Int. J. Plant Sci. 168(6):905–929. 2007. Ó 2007 by The University of Chicago. All rights reserved. 1058-5893/2007/16806-0010$15.00 SPORE MORPHOLOGY IN RELATION TO PHYLOGENY IN THE FERN GENUS ELAPHOGLOSSUM (DRYOPTERIDACEAE) Robbin C. Moran,1 Judith Garrison Hanks,2 and Germinal Rouhan3 New York Botanical Garden, Bronx, New York 10458-5126, U.S.A.; Marymount Manhattan College, 221 East Seventy-First Street, New York, New York 10021, U.S.A.; and Institut de Recherche pour le Développement US084, Muséum National d’Histoire Naturelle, Département Systématique et Evolution, Herbier National, 16 rue Buffon, 75005 Paris, France The perispore structure of Elaphoglossum was studied using a scanning electron microscope. Of the species examined, 119 corresponded to those used in a previously published phylogenetic analysis of the genus based on two chloroplast noncoding DNA regions, trnL-trnF and rps4-trnS. The spores of 102 additional species were examined for comparative purposes. Five perispore characters were scored for each species and optimized onto the previously published molecular tree. The morphology of the perispore and its character state changes are described and discussed in a phylogenetic context. Synapomorphies for major clades within the genus were identified, such as spines for the Neotropical species of sect. Squamipedia and perforations, spines, and cristae for subsect. Pachyglossa and a large subclade within sect. Setosa. This study is the largest done on perispore morphology in relation to phylogeny in a genus of ferns. Spore images of all species studied are available at http://www.plantsystematics.org. Keywords: Elaphoglossum, ferns, perispore, phylogeny, pteridophytes, spores. Introduction phyllopodia, and aerophores. With regard to spores, they showed that considerable diversity exists, and in some cases this diversity corresponded to their proposed subsections or unnamed species groups. Mickel (1980, 1985) published spore images for 24 species of Elaphoglossum, and the spores of 50 species of Elaphoglossum were examined by Tryon and Lugardon (1991). These workers found the spores to be monolete and bilateral, to have a longitudinal laesura, and (depending on the species) to be from 20 to 80 mm long. None of the above studies, however, assessed perispore morphology in a cladistic framework because phylogenetic trees were lacking. Rouhan et al. (2004) and Skog et al. (2004) presented phylogenetic hypotheses for Elaphoglossum based on DNA sequences from the chloroplast genome (trnL-trnF and rps4-trnS by Rouhan et al. [2004]; rbcL, trnL-F, and rps4-trnS by Skog et al. [2004]). The Rouhan et al. (2004) study was the most comprehensive, containing 123 taxa from both the New and Old Worlds (as opposed to 48 species from only the New World in the Skog et al. [2004] study). Rouhan et al. (2004) recovered 11 main clades within the genus (fig. 1). These clades largely corresponded to the sections and subsections proposed by Mickel and Atehortúa (1980). The clades (starting at the bottom of fig. 1) can be characterized as follows. Section Amygdalifolia consists of a single species (E. amygdalifolium) that has long-creeping rhizomes, phyllopodia, and hydathodes—a unique character combination in the genus. Its reddish young leaves are also unique within the genus. Elaphoglossum aemulum (endemic to Hawaii), E. glaucum (native to Central America), and sect. Elaphoglossum are characterized by phyllopodia and usually thick, glabrous to very sparsely scaly laminae. Based on molecular evidence (Rouhan et al. 2004), sect. Elaphoglossum consists of two main clades, subsects. Pachyglossa and Platyglossa, although no known morphological or anatomical characters separate these two groups. Section Squamipedia is Elaphoglossum contains ca. 600 species and ranks as one of the world’s largest fern genera. It is pantropical but most species-rich in the Neotropics, where ca. 80% of the species occur. Nearly 75% of the species are epiphytes. The genus is related to Bolbitis, Teratophyllum, and Lomagramma, as evinced by chloroplast DNA sequence data (E. Schuettpelz, personal communication) and morphological characters such as acrostichoid sori, sterile-fertile leaf dimorphy, an elongated (in cross section) ventral vascular bundle in the rhizome, and this ventral vascular bundle bearing all the roots (thus, the roots appear to be borne only on the ventral surface of the rhizome; Holttum 1978; Kramer 1990). These genera are currently placed in the Dryopteridaceae, a placement well supported by molecular phylogenetic studies (Smith et al. 2006). Unfortunately, Elaphoglossum lacks a fossil record. Fée (1845) and Christ (1899) proposed subgeneric classifications of Elaphoglossum based primarily on frond shape, texture, scale characteristics, and the presence versus absence of hydathodes. More recently, Mickel and Atehortúa (1980) proposed an infrageneric classification based on sporophyte morphology and spore structure as determined from images taken with a scanning electron microscope (SEM). The morphological characteristics they found useful in subdividing the genus included rhizome habit, lamina scale type and distribution, and the presence versus absence of hydathodes, 1 2 3 Author for correspondence; e-mail rmoran@nybg.org. E-mail j2hanks@mmm.edu. E-mail rouhan@mnhn.fr. Manuscript received December 2006; revised manuscript received March 2007. 905 Fig. 1 Phylogeny of Elaphoglossum. Strict consensus of 1008 most parsimonious trees based on a combined analysis using plastid DNA sequence data from the trnL-trnF and rps4-trnS regions, resulting from a heuristic search using the indel coding ID. Numbers above the branches are bootstrap percentage values >50%. Single letters in dark and light gray boxes on branches refer, respectively, to synapomorphic indels in trnLtrnF and rps4-trnS alignments. Species in boldface are from the Indian Ocean area. After Rouhan et al. (2004). MORAN ET AL.—SPORES OF ELAPHOGLOSSUM characterized by long-creeping rhizomes, paired peglike aerophores on the rhizome near the petiole base, pale brownish flaccid rhizome scales, and a lack of phyllopodia. The subulatescaled clade (Subulata; fig. 1) is characterized by leaves with subulate scales, that is, erect hairlike scales enrolled at the base and often all the way to the tip. This clade is composed of two subsections: subsect. Polytrichia, which lacks hydathodes, and subsect. Setosa, which has them. The last section, sect. Lepido- 907 glossa, is characterized by laminar scales with marginal teeth consisting of acicular cells (all other Elaphoglossum species have laminar scales with marginal teeth consisting of glandular or bulbous marginal cells or gland-tipped marginal processes). Given the broad correspondence of sporophyte morphology with the groups recovered from recent molecular phylogenetic studies, the objective of this article is to compare spore morphology of Elaphoglossum with the phylogeny of the genus. Fig. 2 Spores of Elaphoglossum. A, E. amygdalifolium (Costa Rica, Grayum 2373, NY). B, E. aemulum (Hawai, Krajina & Lamoureux 611029059, NY). C, E. glaucum (Mexico, Mickel 9675, NY). D, E. latifolium (Jamaica, Underwood 3146, NY). E, E. grayumii (Costa Rica, Grayum 3017, NY). F, E. terrestre (Costa Rica, Moraga 509, NY). G, E. marquisearum (Marquesas, Word 6390, P). H, E. guatemalense (Mexico, Martinez 14987, NY). I, E. herminieri (no data). J, E. productum (Ecuador, Øllgaard & Navarrete 105872, AAU). K, E. lepervanchei (also as aff. conforme in Rouhan et al. 2004; Mauritius, Rouhan et al. 175, NY). L, E. subsessile (Madagascar, Rouhan 432, P). Scale bars ¼ 10 mm. 908 INTERNATIONAL JOURNAL OF PLANT SCIENCES Material and Methods Spores were obtained from herbarium specimens of Elaphoglossum at AAU, F, GH, MO, NY, P, and US. For 12 species (those with vouchers are underlined in app. A), spores were taken from the DNA vouchers cited in the Rouhan et al. (2004) study. It was not possible to do this for most of the vouchers cited because they lacked fertile leaves. For these species, attempts were made to use specimens collected from the same country as that of the DNA voucher. Generally, one specimen per species and 20–100 spores per stub were examined. In cases where perispore variation was suspected within a species, two or more specimens were examined (e.g., E. erinaceum, E. nigrescens, E. petiolatum; app. A). Besides those species included in the Rouhan et al. (2004) analysis, 102 additional species were imaged to assess the generality of results. The species examined for this study and voucher information are listed in appendix A. Fig. 3 Spores of Elaphoglossum subsect. Pachyglossa. A, E. sartorii (Mexico, Munn-Estrada & Mendoza 1683, NY). B, E. croatii (Costa Rica, Rojas 1535, NY). C, E. cismense (Costa Rica, Rojas 2877, NY). D, E. vieillardii (New Caledonia, Munzinger 1276, P). E, E. malgassicum (Madagascar, Rouhan 427, P). F, E. sieberi (Mauritius, Rouhan 176, NY). G, E. macropodium (Tanzania, DeBoer & Schippe 457, K). H, E. metallicum (Peru, Sagástegui 14459, NY). I, E. decaryanum (Madagascar, Rasolohery 873, P). J, E. hoffmannii (Costa Rica, Hennipman 6986, NY). K, E. biolleyi (Costa Rica, Davidse et al. 28794, NY). L, E. pteropus (Brazil, Steward & Ramos P19692, P). Scale bars ¼ 10 mm. MORAN ET AL.—SPORES OF ELAPHOGLOSSUM The spores were transferred with dissecting needles from herbarium specimens to aluminum SEM stubs and coated with an asphalt adhesive. The stubs were then coated with goldpalladium in a sputter-coater for 2.5 min, and spores were imaged digitally using a JEOL JSM-5410LV SEM equipped with a JEOL Orion 5410 software interface. The accelerating voltage was 15 kV. Spore images of all 221 species examined for this study (app. A) are available at http://www.plantsystematics.org. 909 To assess perispore evolution in a phylogenetic context, five perispore characters were scored (app. B) from the SEM images (figs. 2–11). Their character state changes were then optimized, using MacClade 4.0 (Maddison and Maddison 2000), onto the strict consensus tree from the analysis of the molecular data (Rouhan et al. 2004). In several instances for the presentation of results, we use different names than those used by Rouhan et al. (2004). The Fig. 4 Spores of Elaphoglossum sect. Platyglossa. A, E. flaccidum (Venezuela, Davidse & Miller 26870, NY). B, E. luridum (Guyana, Clarke & McPherson 1949, NY). C, E. hornei (Seychelles, Rouhan 137, NY). D, E. doanense (Panama, McPherson 12844, NY). E, E. ovalilimbatum (Madagascar, Perrier 7860c, P). F, E. mitorrhizum (Ecuador, Aulestia & Aulestia 1526, MO). G, E. aff. latifolium (Jamaica, Underwood 3146, NY). H, E. lingua (Costa Rica, Mickel 2688, NY). I, E. proximum (Costa Rica, Maxon 464, NY). J, E. conspersum (Costa Rica, Seidenschnur 268, NY). K, E. cardiophyllum (Ecuador, Holm-Nielsen, 5915, NY). L, E. angulatum (La Réunion, Rouhan 216, NY). Scale bars ¼ 10 mm. 910 INTERNATIONAL JOURNAL OF PLANT SCIENCES Fig. 5 Spores of Elaphoglossum. A, E. wawrae (Hawaii, Forbes 350, NY). B, E. glabellum (Panama, Stimson 5411, NY). C, E. acrostichoides (Tanzania, De Boer & Schippe s.n., NY). D, E. affine (Mexico, Camp 2682, NY). E, E. gayanum (E. minutum in Rouhan et al. 2004; Peru, Segastegui et al. 8238, NY). F, E. coursii (Comoros, Rouhan 127, P). G, E. coriaceum (Madagascar, Geay 103, P). H, E. marojejyense (Madagascar, Humbert 23105, P). I, E. peltatum (Mexico, Mendez 7931, NY). J, E. squamipes (also as E. craspedariiforme, a synonym, in Rouhan et al. 2004; Venezuela, Rivero & Diaz, 1496 NY). K, E. tripartitum (Ecuador, Øllgaard 99199, AAU). L, E. prestonii (Brazil, Brade 15514, NY). Scale bars ¼ 10 mm. changes were made necessary by new identifications of vouchers and because some names used by Rouhan et al. (2004) are now considered synonyms of older names (based largely on taxonomic studies by the first author). The name changes resulting from reidentification of vouchers are as follows: E. aff. terrestre of Rouhan et al. (2004) has now been confirmed as E. terrestre (Lorence and Rouhan 2004); E. aff. conforme has been identi- fied as E. lepervanchei (R. C. Moran, personal observation); E. sp. nov. aff. hayesii has been described as E. minutissimum (Moran and Mickel 2004); E. sp. has now been identified as E. cf. tovii (D. Lorence, personal communication); and E. aff. petiolatum has now been identified as E. dussii (R. C. Moran, personal observation). Name changes resulting from synonymy are as follows: E. craspedariiforme is now considered MORAN ET AL.—SPORES OF ELAPHOGLOSSUM 911 Fig. 6 Spores of Elaphoglossum, subulate-scale clade. A, E. crinitum (Costa Rica, Angulo 605, NY). B, E. erinaceum (Costa Rica, Gómez 18826, NY). C, E. backhousianum (Panama, Folsom 3576, NY). D, E. auripilum (Costa Rica, Burger & Stolze 5335, NY). E, E. apodum (Haiti, Ekman 4729, NY). F, E. hybridum var. vulcanii (La Réunion, Rouhan 222, P). G, E. hybridum var. hybridum (La Réunion, Rouhan 217, NY). H, E. decoratum (Brazil, Wacket 248, NY). I, E. piloselloides (Ecuador, Fay & Fay 3519, NY). J, E. spatulatum (La Réunion, Rouhan & Grangaud 246, NY). K, E. boryanum var. eutecnum (Venezuela, Luteyn & Lebron-Luteyn 5176, NY). L, E. cardenasii (Bolivia, Kessler et al. 6564, NY). Scale bars ¼ 10 mm. a synonym of E. squamipes; E. palmense and E. wackettii are now considered synonyms of E. nigrescens; and E. nigrocostatum is now considered a synonym of E. rimbachii. Elaphoglossum minutum is a synonym of E. revolutum, and the earliest name available for the species used in the Rouhan et al. (2004) analysis is E. gayanum. Finally, E. lindenii is now considered a synonym of E. squamatum (Rodrı́guez 1995). These name changes have been incorporated in figure 1 and are noted in appendix A. Based on morphology, we consider four species (E. craspedariiforme, E. aff. conforme, E. palmense, and E. wackettii) to be synonyms of other species in the Rouhan et al. (2004) analysis (i.e., these species were represented more than once in the analysis). Thus, the total number of species in the Rouhan et al. (2004) analysis was 119. The spore images of these species and 102 more not in that analysis are available at http:// www.plantsystematics.org. 912 INTERNATIONAL JOURNAL OF PLANT SCIENCES Fig. 7 Spores of Elaphoglossum. A, E. russelliae (Costa Rica, Gómez et al. 22409, NY). B, E. davidsei (Costa Rica, Montgomery & Huttleston 85–135, NY). C, E. setigerum (Ecuador, Holm-Nielsen & Jeppesen 1293, AAU). D, E. eximium (Costa Rica, Moran 2328, NY). E, E. aubertii (La Réunion, Rouhan et al. 226, NY). F, E. samoense (French Polynesia, Rapa, Motley 2608, NY). G, E. rapense (French Polynesia, Rapa, Motley 2677, NY). H, E. squamatum (Ecuador, Øllgaard & Balslev 8410, NY). I, E. pilosius (Mexico, Breedlove & Smith 22060, NY). J, E. pygmaeum (Ecuador, Moran & Rohrbach 5221, MO). K, E. phanerophlebium (Madagascar, Decary 18115, P). L, E. smithii (Ecuador, Øllgaard et al. 1014, AAU). Scale bars ¼ 10 mm. Results The following results and the spore images in figures 2–11 are arranged according to the ascending order of species shown in figure 1. Sect. Amygdalifolia. The single species in this section had nonperforate cristate perispores with the surfaces between the cristae smooth (fig. 2A). Elaphoglossum aemulum. The spores of this species (fig. 2B) have broadly folded, nonperforate perispores. Between the folds are lower, discontinuous folds. Elaphoglossum glaucum. The spores of this species have broadly folded, nonperforate perispores without spines (fig. 2C). The surfaces between the folds are nearly smooth but have scattered, low, irregular deposits. Sect. Elaphoglossum. From this section, 57 species were MORAN ET AL.—SPORES OF ELAPHOGLOSSUM 913 Fig. 8 Spores of Elaphoglossum. A, E. fournierianum (Costa Rica, Davidse 24999, NY). B, E. papillosum (Ecuador, Øllgaard 38438a, AAU). C, E. oblanceolatum (Panama, Porter et al. 4383, NY). D, E. minutissimum (Costa Rica, Moran 6344, NY). E, E. costaricense (Costa Rica, Seidenschnur 199, NY). F, E. siliquoides (Costa Rica, Lloyd 4283, NY). G, E. cuspidatum (Peru, Smith & Albán 5503, NY). H, E. succisaefolium (Amsterdam Island, Martel-Thoumian 2A, P). I, E. richardii (La Réunion, Rouhan 231, NY). J, E. pringlei (Mexico, Mickel 4463, NY). K, E. petiolatum (Mexico, Reveal & Atwood 3572, NY). L, E. rimbachii (E. nigrocostatum in Rouhan et al. 2004; Ecuador, Øllgaard & Balslev 8643, AAU). Scale bars ¼ 10 mm. examined (app. A; figs. 2D–5G). From subsect. Pachyglossa, we examined 31 species (app. A), 25 of which (figs. 2D–4D) were included in the analysis of Rouhan et al. (2004). The species imaged from this subsection but not in the Rouhan et al. (2004) analysis were E. andicola, E. gloeorrhizum, E. luridum, E. odontolepis, E. pachyphyllum, and E. unduaviense (app. A). All species of this subsection had either broadly folded or cristate perispores, with the cristate condition predominating (fig. 12). Except for E. luridum, all species in the subsection had continuous perisporal folds, and spines were generally present (fig. 13). Perforate perispores were often present (fig. 14), but the nonperforate condition characterized a clade of four Old World species consisting of the New Caledonian E. vieillardii, the Mauritian endemic E. sieberi, the Madagascan 914 INTERNATIONAL JOURNAL OF PLANT SCIENCES Fig. 9 Spores of Elaphoglossum sect. Lepidoglossa. A, E. forsythii-majoris (La Réunion, Cadet 1535, NY). B, E. avaratraense (Madagascar, Rakotondrainibe 1456, P). C, E. achroalepis (Madagascar, van der Werff 12837, P). D, E. scolopendriforme (Madagascar, Rouhan 306, P). E, E. humbertii (Madagascar, Humbert 6900, P). F, E. heterolepis (Mauritius, Rouhan 179, NY). G, E. tomentosum (Mauritius, Rouhan 178, NY). H, E. lanatum (Mauritius, Pike s.n., NY). I, E. paleaceum (Madeira, collector unknown s.n., NY). J, E. cf. tovii (French Polynesia, Lorence 9011, NY). K, E. micropogon (Costa Rica, Lloyd 4111, NY). L, E. rufidulum (La Réunion, Cadet 1535, P). Scale bars ¼ 10 mm. endemic E. malgassicum, and E. macropodium from African and Indian Ocean areas (figs. 3D–3G, 14). Nonperforate perispores were also found in the New World species E. doanense (fig. 4D), E. guatemalense (fig. 2H), E. herminieri (fig. 2I), and E. productum (fig. 2J). From subsect. Platyglossa, 22 species were examined (app. A), 15 of which (figs. 4E–5G) were included in the analysis of Rouhan et al. (2004). The species imaged that were not in the Rouhan et al. (2004) analysis were E. ambiguum, E. antisanae, E. ballivianii, E. conforme, E. lastii, E. proximum, and E. yunnanense (app. A). All species of this subsection had broadly folded perispores (fig. 12), the folds being continuous (except in E. wawrae; fig. 5A) and without spines (fig. 13) or perforations (fig. 14). Sect. Squamipedia. From this section, 12 taxa were examined (app. A), four of which (fig. 5H–5K) were included in MORAN ET AL.—SPORES OF ELAPHOGLOSSUM 915 Fig. 10 Spores of Elaphoglossum sect. Lepidoglossa. A, E. splendens (La Réunion, Rouhan 213, NY). B, E. asterolepis (Madagascar, Rouhan 305, P). C, E. ovatum (Ecuador, Ulloa et al. 1064, NY). D, E. stipitatum (La Réunion, Rouhan 212, NY). E, E. randii (Marion Island, Huntly 464, P). F, E. edwallii (Brazil, Rose & Russell 20482, NY). G, E. welwitschii (Tanzania, Taylor 9099, P). H, E. caricifolium (Costa Rica, Rojas 2898, NY). I, E. lancifolium (La Réunion, Rouhan & Grangaud 201, P). J, E. tectum (Colombia, Zarucchi et al. 5800, NY). K, E. killipii (Buchtien 4239, NY). L, E. dussii (also represented in Rouhan et al. 2004 as E. aff. petiolatum; Dominican Republic, Lloyd 351, NY). Scale bars ¼ 10 mm. the analysis of Rouhan et al. (2004). The eight species imaged that were not in the Rouhan et al. (2004) analysis are E. concinnum, E. deltoideum, E. feei, E. lloense, E. moorei, E. peltatum f. standleyi, E. tripartitum, and E. wrightii (app. A). All species had perispores with broad, continuous folds (fig. 12). Spiny, nonperforate perispores were present in the Neotropical species (E. peltatum–E. tripartitum; fig. 13), but unlike these species, the Madagascan endemic E. marojejyense (fig. 5H) had nonspiny, perforate peripores. Subulata clade. From this clade, 63 species were examined (app. A), 31 of which (figs. 5L–8F) were included in the analysis of Rouhan et al. (2004). From sect. Polytrichia, we examined 16 species, eight of which (figs. 5L, 6A–6G) were in the Rouhan et al. (2004) analysis (app. A). Those species 916 INTERNATIONAL JOURNAL OF PLANT SCIENCES Fig. 11 Spores of Elaphoglossum sect. Lepidoglossa. A, E. huacsaro (Venezuela, Ruiz-Teran 6124, NY). B, E. burchellii (Bolivia, Acebey & Tosto 176, NY). C, E. ciliatum (Costa Rica, Ingram & Ingram-Ferral 1308, NY). D, E. nigrescens (also represented in Rouhan et al. 2004 as the synonyms E. wackettii and E. palmense; Brazil, Maas et al. 6858, NY). E, E. deckenii (Comoros, Rouhan & Rakotondrainibe 105, P). F, E. poolii (Madagascar, Perrier de la Bâthie 12487, P). G, E. leucolepis (Madagascar, McPherson 1492, P). H, E. orbignyanum (Peru, Killip & Smith 24413, NY). I, E. furfuraceum (Costa Rica, Davidse 24757, MO). J, E. muscosum (Venezuela, Ruiz Teran 1864, NY). K, E. vestitum (Mexico, Mickel & Leonard 4705, NY). L, E. auricomum (Guatemala, Johnson 932, NY). Scale bars ¼ 10 mm. with spores imaged that were not in the Rouhan et al. (2004) analysis were E. cordifolium, E. cubense, E. latum, E. lindbergii, E. ornatum, E. pallidum, E. raywaense, and E. truncatum. In these species, the perispores were all broadly and continuously folded, nonperforate, and (with the exception of E. prestonii; fig. 5L) nonspiny (figs. 12–14). From sect. Setosa, we examined 48 taxa, 23 of which (figs. 6H–8F) were in the Rouhan et al. (2004) analysis (app. A). Those taxa with spores imaged that were not in the Rouhan et al. (2004) analysis were E. albescens, E. alfredii, E. bakeri, E. boragineum, E. boryanum, E. castaneum, E. delasotae, E. didymoglossoides, E. haynaldii, E. herpestes, E. heteromorphum, E. horridulum, E. mildbraedii, E. omissum, E. organense, E. phoras, E. proliferans, E. pseudoboryanum, E. pteropodum, E. rosettum, E. tabanense, E. villosum, E. wardiae, E. webbii, and E. zebrinum (app. A). Generally, either broad folds or cristae MORAN ET AL.—SPORES OF ELAPHOGLOSSUM were present, but these were absent in E. oblanceolatum and E. papillosum (fig. 8B, 8C) and in the clade E. setigerum–E. aubertii (fig. 7C–7E). Of those species that had broad folds or cristae, the cristate condition was more common (fig. 12). The cristae were discontinuous in E. boryanum var. eutecnum, E. cardenasii, and E. russelliae (fig. 6K, 6L; fig. 7A). Spiny perispores predominated throughout the sect. Setosa (fig. 13), as did perforations (fig. 14). Sect. Lepidoglossa. From this clade, 86 species were examined (app. A), 41 of which (figs. 8G–11L) were included in the analysis of Rouhan et al. (2004). The perispores typically had broad, continuous folds (fig. 12). Spiny perispores originated several times (fig. 13), and perforations were lacking in all species except E. forsythii-majoris and E. leucolepis (figs. 9A, 11G). The perispore of E. muscosum was distinctive in lacking folds and being beset with low spherical or hemispherical deposits (fig. 11J). Elaphoglossum muscosum had the largest spores (ca. 65 mm long) of any species examined. Discussion This study is one of the largest done on perispore morphology in a genus of leptosporangiate ferns (for lycophytes, see Korall and Taylor 2006). Given the size of this study, it was not possible to examine more than one or a few specimens of each species (app. A). The question arises of whether this sample size is adequate to give general applicability to the results. A comparison of our spore images to those published elsewhere (Mickel 1980, 1985; Mickel and Atehortúa 1980; Tryon and Tryon 1982; Tryon and Lugardon 1991) suggests that perispore morphology is generally consistent within species. Of 56 species compared, 52 were considered consistent. In the four instances that were not, we cannot be certain whether the differences are caused by infraspecific variation or by misidentifications of the specimens. The overall correspondence between our images and those previously published suggests that in most cases our findings adequately represent the species in question. Sect. Amygdalifolia. Sister to all other Elaphoglossum is sect. Amygdalifolia, which consists of a single species, E. amygdalifolium. Its cristate perispore (fig. 2A) represents an autapomorphy (fig. 12). The only species examined with similar spores (i.e., with nonperforate cristate perispores with smooth surfaces between cristae) was E. lepidothrix, which belongs to sect. Lepidoglossa (app. A; see image at http://www .plantsystematics.org). The similarity is most parsimoniously explained by an independent origin of the cristae. Elaphoglossum aemulum. This species is endemic to Hawaii (Palmer 2003). In our unpublished phylogenetic analyses based on DNA sequence data of two chloroplast markers (trnL-F, atpB-rbcL), it forms a clade with four other species endemic to Hawaii: E. alatum, E. crassicaule, E. fauriei, and E. parvisquameum. These four species, called the Alatum group (Palmer 2003), have perforate and cristate perispores (app. A). In contrast, E. aemulum has nonperforate, broadly folded perispores. Its spores more closely resemble those of subsect. Platyglossa. All of its character states represent the plesiomorphic condition (figs. 12–14). Elaphoglossum glaucum. The spores of this species (fig. 2C) resemble those of subsect. Platyglossa; namely, they have 917 broad perisporal folds without perforations and spines. All of the character states of this species represent the plesiomorphic condition (figs. 12–14). Sect. Elaphoglossum. This section consists of two wellsupported clades: subsects. Platyglossa and Pachyglossa (fig. 1). No known macromorphological or anatomical characters distinguish these two clades; they are supported solely by DNA sequencing evidence (Rouhan et al. 2004). Their perispore characteristics, however, support their distinction. Three perispore characters are synapomorphic for subsect. Pachyglossa (figs. 4E–5G): the presence of cristae (fig. 12), spines (fig. 13), and perforations (fig. 14). These characters show reversals within the subsection, such as the nonspiny perispores in the clade formed by the Mauritian endemic E. sieberi, the Madagascan endemic E. malgassicum, and E. macropodium from Africa and Indian Ocean areas (figs. 3D–3G, 13), and a reversal to nonperforate perispores in the Neotropical species E. doanense, E. guatemalense, E. herminieri, and E. productum (fig. 14; figs. 4D, 2H, 2I, 3L, respectively). Nevertheless, the characteristics of the perispore support the monophyly of subsect. Pachyglossa and provide the only morphological characters that distinguish it from subsect. Platyglossa. Sect. Squamipedia. The sister relationship of the Madagascan endemic E. marojejyense to this section, whose members are otherwise entirely Neotropical, was an unexpected result of the Rouhan et al. (2004) molecular study (fig. 1). Elaphoglossum marojejyense agrees morphologically with the Neotropical species in long-creeping rhizomes, lack of phyllopodia, and small leaves; however, it lacks the distinctive peglike aerophores on the rhizome near the petiole base, and its perispore (fig. 5H) differs in being perforate (autapomorphic within the section; fig. 14) and lacking spines (plesiomorphic; fig. 13). In contrast, the perispores of the Neotropical species (fig. 5I–5K) are nonperforate (plesiomorphic; fig. 14) and spiny (synapomorphic; fig. 13). Thus, the perispore of E. marojejyense appears to be morphologically unlike those of the Neotropical species in sect. Squamipedia. Subulata clade. The subulate-scaled clade contains two well-supported subclades: sects. Polytrichia and Setosa (fig. 1). Sect. Polytrichia and a clade within sect. Setosa (E. decoratum, E. spatulatum, and E. piloselloides) have nonperforate perispores with broad folds (fig. 6H–6J), both of which are plesiomorphic character states (fig. 14). Mickel and Atehortúa (1980) divided sect. Polytrichia into subsect. Apoda, characterized by laminae sessile or nearly so, and subsect. Polytrichia, characterized by distinct petioles. No differences in spores distinguish the two subsections (our sampling of subsect. Apoda was exhaustive; i.e., it included all species in the subsection as defined by Atehortúa [1983]: E. apodum [fig. 6E], E. auripilum [fig. 6D], E. backhousianum [fig. 6C], E. cubense, E. latum, and E. raywaense; app. A). In sect. Setosa, the clade formed by E. boryanum var. eutecnum–E. siliquoides (fig. 1) was characterized by three synapomorphies: the presence of cristae (fig. 12), spines (fig. 13), and perforations (fig. 14). Reversals, however, occurred in all these character states. A surprise result of the analyses by Rouhan et al. (2004) and Skog et al. (2004) was that E. decoratum belonged to the subulate-scaled clade (fig. 1). This was unexpected on the basis of morphology because the scales of this species are Fig. 12 Optimization mapping of two perispore characters, presence of broad folds or cristae and perispore character, in Elaphoglossum on the strict consensus tree of Rouhan et al. (2004). 918 919 Fig. 13 Optimization mapping of two perispore characters, continutity of broad folds or cristae and presence of spines, Elaphoglossum on the strict consensus tree of Rouhan et al. (2004). 920 921 Fig. 14 Optimization mapping of a perispore character, presence of perforations, in Elaphoglossum on the strict consensus tree of Rouhan et al. (2004). MORAN ET AL.—SPORES OF ELAPHOGLOSSUM broad and flat, unlike typical subulate scales that are hairlike and enrolled at the base. The species appears so distinct that Mickel and Atehortúa (1980) placed it in its own section (sect. Decorata). A reexamination of specimens, however, showed that the scales, although appearing nonsubulate, were in fact enrolled at the base, thus agreeing with the molecular results. In those results, E. decoratum was sister to E. piloselloides and E. spatulatum (bootstrap support 56%; fig. 1). Their spores are similar to each other (fig. 6H–6J) and to those of other subulate-scaled species, such as E. auripilum (fig. 6D) and E. hybridum var. vulcanii (fig. 6F). In all instances, however, the similarities represent the plesiomorphic condition (figs. 12–14). Sect. Lepidoglossa. The perispores in this section (figs. 8G–11L) were generally characterized by the plesiomorphic states of being broadly and continuously folded, nonperforate, and without spines (figs. 8G–8L, 9–11). The presence of spines appears to be a synapomorphy for smaller clades within the section (fig. 13). One such clade is informally referred to as the ‘‘ciliatum group,’’ consisting of E. burchellii, E. ciliatum, and E. nigrescens (fig. 1; fig. 11B–11D, respectively). Also belonging to this clade on the basis of two chloroplast markers (R. C. Moran, J. G. Hanks, and G. Rouhan, unpublished data) are the Neotropical species E. huacsaro (fig. 11A), E. dussii (fig. 10L), and E. gramineum (app. A) and three endemics of the island of St. Helena in the Atlantic Ocean, E. bifurcatum (app. A), E. dimorphum (app. A), and E. nervosum. All these species except E. dussii have spiny perispores. In sect. Lepidoglossa, the perispore was particularly distinctive in E. muscosum (fig. 11J), a species belonging to subsect. Muscosa (Mickel and Atehortúa 1980). The ca. 15 species in this subsection have laminae densely scaly abaxially, lamina apices acute to obtuse, and elevational ranges above 2000 m (A. Vasco, personal communication). In the seven species examined from the group so far (E. atropunctatum, E. bellermanianum, E. blandum, E. engelii, E. muscosum, E. oculatum, and E. yatesii; app. A), the spores are large, frequently collapsed, and papillate. These characters may even- 923 tually prove to be a synapomorphy for the entire subsection. Within the subsection, perisporal folds are present in some species (E. atropunctatum and E. yatesii) but absent in others (E. oculatum, E. engelii, E. blandum, E. bellermannianum, and E. muscosum; app. A). The presence of large spores suggests polyploidy, but this cannot be assessed because chromosome numbers are lacking for subsect. Muscosa. Mickel and Atehortúa (1980) placed E. ovatum in sect. Squamipedia because of its long-creeping rhizomes and small leaves (<12 cm long); however, it lacks the paired peglike aerophores on the rhizome characteristic of sect. Squamipedia, and it has phyllopodia and setose-margined lamina scales typical of sect. Lepidoglossa. In the analysis of Rouhan et al. (2004), E. ovatum was resolved in sect. Lepidoglossa, and this agrees with its perispore characteristic (fig. 10C) of being smooth, not spiny as in the Neotropical species of sect. Squamipedia (figs. 5H–5K, 13). Although the five perispore characters scored in this study showed considerable homoplasy, synapomorphies were found that were helpful in characterizing clades within Elaphoglossum. The perispore characteristics have proved to be useful in phylogenetic analysis, and we plan to incorporate them in a larger analysis including all morphological characters of the genus. Acknowledgments This work was supported by a grant to R. C. Moran from the U.S. National Science Foundation (DEB-00211969). It was also supported by the generosity of the Lewis B. and Dorothy Cullman Foundation. We thank Dennis W. Stevenson for use of the SEM laboratory in Pfizer Laboratory at the New York Botanical Garden and Lisa Campbell for assistance in the laboratory throughout this study. We also thank Michael Sundue, Alejandra Vasco, Larry Kelly, and Fabian Michelangeli for helpful discussions and two anonymous reviewers for providing many constructive comments on the manuscript. Appendix A Voucher Table Table A1 List of Elaphoglossum Species Whose Spores Were Imaged for This Study Species E. E. E. E. E. E. E. E. E. E. E. achroalepis (Baker) C. Chr. acrostichoides (Hook. & Grev.) Schelpe acutifolium Rosenst. aemulum (Kaulf.) Brack. affine (M. Martens & Galeotii) T. Moore aff. latifolium (Sw.) J. Sm. of Rouhan et al. 2004 alatum (Fée) T. Moore albescens (Sodiro) H. Christ alfredii Rosenst. ambiguum (Mett. ex H. Christ) Alston amygdalifolium (Mett. ex Kuhn) H. Christ Infrageneric group Voucher Figure number Lepidoglossa Platyglossa Platyglossa New section Platyglossa Platyglossa Elaphoglossum Setosa Setosa Platyglossa Amygdalifolia Madagascar, van der Werff 12837 (P) Tanzania, De Boer & Schippe 458 (NY) Bolivia, Kessler et al. 4317 (NY) Hawaii, Topping 2640b (NY) Mexico, Camp 2682 (NY) Guatemala, Molina 21310 (NY) Hawaii, collector unknown (NY) Ecuador, Palacios & Rubio 5275 (MO) Costa Rica, Lloyd 4262 (NY) Ecuador, Navarette & Asimbaya 1767 (NY) Costa Rica, Grayum 2373 (NY) 9C 5C ... 2B 5D 4G ... ... ... ... 2A Table A1 (Continued ) Species E. E. E. E. E. E. E. E. andicola (Fée) T. Moore angulatum (Blume) T. Moore angustius Mickel antisanae (Sodiro) S. Chr. apodum (Kaulf.) Schott ex J. Sm. asterolepis (Baker) C. Chr. atropunctatum Mickel aubertii (Desv.) T. Moore Infrageneric group E. auricomum (Kunze) T. Moore Lepidoglossa auripilum H. Christ avaratraense Rakotondr. backhousianum T. Moore bakeri (Sodiro) H. Christ ballivianii Rosenst. bellermannianum (Klotzsch) T. Moore bifurcatum (Jacq.) Mickel biolleyi H. Christ blandum Rosenst. boragineum (Sodiro) H. Christ boryanum (Fée) T. Moore var. boryanum boryanum (Fée) T. Moore var. eutecnum Mickel E. buchtienii Rosenst. E. burchellii (Baker) C. Chr. Polytricha Lepidoglossa Polytrichia Setosa Platyglossa Lepidoglossa Lepidoglossa Pachyglossa Lepidoglossa Setosa Setosa E. E. E. E. E. E. E. E. E. E. E. E. Voucher Ecuador, Lojtnant et al. 12499 (AAU) La Réunion, Rouhan 216 (NY) Bolivia, Steinbach 9152 (NY) Ecuador, Jaramillo 9887 (AAU) Haiti, Ekman 4729 (NY) Madagascar, Rouhan 305 (P) Ecuador, Holm-Nielsen 1907 (AAU) La Réunion, Rouhan 226 (NY); Madagascar, Rakotondrainibe 2497(P) Guatemala, Johnson 932 (NY); Costa Rica, Horich s.n. (NY) Costa Rica, Burger & Stolze 5335 (NY) Madagascar, Rakotondrainibe 1456 (P) Panama, Folsum 3576 (NY) Costa Rica, Taylor 17696 (NY) Bolivia, Nuñez & Sundue 316 (NY) Colombia, H. Smith 2640 (NY) St. Helena, Cumming s.n. (NY) Costa Rica, Davidse et al. 28794 (NY) Bolivia, Beck 3134 (NY) Panama, Croat 25299 (NY) Guadeloupe, Duss 4151 (NY) Pachyglossa Platyglossa Lepidoglossa Platyglossa Polytrichia Lepidoglossa Lepidoglossa Setosa Setosa Lepidoglossa Lepidoglossa E. E. E. E. E. E. E. cardenasii W. H. Wagner cardiophyllum (Hook.) T. Moore caricifolium Mickel casanense Rosenst. castaneum (Baker) Diels cf. tovii E. Brown (E. sp. of Rouhan et al. 2004) ciliatum (C. Presl) T. Moore Setosa Platyglossa Lepidoglossa Lepidoglossa Setosa Lepidoglossa Lepidoglossa E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. cismense Mickel cocosense Mickel concinnum Mickel conforme (Sw.) Schott conspersum H. Christ cordifolium Rosenst. coriaceum Bonap. costaricense H. Christ coursii Tardieu crassicaule Copel. crassifolium (Gaud.) L. E. Anderson & Crosby crinitum (L.) H. Christ crispipaleaceum M. Kessler & Mickel croatii Mickel cubense (Mett. ex Kuhn) C. Chr. curvans (Kunze) A. Rojas cuspidatum (Willd.) T. Moore davidsei Mickel decaryanum Tardieu deckenii (Kuhn) C. Chr. decoratum Maxon delasotae Mickel deltoideum (Sodiro) H. Christ dendricola (Baker) H. Christ dichroum Mickel didymoglossoides C. Chr. Pachyglossa Lepidoglossa Squamipedia Platyglossa Platyglossa Polytrichia Platyglossa Setosa Platyglossa Elaphoglossum Elaphoglossum Polytrichia Lepidoglossa Pachyglossa Polytrichia Lepidoglossa Lepidoglossa Setosa Pachyglossa Lepidoglossa Setosa Setosa Squamipedia Lepidoglossa Lepidoglossa Setosa 924 Venezuela, Luteyn & Lebron-Luteyn 5176 (NY) Bolivia, Tate 838 (NY) Ecuador, Holm-Nielsen et al. 35187 (NY); Bolivia, Acebey & Tosto 176 (NY) Bolivia, Kessler et al. 6564 (NY) Ecuador, Holm-Nielsen 5915 (NY) Costa Rica, Rojas 2898 (NY) Bolivia, Steinbach 8964bis (NY) Peru, Leon 1961 (NY) Marquesas, Lorence 9011 (NY) Costa Rica, Ingram & Ingram-Ferral 1308 (NY); Ecuador, Balslev & Madson 10402 (NY) Costa Rica, Rojas 2877 (NY) Costa Rica, Rojas 3642 (NY) Peru, Young 1301 (NY) South Africa, Taylor 1456 (NY) Costa Rica, Seidenschnur 268 (NY) Bolivia, Croat 51337 (NY) Madagascar, Geay 103 (P) Costa Rica, Seidenschnur 199 (NY) Madagascar, Rouhan 427 (P) Hawaii, Annable & Reinard 3793 (NY) Hawaii, Degener 9940 (NY) Costa Rica, Angulo 605 (NY) Bolivia, Nuñez 521 (NY) Costa Rica, Rojas 1535 (NY) Cuba, Jack 8058 (NY) Ecuador, Valencia 129 (NY) Peru, Smith & Albán 5503 (NY) Costa Rica, Montgomery & Huttleson 85135 (NY) Madagascar, Rasolohery 873 (P) Comoros, Rouhan & Rakontondrainibe 105 (NY) Brazil, Wackett 284 (NY) Bolivia, Kessler et al. 7123 (NY) Ecuador, Harley & Andersson 18092 (NY) Ecuador, Jorgensen et al. 1827 (NY) Ecuador, Bogh 47862 (AAU) Bolivia, Beck 3615 (NY) Figure number ... 4L ... ... 6E 10B ... 7E 11L 6D 9B 6C ... ... ... ... 3K ... ... ... 6K ... 11B 6L 4K 10H ... ... 9J 11C 3C ... ... ... 4J ... 5G 8E 5F ... ... 6A ... 3B ... ... 8G 7B 3I 11E 6H ... ... ... ... ... Table A1 (Continued ) Species Infrageneric group Voucher Figure number E. dimorphum (Hook. & Grev.) T. Moore E. doanense L. D. Gómez E. dussii Underw. & Maxon (E. aff. petiolatum of Rouhan et al. [2004]) E. edwallii Rosenst. E. engelii (H. Karst.) H. Christ E. erinaceum (Fée) T. Moore Lepidoglossa Pachyglossa St. Helena, collector unknown (P) Panama, McPherson 12844 (NY) Lepidoglossa Lepidoglossa Lepidoglossa Polytrichia 10L 10F ... 6B eximium (Mett.) H. Christ fauriei Copel. feei (Bory ex Fée) T. Moore flaccidum (Jenm.) Alston forsythii-majoris H. Christ fournierianum L. D. Gómez furfuraceum (Mett. ex Kuhn) H. Christ gayanum (Fée) T. Moore (E. minutum [Pohl ex Fée] T. Moore of Rouhan et al. 2004) E. glabellum J. Sm. E. glaucum T. Moore E. gloeorrhizum Mickel E. gramineum (Jenman) Urb. E. grayumii Mickel E. guamanianum (Sodiro) C. Chr. E. guatemalense (Klotzsch) T. Moore E. guentheri Rosenst. E. hartwegii (Fée) T. Moore E. haynaldii (Sodiro) I. Losch E. herminieri (Bory & Fée) T. Moore E. herpestes Mickel E. heterolepis (Fée) T. Moore E. heteromorphum (Klotzsch) T. Moore E. hieronymi (Sodiro) C. Chr. E. hoffmannii (Mett. ex Kuhn) H. Christ E. hornei C. Chr E. horridulum (Kaulf.) J. Sm. E. huacsaro (Ruı́z) H. Chirst E. humbertii C. Chr. E. hybridum (Bory) Brack. E. hybridum (Bory) Brack. var. vulcanii E. inquisitivum M. Kessler & Mickel E. killipii Mickel E. laminarioides (Fée) T. Moore E. lanatum (Bojer ex Baker) Lorence E. lancifolium (Desv.) C. V. Morton Setosa Elaphoglossum Squamipedia Pachyglossa Lepidoglossa Setosa Lepidoglossa Dominican Republic, Lloyd 351 (NY) Brazil, Rose & Russell 20482 (NY) Ecuador, van der Werff & Guidino 11005 (MO) Peru, Gentry et al. 22977 (NY); Bolivia, Solomon 16391 (MO) Costa Rica, Moran 2328 (NY) Hawaii, Baldwin 100 (NY) Guadeloupe, Questel 3267 (NY) Ecuador, Moran 6278 (NY) La Réunion, Cadet 1535 (P) Costa Rica, Davidse 24999 (NY) Costa Rica, Davidse 24757 (MO) 5E 5B 2C ... ... 2E ... 2H ... ... ... 2I ... 9F ... ... 3J 4C ... 11A 9E 6G 6F ... 10K ... 9H 10I Lepidoglossa Platyglossa Pachyglossa Polytrichia Lepidoglossa Peru, Segastegui et al. 8238 (NY) Panama, Stimson 5411 (NY) Mexico, Mickel 9675 (NY) Costa Rica, Rojas 3304 (NY) Dominican Republic, Valeur 634 (NY) Costa Rica, Grayum 3017 (NY) Peru, Barbour 2564 (MO) Mexico, Martinez 14987 (NY) Bolivia, Beck 479 (NY) Venezuela, Ruiz Teran & Lopez Figueiras 2130 (NY) Colombia, Killip 6853 (US); Grant 9846 (US) Suriname, Inghamon & Inghamon 34007 (NY) Ecuador, Madison 6866 (SEL) Mauritius, Rouhan 179 (NY) Ecuador, Croat 58771 (MO) Ecuador, Øllgaard & Balslev 10206 (AAU) Costa Rica, Hennipman et al. 6986 (NY) Seychelles, Rouhan 137 (NY) Brazil, Edwall & Toledo 21187 (NY) Venezuela, Ruiz-Terán 6124 (NY) Madagascar, Humbert 6900 (P) La Réunion, Rouhan 217 (NY) La Réunion, Rouhan 222 (NY) Bolivia, Kessler et al. 11812 (NY) Bolivia, Buchtien 4239 (NY) Ecuador, Palacios 4312 (MO) Mauritius, Pike s.n. (NY) La Réunion, Rouhan 201 (NY); Grande Comore, Rouhan 130 (P) Peru, Segastegui 3250 (NY) Tanzania, DeBoer & Schippe 460 (K) Jamaica, Underwood 3146 (NY) Costa Rica, Lloyd 4162 (NY) Bolivia, Kessler et al. 7061 (NY) Pachyglossa Lepidoglossa Lepidoglossa Polytrichia Platyglossa Lepidoglossa Squamipedia Pachyglossa Lepidoglossa Mauritius, Rouhan et al. 175 (NY) Costa Rica, Rojas et al. 2836 (NY) Madagascar, McPherson 1492 (P) Argentina, Venturi 3256 (US) Costa Rica, Mickel 2688 (NY) Ecuador, Tipaz et al. 2179 (M) Bolivia, Nuñez 51 (NY) Guyana, Clarke & McPherson 1949 (NY) Venezuela, Luteyn et al. 6135 (NY) 2K ... 11G ... 4H ... ... 4B ... E. E. E. E. E. E. E. E. lasioglottis Mickel lastii (Baker) C. Chr. latifolium (Sw.) J. Sm. latum (Mickel) Atehortúa ex Mickel lechlerianum (Mett.) T. Moore lepervanchei (Bory ex Fée) T. Moore (E. aff. conforme [Sw.] J. Sm. of Rouhan et al. 2004) E. leporinum L. D. Gómez E. leucolepis (Baker) Krajina ex Tardieu E. lindbergii (Mett. ex Kuhn) Rosenst. E. lingua (C. Presl) Brack. E. litanum (Sodiro) C. Chr. E. lloense (Hook.) T. Moore E. luridum (Fée) H. Christ E. luteynii Mickel E. E. E. E. E. E. Platyglossa Platyglossa New section Pachyglossa Lepidoglossa Pachyglossa Lepidoglossa Pachyglossa Lepidoglossa Lepidoglossa Setosa Pachyglossa Setosa Lepidoglossa Setosa Pachyglossa Pachyglossa Pachyglossa Setosa Lepidoglossa Lepidoglossa Polytrichia Polytrichia Lepidoglossa Lepidoglossa Lepidoglossa Lepidoglossa Lepidoglossa 925 ... ... 7D ... ... 4A 9A 8A 11I ... ... 2D ... ... Table A1 (Continued ) Species Infrageneric group E. macropodium (Fée) T. Moore Pachyglossa malgassicum C. Chr. marojejyense Tardieu marquisearum Bonap. mathewsii (Fée) T. Moore metallicum Mickel micropogon Mickel minutissimum R. C. Moran & Mickel (E. sp. nov. cf. hayesii [Mett. ex Kuhn] Maxon of Rouhan et al. 2004) E. mildbraedii Hieron. E. mitorrhizum Mickel E. molle (Sodiro) C. Chr. E. moorei (E. Britton) H. Christ E. muscosum (Sw.) T. Moore E. nigrescens (Hook.) T. Moore ex Diels (including E. palmense H. Christ and E. wackettii Rosenst. of Rouhan et al. 2004) E. E. E. E. E. E. E. 3E 5H 2G ... 3H 9K Setosa Setosa Platyglossa Lepidoglossa Squamipedia Lepidoglossa Costa Rica, Moran 6344 (NY) Tanzania, Rouhan 534 (P) Ecuador, Aulestia & Aulestia 1526 (MO) Ecuador, Mille s.n. (P) Colombia, van der Werff & Giraldo 9726 (MO) Venezuela, Ruiz-Terán 1864 (NY) 8D ... 4F ... ... 11J Lepidoglossa Brazil, Maas et al. 6858 (NY); Costa Rica, Ingram & Ferrell-Ingram 1925 (NY) Panama, Porter et al. 4383 (NY) Ecuador, Holm-Nielsen 6246 (AAU) Ecuador, Navarrete 387 (QCA) Ecuador, Øllgaard et al. 945 (AAU) Peru, Killip & Smith 24413 (NY) Brazil, Brade 9647 (NY) Brazil, Jurgens 75 (NY) Madagascar, Perrier 7860c (P) Ecuador, Ulloa et al. 1064 (NY) Bolivia, Beck 23090 (NY) Madeira, collector unknown (NY); Mexico, Lorea 3144 (NY) Mexico, Breedlove 32046 (NY) Ecuador, Øllgaard 38438a (AAU) Hawaii, Degener 31707 (NY) Hawaii, Degener 31704 (NY); Heller 2621 (NY) Costa Rica, Lent 3699 (AAU) Mexico, Mendez 7931 (NY) Madagascar, Perrier 16457 (P) Mexico, Carranza & Pérez 5140 (NY), Reveal & Atwood 3572 (NY); Ecuador, Holm-Nielsen et al. 3517 (AAU); Jamaica, Underwood 545, 3141 (NY), Jenman s.n. (NY); Costa Rica, Moran 3079 (NY); Dominican Republic, Lloyd 753 (NY); Honduras, Molina 22621 (NY) Madagascar, Decary 18115 (P) Costa Rica, Lloyd 4177 (NY); Colombia, Rodrı́gues et al. 4124 (NY) Ecuador, Fay & Fay 3519 (NY) Mexico, Breedlove & Smith 22060 (NY) Brazil, Thomas et al. 9371 (NY) Madagascar, Perrier de la Bâthie 12487 (P) Mexico, Parry & Palmer 1007 (NY) Brazil, Brade 15514 (NY) Mexico, Mickel 4463 (NY) Ecuador, Øllgaard & Navarrete 105872 (AAU) Panama, Herrera 790 (NY) Costa Rica, Maxon 464 (NY) Ecuador, Fay & Fay 3720 (NY) Ecuador, Øllgaard et al. 37834 (AAU) Brazil, Steward & Ramos P19692 (NY) Ecuador, Luteyn & Cotton 11281 (NY) 11D oblanceolatum C. Chr. oculatum Mickel odontolepis Mickel omissum Mickel orbignyanum (Fée) T. Moore organense Brade ornatum (Mett. ex Kuhn) H. Christ ovalilimbatum Bonap. ovatum (Hook. and Grev.) T. Moore pachyphyllum (Kunze) C. Chr. Setosa Lepidoglossa Pachyglossa Setosa Lepidoglossa Setosa Polytrichia Platyglossa Lepidoglossa Pachyglossa E. E. E. E. E. E. E. E. E. paleaceum (Hook. and Grev.) Sledge pallidum (Baker ex Jenman) C. Chr. papillosum (Baker) H. Christ parvisquameum Skottsb. pellucidum (Sodiro) C. Chr. peltatum (Sw.) Urb. forma standleyi peltatum (Sw.) Urb. forma peltatum perrierianum C. Chr. petiolatum (Sw.) Urb. Lepidoglossa Polytrichia Setosa Elaphoglossum Lepidoglossa Squamipedia Squamipedia Lepidoglossa Lepidoglossa E. E. E. E. E. E. E. E. E. E. E. E. E. E. piloselloides (C. Presl) T. Moore pilosius Mickel plumosum (Fée) T. Moore poolii (Baker) H. Christ potosianum H. Christ prestonii J. Sm. pringlei (Davenp.) C. Chr. productum Rosenst. proliferans Maxon & C. V. Morton proximum (J. Bommer) H. Christ pseudoboryanum Mickel pteropodum (Sodiro) C. Chr. pteropus C. Chr. pumilio Mickel Figure number Elaphoglossum Squamipedia Pachyglossa Lepidoglossa Pachyglossa Lepidoglossa E. E. E. E. E. E. E. E. E. E. E. phanerophlebium C. Chr. E. phoras Mickel Voucher La Réunion, Rouhan & Grangaud 207 (NY); Tanzania, DeBoer & Schippe 457 (K) Madagascar, Rouhan 427 (P) Madagascar, Humbert 23105 (P) Marquesas, Wood 6390 (P) Ecuador, Balslev & Mino 69231 (NY) Peru, Sagastegui et al. 14459 (NY) Costa Rica, Lloyd 4111 (NY) Setosa Setosa Setosa Setosa Lepidoglossa Lepidoglossa Lepidoglossa Polytrichia Lepidoglossa Pachyglossa Setosa Platyglossa Setosa Setosa Pachyglossa Lepidoglossa 926 3G 8C ... ... ... 11H ... ... 4E 10C ... 9I ... 8B ... ... ... 5I ... 8K 7K ... 6I 7I ... 11F ... 5L 8J 2J ... 4I ... ... 3L ... Table A1 (Continued ) Species pygmaeum (Mett. ex Kuhn) H. Christ randii Alston & Schelpe rapense Copel. raywaense (Jenman) Alston 3 revaughanii Lorence richardii (Bory ex Fée) H. Christ rimbachii (Sodiro) H. Christ (E. nigrocostatum Mickel of Rouhan et al. 2004) E. rosenstockii H. Christ ex Rosenst. E. rosettum R. C. Moran & Mickel E. rufidulum (Willd. ex Kuhn) C. Chr. E. russelliae Mickel E. rzedowskii Mickel E. samoense Brack. E. sartorii (Liebm.) Mickel E. scolopendriforme Tardieu E. setigerum (Sodiro) Diels E. sieberi (Hook. and Grev.) T. Moore E. siliquoides (Jenm.) C. Chr. E. smithii (Baker) H. Christ E. spatulatum (Bory) T. Moore E. splendens (Bory ex Willd.) Brack. E. squamatum (Sw.) T. Moore (E. lindenii [Bory ex Fée] T. Moore of Rouhan et al. 2004) E. E. E. E. E. E. E. E. squamipes (Hook.) T. Moore (including E. craspedariiforme [Fée] Brade ex Alston of Rouhan et al. 2004) E. stenophyllum (Sodiro) Diels E. stipitatum (Bory ex Fée) T. Moore E. E. E. E. E. E. subcinnamomeum (H. Christ) Hieron. subsessile (Baker) C. Chr. succisaefolium (Thouars) T. Moore tabanense André ex H. Christ tectum (Humb. & Bonpl. ex Willd.) T. Moore tenuiculum (Fée) T. Moore ex C. Chr. E. terrestre A. Rojas (E. sp. nov. cf. terrestre of Rouhan et al. 2004) E. tomentosum (Bory ex Willd.) H. Christ E. tonduzii H. Christ E. tripartitum (Hook. & Grev.) Mickel E. trivittatum (Sodiro) H. Christ E. truncatum Rosenst. E. unduaviense Rosenst. E. vestitum (Schltdl. & Cham.) T. Moore E. vieillardii (Mett.) T. Moore E. villosum (Sw.) J. Sm. E. wardiae Mickel E. wawrae (Luerss.) C. Chr. E. webbii T. Moore E. welwitschii (Baker) C. Chr. E. wrightii (Mett.) T. Moore E. yatesii (Sodiro) H. Christ E. yungense de la Sota E. yunnanense (Baker) C. Chr. E. zebrinum Mickel Infrageneric group Voucher Figure number Setosa Lepidoglossa Setosa Polytrichia Lepidoglossa Lepidoglossa Ecuador, Moran & Rohrbach 5221 (MO) Marion Island, Huntley 464 (P) French Polynesia, Motley 2677 (NY) Ecuador, Palacios 394 (MO) Mauritius, Rouhan et al. 190 (NY) La Réunion, Rouhan 231 (P) 7J 10E 7G ... ... 8I Lepidoglossa Lepidoglossa Setosa Lepidoglossa Setosa Lepidoglossa Setosa Pachyglossa Lepidoglosssa Setosa Pachyglossa Setosa Setosa Setosa Lepidoglossa Ecuador, Øllgaard & Balslev 8643 (AAU) Ecuador, Øllgaard et al. 37940 (AAU) Bolivia, Tate 618 (US) La Réunion, Cadet 1535 (P) Costa Rica, Gómez et al. 22409 (NY) Mexico, Rzedowski 26032 (NY) French Polynesia, Motley 2875 (NY) Mexico, Munn-Estrada & Mendoza 1683 (NY) Madagascar, Rouhan 306 (P) Ecuador, Holm-Nielsen & Jeppesen1293 (AAU) Mauritius, Rouhan et al. 176 (NY) Costa Rica, Lloyd 4282 (NY) Ecuador, Øllgaard et al. 1014 (AAU) La Réunion, Rouhan & Grangaud 246 (NY) La Réunion, Rouhan 213 (NY) 8L ... ... 9L 7A ... 7F 3A 9D 7C 3F 8F 7L 6J 10A Setosa Ecuador, Øllgaard & Balslev 8410 (NY); Colombia, Killip & Smith 18607 (NY) Squamipedia Lepidoglossa Lepidoglossa Venezuela, Rivero & Diaz 1496 (NY) Ecuador, Grijalva 568b (NY) Madagascar, Pichi Sermolli 5120 (P); La Réunion, Rouhan 212 (NY) Kenya, Schelpe 2605 (P) Madagascar, Rouhan 432 (P) Amsterdam Island, Martel-Thoumian 2A (P) Ecuador, Øllgaard & Navarette 2835 (AAU) Colombia, Zarucchi et al. 5800 (NY) Peru, van der Werff et al. 15697 (NY); Ecuador, Pitman & Romero 353 (NY) 5J ... 10D Costa Rica, Moraga & Rojas 509 (NY) Mauritius, Rouhan 178 (NY) Costa Rica, Herrera 3511 (NY) Ecuador, Øllgaard 99199 (AAU) Ecuador, Øllgaard & Balslev 8272 (AAU) Bolivia, Williams 1031 (NY) Bolivia, Tate 345 (NY) Mexico, Mickel & Leonard 4705 (NY) New Caledonia, Munzinger 1276 (P) Jamaica, Yuncker 17696 (NY) Peru, Foster et al. 10890 (NY) Hawaii, Forbes 350 (NY) Colombia, Lehmann BT760 (NY) Tanzania, Taylor 9099 (P) Cuba, Ekman 3882 (NY) Ecuador, Øllgaard & Balslev 8411 (AAU) Bolivia, Kessler et al. 6444 (NY) China, Henry 13110 (NY) Peru, Foster 10891 (NY) 2F 9G ... 5K ... ... ... 11K 3D ... ... 5A ... 10G ... ... ... ... ... Lepidoglossa Pachyglossa Lepidoglossa Setosa Lepidoglossa Lepidoglossa Pachyglossa Lepidoglossa Lepidoglossa Squamipedia Lepidoglossa Polytrichia Pachyglossa Lepidoglossa Pachyglossa Setosa Setosa Platyglossa Setosa Lepidoglossa Squamipedia Lepidoglossa Lepidoglossa Platyglossa Setosa 7H ... 2L 8H ... 10J ... Note. Species with figure numbers given were included in the Rouhan et al. (2004) analysis. Infrageneric groups were assigned on the basis of the Rouhan et al. (2004) analysis, and for those species not in that analysis, on the basis of morphology according to Mickel and Atehortúa (1980). All images are available at http://www.plantsystematics.org. The vouchers used for both this study and that of Rouhan et al. (2004) are underlined. 927 INTERNATIONAL JOURNAL OF PLANT SCIENCES 928 Appendix B Character States Table B1 List of Spore Characters and States Scored for Those Species of Elaphoglossum Included in the Analysis of Rouhan et al. (2004) Species Outgroups: Athyrium filix-femina Bolbitis auriculata Lomariopsis marginata Rumohra adiantiformis Elaphoglossum: E. achroalepis E. acrostichoides E. aemulum E. affine E. amygdalifolium E. angulatum E. apodum E. asterolepis E. aubertii E. auricomum E. auripilum E. avaratraense E. backhousianum E. biolleyi E. boryanum var. eutecnum E. burchellii E. cardenasii E. cardiophyllum E. caricifolium E. ciliatum E. cismense E. conspersum E. coriaceum E. costaricense E. coursii E. crinitum E. croatii E. cuspidatum E. davidsei E. decaryanum E. deckenii E. decoratum E. doanense E. dussii E. edwallii E. erinaceum Characters 12345 00011 00001 1--01 00011 01011 00*11 00011 00011 01011 00011 00011 00011 1--01 00011 00011 00011 00011 01000 01100 00001 01100 00011 00011 00001 01000 00011 00011 01000 00011 00011 01100 00111 01000 01010 00011 00011 00001 00011 00011 00011 Species Characters 12345 Species Characters 12345 E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. 1--01 01000 00010 01*00 00011 00011 00011 00011 01000 00011 00111 00001 01000 00*10 00001 00001 00011 00011 00011 00001 00011 00011 01000 01000 00010 00011 01100 00011 00011 00010 01*00 01000 00011 00011 00011 1--11 00001 1--01 00011 00011 00011 00011 E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. E. 1--01 00001 00011 01000 00011 00000 00011 00001 00011 00011 00011 01000 1--01 00001 01010 00011 00011 00011 01100 01000 01000 00001 1--01 00011 00010 01000 00011 00011 01000 00001 00001 01000 00011 00011 01000 00011 00011 00001 00011 00001 00111 00011 eximium flaccidum forsythii-majoris fournierianum furfuraceum gayanum glabellum glaucum grayumii guatamalense herminieri heterolepis hoffmannii hornei huacsaro humbertii hybridum hybridum var. vulcanii killipii lanatum lancifolium aff. latifolium latifolium lepervanchei leucolepis lingua luridum macropodium malgassicum marojejyense marquisearum metallicum micropogon minutissimum mitorrhizum muscosum nigrescens oblanceolatum orbignyanum ovalilimbatum ovatum paleaceum papillosum peltatum petiolatum phanerophlebium pillosellioides pilosius poolii prestonii pringlei productum proximum pteropus pygmaeum randii rapense richardii rimbachii rufidulum russelliae samoense sartorii scolopendriforme setigerum sieberi siliquoides smithii spatulatum splendens squamatum squamipes stipitatum subsessile succisaefolium tectum terrestre tomentosum tovii tripartitum vestitum vieillardii wawrae welwitschii Note. Five-digit numbers indicate states for characters 1–5, from left to right. Characters and states are as follows. (1) Broad folds or cristae: present ¼ 0, absent ¼ 1. (2) Perispore: broadly folded ¼ 0, cristae ¼ 1. (3) Broad folds or cristae: continuous ¼ 0, discontinuous ¼ 1. (4) Spines: present ¼ 0, absent ¼ 1. (5) Perforations: present ¼ 0; absent ¼ 1. A dash in place of a number indicates a nonapplicable character state, and an asterisk indicates a polymorphism. Literature Cited Atehortúa L 1983 Taxonomic study of the Elaphoglossum apodum complex (Elaphoglossaceae). PhD diss. City University of New York. Christ KHH 1899 Monographie des Genus Elaphoglossum. Neue Denkschr Allg Schweiz Ges Gesammten Naturwiss 36:1–159. Fée ALA 1845 Histoire des acrostichées mémoires fougeres. Vol 2. Berger-Levrault, Strasbourg. Holttum RE 1978 Lomariopsis group. Flora Malesiana Ser II Pteridophyta 1:255–330. MORAN ET AL.—SPORES OF ELAPHOGLOSSUM Korall P, WA Taylor 2006. Megaspore morphology in the Selaginellaceae in a phylogenetic context: a study of the megaspore surface and wall structure using scanning electron microscopy. Grana 45:22–60. Kramer KU 1990 Lomariopsidaceae. Pages 164–170 in KU Kramer, PS Green, eds. The families and genera of vascular plants. Vol 1. Pteridophytes and gymnosperms. Springer, New York. Lorence DH, G Rouhan 2004 A revision of the Mascarene species of Elaphoglossum (Elaphoglossaceae). Ann Mo Bot Gard 91:536– 565. Maddison DR, WP Maddison 2000 MacClade 4. Sinauer, Sunderland, MA. Mickel JT 1980 Relationships of the dissected elaphoglossoid ferns. Brittonia 32:109–117. ——— 1985 The proliferous species of Elaphoglossum (Elaphoglossaceae) and their relatives. Brittonia 37:261–278. Mickel JT, L Atehortúa 1980 Subdivision of the genus Elaphoglossum. Am Fern J 70:47–68. Moran RC, JT Mickel 2004 Three new Neotropical species of Elaphoglossum (Elaphoglossaceae) with subulate scales. Brittonia 56:200–204. 929 Palmer DD 2003 Hawai’i’s ferns and fern allies. University of Hawai’i Press, Honolulu. Rodrı́guez R 1995 Pteridophyta. Pages 119–351 in C Marticorena, R Rodrı́guez, eds. Flora de Chile. Universidad de Concepción. Rouhan G, J-Y Dubuisson, F Rakotondrainibe, TJ Motley, JT Mickel, J-N Labat, RC Moran 2004 Molecular phylogeny of the fern genus Elaphoglossum (Elaphoglossaceae) based on chloroplast noncoding DNA sequences: contributions of species from the Indian Ocean area. Mol Phylogenet Evol 33:745–763. Skog JE, JT Mickel, RC Moran, M Volovsek, EA Zimmer 2004 Molecular studies of representatives species in the fern genus Elaphoglossum (Dryopteridaceae) based on cpDNA sequences rbcL, trnL-F, and rps4-trnS. Int J Plant Sci 165:1063–1075. Smith AR, KM Pryer, E Schuettpelz, P Korall, H Schneider, PG Wolf 2006 A classification for extant ferns. Taxon 55:705–731. Tryon AF, B Lugardon 1991 Spores of the Pteridophyta: surface, wall structure, and diversity based on electron microscope studies. Springer, New York. Tryon RM, AF Tryon 1982 Ferns and allied plants with special reference to tropical America. Springer, Berlin.