A phylogeny of () based on morphological and anatomical data

2 3 YA-YI HUANG!, SCOTT A. MORI , AND GHILLEAN T. PRANCE

1 The New York Botanical Gardenffhe City University of New York, Bronx, New York, NY 10458-5126, USA; e-mail: [email protected] zInstitute of Systematic Botany, The New York Botanical Garden, Bronx, New York, NY 10458­ 5126, USA; e-mail: [email protected] 3 The Old Vicarage, Silver Street, Lyme Regis, Dorset, UK; e-mail: [email protected]

Abstract. Cariniana as previously circumsclibed is a genus of 16 species restricted to neotropical forest habitats on well-drained sites. A phylogenetic analysis of the genus based on 33 morphological and anatomical characters was undertaken. The results show that Cariniana consists of two clades: the AllantomalCariniana decandra clade includes lineata and seven species of actinomorphic-flowered Cariniana and is characterized by 5-merous , camose petals, incurved petal apex, scarcely lobed calyces, eucamptodromous secondary veins, dichotomizing venation, and poorly developed areolation; the C. legalis clade is made up of nine species and is characterized by an obliquely zygomorphic androecium, reticulate tertialy venation, and anomocytic stomata. The actinomorphic-flowered Cariniana are more closely re­ lated to the monotypic Allantoma lineata than they are to the species of the C. legalis clade. In order to reflect these relationships, Cariniana is divided into two genera: species in the C. legalis clade, which includes the generic type C. legalis, remain as Cariniana while species of Cariniana in the AllantomalCariniana decandra clade are transferred to Allantoma. The following new combinations are proposed: AIIantoma decandra, A. integrifolia, A. kuhImannii, A. pluriftora (a nomen novum for Cariniana multiflora because Allantoma multiflora is a synonym of multiflora), A. pachyantha, A. pauciramosa, and A. uaupensis. Key Words: Allantoma lineata, Cariniana, , floral symmetly, Lecythidoideae.

Cariniana Casar. is a Neotropical genus of Miers reinstated the genus Cariniana and the pantropical family Lecythidaceae. This pointed out the striking differences between genus was named after Prince Eugene de Cariniana and Couratari in the androecium Savioe-Carignan who sponsored Casaretto's and seed wings. The definition of the genus trip to . The genus Cariniana was has not changed since Miers. Fifteen species recognized by only a few botanists after it were recognized in the most recent mono­ was published and species of Cariniana were graph (Prance, 1979b), and one species (c. generally published in Couratari Aubi. The parvifolia) has been descIibed since publica­ distinction between Cariniana and Couratari tion of that monograph (Mori, 1995). was not established for a long time because Species of Cariniana have the smallest species of both genera have cylindric or flowers in Lecythidaceae, 3-10cular ovaries, campanulate and winged seeds. Berg cylindric fruits, and seeds with unilateral (1856) placed Cariniana in synonymy under wings. Species of the genus are either canopy Couratari, and his classification was followed or emergent distributed in non-flooded by Bentham and Hooker (1865). In 1874, habitats (terra firme) in northern Colombia,

Brittonia, 60(1), 2008, pp. 69-81. ISSUED: 30 April 2008 © 2008, by The New York Botanical Garden Press, Bronx, NY 10458-5126 U.S.A. 70 BRITTONIA [VOL. 60

Venezuela, Brazil, the coastal forests of eastern did not formally recognize these differences, he Brazil, and the cerrado vegetation of central was aware that there were two different groups Brazil. Representatives of the genus are absent in his concept of Cariniana. in the Guianas and eastern Amazonia. In recent Ducke (1925, 1948) was the first to point keys to the genera of Lecythidaceae (Mori & out the similarity of Allantoma lineata Miers Prance, 1990; Prance & Mori, 1979), Carini­ to some species of Cariniana (e.g., C. ana has been included under both "androecium uaupensis). In his paper, he compared species actinomorphic" and "androecium zygomor­ of different families growing in periodically phic" leads in the keys because the flowers of flooded habitats along Amazonian rivers with some of the species are actinomorphic related species growing in non-flooded hab­ (Fig. lA) while others are zygomorphic itats, and stated that the only difference (Fig. IB). Prance (1979b) placed species of between the two genera of Lecythidaceae Cariniana with actinomorphic flowers and a studied was that the seeds of Cariniana relatively low number of stamens together possess a wing that facilitates dispersal by in his sequence of species numbered 5-11 the wind whereas those of Allantoma have (c. decandra Ducke, C. integr[folia Ducke, C. elongated seeds without wings. The seeds of kuhlmannii Ducke, C. multiflora Ducke, C. A. lineata fall into the water when the fruits pachyantha A. C. Sm., C. pauciramosa W. A. open and are dispersed by currents. Ongoing Rodrigues and C. uaupensis (Spruce ex O. molecular studies (Mori et al., 2007) show Berg) Miers). Species with zygomorphic flow­ that a least one species of actinomorphic­ ers usually have more than 40 stamens inserted flowered Cariniana (c. decandra) fonns a over the interior of the androecium, and group with the monotypic Allantoma lineata, form another group including C. domestica which also has actinomorphic androecia. (Mart.) Miers, C. estrellensis (Raddi) Kuntze, Additional features shared between Allan­ C. ianeirensis R. Knuth, C. legalis (Mart.) toma and actinomorphic-flowered Cariniana Kuntze, C. micrantha Ducke, C. parvifolia S. are 5-merous flowers, camose petals hooked A. Mori, Prance & Menandro, C. pendultflora at the apex, and percurrent tertiary venation. Prance, C. pyriformis Miers, and C. rubra The name Allantoma comes from the Gardner ex Miers. Although Prance (1979b) Greek cx.Mcx.<; (= sausage) and 51-1-0<; (= like)

A [

3mm.

Cariniana integrifolia Cariniana micrantha FIG. 1. Androecia of Cariniana. A. Actinomorphic androecium of a species of the AllantomalG. decandra clade. B. Zygomorphic androecium of a species of the G. legalis clade. 2008] HUANG ET AL.: PHYLOGENY OF CARINIANA (LECYTHIDACEAE) 71 in reference to the cylindrical fiuit. When and small watercourses of the upper Orinoco, Miers (1874) first published this genus to Negro, and Amazon River basins. In the latter accommodate 12 species, he included the it occurs from the mouth of the Rio Negro flowers of species of Couratari and the fruits downstream to the mouth of the Amazon and seeds of Allantoma lineata in his generic River. It is characterized by actinomorphic concept. Using this concept, Huber (1902) androecia, percurrent tertiary venation, car­ described the new genus Goeldinia based on nose petals with hooked apices, cylindrical a flowering collection of Allantoma lineata. , and non-winged seeds. Later Ducke (1925) recognized that Huber's The purpose ofthis paper was to address the species of Goeldinia represented the flowers following questions: 1) what is the relationship of the part of Allantoma defined by fiuits and between species of Cariniana with actino­ seeds in Mier's protologue, and placed morphic androecia and species of Cariniana Huber's two species of Goeldinia in synony­ with zygomorphic androecia, and 2) what is my under A. lineata. It was Eyma (1932) who the relationship between Cariniana and first realized that Miers had included two Allantoma lineata? We addressed these two genera in Allantoma, one corresponding to questions by constructing a phylogeny based flowering material (= Couratari) and the other on anatomical and morphological characters. represented by fruiting material (= Allan­ toma). The fact that Miers did not select a type species further complicated the issue. Materials and methods Eyma pointed this problem out to Sprague Morphological and anatomical characters (1932) who selected A. torulosa Miers as the are based on original observations from her­ lectotype fi'om among those species recog­ barium specimens archived at The New York nized by Miers that were based on fruit Botanical Garden (NY) and the Missouri collections. He argued that the name Allan­ Botanical Garden (MO), or are derived from toma referred to the shape of the fiuit, and, the pickled collection of Lecythidaceae at NY. thus, the generic type should be based on a The ingroup consists of Allantoma lineata species typified by a fruiting collection. To (Prance, 1979a), all 15 species of Cariniana have lectotypified Allantoma with a flowering recognized by Prance in the most recent collection would have created a synonym of monograph (Prance, 1979b), and one species the zygomorphic-flowered Couratari. desclibed after the publication of the mono­ Following Eyma's definition, Knuth (1939) graph (Mori, 1995). Because the relationship recognized 12 species ofAllantoma and placed of Cariniana and Allantoma with other those with 4-chambered fruits in section Neotropical Lecythidaceae is uncertain, six Tetrakolpos and those with 5-chambered fruits species were chosen as outgroups: two species in section Pentakolpus. Most species recog­ from two actinomorphic-flowered genera nized by Knuth were distinguished by minor ( cauliflora L. and Gustavia hexapetala fruit differences. Prance (1 979a), the most (AubI.) Sm.), two species (Couroupita guia­ recent monographer of the genus, stated that nensis Aubi. and Couroupita nicaraguarensis different stages of fruit development, the DC.) from the basal genus of the zygomor­ effect of growing in different habitats, natural phic-flowered group as indicated by molecular fruit variation even within the same individ­ studies (Mori et aI., 2007), and two species ual, and the different methods of drying the ( Aubi. and Couratari fiuits result in considerable variation in size, stellata A.C.M.) from Couratari, a genus that shape, and surface texture. The number of shares morphological features with Cariniana locules in the ovary is also too variable to be a such as cylindrical fiuits, seed wings, and criterion for delimiting species. Therefore, foliaceous cotyledons. Cariniana and Coura­ Prance (l979a) reduced all of Miers and tari have been confused in the past, and one Kunth's species to synonymy under Allan­ recent analysis of molecular data suggests that toma lineata. Couratari is closer to Cariniana than it is to Allantoma lineata is a small to medium­ other zygomorphic-flowered genera (Mori sized found in wet habitats along large et aI., 2007). 72 BRITTONIA [VOL. 60

CHARACTERS todromous secondary veins toward the base and fragments used for study of venation brochidodromous veins for the rest of the leaf patterns and stomatal types were put in tubes blade, and are classified as brochidodromous. with 50% alcohol and the tubes were submerged 4. Domatia in axiis ofsecondary veins (Fig. 2).0= absent, 1= present. Cariniana domestiea, C. in boiling water for 10 min. This procedure was repeated two or three times until the color ofthe estrellensis, C. mierantha, C. pyriformis, c. pendulifiora, and C. rubra and two outgroup solution did not change. The san1ples were then cooled to room temperature, washed with water species, and Couroupita two or three times, and changed to 2% NaOH niearaguarensis, possess small cavities (doma­ solution. The NaOH solution was replaced tia) at the junction of the secondary vein with every day until the mesophyll ofthe leafbecame the primary vein. These cavities are often lined transparent. After deming, the samples were by hairs. The domatia are probably associated washed with water until there was no NaOH with predatory and fungivorous mites. residue, and the specimens were then mounted 5. Tertiary venation. O=percurrent, 1=reticulate. in Hoyer's mounting medium. Anatomical and Allantoma lineata, the actinomorphic-flowered morphological leaf features follow the tem1i­ species of Cariniana, Couratari stellata, Cour­ nology of Hickey (1973) and the Leaf Archi­ oupita, and Grias eaulifiora possess percurrent tecture Working Group (MLA) at http://www. teliiary venation (Figs. 35.1-35.3 in MLA, peabody.yale.edu/collections/pb/MLA.pdt: 1999), in which the tertiary veins cross between adjacent secondaries in parallel paths Pollen, used for scanning electron micros­ without branching. The zygomorphic-flowered copy (SEM) study, was taken from herbarium species of Cariniana, Couratari guianensis, specimens with mature flowers or from and Gustavia hexapetala have reticulate tertia­ buds at an advanced stage of develop­ ry veins (Fig. 35.4 in MLA, 1999) in which the ment. The pollen was mounted directly onto a teliiaries anastomose (rejoin) with other ter­ SEM stub and then coated with gold for tiaries or secondaries at random angles. observation with a Hitachi S2700-SEM. The 6. Quatemary venation. O=dichotomizing, 1= same methodology was used to observe the regular polygonal reticulate. In the dichoto­ adaxial and abaxial cuticular surfaces of mizing type the fomih order veins branch the leaf blade. freely, i.e., they do not unite with other veins The following 33 characters were used in and are the finest order of venation that the leaf the analysis. All multistate characters were exhibits (Fig. 39.4 in MLA, 1999). Allantoma coded as nonadditive. The data matrix is lineata and the actinomorphic-flowered species shown in Table I. of Cariniana possess dichotomizing venation. In the regular polygonal reticulate type, the 1. Leaf base. O=revolute, 1=not revolute. The fomih order veins anastomose to fmID poly­ leaf blade base is clearly revolute only in gons of similar size and shape (Fig. 39.3 in Cariniana legalis and C. parvifolia. MLA, 1999). Fifth order veins are the finest 2. Abaxial leaf surface. O=glabrous, l=stellate venation in this type. The zygomorphic-flow­ hairs present, 2=simple hairs present. ered species of Cariniana and all of the 3. Venation type. O=eucamptodromous, 1=bro­ outgroup taxa possess this type of quatemary chidodromous. Brochidodromous (secondaries veins. joined together in a sel1es of prominent arches) 7. Areolation. O=not well developed, I =4-sided, and eucamptodromous (secondaries uptumed 2=5- or more sided. Areoles are the smallest and gradually diminishing apically inside the areas of leaf tissue surrounded by veins of any margin without forming prominent arches, order (p. 42 in MLA, 1999). The areoles may MLA, 1999, p. 30) venation pattems are the be poorly developed (Allantoma lineata and only types found in Lecythidaceae. The the actinomorphic-f1owered species of Carini­ eucamptodromous venation pattem is found ana) or well-developed with four (C. domes­ in Allantoma lineata and the actinomorphic­ tiea, C. ianeirensis, C. mierantha, and C. flowered species of Cariniana. Most other pendulifiora) or five or more sides (Cariniana species (i.e., zygomorphic-flowered species of estrellensis, C. legalis, C. parvifolia, C. pyr­ Cariniana and outgroups) have a few eucamp­ iformis, c. rubra, and all of the outgroup taxa). 2008] HUANG ET AL.: PHYLOGENY OF CARINIANA (LECYTI-IIDACEAE) 73

TABLE I DATA MATRIX FOR PHYLOGENETIC ANALYSIS.

Characters Taxon II 21 31

Cariniana deeandra 1000000011 0011101000 00-0011100 102 domestiea 1011111 J 11 1001210120 00-1001100 102 estrellensis 1011112111 1001210120 00-1011100 102 ianeirensis 1010111111 1001210020 00-1001100 102 integrifolia 1000000011 1011101000 00-0001100 102 kuhlmannii 1000000011 0011101000 00-0011100 102 legalis 0010112111 1001210120 00-1011100 102 mierantha 1011111111 1001210120 00-1002100 102 multiflora 1000000011 0011101000 00-0001100 102 paehYalltha 1000000011 0011101000 00-0001100 102 parvifolia 0010112111 1001210120 00-1011100 102 pauciramosa 1000000011 0011101000 00-0001100 102 penduliflora 1011111111 1001210120 Oo-IOO?IOO 102 pyriformis 1011111211 1001210120 00-1002100 102 rubra 1011111211 1001210120 00-1002100 102 uaupensis 1000000011 0011101000 00-00?1100 102 Allantoma lineata 1000000010 1011101000 00-1201100 000 Couratari guianensis 1110]]2[0,1] I[0, I] ]]00210111 1100011100 012 stellata 1110012010 ]]00210]]1 10010111 00 012 Couroupita guianensis 1111012100 1200210111 10121000 II 002 niearaguarensis 1011012100 1200210111 1012100011 002 Grias eauliflora 10100[2102 1111000000 10-120[2,3]000 000 Gustavia hexapetala 1010112100 1201210000 10-2100000 001

Polymorphisms are enclosed in brackets, inapplicable characters are indicated with dashes ('-'), and unknown character states are indicated with question marks.

8. Stomatal type. O=brachyparacytic, 1=anomo­ completely enclosing them (Fig. 55.12 in cytic. Allantoma lineata and the actino­ MLA, 1999). The zygomorphic-flowered spe­ morphic-flowered species of Cariniana have cies of Cariniana and all of the outgroup taxa brachyparacytic stomata with two cells parallel have anomocytic stomata with 5 or more cells to the long axis of the guard cells but not surrounding the guard cells and the cells adjacent to the guard cells are not differentiat­ ed in any way from the normal epidermal cells (Fig. 55.2 in MLA, 1999). 9. Inflorescence position. O=cauline, I =not cau­ line. As used in this study, cauline applies to inflorescences that arise from the main trunk (e.g., species of Couyoupita and Grias) and those inflorescences that do not arise from the main trunk are not cauline (the remaining species). 10. Inflorescence type. O=raceme, l=panicle, 2= fascicle. 11. Pedicel. O=absent, 1=present. 12. Flower size. 0=<2 em, 1=2:2 cm<6 em, 2= >6 em. Species of Cariniana and Allantoma lineata have small flowers with a maximum FIG. 2. Cariniana domestiea, leaf domatium. (From Eiten 4304, NY.) diameter no larger than 2 em. In contrast, the 74 BRTTTONIA [VOL. 60

outgroup species have flowers no smaller than androecia that are not clearly separated into a 2 cm in diameter. The flowers of Couratari hood and staminal ring by a stamen-free area guianensis and C. stellata are up to 3.5 cm. (Fig. 30 in Mori & Prance, 1990). We call this The flowers of Couroupita guianensis and C. type of androecium obliquely zygomOlphic nicaraguarensis range from 3.5 to 6 cm, those and it is not found in any other genus of of 2.5 to 5 cm, and those of Lecythidaceae. Gustavia hexapetala from 6 to 9 cm in 20. Hood presence. O=absent, I=present. In Lecy­ diameter. thidaceae, all species with actinomOlphic an­ 13. Calyx lobes. O=with length more than 1/3 the droecia do not have a hood (a strap-like calyx, I =with length no more than 1/3 the structure expanded from one side of the calyx. Species of actinomorphic-flowered androecium, curved over the summit of the Cariniana, Allantoma lineata, and Grias cauli­ ovary, and modified at the apex, see Fig. 9 in flora have scarcely lobed calyces while those of Prance & MOli, 1979 and Fig. 31 in Mori & the zygomorphic-flowered Cariniana, Coura­ Prance, 1(90). The zygomOIphic-flowered spe­ tari, Couroupita, and Gustavia hexapetala have cies of Carillialla have zygomOlphic androecia deeply lobed calyces. but no hood, and species of all other genera 14. Calyx imbrication. O=imbricatc, 1=not imbri­ with zygomorphic androecia have a hood. cate. The calyx lobes of Couratari and 21. Stamen attachment. O=inside a tube, 1=the Couroupita are imbricate while those of Grias, upper margin of a ring or from the surface of a Gustavia, Allantoma, and Cariniana are not cushion. The stamens of species of Grias (Fig. imbricate. 9 in Prance & Mori, 1979) and Gustavia (Fig. 15. Petal number. 0= four, 1=five, 2=six. Four 31 in Mori & Prance, 1990) arise from the rim petals are characteristic of species of Grias, five of a raised ring and those of Couratari and petals are a feature of Allantoma lineata and the Couroupita arise from an annular staminal actinomorphic-flowered species of Cariniana, cushion that does not fonn an elevated rim or and six petals are found in the zygomorphic­ tube. The stamens of Allantoma lilleata (Fig. 9 flowered species of Cariniana, Couratari, in Prance & Mori, 1979) and species of Couroupita, and Gustavia hexapetala. Cariniana (Fig. 9 in Prance & Mori, 1979) 16. Petal texture. O=camose, 1=thin. The actino­ arise entirely or in part from the inner surface mOlphic-flowered species of Cariniana, Allan­ of a staminaI tube. We differentiate between a toma lineata, and Grias cauliflora possess ling, cushion, and tube as follows: the ring is camose petals. The petals of the remaining as wide in diameter as it is tall, a cushion is a taxa are thin. flat or slightly raised area upon which the 17. Petal apex. O=flat, 1=incurved. The petals of stamens arise, and a tube is nan-ower in Allantoma lineata and the actinomOlphic-flow­ diameter than it is tall. ered species of Cariniana are tumed inward 22. Staminodia. O=absent, 1=present. Staminodia are (hooked) at the apices whereas the petal apices found in Couratari and Couroupita. The stami­ of all remaining genera in the family are flat. nodia of Couroupita produce fodder pollen. 18. Petal margins. O=entire, l=ciliate. In Allall­ 23. Fodder pollen. o= absent, 1=present. In spe­ toma lineata, the actinomorphic-flowered spe­ cies of Lecythidaceae with actinomorphic cies of Carinialla, one species from the flowers all of the pollen is the same, Le., zygomOlphic-flowered species of Cariniana pollen serving as the pollinator reward is the (Cariniana ialleirensis), and Grias cauliflora same as fertile pollen and both genninate. the petals have entire margins. The species of Species of Lecythidaceae with zygomorphic the remaining genera are ciliate. flowers may have both white anthers contain­ 19. Androecial symmetry. O=actinomorphic, 1= ing fertile pollen and yellow anthers containing zygomorphic, 2=obliquely zygomOlphic. In pollen that does not gelminate and serves as a New World Lecythidaceae, Allantoma lilleata, reward for pollinators (i.e., fodder pollen the actinomOlphic-flowered species of Carilli­ produced in staminodia). Among the species alla, Grias cauliflora, and species of Gustavia studied, only the two species of Couroupita have actinomorphic androecia while the have fodder pollen (Mori & Orchard 1980). remaining genera are zygomorphic. The zygo­ 24. Fertile stamen number. 0=<25, 1=225<200,2= morphic-flowered species of Carilliana have >200. The actinomOlphic-flowered species of 2008] HUANG ET AL.: PHYLOGENY OF CARINIANA (LECYTHIDACEAE) 75

Cariniana possess fewer than IS stamens (ex­ from the fruit without pulp adhering to them. cept C. pachyantha, which has 17-25 stamens) 31. Unilateral seed wings. O=absent, I =present. and Allantoma lineata has ca. 30 stamens. The Cariniana and Couratari are the only two zygomorphic-flowered species of Cariniana and genera of Lecythidaceae that have seed wings. the two species of Couratari studied possess Tsou and Mori (2002) have demonstrated that 25 to 50 stamens (except , seed wings in Cariniana and Couratari are which has ca. ISO stamens), and Grias cauliflora different, both in origins and structure. Thus, has 85 to ISO fertile stamens. Species of the character of seed wing is coded as two Couroupita and Gustavia hexapetala have more different characters. than 600 stamens. 32. Circumferential seed wings. O=absent, 1= 25. Ovary locules. O=three, I=six, 2=four or five. present. As described in character 32, Coura­ The number of locules is useful in separating tari is the only genus that possesses circum­ species of different genera, e.g., species of ferential seed wings. Cariniana and Couratari have three locules 33. Cotyledons. O=undifferentiated or reduced to and species of Couroupita and Gustavia small scales, I =fleshy plano-convex, 2=leaf­ hexapetala have six locules. The number of like. Allantoma lineata and Grias cauliflora locules in the ovaries of Allantoma lineata and have undifferentiated cotyledons, or reduced to Grias are four or five. scale if present. Gustavia hexapetala is the only 26. Style presence. O=stigma sessile, I =style present. species in this study with fleshy plano-convex 27. Fruit shape. O=globose, I = cylindrical, 2= cotyledons. Species of Cariniana, Couratari, pyrifOlm, 3=fusifOlm. The fruits of Allantoma and Couroupita have leaf-like cotyledons. lineata and the actinomorphic-flowered Car­ iniana and Couratari are cylindric, those of zygomorphic-flowered Cariniana are either EXCLUDED CHARACTERS cylindlic or pylifOlm, and those of Couroupita and Gustavia are globose. The fruits of Grias Pollen characters, abaxial leaf blade surfa­ cauliflora are fusiform or globose. ces observed using the SEM, and leaf 28. Fruit dehiscence. 0= indehiscent, 1= dehiscent. measurements were explored for possible The fruits of Lecythidaceae are indehiscent or use in the phylogenetic analysis, but were dehiscent. In order to be scored as dehiscent, excluded because they did not provide dis­ the fiuit must have an operculum that stays crete characters needed for cladistic analysis. intact when it falls from the fruit. Allantoma, Although there is a tendency for the Car­ Cariniana, and Couratari belong to this type. iniana legalis group to have larger pollen In Gustavia hexapetala, the opercular area with a more finely perforate tectum (Fig. 3A­ breaks down and the fruit appears dehiscent B) and that of the C. decandra clade and but there is no intact operculum and the seeds Allantoma lineata to have smaller pollen with remain in the fiuit. This represents an inter­ a more reticulate tectum (Fig. 3C-D), the mediate step between indehiscent and dehis­ variation is continuous and could not be cent fiuits, but because the seeds remain in the clearly divided into discrete states. The fiuit it is coded as indehiscent. Grias cauliflora cuticle structure of the leaf blade surface is and Couroupita also have indehiscent fruits. not consistent and variation in this feature is 29. Seed trichomes. O=absent, I =present. The only found both within and among the species such species of Lecythidaceae with seed trichomes that it cannot be used in cladistic analysis. Leaf are found in Couroupita. This feature has been measurements are also continuous and could illustrated by Tsou and Mori (2002). not be divided into discrete states, but the 30. Seeds embedded in pulp. O=absent, I =pres­ ofAllantoma lineata and the species of ent. Seeds that can not be easily separated the Cariniana decandra group are often from the surrounding pulp are found only in larger than those of the C. legalis group. species of Couroupita (see section of fruit in Fig. 36 in Mori & Prance, 1990). In species of PHYLOGENETIC ANALYSES Grias and Gustavia, the pericarp tightly surrounds the seeds but it is not juicy and WinClada (Nixon, 19(9) was used to pulpy and the seeds can be easily removed create a data matrix that was submitted to 76 BRITTONIA [VOL. 60

FIG. 3. Pollen of the Cariniana legalis clade and the A//antomalC. decandra clade. A. C. domestica (Walter et al. 00055, NY), polar view. The tectum is finely perforated and colpus membranes are more or less smooth. B. C. legalis (Jardinz 371, NY), equatorial/polar view. The colpus membranes are velTucose, and the tectum is finely perforated. C. C. decandra (KrukofJ 8717, NY), polar vicw. The tcctum is coarsely reticulate. D. C. multiflora (Soria 29, NY), polar view. The pollen grain is spherical and the colpus membranes are ven1Jcose. The tectum is coarsely reticulate with verrucae in the lumina.

Nona (Goloboff, 1993) for phylogenetic index (Cl) of 0.64 and a retention index (Rl) analyses. Heuristic searches with 100,000 of 0.86 (Fig. 4). Ten nodes collapsed in a maximum trees retained, 1000 replications, strict consensus of 84 most parsimonious and 10 starting trees per replication were trees (Fig. 5). Cariniana is not resolved as employed using parsimony with the character monophyletic, but Cariniana and Allantoma states nonadditive and equally weighted. A together form a monophyletic group with two strict consensus tree was calculated in Win­ major clades: 1) the Allantoma/C. decandra Clada. Character optimizations were calculat­ clade with Allantoma lineata and the seven ed using the fast option of Winclada. Branch species of actinomorphic-flowered Cariniana, support was constructed by Bootstrap analy­ and 2) the C. legalis clade with the nine sis with 1000 replications, 10 random taxon species of zygomorphic-flowered Cariniana. entry sequences per replication, and one tree saved per replication. Discussion The results indicate that Cariniana is not monophyletic and that Cariniana and Allan­ Results toma together form a monophyletic group. The Heuristic analysis generated 84 most parsi­ actinomorphic-flowered species of Cariniana monious trees of 65 steps with a consistency are more closely related to the monotypic genus 2008] HUANG ET AL.: PHYLOGENY OF CARINIANA (LECYTHIDACEAE) 77

101325 Z1

12 Z3 24 2S

Outgroup 5 24 151633

102S 31 33

36113151817 892728

10122131

5 8 19

FIG. 4. One of 84 equally parsimonious trees (length=65, CI=O.64, RI=O.86). Character optimizations were obtained using the fast option of Winclada. Characters changes are labcled above with the numbers corresponding to those in the text. The arrows indicate the collapsed nodes after consensus analysis. Black circles indicate 11011­ homoplasious changes, and white circles indicate homoplasious changes.

Allantoma than they are to the zygomorphic­ characters are convergent in Grias cauliflora flowered species of Cariniana. The monophyly and the AllantomalCariniana decandra clade of Cariniana and Allantoma is supported by (Fig. 4). four characters: paniculate inflorescence (10), There are three characters supporting species flower size smaller than 2 cm in diameter (12), of the zygomorphic-flowered Cariniana clade stamens that arise from the inner part of a (refen'ed to as the C. legalis clade): reticulate stamina1 tube (21), and unilateral seed wings tertiary veins (5), anomocytic stomata (8), and (31). The presence of a racemose inflorescence obliquely zygomorphic androecium (19). Re­ (10) and unilateral seed wings (31) in Allantoma ticulate tertiary veins evolved independently in lineata represent reversals. The monophyly of the C. legalis clade, Couratari guianensis, and the actinomorphic-flowered Cariniana plus A. Gustavia hexapetala. Anomocytic stomata are lineata is supported by seven characters: a reversal in the C. legalis clade. eucamptodromous venation (3), dichotomizing Within the Cariniana legalis clade, C. quatemmy venation (6), poorly developed legalis and C. parvifolia are the only species areolation (7), scarcely lobed calyces with with revolute leaf bases (1) among the length no more than one-third the calyx (13), neotropical Lecytllidaceae. These two species five petals (15), camose petals (16), and hooked possess the smallest leaves in Cariniana (4­ petal apices (17). Scarcely lobed calyces (13) 5 cm average length) and, along with C. and camose petals (16) can also be found in ianeirensis, are tlle only species tllat do not Grias cauliflora indicating that these two have leaf domatia (4, Fig. 2) in the C. legalis 78 BRITTONIA [VOL. 60

Grias caulijIora Gustavla IIexapetala 57 I Couroupita guiallensls I 93 I I - Courouplta Ilicaraguarellsis

94 I Couratari gulallellsis I Couratarl stellata Allalltoma lllleata 82 Carillialla illtegrifolia

~ 75 Carillialla decalldra - ~ Carillialla kuhlmallllil 66 ~ Carillialla multijlora ""'­ I-­ Carillialla pachyalltha I-­ Carilliana pauciramosa "-Carillialla uaupe/lsis Carillialla domestica Carillialla estrellellsls Carillialla ialleirellsis

58 Carilliana micralltlla Carillialla pelldulijlora Carillialla rubra 66 Carillialla legalis Carillialla parvlfolia FIG. 5. Strict consensus of 84 equally parsimonious trees. Bootstrap values of over 50% are shown above the branches. clade. Leaf domatia are small cavities usually transfelTing the species of Cariniana in this located in the secondary vein axils on the clade to Allantoma. New combinations are abaxial leaf surface may be associated with made for the transfers of the actinomorphic­ predatory and fungivorous mites of potential flowered species of Cariniana to Allantoma. benefit to their host (Pemberton & Turner, The combination, Allantoma multiflora, 1989; Walter & O'Dowd, 1992; Rozario, would be a later homonym of A. multiflora 1995; O'Dowd & Pemberton, 1998). We (Sm.) Miers, a synonym of Couratari multi­ hypothesize that the revolute leaf bases of flora (Sm.) Eyma, so we have coined A. C. legalis and C. parvifolia may also harbor plur!flora S. A. Mori, Y.-Y. Huang & Prance mites thereby eliminating the need for doma­ as the new name for this species. In vegeta­ tia in the axils of the secondary veins. This tive, floral, and molecular features these character is also found in two of the outgroup species are consistent with the characters of taxa, Couroupita guianensis and Couroupita Allantoma and their close relationship, first nicaraguarensis. recognized by Ducke (1925, 1948), is dem­ In order to reflect these relationships, we onstrated in our tree (Figs. 4, 5). The major propose retaining the clade with the zygo­ difference lies in seed structure. The seeds of morphic-flowered species in Cariniana as it Allantoma lack a unilateral wing (31), lack includes the type of the genus (c. legalis). leaf-like cotyledons (33), and are rich in oil Because the actinomorphic-flowered clade content, which allows them to float for at includes Allantoma lineata, we propose least six months (Ducke, 1948). The seeds 2008] HUANG ET AL.: PHYLOGENY OF CARINIANA (LECYTI-IIDACEAE) 79 float away and become stranded at the edges metry (19) evolved three times in neotropical of rivers where they germinate when the Lecythidaceae based on the limited selection water recedes. Mori and Prance (1990) of taxa included here (Fig. 4). Species of Old suggest that Allantoma lineata has evolved World Lecythidaceae possess only actino­ from Cariniana-like ancestors in response to morphic androecia. In the New World, four the selective pressures afforded by the peri­ out of ten genera have actinomorphic androe­ odically flooded forest habitats of Amazonia. cia (Allantoma, species of Cariniana in the Character optimization on one of the most AllantomalCariniana decandra clade, Gusta­ parsimonious tree shows that the seed wing via, Grias), while seven other genera have has been lost in A. lineata, and that undiffer­ zygomorphic androecia (Bertholletia, Carini­ entiated cotyledons represent convergent evo­ ana legalis clade, Corythophora, Couratari, lution in A. lineata and Grias cauliflora Couroupita, Eschweilera, and Lecythis) with (Fig. 4). Both species grow in riverine habitats one side of the androecium expanded into a and are dispersed by water current, although strap-like structure that curves over the those of G. caul[flora may be primarily summit of the ovary (Prance & Mori, 1979; dispersed by animals seeking the edible Mori & Prance, 1990). The genus Cariniana pericarp. Thus the hypothesis by Mori and as previously circumscribed has been includ­ Prance (1990) is supported in our analysis. ed in both groups because some species have The evolution of androecial symmetry and actinomorphic and others have zygomorphic the relationship of Cariniana to other New androecia. World Lecythidaceae, especially with Coura­ The actinomorphic, fertile pollen producers tari have not yet been resolved. Cariniana are considered to be the most primitive of the was initially confused with Couratari be­ neotropical Lecythidaceae, whereas the zygo­ cause both genera have cylindrical fruits, 3­ morphic-flowered group is more advanced locular ovaries, and seed wings. Although (Mori et aI., 1978; Knudsen & Mori, 1996). both Cariniana and Couratari possess Mori and Boeke (1987) suggested that the winged seeds (31, 32), the seed wings evolution of pollinator rewards in New World evolved independently in these two genera. Lecythidaceae goes from all fertile pollen, to Tsou and Mori (2002) pointed out that the both fertile and fodder pollen, and then to seed wing type, seed wing origin, and seed nectar. Our results support this hypothesis coat structure of Cariniana are different from and show that zygomorphy is derived from Couratari. Species of Cariniana have a actinomorphy on three separate occasions unilateral seed wing that originates from the based on the tree. Nevertheless, it is interest­ mesophyll of the outer integument of the ing to note that actinomorphy in the Allan­ ovule, there is no exotesta in the seed wing, tomale. decandra clade is different in some and the seed coat has branched vascular respects from the actinomorphy that is found bundles but no fibers. On the other hand, in other genera of the family. In Grias and species of Couratari have circumferentially Gustavia, the androecium has a fused basal winged seeds that originate from both the portion, the staminal ring, and the stamens exotesta and the outer layers of mesotesta of arise from the upper margin of the ring the seed body, the seed wing has an exotesta, (Fig. 2A in Mori & Lepsch-Cunha, 1995 and and the seed coat has unbranched vascular Fig. 9 in Prance & Moli, 1979). In contrast, the bundles and fibers (Tsou & Mori, 2()02). Tsou fused basal portion elongates to fOlm a tube and Mori (2002) suggest that the seed wings and at least some of the stamens alise from the of Cariniana and Couratari are caused by inner surface of the tube in the e. decandra convergence for wind dispersal and do not clade (Fig. 9 in Mori & Lepsch-Cunha, 1995 necessarily indicate recently common ancestry. and Fig. 9 in Prance & Moli, 1979). The Floral evolution in New World Lecythida­ species that produce only fertile pollen have ceae is mostly related to androecial symmetry large numbers of stamens (up to 1210 in some (Prance & Mori, 1979). Character optimiza­ species of Gustavia), whereas those that tion using the fast option of Winclada produce nectar often have a reduced number indicates that differences in androecial sym­ of stamens (as few as lOin some species of 80 BRITTONIA [VOL. 60

Couratari). The feltile stamens are numerous relationship that has not previously been in Grias (85-210; Mori, 197911) and Gustavia suggested. Combining mOll'hology with mo­ (500-1,210; Mori, 1979a) whereas there are lecular data for an expanded sample of taxa no more than 30 fertile stamens in the may shed more light on the evolution of Allantoma and no more than 15 in the C. androecial features in the family. decandra clade (except C. pachyantha, which has 17-25 stamens). New combinations and new name On the other hand, the androecia of the for the actinomorphic species of Cariniana species of the C. legalis clade are also 1. Allantoma dccandra (Ducke) S. A. Mori, Y-Y different from other zygomorphic-flowered Huang & Prance, comb. nov. Cariniana decandra genera. Species of the C. legalis clade do Ducke, Arch. Jard. Bot. Rio de Janeiro 4: 153, t. not have hoods (Figs. 2C-D in Mori & 16. 1925. Lepsch-Cunha, 1995) nor do they have a 2. Allantoma integrifolia (Ducke) S. A. Mori, Y­ stamen free region that separates the hood Y Huang & Prance, comb. nov. Cariniana from the stamina1 ring. The stamens of integrifolia Ducke, Trop. Woods 31: 18. 1932. species of the C. legalis clade are spread all 3. Allantoma kuhImannii (Ducke) S. A. MOli, Y­ over the wall of a slightly elongated, oblique­ Y. Huang & Prance, comb. nov. Cariniana ly oriented tube (Figs. 68D, 70D in Prance, kuhlmannii Ducke, Arch. Jard. Bot. Rio de 1979b). For this reason, we consider this type Janeiro 4: 154, t. 16. 1925. of androecium as oblique zygomorphy. 4. Allantoma pIuriflora S. A. Mori, Y.-Y. Huang Analysis of Lecythidaceae based on com­ & Prance, nom. nov. Cariniana multiflora bined ndhF and trnL-F gene sequences (Mori Ducke, Arq. mst. BioI. Veg. 2: 65. 1935. et aI., 2007) indicate that an actinomorphic­ 5. Allantoma pachyantha (A. C. Sm.) S. A. Mori, flowered group (Gustavia & Grias) is sister to Y-Y Huang & Prance, comb. nov. Cariniana a clade consisting of all of the zygomorphic­ pachyantha A. C. Sm., Amer. J. Bot. 26: 411. flowered taxa. Within the zygomorphic­ 1939. flowered clade, Couroupita is sister to a large 6. Allantoma pauciramosa (W. A. Rodrigues) S. clade that includes all other zygomorphic­ A. Mori, Y-Y Huang & Prance, comb. nov. flowered genera, the actinomorphic-flowered Cariniana paucirasoma W. Rodrigues., Acta Allantoma/C. decandra clade, and the zygo­ Amazonica 4(1): 8-9. 1974. morphic-flowered C. legalis clade. The mono­ 7. Allantoma uaupensis (Spruce ex O. Berg) S. phyly of the Allantoma/c. decandra clade is A. Mori, Y.-Y Huang & Prance, comb. nov. well supported. However, the relationships of Couratari uaupensis Spruce ex O. Berg, Mart. the Allantoma/C. decandra clade (represented n Bras. 14 (1): 508, t. 78. 1858; Cariniana only by C. decandra & A. lineata), the C. uaupensis (Spruce ex O. Berg) Mel'S, Trans. legalis clade (represented by C. domestica, C. Linn. Soc. 30: 288, t. 63, Fig. 9. 1874. estrellensis, and C. ianeirensis), Couratari, and other zygomorphic-flowered genera (Lecythis, Bertholletia, Eschweilera, and Corythophora) Acknowledgments are not resolved. In this scenario, the most parsimonious evolution of floral symmentry We thank Dominick Basile and Mike suggests reversal from zygomorphy to actino­ Baxter for teaching techniques used in study­ morphy in the Allantoma/C. decandra clade. ing anatomy and electron microscopy to Our results do not agree with this study because the first author, respectively; Larry Kelly and optimization ofthis character on one ofour most Fabian Michelangeli for sharing their knowl­ parsimonious trees indicates that zygomorphy edge of phylogenetic analyses; Douglas Daly evolved three times: one led to the basal genus for his aid in selecting and understanding leaf of the zygomorphic-flowered genera, Courou­ venation characters; Lisa Campbell for her pita, another line resulted in Couratari, and assistance in the anatomy lab; Chih-Hua Tsou another led to the Cariniana legalis clade for advice and comments during the course of (Fig. 4). In our tree, the actinomorphic­ this study; and several staff members of the flowered Gustavia is sister to Couroupita, a Missouri Botanical Garden for their help 2008] HUANG ET AL.: PHYLOGENY OF CARINIANA (LECYTHIDACEAE) 81

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