A peer-reviewed open-access journal PhytoKeys 7: 27–36 (2011) Coccinia intermedia – a new Cucurbitaceae species from West Africa doi: 10.3897/phytokeys.7.2032 reseArCh ArtICle www.phytokeys.com 27 Launched to accelerate biodiversity research Coccinia intermedia – a new Cucurbitaceae species from West Africa Norbert Holstein, Susanne S. Renner Systematic Botany and Mycology, Menzinger Str. 67, 80638 Munich, Germany Corresponding author: Norbert Holstein (holstein@lrz.uni-muenchen.de) Academic editor: H. Schaefer | Received 8 September 2011 | Accepted 9 November 2011 | Published 29 November 2011 Citation: Holstein N, Renner SS (2011) Coccinia intermedia – a new Cucurbitaceae species from West Africa. PhytoKeys 7: 27–36. doi: 10.3897/phytokeys.7.2032 Abstract Nuclear and plastid sequences from two individuals of a suspected new species of Coccinia from West Africa were added to an available molecular phylogeny for the remaining 27 species of the genus. Phylogenetic analyses of these data indicate the new species' monophyletic status and closest relatives. Based on four fertile collections, we here describe and illustrate Coccinia intermedia Holstein. We also provide a key to the Coccinia species of West Africa and map their distributions. Keywords Benin, Ivory Coast, Ghana, leaky dioecy, molecular phylogenetics, species monophyly, Togo Introduction he genus Coccinia Wight et Arn. so far consisted of 27 species distributed mainly in Sub-Saharan Africa, with centers of diversity in East Africa and southern Africa (Holstein, ongoing monograph). Only four species were known from West Africa, including C. longicarpa Jongkind, C. keayana R. Fern., and C. barteri (Hook. f.) Keay, which apparently evolved during Pliocene-Pleistocene climatic oscillations (Holstein and Renner 2011). he fourth species, C. grandis (L.) Voigt, is much more widespread, occurring not only in Africa but also in South and South East Asia, and being naturalized on several Paciic islands, Australia, and in the Neotropics. During a study of Copyright N. Holstein, S.S. Renner SS. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 28 Norbert Holstein & Susanne S. Renner / PhytoKeys 7: 27–36 (2011) the evolution and biogeography of the genus (Holstein and Renner 2011), we came across a male specimen from the northeastern Ivory Coast that in its plastid sequences difered suiciently from all other sequenced material for us to suspect it might represent a new species. We therefore provisionally labeled it Coccinia sp. nov. We have since found three more specimens of the new species, all of them with fruits, and two with lowers, and based on their morphology as well as additional nuclear and plastid sequences, we here describe the new species C. intermedia. Methods We produced new sequences of the plastid rpl20–rps12 intergenic spacer (JN653687), trnSGCU–trnGUCC intergenic spacer (JN653686) and the nuclear LEAFY-like second intron (JN653688) from the female specimen A. Akoègninou et al. 2625 (WAG0278370) of the new species, following standard procedures (Holstein and Renner 2011). We added the new sequences, named “C. intermedia 2”, to our published matrices and carried out maximum likelihood tree searches, using the approaches described in Holstein and Renner (2011). results Phylogenetic placement he two Coccinia intermedia accessions in the plastid tree form a clade (Fig. 1) within the barteri clade. In the nuclear LEAFY phylogeny, C. intermedia groups with C. barteri, C. heterophylla (Hook.f.) Holstein, C. keayana, C. longicarpa, C. mildbraedii Gilg, and C. racemilora Keraudren (Fig. 2), albeit without bootstrap support. Morphological description Coccinia intermedia sp. nov. urn:lsid:ipni.org:names:77115897-1 http://species-id.net/wiki/Coccinia_intermedia A Coccinia longicarpa difert calycis dentibus angustis, corolla campanulata et fructu elliptico ad oblongo. A C. keayana et C. grandis difert calycis dentibus ad corollam adpressis vel apicem versus leviter recurvatis et lamina foliorum subtus glandibus fuscis provisa. A C. barteri difert loribus femineis 1–3 fasciculatis non racemosis, corolla campanulata. Coccinia intermedia – a new Cucurbitaceae species from West Africa 29 Cucumis sativus 100 Cucumis hirsutus Scopellaria marginata Peponium vogelii 100 Coccinia barteri 7 Coccinia barteri 1 Coccinia barteri 3 Coccinia barteri 6 Coccinia racemiflora 2 Coccinia racemiflora 1 Coccinia barteri 4 Coccinia barteri 2 Coccinia barteri 5 Coccinia subsessiliflora Coccinia heterophylla Coccinia longicarpa Coccinia keayana 1 93 Coccinia keayana 2 Diplocyclos schliebenii Diplocyclos palmatus } Coccinia intermedia 2 100 Coccinia intermedia 1 Coccinia mildbraedii 99 Coccinia mildbraedii 2 Coccinia mackenii 98 Coccinia quinqueloba Coccinia hirtella 2 97 Coccinia hirtella 1 Coccinia sessilifolia Coccinia senensis 5 Coccinia senensis 6 Coccinia senensis 3 Coccinia senensis 4 Coccinia senensis 2 Coccinia senensis 1 Coccinia aurantiaca 3 Coccinia aurantiaca 4 Coccinia aurantiaca 2 Coccinia aurantiaca 1 Coccinia samburuensis Coccinia schliebenii 1 Coccinia schliebenii 2 Coccinia grandiflora 1 99 Coccinia grandiflora 2 Coccinia adoensis 2 80 85 Coccinia adoensis 5 Coccinia adoensis 8 Coccinia adoensis 3 Coccinia adoensis 7 86 Coccinia adoensis 4 Coccinia adoensis 9 Coccinia adoensis 1 94 100 Coccinia adoensis 6 Coccinia grandis 3 Coccinia grandis 1 98 Coccinia grandis 2 Coccinia ogadensis Coccinia abyssinica 1 85 Coccinia abyssinica 2 Coccinia megarrhiza 2 86 Coccinia megarrhiza 3 Coccinia megarrhiza 1 Coccinia megarrhiza 4 Coccinia microphylla 2 Coccinia microphylla 1 Coccinia trilobata Coccinia rehmannii 1 90 Coccinia rehmannii 2 var. littoralis Coccinia rehmannii 3 var. rehmannii Coccinia rehmannii 4 „ovifera“ } 100 81 Figure 1. Maximum likelihood phylogeny for Coccinia based on plastid DNA sequences analyzed under GTR+Γ model of substitution. he tree is based on 4,551 nucleotides (140 parsimony-informative sites) from the trnSGCU–trnGUCC intergenic spacer (IS), the rpl20–rps12 IS, the ndhF–rpl32 IS, trnLUAA intron, trnLUAA–trnFGAA IS, and the matK gene (expanded matrix from Holstein and Renner 2011). Numbers below branches represent bootstrap support ≥ 80% from 1000 replicates. Dots on branches and behind brackets refer to uniquely shared insertions or deletions. Species names follow Holstein and Renner (2011) except for the new species C. intermedia 1, earlier called Coccinia sp. nov. Type. BENIN. Atakora: Natitingou, Kouaténa (Perma), 10°12.00'N; 1°30.18'E, river bed, female, l, fr, 3 Oct 2000, A.Akoègninou et al. 3625 (Holotype: WAG0278370!; isotype: WAG0278369!). Description. Perennial, diclinous climber. Shoot length unknown, but likely several meters. Shoots lignify with whitish bark and up to 1 cm diam. Fresh shoots green, glabrous, older shoots with clear to white pustules. Petioles 2.8–10.8 cm, glabrous, when older with clear to white pustules (Fig. 3a). Leaves 6–15 × 7–18 cm, shallowly to profoundly 5-lobate, more or less auriculate (Fig. 4). Upper lamina glabrous with clear to whitish pustules. Lower lamina paler than upper lamina, glabrous, often with small dark glands near the leaf base (Fig. 3a). Tendrils simple or 30 Norbert Holstein & Susanne S. Renner / PhytoKeys 7: 27–36 (2011) Peponium vogelii 100 Diplocyclos schliebenii Diplocyclos palmatus Coccinia hirtella 1 Coccinia quinqueloba Coccinia sessilifolia Coccinia microphylla 2 Coccinia rehmannii 3 var. rehmannii Coccinia rehmannii 1 Coccinia microphylla 1 94 Coccinia rehmannii 4 "ovifera" Coccinia megarrhiza 2 99 Coccinia megarrhiza 4 Coccinia samburuensis Coccinia adoensis 9 Coccinia adoensis 2 95 Coccinia aurantiaca 3 Coccinia aurantiaca 2 Coccinia senensis 1 91 Coccinia senensis 2 Coccinia mildbraedii 2 Coccinia keayana 2 Coccinia barteri 4 98 Coccinia racemiflora 1 Coccinia racemiflora 2 Coccinia barteri 6 Coccinia heterophylla 99 Coccinia barteri 2 Coccinia longicarpa Coccinia intermedia 2 Coccinia schliebenii 2 Coccinia grandiflora 2 Coccinia grandiflora 1 Coccinia grandis 1 99 Coccinia grandis 3 Coccinia grandis 2 Figure 2. Maximum likelihood phylogeny for Coccinia based on nuclear DNA sequences from the LEAFY-like 2nd intron analyzed under the GTR+Γ model of substitution. he tree is based on 505 nucleotides (56 parsimony-informative sites). Numbers below branches represent bootstrap support ≥ 80% from 100 replicates. Dots on branches and behind brackets refer to uniquely shared insertions or deletions. Species names follow Holstein and Renner (2011) except for the new species C. intermedia. biid. Probracts up to 2.5 mm long, glabrous, apex rounded (Fig. 3a). Male lowers in few-lowered racemes (Fig. 5), likely sometimes accompanied by a single lower. Common peduncle up to 1 cm, pedicels in racemose lowers 2–4 mm, glabrous. Bracts up to 1.5 mm long, round to obovoid. Receptacle pale green, glabrous. Calyx teeth 1.5 mm long, lineal to narrow triangulate, erect with slightly recurved tips (Figs. 3–5). Corolla campanulate, 1.6 cm long, pale reddish-yellow to yellow, lobes 0.7 cm long (Fig. 5). Anthers sinuate, in a globose head (Fig. 3c). Pollen unknown. Female lowers 1–3 clustered (strongly reduced raceme; Fig. 4). Pedicels 0.6–1.2 cm, glabrous. Perianth like in males. Ovary fusiform, glabrous. Stigma and staminodes unknown. Fruit 4.5 × 2.5 cm, elliptical to oblong, smooth. Unripe green with pale green longitudinal mottling. Ripe orange?, more likely becoming red via orange ripening stage. Fruit with waxy cover. Size of mature seeds unknown (≥ 5.5 × 3.5 × 1.3 mm), symmetrical (to slightly asymmetrical), face lat (Fig. 3b). Distribution. (Fig. 6). NE Ivory Coast, SE Ghana (likely also in the north), S Togo (likely also in the north), NW Benin. Based on the current collections, Coccinia intermedia is likely to occur in the Dahomey Gap region and the Isoberlinia woodlands of West Africa. Ecology. Wooded grasslands (semi-humid savanna), woodlands, dry forests, and along rivers. Flowering specimens have been collected during May, August, and October, which in each site was during or shortly after the rainy season. Coccinia intermedia – a new Cucurbitaceae species from West Africa 31 Figure 3. a Coccinia intermedia leaf basis and node with lowers b seeds from late, but immature fruit c node with young fruit and male lower bud with sinuate anthers; all from J.B.Hall & J.M.Lock GC46016 (K). Etymology. he epithet refers to the species' status as the only Coccinia from West Africa that occurs in habitats intermediate between semi-arid and humid conditions. Morphologically, C. intermedia combines characters also found in the other four West African species although not in this combination. List of specimens examined. Benin: Atakora, Natitingou, Kouaténa (Perma), 10°12.00'N; 1°30.18'E, river bed, female, l, fr, 3 Oct 2000, A.Akoègninou et al. 3625 (WAG 2 sheets). Ghana: Shai Hills Game Reserve, monoecious, l, fr, 25 May 1976, J.B.Hall & J.M.Lock GC 46016 (K 4 sheets, MO). Ivory Coast: Bouna, male, l, 10 Aug 1967, C.Geerling & J.Bokdam 662 (MO, WAG). Togo: between Lomé and Aného, female, fr, 25 Jun 1994, L.Aké Assi 18982 (MO). 32 Norbert Holstein & Susanne S. Renner / PhytoKeys 7: 27–36 (2011) Figure 4. Habitus of Coccinia intermedia as reconstructed from J.B.Hall & J.M.Lock GC46016 (K). Key to West African Coccinia species 1 1' 2 2' Plant glabrous. Leaves with few large pale glands between main nerves of lower lamina. Nerves on lower lamina with or without white pustules. Leaf margin dentate, in mature plants often red to brown (black when dry). Tendrils always simple. Male and female lowers 1 solitary (rarely male lowers clustered or in short-peduncled racemes). Calyx teeth spreading to relexed, tips red to brown. Corolla campanulate, white or buf. Fruit ovoid. Plant of semi-arid habitats......................................................................... C. grandis Plant glabrous or with hairs, especially on adaxial petiole. Leaves with small blackish glands (often many) centered towards the leaf base or without glands on lower lamina. Tendrils simple or biid. Male and female lowers in racemes or solitary. Corolla in yellowish tones, never white. .....................................2 Plant glabrous. Leaves with small blackish glands centered towards the leaf base (Fig. 3). Nerves on lower lamina with or without white pustules. Leaf margin at maturity with colored teeth (color in living plants unknown, black when dry). Tendrils simple or biid. Male lowers (Fig. 5) bracteate, in fewlowered racemes, female lowers 1–3 solitary/clustered (Fig. 3 and 4). Calyx teeth erect with recurved tips (Figs 3–5). Corolla campanulate. Fruit ovoid to short cylindrical. Plant of wooded grasslands (tree savanna), woodlands, or dry forests. ............................................................................. C. intermedia Plant glabrous or with hairs, esp. on adaxial petiole. Leaves with small blackish glands centered towards the leaf base or without glands. Nerves on lower Coccinia intermedia – a new Cucurbitaceae species from West Africa 33 Figure 5. Male inlorescence of Coccinia intermedia from C.Geerling & J.Bokdam 662 (WAG). 3 leaf lamina without white pustules. Tendrils simple or biid. Male lowers in few to many-lowered racemes, rarely accompanied by a solitary lower. Female lowers in few- to many-lowered racemes or solitary. Flowers bracteate or ebracteate. Corolla urn-, cup- to funnel-shaped. Plant of humid climates (rainforests, gallery forests, etc.) ..................................................................3 Leaf margin with pale (when dry blackening) glandular teeth. Tendrils simple. Flowers without bracts, calyx teeth erect, > 1.5 mm at base. Fruits long cylindrical. .............................................................................. C. longicarpa Norbert Holstein & Susanne S. Renner / PhytoKeys 7: 27–36 (2011) 34 3' 4 4' Leaf margin without conspicuously colored teeth. Tendrils simple or biid. Flowers with or without bracts. Calyx teeth erect, spreading, or relexed, but narrow (< 1.2 mm at base). Fruits ovoid. ....................................................4 Tendrils simple. Male lowers in lax racemes, female lowers solitary or in few-lowered racemes. Flowers without bracts. Calyx teeth in buds spreading, later relexed. ..............................................................................C. keayana Tendrils simple or biid. Male lowers in dense few- to many-lowered racemes, with or without bracts. Female lowers in racemes, rarely solitary. Flowers with or without bracts. Calyx teeth variable. .................... C. barteri Discussion Coccinia intermedia is morphologically similar to the other West African species. From C. grandis, it difers most readily in the glands on the lower lamina and in its calyx teeth (erect with recurved tips in C. intermedia and spreading to relexed in C. grandis). From C. longicarpa, it difers in its ovoid fruits (instead of long cylindrical fruits in C. longicarpa). Additionally, C. longicarpa has ebracteate racemes and much broader (> 1.5 mm at the base) erect calyx teeth, and an urn-shaped corolla. From C. keayana, it difers in having bracteate male lowers in denser racemes, a campanulate corolla and calyx teeth that are adpressed to the corolla with recurved tips, instead of spreading (in buds) to relexed calyx teeth. Secure distinction of C. intermedia from C. barteri requires fertile material with lowers (see the key above). Ecologically, the new species is a member of White's (1983) Sudanian center of endemism and his Guinea-Congolia/Sudania regional transition zone (Fig. 6). he only species with a similar habitat as C. intermedia is C. adoensis, the most western known occurrence of which is Adamawa State (eastern Nigeria). Whether the species co-occur is not known. hey could be distinguished by fruit shape (not beaked in C. intermedia, beaked in C. adoensis, although this character can vary in the latter). Additionally, C. adoensis has inlorescence peduncles that are longer than 1 cm (in its male racemes) and petioles that are often hairy. Two DNA characters, namely base pairs 310 and 323 in the trnSGCU–trnGUCC intergenic spacer region, suggest the placement of C. intermedia as sister to a clade that we have earlier referred to as the Coccinia barteri clade (Holstein and Renner 2011). If this placement is correct, then the Coccinia species occurring in the rain or mist forests of West and Central African are monophyletic and probably evolved in situ. One of the four collections, J.B.Hall & J.M.Lock GC 46016, bears male and female lowers/fruits on the same node (Fig. 3c). he male lowers are buds, and it is not clear, whether they are fertile. Kumar and Vishveshwaraiah (1952) report a “gynodioecious form” of C. grandis in which the male lowers of the hermaphrodite (monoecious) plants are sterile. An occasional occurrence of bisexual plants in otherwise dioecious species, sometimes called “leaky dioecy” (Baker and Cox 1984), has also been observed in other Cucurbitaceae (Schaefer and Renner 2010). Coccinia intermedia – a new Cucurbitaceae species from West Africa 35 Figure 6. Map of West African Coccinia species. Pale yellow circles = C. intermedia, cyan circles = C. grandis, dark blue circles = C. barteri, pale blue squares = C. keayana, bright blue triangles = C. longicarpa. hick dark grey lines are phytochoria drawn after White (1983), I = Guineo-Congolian regional center of endemism, III = Sudanian regional center of endemism, XI = Guinea-Congolia/Sudania transition zone, XVI = Sahel regional transition zone. hin dark grey lines (after White (1983)) diferentiate between White’s vegetation types of zone III: 27 = Sudanian woodland with abundant Isoberlinia; 29a = undifferentiated Sudanian woodland. Location of C. intermedia in Ivory Coast estimated (only the department is given on the herbarium sheet). However, true monoecy in C. intermedia would be surprising as none of ca. 1,500 specimens of other Coccinia species studied is bisexual (Holstein, ongoing monograph). Acknowledgements We thank Dietrich Podlech for help with the diagnosis. We also thank the curators of K, MO, and WAG for sending material on loan, and permission to dissect material (K) and extract DNA (MO, WAG). he work was supported by German Research Council (RE 603/6–1 and 6–2). references Baker HG, Cox PA (1984) Further thoughts on dioecism and islands. Annals of the Missouri Botanical Garden 71: 244–253. Holstein N, Renner SS (2011) A dated phylogeny and collection records reveal repeated biome shifts in the African genus Coccinia (Cucurbitaceae). BMC Evolutionary Biology 11: 28. doi: 10.1186/1471-2148-11-28 36 Norbert Holstein & Susanne S. Renner / PhytoKeys 7: 27–36 (2011) Kumar LSS, Vishveshwaraiah S (1952) Sex mechanism in Coccinia indica Wight and Arn. Nature 170: 330–331. doi: 10.1038/170330a0 Schaefer H, Renner SS (2010) A three-genome phylogeny of Momordica (Cucurbitaceae) suggests seven returns from dioecy to monoecy and recent long-distance dispersal to Asia. Molecular Phylogenetics and Evolution 54: 553–560. doi: 10.1016/j.ympev.2009.08.006 White F (1983) he vegetation of Africa. Unesco, Paris, 356 pp.