Enlarging the monotypic Monocarpieae (Annonaceae, Malmeoideae): recognition of a second genus from Vietnam informed by morphology and molecular phylogenetics Author Chaowasku, Tanawat Author Damthongdee, Anissara Author Jongsook, Hathaichanok Author Ngo, Dung T. Author Le, Hung T. Author Tran, Duc M. Author Suddee, Somran text Candollea 2018 2018-11-23 73 2 261 275 journal article 28564 10.15553/c2018v732a11 a6c2daaa-106b-4373-9e9b-22b88fa2276e 2559368 Leoheo Chaowasku , gen. nov. Typus: Leoheo domatiophorus Chaowasku, D.T. Ngo & H.T. Le. Medium-sized to large trees ; indumentum of simple hairs; intramarginal leaf veins absent, tertiary leaf venation percurrent; inflorescences 1- or few-flowered, axillary; bracts present; flowers bisexual, both petal whorls of ± equal size; stamens 70– 76, connective truncate and dilated, covering thecae; carpels 3– 4 per flower, free in flower and fruit; stigmas ± peltate and lobed; ovules many and arranged in two rows, placentation lateral; monocarps subsessile, cylindrical, monocarp abscission basal, pericarp thick and hardened when dry; aril absent; endosperm ruminations spiniform. Etymology. – The generic epithet Leoheo is from the local Vietnamese name of “Lèo Heo” for this plant and is designated as a masculine noun of nominative singular in third declension with genitive singular “Leoheonis”. Fig. 1. – 50% majority-rule consensus phylogram derived from Bayesian inference of combined seven plastid DNA regions. Bayesian posterior probabilities (PP) indicated on the right; maximum likelihood bootstrap (BS) percentages in the middle; parsimony symmetric resampling (SR) percentages on the left [** denotes BS/SR <50%]. DEN. = Dendrokingstonieae ; MAL. = Malmeeae ; MIL. = Miliuseae ; MON. = Monocarpieae ; PIP. = Piptostigmateae . Scale bar unit = substitutions per site. Notes . – The principal morphological differences between Leoheo and Monocarpia are highlighted in Table 1 . Furthermore, in view of molecular phylogenetics, the genera Leoheo and Monocarpia each is characterizable not only by nucleotide substitutions, but also by an indel structure in the psbA-trnH intergenic spacer for Leoheo and another indel structure in the trnL-trnF intergenic spacer for Monocarpia . Although the other two species of Monocarpia , M. borneensis Mols & Kessler and M. kalimantanensis Kessler , have not been included in the present molecular phylogenetic analyses due to the failure in DNA amplification, their morphologies (e.g. Fig. 2A, 3B ) substantially coincide with those of M. euneura Miq. and M. maingayi (Hook. f. & Thomson) I.M. Turner ( Fig. 2B ; TURNER, 2012 ), hence we are convinced that such two missing species (both or any of them) will not retrieve as the sister group of Leoheo or the Leoheo-Monocarpia clade. It is noteworthy that each of the axillary inflorescences of the genus Leoheo often contains leaf-like bract(s) at the top of peduncle ( Fig. 3A, 6 ), probably this feature is a transition to the terminal inflorescences characteristic for the genus Monocarpia ( Fig. 3B ). The presence of domatia on the lower leaf surface is a rare phenomenon in Annonaceae . It occurs only in a limited number of genera and species, for examples, Annona L. ( VAN DEN BOS et al., 1989 ), Dendrokingstonia Rauschert ( CHAOWASKU et al., 2012b ), Huberantha Chaowasku ( CHAOWASKU et al., 2012a ), Mitrephora (Blume) Hook. f. & Thomson ( WEERASOORIYA & SAUNDERS, 2010 ), and Tridimeris Baill. ( ORTIZ-RODRIGUEZ et al., 2016 ). Of these genera, the hairy type of domatium similar to that of Leoheo ( Fig. 4A ) can be found in Annona , Huberantha , and Mitrephora ( CHAOWASKU et al., 2012a ). The well-supported sister relationship of Fenerivieae and Maasieae is reported herein for the first time ( Fig. 1 ). The two tribes share a number of features, e.g., axillary inflorescences, generally one ovule per ovary, and spiniform endosperm ruminations ( MOLS et al., 2008 ; SAUNDERS et al., 2011 ). Their possible closest relationships were previously discussed ( SCHATZ & LE THOMAS, 1990 ; SAUNDERS et al., 2011 ). The tribes Phoenicantheae , Monocarpieae , Dendrokingstonieae , and Miliuseae constitute a strongly supported clade substantially composed of Asian-Pacific species. The fact that the Sri Lankan endemic tribe Phoenicantheae is the sister group of the remainder of this clade coupled with the restricted distribution of the tribes Monocarpieae and Dendrokingstonieae might have some biogeographic implication, especially on the geographic origin of Miliuseae , the most diverse tribe of Malmeoideae ( CHATROU et al., 2012 ), but it is currently not possible to perform an in-depth biogeographic analysis because there are still unignorable phylogenetic uncertainties, especially the unsupported sister relationships of Dendrokingstonieae and Miliuseae , as well as of Malmeeae and a clade composed of Fenerivieae and Maasieae ( Fig. 1 ). So far, no macromorphological features have yet been found to be synapomorphic for the Phoenicantheae-Monocarpieae-Dendrokingstonieae- Miliuseae clade. Nevertheless, some palynological correlations have been observed, i.e., any taxa of Malmeoideae recovered outside Miliuseae possess monosulcate pollen ( CHAOWASKU et al., 2012b , 2014 ). Currently the pollen data of Phoenicanthus and Leoheo are unavailable, but they are anticipated to exhibit monosulcate pollen based on such correlations. In addition, it is worthwhile to note that Phoenicantheae , Monocarpieae , and Dendrokingstonieae all possess a highly reduced carpel number to 1–4 per flower ( HUBER, 1985 ; CHAOWASKU et al., 2012b ). It is likely that the reduction in carpel number per flower is the ancestral trait of the Phoenicantheae-Monocarpieae- Dendrokingstonieae-Miliuseae clade. Table 2 compares the important macromorphological and pollen morphological characters of the amended Monocarpieae and the other three closely related tribes: Phoenicantheae , Dendrokingstonieae , and Miliuseae . Monocarpia , the sister group of Leoheo , occurs in southern Thailand, Peninsular Malaysia, Sumatra, and Borneo ( TURNER, 2012 ); the shortest distance of the two genera is about 1300 km away. This disjunct distribution pattern is, however, not unprecedented. In Phaeanthus Hook. f. & Thomson ( Malmeoideae , Miliuseae ; CHATROU et al., 2012 ), P. vietnamensis Bân is the only species occurring in Indochinese Peninsula; the remaining species occur in southern Thailand Provinces bordering Malaysia ( GARDNER et al., 2015 ), Malay Peninsula to the Philippines and New Guinea ( MOLS & KESSLER, 2000a ). In Neo-uvaria Airy Shaw ( Malmeoideae , Miliuseae ; CHATROU et al., 2012 ), the recently described N. laosensis Tagane & Soulad. hitherto endemic to central Laos ( TAGANE et al., 2018 ) is the only species disjunctly occurring north of Peninsular Thailand and Table 1. – Principal morphological differences between Leoheo Chaowasku and Monocarpia Miq.

Characters	Leoheo	Monocarpia
Intramarginal leaf veins	Absent	Present
Domatia on lower leaf surface	Present	Absent
Inflorescence position	Axillary	Terminal
Monocarp surface	With longitudinal ridges	Without longitudinal ridges
Monocarp base	Contracted into a short and stout stipe	Not contracted into a stipe

Fig. 2. – A . Leaf of Monocarpia kalimantanensis Kessler , showing conspicuous intramarginal veins; B . Fruit of Monocarpia maingayi (Hook. f. & Thomson) I.M. Turner, showing monocarps without longitudinal ridges; C–H: Leoheo domatiophorus Chaowasku , D.T. Ngo & H.T. Le ; C. Leaf without intramarginal veins; D. Fruit, showing monocarps with longitudinal ridges; E. Flowering branches; F. Dissected flower and young fruit; G . Dissected flower, showing detached stamens and stigmas; H. Flower, showing enlarged and irregularly lobed stigmas. [ A: Sidiyasa et al. 3469 , L; B: Gardner & Sidisunthorn ST0541a , L; C–D: Chaowasku 131 , CMUB; E–H: HUAF collectors 2009-0 3-19-ND , CMUB] [Photos: A: Arbainsyah; B: S. Gardner & P. Sidisunthorn; C–H: D.T. Ngo] Table 2. – Comparisons of important macromorphological and pollen morphological features of four closely related tribes in Malmeoideae .

Characters	Phoenicantheae	Monocarpieae	Dendrokingstonieae	Miliuseae
Tertiary leaf venation	Reticulate	Percurrent	Percurrent	Reticulate to percurrent
Inflorescence position	Axillary or terminal	Axillary or terminal	Axillary	Axillary or terminal
Stamen number per flower	6 or 9	35–76	9–56	3 to ≥ 200
Stamen connective tissue	With a minute prolongation, not covering thecae	Truncate and dilated, covering thecae	Truncate and dilated, covering thecae	Without an apical prolongation; or with a minute prolongation, not covering thecae; or truncate and dilated, covering thecae; or +/– truncate, not covering thecae
Carpel number per flower	1–3	1–4	1–2	1 to ≥ 70
Pollen apertural system	Monosulcate	Monosulcate	Monosulcate	Cryptoaperturate/disulculate

Malaysia, the nearest area where three other species of Neouvaria can be found ( CHAOWASKU et al., 2011 ). Additionally, in Disepalum Hook. f. ( Annonoideae , Annoneae ; CHATROU et al., 2012 ) subg. Enicosanthellum (Bân) P.S. Li, D.C. Thomas & R.M.K. Saunders , a clade composed of two particular species: D. petelotii (Merr.) D.M. Johnson and D. plagioneurum (Diels) D.M. Johnson , both occurring in China and Vietnam (and Laos for the former; JOHNSON, 1989 ) is the sister group of D. pulchrum (King) J. Sinclair ( LI et al., 2015 , 2017 ) which can only be found in southern Thailand Provinces bordering Malaysia ( CHAMCHUMROON et al., 2017 ) and Malay Peninsula ( JOHNSON, 1989 ). It is interesting to understand the plausible biogeographic scenarios and other biotic/abiotic factors shaping the mentioned disjunct distribution pattern within Southeast Asia.