. This evidence, yunnanensis, 46 accessions representing 27 genera together with the moniliform shape of the monocarps, led from subfam. Malmeoideae, and 13 accessions repre- them to recognize both P. yunnanensis and P. saccope- senting 11 genera from subfam. Annonoideae. The taloides in Desmos (Tsiang & Li, 1979; Li, 1993), a genus outgroups consisted of three species from subfam. that has moniliform fruits that resemble those of Ambavioideae and one species of Anaxagorea A. Dasymaschalon, but which, unlike Dasymaschalon,has St.‐Hil. (subfam. Anaxagoreoideae). Preliminary phy- flowers with six petals in two whorls. logenetic analyses indicated that D. saccopetaloides Desmos yunnanensis in Thailand was described was nested within tribe Miliuseae (subfam. Malmeoi- and illustrated by Chalermglin (2001: 118–119) as deae), and we accordingly focused taxon sampling on “Dasymaschalon yunnanense (Hu) Bân.” Wang et al. this tribe, including one to four species for each genus. (2009) examined Thai specimens and found that this Sequences of five chloroplast DNA regions (matK, species only has three petals; on the basis of previous ndhF, rbcL, psbA‐trnH, and trnL‐F) which are descriptions by Hu (1940) and Tsiang & Li (1979) of six frequently used in Annonaceae phylogenetic studies petals per flower, Wang et al. (2009) believed that the were newly generated or downloaded from GenBank Thai specimens differed from “true” Desmos yunna- (http://www.ncbi.nlm.nih.gov). In order to better nensis in China. They therefore described a new resolve the tribe Miliuseae, which is notoriously species, Dasymaschalon obtusipetalum Jing Wang, recalcitrant (Mols et al., 2004b; Couvreur et al., Chalermglin & R. M. K. Saunders. The two enigmatic 2011; Xue et al., 2011, 2012; Chaowasku et al., species, currently recognized in the Flora of China (Li 2012; Chatrou et al., 2012; Thomas et al., 2012), we et al., 2011) as Desmos saccopetaloides and Desmos incorporated three additional regions (ndhF‐rpl32, yunnanensis, have therefore been variously associated rpl32‐trnL, and ycf1) that have been shown to be with several morphologically and phylogenetically highly variable and phylogenetically useful in the tribe distinct genera. Desmos and Dasymaschalon (both in Miliuseae (Thomas et al., 2012). In total, 59 sequences subfam. Annonoideae) share the unusual characteristic were newly generated for this study (voucher informa- of moniliform monocarps, but differ markedly in floral tion and GenBank accession numbers listed in structure: Desmos flowers have six petals in two whorls, Appendix I). Percentages of missing data are 33.8% with both whorls basally constricted around the in the total dataset and 28.1% in subfamily reproductive organs; whereas Dasymaschalon flowers Malmeoideae. have only three petals (regarded as homologous with the outer petals of other Annonaceae: Wang et al., 2012) 1.2 DNA extraction, amplification, and sequencing that are apically connivent over the reproductive Total DNA was isolated from herbarium material organs. Phaeanthus (subfam. Malmeoideae) is charac- using the innuPrep Plant DNA Kit (Analytik Jena, Jena,

Germany) following the manufacturer’s instructions. A identity and run under the general time reversible model single amplification protocol was used for amplification with rate heterogeneity modeled by a gamma distribu- of the chloroplast regions: template denaturation at tion (GTR þ G), which is stricter than the other model 94 °C for 5 min, followed by 35 cycles of denaturation (GTRCAT) provided by the CIPRES Science Gateway. at 95 °C for 30 s; primer annealing at 50 °C for 1 min; Subsequently, support for branches was estimated with and primer extension at 72 °C for 1 min, followed by a 1000 non‐parametric bootstraps under the partition data final extension step at 72 °C for 10 min. The primers model. used to amplify the psbA‐trnH intergenic spacer were Bayesian analysis was undertaken using MrBayes psbAF (Sang et al., 1997) and trnH2 (Tate & Simpson, version 3.1.2 (Huelsenbeck & Ronquist, 2001; Ron- 2003); other primers and the polymerase chain reaction quist & Huelsenbeck, 2003). Two partition strategies (PCR) mixture were performed following the methods were used for this study: eight partitions based on DNA described by Thomas et al. (2012). PCR products were region identity; and eight regions concatenated without purified, amplified using the BigDye Terminator Cycle partitioning. For the partitioned dataset, the parameters Sequencing Kit (Applied Biosystems, Foster City, CA, for each locus were allowed to evolve independently USA), and sequenced on an ABI 3730 DNA Analyzer using the unlinked setting. MrModeltest version 2.3 (Applied Biosystems) by BGI (Hong Kong, China). (Nylander, 2004) was used to determine the best‐fit likelihood model for each locus and the concatenated 1.3 Alignment and phylogenetic analyses matrix using the Akaike Information Criterion: the Sequence fragments were assembled and checked general time reversible model with a gamma distribu- using Sequencher version 4.5 (Gene Codes, Ann Arbor, tion of substitution rates (GTR þ G) was chosen for MI, USA). Individual regions were subsequently aligned matK, ndhF‐rpl32, rpl32‐trnL, and trnL‐F regions; the automatically in BioEdit version 7.0.1 (Hall, 1999) and GTR þ I þ G model with a proportion of invariant sites then further optimized manually using Se‐Al version 2.0 was selected for the ndhF, rbcL, and ycf1 partitions and (Rambaut, 1996). A total of 202 ambiguously aligned the non‐partitioned nucleotide dataset; and the GTR þ I positions were excluded from the analyses because of model was used for psbA‐trnH. For analyses using both difficult homology assessment (ndhF‐rpl32, 32 positions partitioned and the non‐partitioned datasets, two from a single region; rpl32‐trnL, 121 positions from four independent Metropolis‐coupled Markov chain Monte regions; and trnL‐F, 49 positions from two regions). Two Carlo analyses were run. Each search used three inversions (15 positions in psbA‐trnH and six positions in incrementally heated and one cold Markov chain, and ndhF‐rpl32)wereidentified and reverse‐complemented was run for 10 million generations and sampled every in the alignment, following a strategy previously applied 1000th generation. The temperature parameter was set by Pirie et al. (2006), to retain substitution information in to 0.08. The mean branch length prior was set from the the fragments. default mean (0.1) to 0.01 (brlenspr ¼ unconstrained: Maximum parsimony (MP), maximum likelihood exponential (100.0)) to reduce the likelihood of (ML), and Bayesian inference methods were used for stochastic entrapment in local tree length optima phylogeny reconstruction. For the MP analyses, all (Brown et al., 2010; Marshall, 2010). Convergence characters were treated as independent and of equal was assessed using the standard deviation of split weight. A heuristic search was performed in PAUP frequencies, with values <0.01 interpreted as indicating version 4.0b10 (Swofford, 2002) with 2000 random good convergence. The first 25% of samples (2500 addition sequence replicates and tree bisection–recon- trees) were discarded as burn‐in, and the post‐burn‐in nection branch‐swapping, saving 10 trees per replicate. samples summarized as a 50% majority‐rule consensus The most parsimonious trees were summarized using a tree. Overall performance of analyses of non‐parti- strict consensus tree. Clade support was evaluated using tioned and partitioned nucleotide datasets was assessed the jackknife (JK) method (Farris et al., 1996) with the with Bayes factor comparison in Tracer version 1.5 removal probability set to approximately e1 (Rambaut & Drummond, 2009). The standard criterion (36.7879%), and “jac” resampling emulated. One of 2ln Bayes factor over 10 was used as a benchmark, thousand JK replicates were performed with 100 indicating very strong evidence against an alternative random addition tree bisection–reconnection searches strategy (Kass & Raftery, 1995; Nylander et al., 2004). (each with a maximum of 10 trees held) per replicate. Maximum likelihood analyses were performed in 1.4 Morphological studies RAxML (Stamatakis, 2006) provided by the CIPRES Comparative morphological data were obtained Science Gateway (Miller et al., 2010). The dataset was from specimens deposited in A and PE herbaria and divided into eight partitions based on DNA region from the literature. Floral samples were mounted on

© 2013 Institute of Botany, Chinese Academy of Sciences 4 Journal of Systematics and Evolution Vol. 52 No. 1 2014 aluminum stubs, coated with gold‐palladium and as the “phaeanthoid” clade in Fig. 1) inclusive of examined using a Hitachi S4800 (Tokyo, Japan) representative species from the genera Monoon Miq., scanning electron microscope at 10–15 kV. Neo‐uvaria Airy Shaw, Phaeanthus, Sageraea Dalzell, p.p., and Stelechocarpus Hook. f. & Thomson; this clade is well supported in the Bayesian analysis 2 Results (PP ¼ 1.00), but only has weak support in the MP analysis (JK ¼ 50) and ML analysis (BS ¼ 63). The The concatenated alignment of the 66‐terminal Bayesian analysis furthermore indicates a possible dataset consisted of 9183 characters. The statistics of relationship between D. saccopetaloides and the genera each data matrix and the corresponding parameters are Monoon and Neo‐uvaria, although support is very low presented in Table 1. The MP heuristic search retrieved (PP ¼ 0.62) and lacking in the MP and ML analyses. 13 most parsimonious trees of 4121 steps (consistency Morphological observations are illustrated in index ¼ 0.68; retention index ¼ 0.74). Figs. 2–4 and treated in detail in Section 3. A considerable improvement in mean –ln L value was observed in the Bayesian analyses (mean –lnLnon‐ partitioned ¼ 4157; mean –lnLpartitioned ¼ 2797). Bayes 3 Discussion factor comparison indicated the better fit of the partitioned analyses than the non‐partitioned model: 3.1 Phylogenetic relationships and taxonomic 2lnB (eight partitions over non‐partitioned) ¼ 728, treatment of Desmos saccopetaloides significantly above the threshold value of 10. The The phylogenetic position of D. saccopetaloides, 50% majority‐rule consensus tree derived from the firmly nested within subfam. Malmeoideae tribe Mil- analyses using the partitioned strategy was therefore iuseae (JK ¼ 100, BS ¼ 100, PP ¼ 1.00; Fig. 1), clearly selected to present the results of the Bayesian analyses. contradicts any association with subfam. Annonoideae. In The MP, ML, and Bayesian analyses are topologi- previous phylogenetic studies, the relationships within cally similar, differing mainly in the relative MP the tribe Miliuseae have been poorly resolved, particular- jackknife, ML bootstrap (BS), and posterior probability ly at deeper nodes (Mols et al., 2004b; Couvreur (PP) values for particular groups (Fig. 1). The results are et al., 2011; Xue et al., 2011, 2012; Chaowasku consistent with previous phylogenetic analyses of the et al., 2012; Chatrou et al., 2012; Thomas et al., 2012). family, with the majority of species forming two large, Resolution of the topology has recently been improved well‐supported clades (JK ¼ 100; BS ¼ 100; PP ¼ following the inclusion of the phylogenetically more 1.00) corresponding with subfamilies Annonoideae informative regions ndhF‐rpl32, rpl32‐trnL,andycf1 and Malmeoideae. (Chaowasku et al., 2012; Thomas et al., 2012; and in the Desmos yunnanensis and D. saccopetaloides are present analyses), enabling characterization of the shown to be only distantly related to each other, and “phaeanthoid” clade, consisting of D. saccopetaloides, neither species is associated with Desmos. Desmos Monoon, Neo‐uvaria, Phaeanthus, Sageraea, Stelecho- yunnanensis is unambiguously retrieved as sister to carpus, and a recently described genus, Winitia Dasymaschalon obtusipetalum Jing Wang, Chalerm- Chaowasku (Chaowasku & Van der Ham, 2013). glin & R. M. K. Saunders (JK ¼ 100; BS ¼ 100; The phylogenetic relationships of the phaeanthoid PP ¼ 1.00), within subfam. Annonoideae. Desmos genera are supported by endosperm rumination patterns saccopetaloides, in contrast, is nested within subfam. in the seed. Van Setten & Koek‐Noorman (1992) Malmeoideae tribe Miliuseae, forming a clade (labeled identified two main types of endosperm rumination in

Table 1 Data‐matrix statistics and parameters for each of the phylogenetic analyses Matrix Terminals Characters Variable Parsimony‐informative Best‐fitting model analyzed characters (%) characters (%) for Bayesian analyses matK 66 789 308 (39.0) 141 (17.8) GTR þ G ndhF 61 2093 818 (39.1) 493 (23.6) GTR þ I þ G ndhF‐rpl32 23 636 117 (18.4) 41 (6.4) GTR þ G psbA‐trnH 33 462 123 (26.6) 49 (10.6) GTR þ I rbcL 59 1333 225 (16.9) 120 (9.0) GTR þ I þ G rpl32‐trnL 17 1229 184 (15.0) 25 (2.0) GTR þ G trnL‐F 65 1006 321 (31.9) 142 (14.1) GTR þ G ycf1 29 1635 239 (14.6) 69 (4.2) GTR þ I þ G Combined data 66 9183 2335 (25.4) 1080 (11.8) GTR þ I þ G

Fig. 1. Bayesian 50% majority‐rule consensus tree under partitioned models (chloroplast DNA data: matK, ndhF, ndhF‐rpl32, psbA‐trnH, rbcL, rpl32‐ trnL, trnL‐F, and ycf1; 66 taxa). Samples of Desmos saccopetaloides and Desmos yunnanensis shown in bold. Numbers at the nodes indicate Bayesian posterior probabilities, maximum parsimony jackknife values, and maximum likelihood bootstrap values (>50%), in that order. Scale bar ¼ 0.02 substitutions per site.

Fig. 2. Flower, fruit, and seed morphology of Wangia (¼Desmos) saccopetaloides. A, Abaxial view of leaf, showing eucamptodromous leaf venation with percurrent tertiary veins. B, A branch, showing leaf‐opposed inflorescence position. C, Lateral view of flower, showing shorter outer petals and longer inner petals. D, Basal view of flower, showing slightly saccate inner petal base, sepaloid outer petals, and slightly connate sepals. E, Dissected flower, showing pigmented glandular tissue at base of inner petal. F, Fruit with torulose monocarps. G, Single monocarp, dissected to show uniseriate seed arrangement. H, Flattened‐ellipsoid seed. I, Cross‐section of a seed, showing lamelliform endosperm rumination. Scale bar (H, I) ¼ 5 mm. Photographs by Yun‐Hong Tan. the Annonaceae, viz. lamelliform (or lamellate) and uvaria. On the basis of patterns of secondary and tertiary spiniform, although intermediate forms also occur. The leaf venation, it appears that D. saccopetaloides is likely majority of lineages in the tribe Miliuseae consist of to be more closely related to Monoon, Neo‐uvaria, and/ species with spiniform endosperm ruminations, where- or Phaeanthus than to Sageraea, Stelechocarpus,or as members of the phaeanthoid clade (together with Winitia. Desmos saccopetaloides, Monoon, Neo‐uvaria, Miliusa Lesch. ex A. DC. and Sapranthus Seem.) have and Phaeanthus share eucamptodromous leaf venation lamelliform endosperm ruminations (Van Setten & with percurrent tertiary veins (sensu Hickey, 1979), with Koek‐Noorman, 1992; Mols et al., 2004a). The parallel secondary veins (with smaller veins that bridge rumination pattern in D. saccopetaloides is lamelliform adjacent secondaries) that lack prominent marginal (Fig. 2: I) suggesting that this endosperm rumination loops (Fig. 2: A; Klucking, 1986). In contrast, Sageraea, may be synapomorphic for the entire phaeanthoid clade, Stelechocarpus, and Winitia have “festooned brochi- and possibly convergent with lamelliform rumination in dodromous” venation with (weakly) reticulate tertiary subfam. Annonoideae. veins (sensu Hickey & Wolfe, 1975), in which the The position of D. saccopetaloides within the secondary veins merge to form brochidodromous loops, phaeanthoid clade is unresolved in the MP and ML with secondary loops outside the main brochidodro- analyses; the results of the Bayesian analyses indicate mous arch (Klucking, 1986; Chaowasku & Van der low PP (0.62) for a relationship with Monoon and Neo‐ Ham, 2013). The following morphological characters

© 2013 Institute of Botany, Chinese Academy of Sciences GUO et al.: Wangia, gen. nov. (Annonaceae) 7 were found to be particularly useful for distinguishing D. saccopetaloides from its close relatives: (i) position 11 – shaped ‐ of inflorescence; (ii) shape of outer petal; (iii) shape of Winitia cryptoaperturate ellipsoid petals inner petal base; (iv) pollen aperture type; and & Inaperturate or Subglobose or (v) arrangement of seeds. The diagnostic morphological Similar to inner characters are summarized in Table 2. Inflorescences within Annonaceae can be either terminal or axillary (Fries, 1959), a distinction that is very important as they rarely coexist within genera ‐ fl

(Koek Noorman et al., 1990). Terminal in orescences shaped Spoon “ ” ‐ asku & Van der Ham (2013), and personal cryptoaperturate ellipsoid can become overtopped by the uppermost axillary petals 12 2

‐ – Inaperturate or shoot and consequently appear leaf opposed. Terminal Subglobose or and axillary inflorescences may furthermore become slightly supra‐ or extra‐axillary as a result of “metatopic displacement” (Weberling & Hoppe, 1996). The flowers of D. saccopetaloides are leaf‐opposed (Fig. 2: B), differing from other members of the phaeanthoid clade, shaped Spoon ‐ which are axillary in Monoon, Neo‐uvaria, and Sager- axillary Axillary Cauliflorous ‐ cryptoaperturate aea, extra‐axillary in Phaeanthus, and at the base of the ellipsoid Globose or trunk in Stelechocarpus and Winitia.MostAnnonaceae Similar to sepals Similar to inner possess a tripartite perianth consisting of morphologically distinct sepals, outer petals, and inner petals, although the distinctions between the two whorls of petals are slight or lacking in some genera. The outer fl petals in D. saccopetaloides owers are considerably uvaria Phaeanthus Sageraea Stelechocarpus shaped Spoon ‐ ‐ and its close relatives ellipsoid smaller than the inner petals (Fig. 2: C, D; Fig. 3: A; Fig. petals – Inaperturate Inaperturate or Globose or 4: D, E) and have a similar indument to sepals (Fig. 3: B Similar to inner D). Such sepaloid outer petals are relatively common in the tribe Miliuseae, occurring, for example, in some species of Miliusa, Meiogyne Miq. (species previously saccopetaloides classified as Fitzalania F. Muell.), Orophea Blume, and ) Pseuduvaria Miq. (Mols et al., 2004a). Within the shaped Spoon Wangia phaeanthoid clade, sepaloid outer petals are only ‐ ¼ Monoon Neo ( cryptoaperturate ellipsoid observed in D. saccopetaloides and Phaeanthus. petals The base of the petals can either be spoon‐shaped, Inaperturate or clawed, or saccate in species of the tribe Miliuseae. Desmos Although not as obvious as in Alphonsea Hook. f. & Thomson and Miliusa, the base of the inner petals is slightly saccate in D. saccopetaloides (Fig. 2: C, D). ‐

Other genera within the phaeanthoid clade possess 8 1 1 (rarely 2) 1 (rarely 2) 2 – þ þ þ þ spoon‐shaped petals, common in most Annonaceae 5 Desmos opposed Axillary Axillary Extra ‐ (Mols et al., 2004a). The base of the adaxial surface of cryptoaperturate saccopetaloides the inner petals of D. saccopetaloides is glandular and Slightly saccate Spoon distinctly pigmented (Fig. 2: E); this is also observed in Meiogyne, Orophea, Pseuduvaria, and Sapranthus , absent; n/a, not applicable.

(Van Heusden, 1992). Within the phaeanthoid clade, only Monoon has been reported to have glandular inner petals (Van Heusden, 1992, as Enicosanthum Becc.), although this was not present in species examined , Present; þ during the present study (e.g., Monoon fuscum (King) Diagnostically important morphological characters of B. Xue & R. M. K. Saunders). monocarp inner petal (adaxially) inner petal (adaxially) Pollen morphology has been extensively used in petal ﬁ Data sources: Waha & Hesse (1986), Van Heusden (1995, 1997), Mols & Keßler (2000), Mols et al. (2004a), Chaowasku et al. (2011), Xue et al. (2012), Chaow observations. Seed arrangementPollen shapePollen apertures Uniseriate Disulculate Rectangular n/a Globose n/a Globose Globose n/a Globose Biseriate Globose Biseriate Seed number per Glandular tissue at base of Pigmentation of base of Monocarp shape Torulose Globose or Character Shape of base of inner Table 2 Shape of outer petals Similar to sepals Similar to inner the classi cation of Annonaceae, especially pollen Position of inflorescence Leaf

Fig. 3. Flower and pollen morphology in Wangia (¼Desmos) saccopetaloides. A, Flower with carpels and inner petals removed showing tiny sepals (S) and sepaloid outer petals (OP). B, Glabrous adaxial surface of sepal. C, Glabrous adaxial surface of outer petal. D, Densely hairy adaxial surface of inner petal. E, F, Pollen grains showing rectangular shape and depressed cryptoapertures. G, Carpel, dissected to show uniseriate ovule arrangement. H, Abaxial view of stamen. I, Rugulate exine ornamentation. Scale bars: (A, G, H) ¼ 0.5 mm; (B–D) ¼ 100 mm; (E, F) ¼ 10 mm; (I) ¼ 2 mm. grain shape and aperture type. Monoon, Neo‐uvaria, rate” genera (e.g., Alphonsea, Meiogyne, Miliusa, Phaeanthus, Sageraea, Stelechocarpus, and Winitia Monoon, Phaeanthus, Platymitra Boerl., Popowia were described by Walker (1971) and Chaowasku & Endl., and Sageraea) are functionally aperturate, Van der Ham (2013) as having inaperturate, globose generally with two clearly recognizable germination pollen grains. Desmos saccopetaloides, however, has zones at the intine layers, they lack the remarkable rectangular pollen grains with two conspicuous sunken modiﬁcations observed in D. saccopetaloides. depressed cryptoapertures (Fig. 3: E, F). Although a Desmos saccopetaloides is characterized by transmission electron microscope study by Mols et al. torulose monocarps with uniseriate seeds; in marked (2004a) suggested that several seemingly “inapertu- contrast, Monoon, Neo‐uvaria, and Phaeanthus have

Fig. 4. Wangia saccopetaloides, comb. nov. A, Flowering branch. B, Flower bud. C, Sepal. D, Outer petal. E, Inner petal (originally lanceolate; shape shown here is due to shrinkage after specimen preservation). F, Carpel. G, Stamen. H, Fruit with ﬁve monocarps. Scale bars: (A, E, H) ¼ 1 cm; (B) ¼ 4 mm; (C, D, F, G) ¼ 1 mm. globose or ellipsoid monocarps, each with only one or Winitia; our examination of specimens reveals that it is two seeds. Desmos saccopetaloides was originally actually uniseriate (Fig. 2: G; Fig. 3: G). described as having numerous seeds in two rows (Wu & Our molecular phylogenetic analyses indicate that Wang, 1957), as in Sageraea, Stelechocarpus and D. saccopetaloides is nested within the phaeanthoid

© 2013 Institute of Botany, Chinese Academy of Sciences 10 Journal of Systematics and Evolution Vol. 52 No. 1 2014 clade, and is not sister to any of the genera Monoon, hairy; with 1–4 bracts from base to middle, 1–2 mm long, Neo‐uvaria, Phaeanthus, Sageraea,orStelechocarpus. 0.4–1 mm wide, hairy. Sepals ovate‐triangular, slightly The morphological data furthermore provide strong connate at base (Fig. 2: D), 1–3.2 mm long, 1.2–2.5 mm support for distinguishing D. saccopetaloides from its wide, glabrous adaxially, ferruginous‐pubescent abax- related genera. A new genus is accordingly warranted, ially. Petals 6, valvate, free, in 2 whorls; outer petals and is described below as Wangia. ovate‐triangular, 4–7 mm long, 3.5–4 mm wide, adaxially glabrous, abaxially pubescent; inner petals ovate‐oblong Wangia X. Guo & R. M. K. Saunders, gen. nov. to lanceolate, greenish‐yellow, 27–52 mm long, 10– (Figs. 2–4) 18 mm wide, adaxially densely pubescent, glandular and Type species: Wangia saccopetaloides (W. T pigmented at the base, abaxially pubescent. Stamens Wang) X. Guo & R. M. K. Saunders. numerous, oblong, 1.6–2.4 mm long, ca. 0.8 mm wide, Small trees. Leaves ovate‐oblong, secondary veins connectives apically truncate (Fig. 3: H); filament ca. parallel, tertiary veins percurrent. Inflorescences leaf‐ 0.8 mm long, 0.6 mm wide. Carpels 12–20 per flower, opposed, 1–3(–4) flowered; pedicel long; pedicel bract 2.2–5 mm long, 0.7–1 mm wide, tomentose; ovules 5–10 lanceolate. Flowers bisexual; sepals slightly connate, per carpel, uniseriate; stigmas sessile, globose. Torus ovate‐triangular; outer petals ovate‐triangular, shorter flattened. Monocarp stipes 0.4–1.4 cm long; monocarps than inner petals; inner petals spreading at anthesis. torulose(Fig.2:F;Fig.4:H),1.8–5.6 cm long, 1–1.8 cm Stamens numerous; connectives apically truncate. wide, with 4–9 constrictions, glabrous. Seeds 5–10 per Carpels 12–20 per flower; ovaries densely hairy; stigma monocarp, flattened‐ellipsoid(Fig.2:G,H;Fig.3:G; globose; ovules 5–10, uniseriate. Monocarps with short Fig. 4: H). Pollen grains solitary, symmetrical, rectangu- stipe, torulose with 4–9 constrictions; seeds 5–10 per lar, disulculate‐cryptoaperturate with two depressed areas monocarp, flattened‐ellipsoid (Fig. 2: G, H). indicating two intinous germination zones, long axis 28– Distribution: A single species, known from 36 mm, short axis 17–24 mm, rugulate (Fig. 3: I). Yunnan province, China. Distribution and habitat: Known from three Etymology: Named after Wang Wentsai (¼Wang localities in tropical parts of Yunnan province Wen‐Cai, of the Institute of Botany, Chinese Academy (Fig. 5): Mang‐lung region in Chen‐kang (¼Zhen‐ of Sciences), who published the basionym of the type kang) county, Hila region in Shun‐ning (¼Feng‐qing) species, in honor of his contributions to plant taxonomy. county and Lan‐cang county. Growing under closed canopy in a primary forest, at high elevations (1700– Wangia saccopetaloides (W. T. Wang) X. Guo & 2300 m). R. M. K. Saunders, comb. nov.—Phaeanthus Phenology: Mature flowers collected in August; saccopetaloides W. T. Wang in C. Y. Wu & W. T. fruits collected in June. Wang, Acta Phytotax. Sin. 6: 198. 1957—Desmos Paratype: China. Yunnan: Shun‐ning, Hila, 1938‐ saccopetaloides (W. T. Wang) P. T. Li, Guihaia 13: 06‐26, T. T. Yu 16484 (PE!). 314. 1993. Type: China. Yunnan: Chen‐kang, Mang‐ Additional specimens examined. China. lung, 1938‐08‐16, T. T. Yu 17336 (holotype, PE!; Yunnan: Lan‐cang, 2012‐09‐08, Yun‐hong Tan isotypes, A!, PE!). 6940; 2012‐09‐18, Yun‐hong Tan 6931 (HITBC).

Trees, 6–8 m tall. Twigs appressed ferruginous‐ pubescent, soon glabrate, densely lenticellate; lenticels distinct, elliptic to orbicular. Petioles 3.5–10 mm long, 1–2 mm in diameter, slightly hairy, soon glabrate; leaf blade elliptic, oblong, or ovate‐oblong, 4–16 cm long, 1.8–5 cm wide, length/width 2.2–3.7, base cuneate, apex acuminate, thinly leathery, abaxially sparsely hairy to glabrescent, adaxially glabrous; midrib impressed and puberulent above, raised and sparsely hairy to glabrous below; secondary veins 7–11 on each side, parallel, diverging at 45–60° from midrib, prominent abaxially; tertiary veins percurrent. Inﬂorescences on young growth, leaf‐opposed, 1–3(–4) ﬂowered. Peduncles 5–6 mm long, sparsely hairy. Flowering pedicels 2–3.5 cm long, 1–1.5mmindiameter,sparsely Fig. 5. Distribution of Wangia saccopetaloides.

IUCN conservation status: CR B1ab(i,iii,iv)D obtusipetalum must be regarded as a heterotypic (IUCN, 2012). Wangia saccopetaloides was not synonym. collected for over 70 years after it was ﬁrst described Dasymaschalon yunnanense (Hu) Bân, Bot. Zhurn. 60 in 1938. Over the past 5 years, one of the authors (Yun‐ (2): 230. 1975—Phaeanthus yunnanensis Hu, Bull. Fan Hong Tan) has searched for this species in the type Mem. Inst. Biol. Bot. ser. 10: 125. 1940—Desmos localities, Feng‐qing and Zhen‐kang, but found that the yunnanensis (Hu) P. T. Li, Fl. Reipubl. Popularis Sin. forest had been degraded in these areas and did not 30(2): 51–53. 1979. Type: China. Yunnan: Jah‐leei, relocate the species. Yun‐Hong Tan discovered the Che‐li Hsien, 1936‐10, C. W. Wang 79167 (holotype, species in 2012, however, in Lan‐cang county. The PE!; isotypes, A!, PE!). distribution area is very restricted, being concentrated in Dasymaschalon obtusipetalum Jing Wang, Cha- less than 16 km2, with fewer than 30 individuals. lermglin & R. M. K. Saunders, Syst. Bot. 34: 262–263. Wangia saccopetaloides is at a very high risk of 2009, syn. nov. Type: Thailand. Northern: Doi Tung, extinction in the wild due to intense and increasing Chiang Rai Province, 1991‐05‐21, R. Pooma 487 levels of deforestation. (holotype: BKF!).

3.2 Phylogenetic relationships and taxonomic treatment of Desmos yunnanensis Acknowledgements This research was supported by Our phylogenetic analyses indicate that Desmos a grant from the Hong Kong Research Grants Council yunnanensis is unequivocally associated with Dasy- (Grant No. HKU 7578/05M), awarded to RMKS and maschalon obtusipetalum (JK ¼ 100; BS ¼ 100; PP ¼ YCFS. We are grateful to Jim DOYLE and an 1.00), a species recently described from Thailand anonymous reviewer for helping to improve the (Wang et al., 2009). We believe that these two names manuscript, the curators of A and PE herbaria for refer to the same species as the chloroplast DNA providing leaf material, Caren Pearl SHIN for drawing sequences of the two accessions (matK and ndhF) are Fig. 4, staff of the Electron Microscope Unit at The identical. There are two distinct loci that can distinguish University of Hong Kong for their assistance, and Laura Desmos yunnanensis and Dasymaschalon obtusipeta- WONG for general technical assistance. lum from other Dasymaschalon species (i.e., “T” instead of “C” in position 441 bp in the alignment; and “A” instead of “T” in position 599 bp). DNA References sequences of Dasymaschalon obtusipetalum and Bân NT. 1975. Notes on the genus Dasymaschalon (Hook. f. et Desmos yunnanensis were generated independently Thoms.) Dalle Torre et Harms (Annonaceae). Botaniceskij by the second and first authors in 2008 and 2012, Žurnal (Moscow & Leningrad) 60: 223–233. (in Russian) respectively, which suggests that our finding is not an Brown JM, Hedtke SM, Lemmon AR, Lemmon EM. 2010. artifact resulting from contamination. The protologue of When trees grow too long: Investigating the causes of “ ” highly inaccurate Bayesian branch‐length estimates. Sys- Phaeanthus yunnanensis included the erroneous – fl tematic Biology 59: 145 161. statement that the owers had two whorls of three Chalermglin P. 2001. Family Annonaceae. Bangkok: Ban & petals (Hu, 1940); Bân (1975) correctly asserted that Suan. (in Thai) this was a mistake, but Hu’s error was apparently Chaowasku T, Johnson DM, Van der Ham RWJM, Chatrou LW. corroborated by Tsiang & Li (1979). The perpetuation 2012. Characterization of Hubera (Annonaceae), a new of this error led Wang et al. (2009) to overlook the true genus segregated from Polyalthia and allied to Miliusa. taxonomic affinities of the species, and they according- Phytotaxa 69: 33–56. ly described the same species under a new name, Chaowasku T, Keßler PJA, Punnadee S, Van der Ham RWJM. 2011. Taxonomic novelties and pollen morphological study Dasymaschalon obtusipetalum. Following our molec- ‐ – ‐ in the genus Neo uvaria (Annonaceae). Phytotaxa 32: 27 42. ular results, we re examined the type material of Chaowasku T, Van der Ham RWJM. 2013. Integrative “Phaeanthus yunnanensis” and confirmed that the systematics supports the establishment of Winitia, a new flowers do indeed only have three petals in one whorl. genus of Annonaceae (Malmeoideae, Miliuseae) allied to Desmos yunnanensis and Dasymaschalon obtusipeta- Stelechocarpus and Sageraea. Systematics and Biodiversi- lum are conspecific, characterized by long flowering ty 11: 195–207. pedicels, wide flowers with petal length/width ratios in Chatrou LW, Pirie MD, Erkens RHJ, Couvreur TLP, Neubig KM, Abbott JR, Mols JB, Maas JW, Saunders RMK, Chase the range 1.5–2.1, monocarps that are sparsely hairy, fi – MW. 2012. A new subfamilial and tribal classi cation of and seeds that are ellipsoid (length/width ratio 1.8 2.5). the pantropical flowering plant family Annonaceae The correct name for the species is therefore Dasyma- informed by molecular phylogenetics. Botanical Journal schalon yunnanense (Hu) Bân, and the name D. of the Linnean Society 169: 5–40.

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Walker JW. 1971. Pollen morphology, phytogeography, and —, —, AY841602, —, AY841680, —; Cleistopholis phylogeny of the Annonaceae. Contributions of the Gray glauca Pierre ex Engl. & Diels, AY841395, – Herbarium of Harvard University 202: 1 131. AY841404, —, —, AY841603, —, AY841681, —; Wang J, Chalermglin P, Saunders RMK. 2009. The genus Dasymaschalon dasymaschalum (Blume) I. M. Turn- Dasymaschalon (Annonaceae) in Thailand. Systematic — — — Botany 34: 252–265. er, JQ768549, JQ768588, , , JQ768709, , Wang J, Thomas DC, Su YCF, Meinke S, Chatrou LW, Saunders JQ768669, —; Dasymaschalon macrocalyx Finet & RMK. 2012. A plastid DNA phylogeny of Dasymaschalon Gagnep., EF179277, EF179290, —, —, AY841610, (Annonaceae) and allied genera: Evidence for generic non‐ —, AY841688, —; Dasymaschalon yunnanense (Hu) monophyly and the parallel evolutionary loss of inner Bân [¼Dasymaschalon obtusipetalum Jing Wang, P. – petals. Taxon 61: 545 558. Chalermglin & R. M. K. Saunders], JQ768560, Weberling F, Hoppe JR. 1996. Comparative morphological — — — — ﬂ JQ768598, , , JQ768720, , JQ768680, ; evaluation of in orescence characters in Annonaceae. In: ¼ Morawetz W, Winkler H eds. Reproductive morphology in Dasymaschalon yunnanense (Hu) Bân [ Desmos Annonaceae (Biosystematics and Ecology Series 10). yunnanensis (Hu) P. T. Li], C. W. Wang 79167 (A), Vienna: Österreichische Akademie der Wissenschaften. China, Yunnan, KF680919 , KF680922 , —, —, —, 29–53. —, —, —; Desmopsis microcarpa R. E. Fr., Wu C‐Y, Wang W‐T. 1957. Preliminary reports on studies of the AY518804, JX544771, —, AY841461, AY319059, plants of tropical and subtropical regions of Yunnan. Acta —, AY319173, JX544758; Desmopsis schippii Standl., Phytotaxonomica Sinica 6: 183–254. AY518805, JQ723786, —, —, AY319060, —, Xue B, Su YCF, Mols JB, Keßler PJA, Saunders RMK. 2011. — Further fragmentation of the polyphyletic genus Polyalthia AY319174, ; Desmos cochinchinensis Lour., (Annonaceae): Molecular phylogenetic support for a JQ768568, JQ768604, —, —, JQ768688, —, broader delimitation of Marsypopetalum. Systematics and JQ768728, —; Fenerivia angustielliptica (G. E. Schatz Biodiversity 9: 17–26. & A. Le Thomas) R. M. K. Saunders, O. Poncy 1540 Xue B, Su YCF, Thomas DC, Saunders RMKS. 2012. Pruning (P), Madagascar, Toamasina, JF810373, KF682115, the polyphyletic genus Polyalthia (Annonaceae) and —, KF709054, JF810385, —, JF810397, —; Feneri- resurrecting the genus Monoon. Taxon 61: 1021–1039. via chapelieri (Baill.) R. M. K. Saunders, JF810375, JQ723788, —, —, JF810387, —, JF810399, —; Friesodielsia desmoides (Craib) Steenis, JQ768577, Appendix I JQ768612, —, —, JQ768696, —, JQ768738, —; Goniothalamus grifﬁthii Hook. f. & Thomson, Voucher information and GenBank accession AY743484, EF179296, —, —, AY743446, —, numbers for samples used in this study (—, missing AY743465, —; Greenwayodendron oliveri (Engl.) data; , newly generated sequences). Voucher data are Verdc., AY743489, AY841408, —, AY841465, given for accessions for which DNA sequences were AY743451, —, AY743470, —; Guatteria anomala newly obtained, using the following format: taxon R. E. Fr., AY740913, EF179298, —, —, AY740962, name, country, largest political subdivision, collector(s) —, AY741011, —; Hubera cerasoides (Roxb.) and collection number, herbarium acronym, and matK, Chaowasku, B. Xue & P. Chalermglin XB11 (HKU), ndhF, ndhF‐rpl32, psbA‐trnH, rbcL, rpl32‐trnL, trnL‐ Thailand, Saraburi Province, AY518854, JQ723810, F, and ycf1 GenBank accession numbers. For DNA JQ723843, KF709055, AY319017, JQ723896, sequences published in previous studies, voucher AY319131, JQ723950; Hubera nitidissima (Dunal) information is available from GenBank. Chaowasku, Ford & Metcalfe 4708 (HKU), Australia, Alphonsea boniana Finet & Gagnep., AY518809, Queensland, KF682110, KF682116, —, KF709056, JQ723785, JQ723816, —, AY318965, —, AY319077, KF682103, —, KF682105, —; Maasia discolor —; Ambavia gerrardii (Baill.) Le Thomas, AY220435, (Diels) Mols, P. J. A. Kessler & Rogstad, AY518872, AY218168, —, —, —, —, AY220411 (intron) AY841416, —, AY841500, AY319021, —, AY220358 (spacer), —; Anaxagorea silvatica R. E. AY841584, —; Maasia glauca (Hassk.) Mols, P. J. Fr., AY743477, EF179280, —, —, AY743439, —, A. Kessler & Rogstad, P. Chalermglin 530522 (HKU), AY743458, —; Annickia chlorantha (Oliv.) Setten & Thailand, Bangkok, AY518871, KF682117, Maas, AY841393, AY841401, —, —, AY841594, —, KF709049, AY841501, AY319023, —, AY319137, AY841671, —; Annona glabra L., DQ125050, —; Maasia sumatrana (Miq.) Mols, P. J. A. Kessler & EF179281, —, —, AY841596, —, AY841673, —; Rogstad, AY518873, AY841418, —, AY841503, Asimina triloba (L.) Dunal, AY743479, EF179287, —, AY319039, —, AY319153, —; Marsypopetalum —, AY743441, —, AY743460, —; Cananga odorata crassum (R. Parker) B. Xue & R. M. K. Saunders, P. (Lam.) Hook. f. & Thomson, AY841394, AY841403, Chalermglin 521212‐1 (HKU), Thailand, Cha Choeng

Sao Province, HQ286571, JQ723792, JQ723822, ophthalmicus (Roxb. ex G. Don) J. Sinclair [as KF709057, HQ286577, JQ723875, HQ286583, Phaeanthus ebracteolatus (C. Presl) Merr. in Gen- JQ723929; Meiogyne cylindrocarpa (Burck) Heusden Bank], AY518863, —, —, —, AY319012, —, subsp. cylindrocarpa, AY518796, JQ723794, AY319125, —; Phaeanthus splendens Miq., JQ723824, —, AY318981, JQ723877, AY319093, AY518864, JX544777, —, JX544790, JX544754, —, JQ723931; Meiogyne hainanensis (Merr.) Bân, AY319126, JX544765; Piptostigma mortehani De JQ723773, —, JQ723829, —, JQ723860, JQ723882, Wild., AY743492, AY841415, —, —, AY743454, —, JQ723913, JQ723936; Meiogyne mindorensis (Merr.) AY743473, —; Platymitra macrocarpa Boerl., Ardi Heusden, JQ723776, JQ723800, JQ723832, —, WI 52 (HKU), cultivated in Kebun Raya Bogor, JQ723863, JQ723885, JQ723916, JQ723939; Meio- Indonesia, AY518812, JQ723809, JQ723842, gyne virgata (Blume) Miq., AY518798, JQ723805, KF709062, AY319013, JQ723895, AY319127, JQ723838, JX544784, AY318982, JQ723891, JQ723949; Polyalthia johnsonii (F. Muell.) B. Xue AY319094, JQ723945; Miliusa mollis Pierre, & R. M. K. Saunders, B. P. M. Hyland 7142 (L), AY518851, JQ690503, —, JQ690504, —, —, Australia, Queensland, JQ723767, JQ723791, AY319102, JQ690505; Miliusa velutina (Dunal) JQ723821, KF709063, JQ723854, —, JQ723907, Hook. f. & Thomson, AY518847, JQ690536, —, JQ723928; Polyalthia lateritia J. Sinclair, P. Chalerm- JQ690537, AY318993, —, AY319105, JQ690538; glin 510528 (HKU), Thailand, Songkhla Province, Mitrephora alba Ridl., P. Chalermglin 530706 (HKU), JX227890, KF682121, KF709052, KF709064, Thailand, Bangkok, AY518855, JQ723807, JQ723840, JX227915, —, JX227866, KF682104; Popowia KF709058, AY318994, JQ723893, AY319106, pisocarpa (Blume) Endl., B. Xue XB13 (HKU), China, JQ723947; Mkilua fragrans Verdc., DQ125060, Hong Kong, AY518862, JQ723812, JQ723846, EF179303, —, —, AY841634, —, AY841712, —; KF709065, AY319044, JQ723899, AY319158, Monanthotaxis whytei (Stapf) Verdc., EF179278, JQ723953; Pseuduvaria fragrans Y. C. F. Su, EF179304, —, —, AY841635, —, AY841713, —; Chaowasku & R. M. K. Saunders, T. Chaowasku 27 Monocarpia euneura Miq., AY518865, AY841412, (HKU), Thailand, Surat Thani Province, JQ723784, —, AY841477, AY318998, —, AY319111, —; JQ723813, JQ723847, KF709066, JQ723871, Monocarpia marginalis (Scheff.) J. Sinclair, P. JQ723900, JQ723924, JQ723954; Sageraea elliptica Chalermglin 540103 (HKU), Malaysia, KF682111, (A. DC.) Hook. f. & Thomson, W. H. Ardi WI 48 KF682118, KF709050, —, —, —, KF682106, —; (HKU), cultivated in Kebun Raya Bogor, Indonesia, Monoon fuscum (King) B. Xue & R. M. K. Saunders, KF682114, KF682122, —, —, —, —, KF682109, P. Chalermglin & B. Xue XB6 (HKU), Thailand, —; Sageraea lanceolata Miq., AY518799, JX544774, Phetchaburi Province, AY518787, JQ723787, —, JX544787, AY319050, —, AY319164, JX544762; JQ723817, KF709059, AY318973, —, AY319085, Sapranthus viridiflorus G. E. Schatz, AY743493, JX544767; Monoon lateriflorum (Blume) Miq., A. M. AY841422, JQ723848, AY841515, AY319051, A. S. Attanayake XX.D49a (HKU), cultivated in Kebun JQ723901, AY319165, JQ723955; Stelechocarpus Raya Bogor, Indonesia, JQ723783, JQ723811, burahol (Blume) Hook. f. & Thomson, Karim s.n. JQ723844, KF709060, JQ723870, JQ723897, (HKU), cultivated in Singapore Botanic Garden, JQ723923, JQ723951; Neo‐uvaria acuminatissima Singapore, AY518803, JQ723814, JQ723849, (Miq.) Airy‐Shaw, AY518793, —, —, —, KF709067, AY319053, JQ723902, AY319167, AY318999, —, AY319112, —; Neo‐uvaria paralleli- JQ723956; Stelechocarpus cauliflorus (Scheff.) venia (Boerl.) H. Okada & K. Ueda, AY518794, —, J. Sinclair, W. H. Ardi WI 57 (HKU), cultivated in —, —, AY319000, —, AY319113, —; Neo‐uvaria Kebun Raya Bogor, Indonesia, AY518800, telopea Chaowasku, JX544751, JX544778, —, KF682123, KF709053, JX544789, AY319054, —, JX544791, JX544755, —, JX544783, JX544766; AY319168, JX544764; Stenanona costaricensis R. E. Orophea cuneiformis King, P. Chalermglin 531108 Fr., AY518801, JX544772, —, AY841516, (HKU), Thailand, Phetchaburi Province, KF682112, AY319069, —, AY319183, JX544759; Tridimeris KF682119, —, —, —, —, KF682107, —; Orophea sp., JX544750, JX544773, —, JX544786, JX544753, sp., P. Chalermglin 530207‐1 (HKU), Thailand, —, JX544782, JX544761; Trigynaea lanceipetala Kanchanaburi Province, JQ723782, JQ723808, D. M. Johnson & N. A. Murray, AY743487, JQ723841, KF709061, JQ723869, JQ723894, EF179309, —, —, AY743449, —, AY743468, —; JQ723922, JQ723948; Phaeanthus sp., P. Chalermglin Trivalvaria costata (Hook. f. & Thomson) I. M. Turner, 540101 (HKU), Malaysia, KF682113, KF682120, M. Y. Wong 0805 (HKU), cultivated in South China KF709051, —, —, —, KF682108, —; Phaeanthus Botanic Garden, China, HQ286574, JQ723815,

JQ723850, KF709068, HQ286580, JQ723903, —, KF680925, KF680927, KF680929, KF680931, HQ286586, JQ723957; Uvaria lucida Benth. subsp. KF680933; Wangia saccopetaloides (W. T. Wang) virens (N. E. Br.) Verdc., AY238966, EF179310, —, X. Guo & R. M. K. Saunders [¼Desmos saccopeta- —, AY238957, —, EF179319, —; Wangia saccope- loides (W. T. Wang) P. T. Li], T. T. Yu 17336 (A), taloides (W. T. Wang) X. Guo & R. M. K. Saunders China, Yunnan, KF680920, KF680923, —, [¼Desmos saccopetaloides (W. T. Wang) P. T. Li], KF680924, KF680926, KF680928, KF680930, T. T. Yu 16484 (PE), China, Yunnan, KF680921, —, KF680932.