Spore Morphology in Relation to Phylogeny in the Fern Genus Elaphoglossum (Dryopteridaceae)

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 De´veloppement US084, Muse´um National d’Histoire Naturelle, De´partement Syste´matique 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 diver- Elaphoglossum contains ca. 600 species and ranks as one sity corresponded to their proposed subsections or unnamed of the world’s largest fern genera. It is pantropical but most species groups. Mickel (1980, 1985) published spore images species-rich in the Neotropics, where ca. 80% of the species oc- for 24 species of Elaphoglossum, and the spores of 50 species cur. Nearly 75% of the species are epiphytes. The genus is re- of Elaphoglossum were examined by Tryon and Lugardon lated to Bolbitis, Teratophyllum,andLomagramma,asevinced (1991). These workers found the spores to be monolete and by chloroplast DNA sequence data (E. Schuettpelz, personal bilateral, to have a longitudinal laesura, and (depending on communication) and morphological characters such as acrosti- the species) to be from 20 to 80 mm long. None of the above choid sori, sterile-fertile leaf dimorphy, an elongated (in cross studies, however, assessed perispore morphology in a cladis- section) ventral vascular bundle in the rhizome, and this ventral tic framework because phylogenetic trees were lacking. vascular bundle bearing all the roots (thus, the roots appear to Rouhan et al. (2004) and Skog et al. (2004) presented phylo- be borne only on the ventral surface of the rhizome; Holttum genetic hypotheses for Elaphoglossum based on DNA sequences 1978; Kramer 1990). These genera are currently placed in the from the chloroplast genome (trnL-trnF and rps4-trnS by Rouhan Dryopteridaceae, a placement well supported by molecular phy- et al. [2004]; rbcL, trnL-F,andrps4-trnS by Skog et al. [2004]). logenetic studies (Smith et al. 2006). Unfortunately, Elaphoglos- The Rouhan et al. (2004) study was the most comprehensive, sum lacks a fossil record. containing 123 taxa from both the New and Old Worlds (as Feé (1845) and Christ (1899) proposed subgeneric classifi- opposed to 48 species from only the New World in the Skog cations of Elaphoglossum based primarily on frond shape, et al. [2004] study). Rouhan et al. (2004) recovered 11 main texture, scale characteristics, and the presence versus absence clades within the genus (fig. 1). These clades largely corre- of hydathodes. More recently, Mickel and Atehortu´ a (1980) sponded to the sections and subsections proposed by Mickel proposed an infrageneric classification based on sporophyte and Atehortuá (1980). The clades (starting at the bottom of fig. morphology and spore structure as determined from images 1) can be characterized as follows. Section Amygdalifolia con- taken with a scanning electron microscope (SEM). The mor- sists of a single species (E. amygdalifolium)thathaslong-creeping phological characteristics they found useful in subdividing rhizomes, phyllopodia, and hydathodes—a unique character the genus included rhizome habit, lamina scale type and dis- combination in the genus. Its reddish young leaves are also tribution, and the presence versus absence of hydathodes, unique within the genus. Elaphoglossum aemulum (endemic to Hawaii), E. glaucum (native to Central America), and sect. Elaphoglossum are characterized by phyllopodia and usually 1 Author for correspondence; e-mail [email protected]. thick, glabrous to very sparsely scaly laminae. Based on mo- 2 E-mail [email protected]. lecular evidence (Rouhan et al. 2004), sect. Elaphoglossum 3 E-mail [email protected]. consists of two main clades, subsects. Pachyglossa and Platy- Manuscript received December 2006; revised manuscript received March glossa, although no known morphological or anatomical char- 2007. acters separate these two groups. Section Squamipedia is

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 trnL- trnF and rps4-trnS alignments. Species in boldface are from the Indian Ocean area. After Rouhan et al. (2004). MORAN ET AL.—SPORES OF ELAPHOGLOSSUM 907 characterized by long-creeping rhizomes, paired peglike aero- glossa, is characterized by laminar scales with marginal teeth phores on the rhizome near the petiole base, pale brownish consisting of acicular cells (all other Elaphoglossum species ﬂaccid rhizome scales, and a lack of phyllopodia. The subulate- have laminar scales with marginal teeth consisting of glandular scaled clade (Subulata; ﬁg. 1) is characterized by leaves with or bulbous marginal cells or gland-tipped marginal processes). subulate scales, that is, erect hairlike scales enrolled at the base Given the broad correspondence of sporophyte morphology and often all the way to the tip. This clade is composed of two with the groups recovered from recent molecular phylogenetic subsections: subsect. Polytrichia, which lacks hydathodes, and studies, the objective of this article is to compare spore mor- subsect. Setosa, which has them. The last section, sect. Lepido- phology 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 from the same country as that of the DNA voucher. Gener- ally, one specimen per species and 20–100 spores per stub Spores were obtained from herbarium specimens of Ela- were examined. In cases where perispore variation was sus- phoglossum at AAU, F, GH, MO, NY, P, and US. For 12 spe- pected within a species, two or more specimens were examined cies (those with vouchers are underlined in app. A), spores (e.g., E. erinaceum, E. nigrescens, E. petiolatum;app.A).Be- were taken from the DNA vouchers cited in the Rouhan sides those species included in the Rouhan et al. (2004) analy- et al. (2004) study. It was not possible to do this for most of sis, 102 additional species were imaged to assess the generality the vouchers cited because they lacked fertile leaves. For of results. The species examined for this study and voucher in- these species, attempts were made to use specimens collected formation 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, Saga´stegui 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 909

The spores were transferred with dissecting needles from her- To assess perispore evolution in a phylogenetic context, barium specimens to aluminum SEM stubs and coated with ﬁve perispore characters were scored (app. B) from the SEM an asphalt adhesive. The stubs were then coated with gold- images (ﬁgs. 2–11). Their character state changes were then palladium in a sputter-coater for 2.5 min, and spores were im- optimized, using MacClade 4.0 (Maddison and Maddison aged digitally using a JEOL JSM-5410LV SEM equipped with a 2000), onto the strict consensus tree from the analysis of the JEOL Orion 5410 software interface. The accelerating voltage molecular data (Rouhan et al. 2004). was 15 kV. Spore images of all 221 species examined for this In several instances for the presentation of results, we use dif- study (app. A) are available at http://www.plantsystematics.org. ferent 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 Re´union, 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 fied as E. lepervanchei (R. C. Moran, personal observation); and because some names used by Rouhan et al. (2004) are now E. sp. nov. aff. hayesii has been described as E. minutissimum considered synonyms of older names (based largely on taxonomic (Moran and Mickel 2004); E. sp. has now been identified as studies by the first author). The name changes resulting from E.cf.tovii (D. Lorence, personal communication); and E. aff. reidentification of vouchers are as follows: E. aff. terrestre of petiolatum has now been identified as E. dussii (R. C. Moran, Rouhan et al. (2004) has now been confirmed as E. terrestre personal observation). Name changes resulting from syn- (Lorence and Rouhan 2004); E. aff. conforme has been identi- onymy 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, Go´ mez 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 Reúnion, Rouhan 222, P). G, E. hybridum var. hybridum (La Reúnion, Rouhan 217, NY). H, E. decoratum (Brazil, Wacket 248, NY). I, E. piloselloides (Ecuador, Fay & Fay 3519, NY). J, E. spatulatum (La Reúnion, 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 Based on morphology, we consider four species (E. craspe- now considered synonyms of E. nigrescens;andE. nigrocostatum dariiforme, E. aff. conforme, E. palmense, and E. wackettii) is now considered a synonym of E. rimbachii. Elaphoglossum to be synonyms of other species in the Rouhan et al. (2004) minutum is a synonym of E. revolutum, and the earliest name analysis (i.e., these species were represented more than once in available for the species used in the Rouhan et al. (2004) analysis the analysis). Thus, the total number of species in the Rouhan is E. gayanum. Finally, E. lindenii is now considered a synonym of et al. (2004) analysis was 119. The spore images of these spe- E. squamatum (Rodrı´guez 1995). These name changes have been cies and 102 more not in that analysis are available at http:// incorporated in figure 1 and are noted in appendix A. www.plantsystematics.org. 912 INTERNATIONAL JOURNAL OF PLANT SCIENCES

Fig. 7 Spores of Elaphoglossum. A, E. russelliae (Costa Rica, Go´mez 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 Re´union, 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 Elaphoglossum aemulum. The spores of this species (fig. 2B) have broadly folded, nonperforate perispores. Between The following results and the spore images in figures 2–11 the folds are lower, discontinuous folds. are arranged according to the ascending order of species shown Elaphoglossum glaucum. The spores of this species have in figure 1. broadly folded, nonperforate perispores without spines (fig. Sect. Amygdalifolia. The single species in this section had 2C). The surfaces between the folds are nearly smooth but nonperforate cristate perispores with the surfaces between have scattered, low, irregular deposits. the cristae smooth (fig. 2A). 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 & Albań 5503, NY). H, E. succisaefolium (Amsterdam Island, Martel-Thoumian 2A, P). I, E. richardii (La Reúnion, 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, folded or cristate perispores, with the cristate condition pre- we examined 31 species (app. A), 25 of which (figs. 2D–4D) dominating (fig. 12). Except for E. luridum, all species in the were included in the analysis of Rouhan et al. (2004). The subsection had continuous perisporal folds, and spines were species imaged from this subsection but not in the Rouhan generally present (fig. 13). Perforate perispores were often present et al. (2004) analysis were E. andicola, E. gloeorrhizum, E. (fig. 14), but the nonperforate condition characterized a clade luridum, E. odontolepis, E. pachyphyllum,andE. unduaviense of four Old World species consisting of the New Caledonian E. (app. A). All species of this subsection had either broadly 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 Reúnion, 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 Reúnion, Cadet 1535, P). Scale bars ¼ 10 mm. endemic E. malgassicum,andE. macropodium from African the Rouhan et al. (2004) analysis were E. ambiguum, E. anti- and Indian Ocean areas (figs. 3D–3G, 14). Nonperforate peri- sanae, E. ballivianii, E. conforme, E. lastii, E. proximum,and spores were also found in the New World species E. doanense E. yunnanense (app. A). All species of this subsection had (fig. 4D), E. guatemalense (fig. 2H), E. herminieri (fig. 2I), broadly folded perispores (fig. 12), the folds being continuous and E. productum (fig. 2J). (except in E. wawrae; fig. 5A) and without spines (fig. 13) or From subsect. Platyglossa, 22 species were examined (app. perforations (fig. 14). A), 15 of which (figs. 4E–5G) were included in the analysis Sect. Squamipedia. From this section, 12 taxa were ex- of Rouhan et al. (2004). The species imaged that were not in amined (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 Reúnion, Rouhan 213, NY). B, E. asterolepis (Madagascar, Rouhan 305, P). C, E. ovatum (Ecuador, Ulloa et al. 1064, NY). D, E. stipitatum (La Reúnion, 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 Reúnion, 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 species, the Madagascan endemic E. marojejyense (fig. 5H) that were not in the Rouhan et al. (2004) analysis are E. con- had nonspiny, perforate peripores. cinnum, E. deltoideum, E. feei, E. lloense, E. moorei, E. pelta- Subulata clade. From this clade, 63 species were exam- tum f. standleyi, E. tripartitum,andE. wrightii (app. A). All ined (app. A), 31 of which (figs. 5L–8F) were included in the species had perispores with broad, continuous folds (fig. 12). analysis of Rouhan et al. (2004). From sect. Polytrichia,we Spiny, nonperforate perispores were present in the Neotropical examined 16 species, eight of which (figs. 5L,6A–6G) were species (E. peltatum–E. tripartitum; fig. 13), but unlike these 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 Baˆthie 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) Those taxa with spores imaged that were not in the Rouhan analysis were E. cordifolium, E. cubense, E. latum, E. lindbergii, et al. (2004) analysis were E. albescens, E. alfredii, E. bakeri, E. ornatum, E. pallidum, E. raywaense,andE. truncatum.In E. boragineum, E. boryanum, E. castaneum, E. delasotae, E. these species, the perispores were all broadly and continuously didymoglossoides, E. haynaldii, E. herpestes, E. heteromor- folded, nonperforate, and (with the exception of E. prestonii; phum, E. horridulum, E. mildbraedii, E. omissum, E. organense, fig. 5L) nonspiny (figs. 12–14). E. phoras, E. proliferans, E. pseudoboryanum, E. pteropodum, From sect. Setosa, we examined 48 taxa, 23 of which (figs. E. rosettum, E. tabanense, E. villosum, E. wardiae, E. webbii, 6H–8F) were in the Rouhan et al. (2004) analysis (app. A). and E. zebrinum (app. A). Generally, either broad folds or cristae MORAN ET AL.—SPORES OF ELAPHOGLOSSUM 917 were present, but these were absent in E. oblanceolatum and broad perisporal folds without perforations and spines. All of E. papillosum (fig. 8B,8C) and in the clade E. setigerum–E. the character states of this species represent the plesiomor- aubertii (fig. 7C–7E). Of those species that had broad folds or phic condition (figs. 12–14). cristae, the cristate condition was more common (fig. 12). The Sect. Elaphoglossum. This section consists of two well- cristae were discontinuous in E. boryanum var. eutecnum, E. supported clades: subsects. Platyglossa and Pachyglossa (fig. cardenasii, and E. russelliae (fig. 6K,6L; fig. 7A). Spiny peri- 1). No known macromorphological or anatomical characters spores predominated throughout the sect. Setosa (fig. 13), as distinguish these two clades; they are supported solely by DNA did perforations (fig. 14). sequencing evidence (Rouhan et al. 2004). Their perispore Sect. Lepidoglossa. From this clade, 86 species were ex- characteristics, however, support their distinction. Three peri- amined (app. A), 41 of which (figs. 8G–11L) were included spore characters are synapomorphic for subsect. Pachyglossa in the analysis of Rouhan et al. (2004). The perispores typi- (figs. 4E–5G): the presence of cristae (fig. 12), spines (fig. 13), cally had broad, continuous folds (fig. 12). Spiny perispores and perforations (fig. 14). These characters show reversals within originated several times (fig. 13), and perforations were lack- the subsection, such as the nonspiny perispores in the clade ing in all species except E. forsythii-majoris and E. leucolepis formed by the Mauritian endemic E. sieberi, the Madagascan (figs. 9A,11G). The perispore of E. muscosum was distinctive endemic E. malgassicum, and E. macropodium from Africa in lacking folds and being beset with low spherical or hemi- and Indian Ocean areas (figs. 3D–3G, 13), and a reversal to spherical deposits (fig. 11J). Elaphoglossum muscosum had the nonperforate perispores in the Neotropical species E. doanense, largest spores (ca. 65 mm long) of any species examined. E. guatemalense, E. herminieri,andE. productum (fig. 14; figs. 4D,2H,2I,3L, respectively). Nevertheless, the characteristics Discussion of the perispore support the monophyly of subsect. Pachyglossa and provide the only morphological characters that distinguish This study is one of the largest done on perispore morphol- it from subsect. Platyglossa. ogy in a genus of leptosporangiate ferns (for lycophytes, see Sect. Squamipedia. The sister relationship of the Madagascan Korall and Taylor 2006). Given the size of this study, it was endemic E. marojejyense to this section, whose members are not possible to examine more than one or a few specimens otherwise entirely Neotropical, was an unexpected result of of each species (app. A). The question arises of whether this the Rouhan et al. (2004) molecular study (fig. 1). Elaphoglos- sample size is adequate to give general applicability to the re- sum marojejyense agrees morphologically with the Neotropi- sults. A comparison of our spore images to those published cal species in long-creeping rhizomes, lack of phyllopodia, and elsewhere (Mickel 1980, 1985; Mickel and Atehortu´ a 1980; small leaves; however, it lacks the distinctive peglike aero- Tryon and Tryon 1982; Tryon and Lugardon 1991) suggests phores on the rhizome near the petiole base, and its perispore that perispore morphology is generally consistent within spe- (fig. 5H) differs in being perforate (autapomorphic within the cies. Of 56 species compared, 52 were considered consistent. section; fig. 14) and lacking spines (plesiomorphic; fig. 13). In In the four instances that were not, we cannot be certain contrast, the perispores of the Neotropical species (fig. 5I–5K) whether the differences are caused by infraspecific variation are nonperforate (plesiomorphic; fig. 14) and spiny (synapo- or by misidentifications of the specimens. The overall corre- morphic; fig. 13). Thus, the perispore of E. marojejyense ap- spondence between our images and those previously published pears to be morphologically unlike those of the Neotropical suggests that in most cases our findings adequately represent species in sect. Squamipedia. the species in question. Subulata clade. The subulate-scaled clade contains two Sect. Amygdalifolia. Sister to all other Elaphoglossum is well-supported subclades: sects. Polytrichia and Setosa (fig. 1). sect. Amygdalifolia, which consists of a single species, E. Sect. Polytrichia and a clade within sect. Setosa (E. decoratum, amygdalifolium. Its cristate perispore (fig. 2A) represents an E. spatulatum,andE. piloselloides) have nonperforate peri- autapomorphy (fig. 12). The only species examined with sim- spores with broad folds (fig. 6H–6J), both of which are plesio- ilar spores (i.e., with nonperforate cristate perispores with morphic character states (fig. 14). Mickel and Atehortuá (1980) smooth surfaces between cristae) was E. lepidothrix, which divided sect. Polytrichia into subsect. Apoda, characterized by belongs to sect. Lepidoglossa (app. A; see image at http://www laminae sessile or nearly so, and subsect. Polytrichia, character- .plantsystematics.org). The similarity is most parsimoniously ex- ized by distinct petioles. No differences in spores distinguish the plained by an independent origin of the cristae. two subsections (our sampling of subsect. Apoda was exhaus- Elaphoglossum aemulum. This species is endemic to tive; i.e., it included all species in the subsection as defined by Hawaii (Palmer 2003). In our unpublished phylogenetic anal- Atehortuá [1983]: E. apodum [fig. 6E], E. auripilum [fig. 6D], yses based on DNA sequence data of two chloroplast markers E. backhousianum [fig. 6C], E. cubense, E. latum,andE. ray- (trnL-F, atpB-rbcL), it forms a clade with four other species waense; app. A). endemic to Hawaii: E. alatum, E. crassicaule, E. fauriei,andE. In sect. Setosa, the clade formed by E. boryanum var. parvisquameum. These four species, called the Alatum group eutecnum–E. siliquoides (fig. 1) was characterized by three syn- (Palmer 2003), have perforate and cristate perispores (app. A). apomorphies: the presence of cristae (fig. 12), spines (fig. 13), In contrast, E. aemulum has nonperforate, broadly folded peri- and perforations (fig. 14). Reversals, however, occurred in all spores. Its spores more closely resemble those of subsect. Platy- these character states. glossa. All of its character states represent the plesiomorphic A surprise result of the analyses by Rouhan et al. (2004) condition (figs. 12–14). and Skog et al. (2004) was that E. decoratum belonged to Elaphoglossum glaucum. The spores of this species (fig. the subulate-scaled clade (fig. 1). This was unexpected on the 2C) resemble those of subsect. Platyglossa; namely, they have 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 923 broad and flat, unlike typical subulate scales that are hairlike tually prove to be a synapomorphy for the entire subsection. and enrolled at the base. The species appears so distinct that Within the subsection, perisporal folds are present in some Mickel and Atehortu´ a (1980) placed it in its own section species (E. atropunctatum and E. yatesii) but absent in others (sect. Decorata). A reexamination of specimens, however, showed (E. oculatum, E. engelii, E. blandum, E. bellermannianum, that the scales, although appearing nonsubulate, were in fact and E. muscosum; app. A). The presence of large spores sug- enrolled at the base, thus agreeing with the molecular results. gests polyploidy, but this cannot be assessed because chromo- In those results, E. decoratum was sister to E. piloselloides some numbers are lacking for subsect. Muscosa. and E. spatulatum (bootstrap support 56%; fig. 1). Their Mickel and Atehortuá (1980) placed E. ovatum in sect. spores are similar to each other (fig. 6H–6J) and to those of Squamipedia because of its long-creeping rhizomes and small other subulate-scaled species, such as E. auripilum (fig. 6D) leaves (<12 cm long); however, it lacks the paired peglike and E. hybridum var. vulcanii (fig. 6F). In all instances, how- aerophores on the rhizome characteristic of sect. Squamipe- ever, the similarities represent the plesiomorphic condition dia, and it has phyllopodia and setose-margined lamina (figs. 12–14). scales typical of sect. Lepidoglossa. In the analysis of Rouhan Sect. Lepidoglossa. The perispores in this section (figs. et al. (2004), E. ovatum was resolved in sect. Lepidoglossa, 8G–11L) were generally characterized by the plesiomorphic and this agrees with its perispore characteristic (fig. 10C)of states of being broadly and continuously folded, nonperforate, being smooth, not spiny as in the Neotropical species of sect. and without spines (figs. 8G–8L, 9–11). The presence of Squamipedia (figs. 5H–5K, 13). spines appears to be a synapomorphy for smaller clades within Although the five perispore characters scored in this study the section (fig. 13). One such clade is informally referred to showed considerable homoplasy, synapomorphies were found as the ‘‘ciliatum group,’’ consisting of E. burchellii, E. cilia- that were helpful in characterizing clades within Elaphoglos- tum, and E. nigrescens (fig. 1; fig. 11B–11D, respectively). sum. The perispore characteristics have proved to be useful Also belonging to this clade on the basis of two chloroplast in phylogenetic analysis, and we plan to incorporate them in markers (R. C. Moran, J. G. Hanks, and G. Rouhan, unpub- a larger analysis including all morphological characters of lished data) are the Neotropical species E. huacsaro (fig. the genus. 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 Acknowledgments E. nervosum. All these species except E. dussii have spiny perispores. This work was supported by a grant to R. C. Moran from In sect. Lepidoglossa, the perispore was particularly dis- the U.S. National Science Foundation (DEB-00211969). It tinctive in E. muscosum (fig. 11J), a species belonging to sub- was also supported by the generosity of the Lewis B. and sect. Muscosa (Mickel and Atehortuá 1980). The ca. 15 Dorothy Cullman Foundation. We thank Dennis W. Stevenson species in this subsection have laminae densely scaly abaxially, for use of the SEM laboratory in Pfizer Laboratory at the lamina apices acute to obtuse, and elevational ranges above New York Botanical Garden and Lisa Campbell for assis- 2000 m (A. Vasco, personal communication). In the seven tance in the laboratory throughout this study. We also thank species examined from the group so far (E. atropunctatum, Michael Sundue, Alejandra Vasco, Larry Kelly, and Fabian E. bellermanianum, E. blandum, E. engelii, E. muscosum, E. Michelangeli for helpful discussions and two anonymous re- oculatum, and E. yatesii; app. A), the spores are large, fre- viewers for providing many constructive comments on the quently collapsed, and papillate. These characters may even- manuscript.

Appendix A

Voucher Table

Table A1 List of Elaphoglossum Species Whose Spores Were Imaged for This Study Species Infrageneric group Voucher Figure number E. achroalepis (Baker) C. Chr. Lepidoglossa Madagascar, van der Werff 12837 (P) 9C E. acrostichoides (Hook. & Grev.) Schelpe Platyglossa Tanzania, De Boer & Schippe 458 (NY) 5C E. acutifolium Rosenst. Platyglossa Bolivia, Kessler et al. 4317 (NY) ... E. aemulum (Kaulf.) Brack. New section Hawaii, Topping 2640b (NY) 2B E. affine (M. Martens & Galeotii) T. Moore Platyglossa Mexico, Camp 2682 (NY) 5D E. aff. latifolium (Sw.) J. Sm. of Rouhan et al. 2004 Platyglossa Guatemala, Molina 21310 (NY) 4G E. alatum (Feé) T. Moore Elaphoglossum Hawaii, collector unknown (NY) ... E. albescens (Sodiro) H. Christ Setosa Ecuador, Palacios & Rubio 5275 (MO) ... E. alfredii Rosenst. Setosa Costa Rica, Lloyd 4262 (NY) ... E. ambiguum (Mett. ex H. Christ) Alston Platyglossa Ecuador, Navarette & Asimbaya 1767 (NY) ... E. amygdalifolium (Mett. ex Kuhn) H. Christ Amygdalifolia Costa Rica, Grayum 2373 (NY) 2A Table A1 (Continued ) Species Infrageneric group Voucher Figure number E. andicola (Feé) T. Moore Pachyglossa Ecuador, Lojtnant et al. 12499 (AAU) ... E. angulatum (Blume) T. Moore Platyglossa La Reúnion, Rouhan 216 (NY) 4L E. angustius Mickel Lepidoglossa Bolivia, Steinbach 9152 (NY) ... E. antisanae (Sodiro) S. Chr. Platyglossa Ecuador, Jaramillo 9887 (AAU) ... E. apodum (Kaulf.) Schott ex J. Sm. Polytrichia Haiti, Ekman 4729 (NY) 6E E. asterolepis (Baker) C. Chr. Lepidoglossa Madagascar, Rouhan 305 (P) 10B E. atropunctatum Mickel Lepidoglossa Ecuador, Holm-Nielsen 1907 (AAU) ... E. aubertii (Desv.) T. Moore Setosa La Reúnion, Rouhan 226 (NY); Madagascar, 7E Rakotondrainibe 2497(P) E. auricomum (Kunze) T. Moore Lepidoglossa Guatemala, Johnson 932 (NY); Costa Rica, 11L Horich s.n. (NY) E. auripilum H. Christ Polytricha Costa Rica, Burger & Stolze 5335 (NY) 6D E. avaratraense Rakotondr. Lepidoglossa Madagascar, Rakotondrainibe 1456 (P) 9B E. backhousianum T. Moore Polytrichia Panama, Folsum 3576 (NY) 6C E. bakeri (Sodiro) H. Christ Setosa Costa Rica, Taylor 17696 (NY) ... E. ballivianii Rosenst. Platyglossa Bolivia, Nun˜ ez & Sundue 316 (NY) ... E. bellermannianum (Klotzsch) T. Moore Lepidoglossa Colombia, H. Smith 2640 (NY) ... E. bifurcatum (Jacq.) Mickel Lepidoglossa St. Helena, Cumming s.n. (NY) ... E. biolleyi H. Christ Pachyglossa Costa Rica, Davidse et al. 28794 (NY) 3K E. blandum Rosenst. Lepidoglossa Bolivia, Beck 3134 (NY) ... E. boragineum (Sodiro) H. Christ Setosa Panama, Croat 25299 (NY) ... E. boryanum (Feé) T. Moore var. boryanum Setosa Guadeloupe, Duss 4151 (NY) ... E. boryanum (Feé) T. Moore var. eutecnum Mickel Setosa Venezuela, Luteyn & Lebron-Luteyn 5176 (NY) 6K E. buchtienii Rosenst. Lepidoglossa Bolivia, Tate 838 (NY) ... E. burchellii (Baker) C. Chr. Lepidoglossa Ecuador, Holm-Nielsen et al. 35187 (NY); 11B Bolivia, Acebey & Tosto 176 (NY) E. cardenasii W. H. Wagner Setosa Bolivia, Kessler et al. 6564 (NY) 6L E. cardiophyllum (Hook.) T. Moore Platyglossa Ecuador, Holm-Nielsen 5915 (NY) 4K E. caricifolium Mickel Lepidoglossa Costa Rica, Rojas 2898 (NY) 10H E. casanense Rosenst. Lepidoglossa Bolivia, Steinbach 8964bis (NY) ... E. castaneum (Baker) Diels Setosa Peru, Leon 1961 (NY) ... E. cf. tovii E. Brown (E. sp. of Rouhan et al. 2004) Lepidoglossa Marquesas, Lorence 9011 (NY) 9J E. ciliatum (C. Presl) T. Moore Lepidoglossa Costa Rica, Ingram & Ingram-Ferral 1308 (NY); 11C Ecuador, Balslev & Madson 10402 (NY) E. cismense Mickel Pachyglossa Costa Rica, Rojas 2877 (NY) 3C E. cocosense Mickel Lepidoglossa Costa Rica, Rojas 3642 (NY) ... E. concinnum Mickel Squamipedia Peru, Young 1301 (NY) ... E. conforme (Sw.) Schott Platyglossa South Africa, Taylor 1456 (NY) ... E. conspersum H. Christ Platyglossa Costa Rica, Seidenschnur 268 (NY) 4J E. cordifolium Rosenst. Polytrichia Bolivia, Croat 51337 (NY) ... E. coriaceum Bonap. Platyglossa Madagascar, Geay 103 (P) 5G E. costaricense H. Christ Setosa Costa Rica, Seidenschnur 199 (NY) 8E E. coursii Tardieu Platyglossa Madagascar, Rouhan 427 (P) 5F E. crassicaule Copel. Elaphoglossum Hawaii, Annable & Reinard 3793 (NY) ... E. crassifolium (Gaud.) L. E. Anderson & Crosby Elaphoglossum Hawaii, Degener 9940 (NY) ... E. crinitum (L.) H. Christ Polytrichia Costa Rica, Angulo 605 (NY) 6A E. crispipaleaceum M. Kessler & Mickel Lepidoglossa Bolivia, Nun˜ ez 521 (NY) ... E. croatii Mickel Pachyglossa Costa Rica, Rojas 1535 (NY) 3B E. cubense (Mett. ex Kuhn) C. Chr. Polytrichia Cuba, Jack 8058 (NY) ... E. curvans (Kunze) A. Rojas Lepidoglossa Ecuador, Valencia 129 (NY) ... E. cuspidatum (Willd.) T. Moore Lepidoglossa Peru, Smith & Albań 5503 (NY) 8G E. davidsei Mickel Setosa Costa Rica, Montgomery & Huttleson 85135 (NY) 7B E. decaryanum Tardieu Pachyglossa Madagascar, Rasolohery 873 (P) 3I E. deckenii (Kuhn) C. Chr. Lepidoglossa Comoros, Rouhan & Rakontondrainibe 105 (NY) 11E E. decoratum Maxon Setosa Brazil, Wackett 284 (NY) 6H E. delasotae Mickel Setosa Bolivia, Kessler et al. 7123 (NY) ... E. deltoideum (Sodiro) H. Christ Squamipedia Ecuador, Harley & Andersson 18092 (NY) ... E. dendricola (Baker) H. Christ Lepidoglossa Ecuador, Jorgensen et al. 1827 (NY) ... E. dichroum Mickel Lepidoglossa Ecuador, Bogh 47862 (AAU) ... E. didymoglossoides C. Chr. Setosa Bolivia, Beck 3615 (NY) ...

924 Table A1 (Continued ) Species Infrageneric group Voucher Figure number E. dimorphum (Hook. & Grev.) T. Moore Lepidoglossa St. Helena, collector unknown (P) ... E. doanense L. D. Go´ mez Pachyglossa Panama, McPherson 12844 (NY) ... E. dussii Underw. & Maxon (E. aff. petiolatum of Rouhan et al. [2004]) Lepidoglossa Dominican Republic, Lloyd 351 (NY) 10L E. edwallii Rosenst. Lepidoglossa Brazil, Rose & Russell 20482 (NY) 10F E. engelii (H. Karst.) H. Christ Lepidoglossa Ecuador, van der Werff & Guidino 11005 (MO) ... E. erinaceum (Feé) T. Moore Polytrichia Peru, Gentry et al. 22977 (NY); Bolivia, 6B Solomon 16391 (MO) E. eximium (Mett.) H. Christ Setosa Costa Rica, Moran 2328 (NY) 7D E. fauriei Copel. Elaphoglossum Hawaii, Baldwin 100 (NY) ... E. feei (Bory ex Feé) T. Moore Squamipedia Guadeloupe, Questel 3267 (NY) ... E. flaccidum (Jenm.) Alston Pachyglossa Ecuador, Moran 6278 (NY) 4A E. forsythii-majoris H. Christ Lepidoglossa La Reúnion, Cadet 1535 (P) 9A E. fournierianum L. D. Go´ mez Setosa Costa Rica, Davidse 24999 (NY) 8A E. furfuraceum (Mett. ex Kuhn) H. Christ Lepidoglossa Costa Rica, Davidse 24757 (MO) 11I E. gayanum (Feé) T. Moore (E. minutum [Pohl ex Feé] T. Moore of Rouhan et al. 2004) Platyglossa Peru, Segastegui et al. 8238 (NY) 5E E. glabellum J. Sm. Platyglossa Panama, Stimson 5411 (NY) 5B E. glaucum T. Moore New section Mexico, Mickel 9675 (NY) 2C E. gloeorrhizum Mickel Pachyglossa Costa Rica, Rojas 3304 (NY) ... E. gramineum (Jenman) Urb. Lepidoglossa Dominican Republic, Valeur 634 (NY) ... E. grayumii Mickel Pachyglossa Costa Rica, Grayum 3017 (NY) 2E E. guamanianum (Sodiro) C. Chr. Lepidoglossa Peru, Barbour 2564 (MO) ... E. guatemalense (Klotzsch) T. Moore Pachyglossa Mexico, Martinez 14987 (NY) 2H E. guentheri Rosenst. Lepidoglossa Bolivia, Beck 479 (NY) ... E. hartwegii (Feé) T. Moore Lepidoglossa Venezuela, Ruiz Teran & Lopez Figueiras 2130 (NY) ... E. haynaldii (Sodiro) I. Losch Setosa Colombia, Killip 6853 (US); Grant 9846 (US) ... E. herminieri (Bory & Feé) T. Moore Pachyglossa Suriname, Inghamon & Inghamon 34007 (NY) 2I E. herpestes Mickel Setosa Ecuador, Madison 6866 (SEL) ... E. heterolepis (Feé) T. Moore Lepidoglossa Mauritius, Rouhan 179 (NY) 9F E. heteromorphum (Klotzsch) T. Moore Setosa Ecuador, Croat 58771 (MO) ... E. hieronymi (Sodiro) C. Chr. Pachyglossa Ecuador, Øllgaard & Balslev 10206 (AAU) ... E. hoffmannii (Mett. ex Kuhn) H. Christ Pachyglossa Costa Rica, Hennipman et al. 6986 (NY) 3J E. hornei C. Chr Pachyglossa Seychelles, Rouhan 137 (NY) 4C E. horridulum (Kaulf.) J. Sm. Setosa Brazil, Edwall & Toledo 21187 (NY) ... E. huacsaro (Ruı´z) H. Chirst Lepidoglossa Venezuela, Ruiz-Terań 6124 (NY) 11A E. humbertii C. Chr. Lepidoglossa Madagascar, Humbert 6900 (P) 9E E. hybridum (Bory) Brack. Polytrichia La Reúnion, Rouhan 217 (NY) 6G E. hybridum (Bory) Brack. var. vulcanii Polytrichia La Reúnion, Rouhan 222 (NY) 6F E. inquisitivum M. Kessler & Mickel Lepidoglossa Bolivia, Kessler et al. 11812 (NY) ... E. killipii Mickel Lepidoglossa Bolivia, Buchtien 4239 (NY) 10K E. laminarioides (Feé) T. Moore Lepidoglossa Ecuador, Palacios 4312 (MO) ... E. lanatum (Bojer ex Baker) Lorence Lepidoglossa Mauritius, Pike s.n. (NY) 9H E. lancifolium (Desv.) C. V. Morton Lepidoglossa La Reúnion, Rouhan 201 (NY); 10I Grande Comore, Rouhan 130 (P) E. lasioglottis Mickel Lepidoglossa Peru, Segastegui 3250 (NY) ... E. lastii (Baker) C. Chr. Platyglossa Tanzania, DeBoer & Schippe 460 (K) ... E. latifolium (Sw.) J. Sm. Pachyglossa Jamaica, Underwood 3146 (NY) 2D E. latum (Mickel) Atehortu´ a ex Mickel Polytrichia Costa Rica, Lloyd 4162 (NY) ... E. lechlerianum (Mett.) T. Moore Lepidoglossa Bolivia, Kessler et al. 7061 (NY) ... E. lepervanchei (Bory ex Feé) T. Moore (E. aff. conforme [Sw.] J. Sm. of Rouhan et al. 2004) Pachyglossa Mauritius, Rouhan et al. 175 (NY) 2K E. leporinum L. D. Go´ mez Lepidoglossa Costa Rica, Rojas et al. 2836 (NY) ... E. leucolepis (Baker) Krajina ex Tardieu Lepidoglossa Madagascar, McPherson 1492 (P) 11G E. lindbergii (Mett. ex Kuhn) Rosenst. Polytrichia Argentina, Venturi 3256 (US) ... E. lingua (C. Presl) Brack. Platyglossa Costa Rica, Mickel 2688 (NY) 4H E. litanum (Sodiro) C. Chr. Lepidoglossa Ecuador, Tipaz et al. 2179 (M) ... E. lloense (Hook.) T. Moore Squamipedia Bolivia, Nun˜ ez 51 (NY) ... E. luridum (Feé) H. Christ Pachyglossa Guyana, Clarke & McPherson 1949 (NY) 4B E. luteynii Mickel Lepidoglossa Venezuela, Luteyn et al. 6135 (NY) ...

925 Table A1 (Continued ) Species Infrageneric group Voucher Figure number E. macropodium (Feé) T. Moore Pachyglossa La Reúnion, Rouhan & Grangaud 207 (NY); 3G Tanzania, DeBoer & Schippe 457 (K) E. malgassicum C. Chr. Elaphoglossum Madagascar, Rouhan 427 (P) 3E E. marojejyense Tardieu Squamipedia Madagascar, Humbert 23105 (P) 5H E. marquisearum Bonap. Pachyglossa Marquesas, Wood 6390 (P) 2G E. mathewsii (Feé) T. Moore Lepidoglossa Ecuador, Balslev & Mino 69231 (NY) ... E. metallicum Mickel Pachyglossa Peru, Sagastegui et al. 14459 (NY) 3H E. micropogon Mickel Lepidoglossa Costa Rica, Lloyd 4111 (NY) 9K E. minutissimum R. C. Moran & Mickel (E. sp. nov. cf. hayesii [Mett. ex Kuhn] Maxon of Rouhan et al. 2004) Setosa Costa Rica, Moran 6344 (NY) 8D E. mildbraedii Hieron. Setosa Tanzania, Rouhan 534 (P) ... E. mitorrhizum Mickel Platyglossa Ecuador, Aulestia & Aulestia 1526 (MO) 4F E. molle (Sodiro) C. Chr. Lepidoglossa Ecuador, Mille s.n. (P) ... E. moorei (E. Britton) H. Christ Squamipedia Colombia, van der Werff & Giraldo 9726 (MO) ... E. muscosum (Sw.) T. Moore Lepidoglossa Venezuela, Ruiz-Terań 1864 (NY) 11J E. nigrescens (Hook.) T. Moore ex Diels (including E. palmense H. Christ and E. wackettii Rosenst. of Rouhan et al. 2004) Lepidoglossa Brazil, Maas et al. 6858 (NY); Costa Rica, 11D Ingram & Ferrell-Ingram 1925 (NY) E. oblanceolatum C. Chr. Setosa Panama, Porter et al. 4383 (NY) 8C E. oculatum Mickel Lepidoglossa Ecuador, Holm-Nielsen 6246 (AAU) ... E. odontolepis Mickel Pachyglossa Ecuador, Navarrete 387 (QCA) ... E. omissum Mickel Setosa Ecuador, Øllgaard et al. 945 (AAU) ... E. orbignyanum (Feé) T. Moore Lepidoglossa Peru, Killip & Smith 24413 (NY) 11H E. organense Brade Setosa Brazil, Brade 9647 (NY) ... E. ornatum (Mett. ex Kuhn) H. Christ Polytrichia Brazil, Jurgens 75 (NY) ... E. ovalilimbatum Bonap. Platyglossa Madagascar, Perrier 7860c (P) 4E E. ovatum (Hook. and Grev.) T. Moore Lepidoglossa Ecuador, Ulloa et al. 1064 (NY) 10C E. pachyphyllum (Kunze) C. Chr. Pachyglossa Bolivia, Beck 23090 (NY) ... Madeira, collector unknown (NY); Mexico, 9I E. paleaceum (Hook. and Grev.) Sledge Lepidoglossa Lorea 3144 (NY) E. pallidum (Baker ex Jenman) C. Chr. Polytrichia Mexico, Breedlove 32046 (NY) ... E. papillosum (Baker) H. Christ Setosa Ecuador, Øllgaard 38438a (AAU) 8B E. parvisquameum Skottsb. Elaphoglossum Hawaii, Degener 31707 (NY) ... E. pellucidum (Sodiro) C. Chr. Lepidoglossa Hawaii, Degener 31704 (NY); Heller 2621 (NY) ... E. peltatum (Sw.) Urb. forma standleyi Squamipedia Costa Rica, Lent 3699 (AAU) ... E. peltatum (Sw.) Urb. forma peltatum Squamipedia Mexico, Mendez 7931 (NY) 5I E. perrierianum C. Chr. Lepidoglossa Madagascar, Perrier 16457 (P) ... E. petiolatum (Sw.) Urb. Lepidoglossa Mexico, Carranza & Pe´rez 5140 (NY), Reveal & Atwood 8K 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) E. phanerophlebium C. Chr. Setosa Madagascar, Decary 18115 (P) 7K E. phoras Mickel Setosa Costa Rica, Lloyd 4177 (NY); Colombia, Rodrı´gues ... et al. 4124 (NY) E. piloselloides (C. Presl) T. Moore Setosa Ecuador, Fay & Fay 3519 (NY) 6I E. pilosius Mickel Setosa Mexico, Breedlove & Smith 22060 (NY) 7I E. plumosum (Feé) T. Moore Lepidoglossa Brazil, Thomas et al. 9371 (NY) ... E. poolii (Baker) H. Christ Lepidoglossa Madagascar, Perrier de la Baˆthie 12487 (P) 11F E. potosianum H. Christ Lepidoglossa Mexico, Parry & Palmer 1007 (NY) ... E. prestonii J. Sm. Polytrichia Brazil, Brade 15514 (NY) 5L E. pringlei (Davenp.) C. Chr. Lepidoglossa Mexico, Mickel 4463 (NY) 8J E. productum Rosenst. Pachyglossa Ecuador, Øllgaard & Navarrete 105872 (AAU) 2J E. proliferans Maxon & C. V. Morton Setosa Panama, Herrera 790 (NY) ... E. proximum (J. Bommer) H. Christ Platyglossa Costa Rica, Maxon 464 (NY) 4I E. pseudoboryanum Mickel Setosa Ecuador, Fay & Fay 3720 (NY) ... E. pteropodum (Sodiro) C. Chr. Setosa Ecuador, Øllgaard et al. 37834 (AAU) ... E. pteropus C. Chr. Pachyglossa Brazil, Steward & Ramos P19692 (NY) 3L E. pumilio Mickel Lepidoglossa Ecuador, Luteyn & Cotton 11281 (NY) ...

926 Table A1 (Continued ) Species Infrageneric group Voucher Figure number E. pygmaeum (Mett. ex Kuhn) H. Christ Setosa Ecuador, Moran & Rohrbach 5221 (MO) 7J E. randii Alston & Schelpe Lepidoglossa Marion Island, Huntley 464 (P) 10E E. rapense Copel. Setosa French Polynesia, Motley 2677 (NY) 7G E. raywaense (Jenman) Alston Polytrichia Ecuador, Palacios 394 (MO) ... E. 3 revaughanii Lorence Lepidoglossa Mauritius, Rouhan et al. 190 (NY) ... E. richardii (Bory ex Feé) H. Christ Lepidoglossa La Reúnion, Rouhan 231 (P) 8I E. rimbachii (Sodiro) H. Christ (E. nigrocostatum Mickel of Rouhan et al. 2004) Lepidoglossa Ecuador, Øllgaard & Balslev 8643 (AAU) 8L E. rosenstockii H. Christ ex Rosenst. Lepidoglossa Ecuador, Øllgaard et al. 37940 (AAU) ... E. rosettum R. C. Moran & Mickel Setosa Bolivia, Tate 618 (US) ... E. rufidulum (Willd. ex Kuhn) C. Chr. Lepidoglossa La Reúnion, Cadet 1535 (P) 9L E. russelliae Mickel Setosa Costa Rica, Go´ mez et al. 22409 (NY) 7A E. rzedowskii Mickel Lepidoglossa Mexico, Rzedowski 26032 (NY) ... E. samoense Brack. Setosa French Polynesia, Motley 2875 (NY) 7F E. sartorii (Liebm.) Mickel Pachyglossa Mexico, Munn-Estrada & Mendoza 1683 (NY) 3A E. scolopendriforme Tardieu Lepidoglosssa Madagascar, Rouhan 306 (P) 9D E. setigerum (Sodiro) Diels Setosa Ecuador, Holm-Nielsen & Jeppesen1293 (AAU) 7C E. sieberi (Hook. and Grev.) T. Moore Pachyglossa Mauritius, Rouhan et al. 176 (NY) 3F E. siliquoides (Jenm.) C. Chr. Setosa Costa Rica, Lloyd 4282 (NY) 8F E. smithii (Baker) H. Christ Setosa Ecuador, Øllgaard et al. 1014 (AAU) 7L E. spatulatum (Bory) T. Moore Setosa La Reúnion, Rouhan & Grangaud 246 (NY) 6J E. splendens (Bory ex Willd.) Brack. Lepidoglossa La Reúnion, Rouhan 213 (NY) 10A E. squamatum (Sw.) T. Moore (E. lindenii [Bory ex Feé] T. Moore of Rouhan et al. 2004) Setosa Ecuador, Øllgaard & Balslev 8410 (NY); Colombia, 7H Killip & Smith 18607 (NY) E. squamipes (Hook.) T. Moore (including E. craspedariiforme [Feé] Brade ex Alston of Rouhan et al. 2004) Squamipedia Venezuela, Rivero & Diaz 1496 (NY) 5J E. stenophyllum (Sodiro) Diels Lepidoglossa Ecuador, Grijalva 568b (NY) ... E. stipitatum (Bory ex Feé) T. Moore Lepidoglossa Madagascar, Pichi Sermolli 5120 (P); La Reúnion, 10D Rouhan 212 (NY) E. subcinnamomeum (H. Christ) Hieron. Lepidoglossa Kenya, Schelpe 2605 (P) ... E. subsessile (Baker) C. Chr. Pachyglossa Madagascar, Rouhan 432 (P) 2L E. succisaefolium (Thouars) T. Moore Lepidoglossa Amsterdam Island, Martel-Thoumian 2A (P) 8H E. tabanense Andre´ ex H. Christ Setosa Ecuador, Øllgaard & Navarette 2835 (AAU) ... E. tectum (Humb. & Bonpl. ex Willd.) T. Moore Lepidoglossa Colombia, Zarucchi et al. 5800 (NY) 10J E. tenuiculum (Feé) T. Moore ex C. Chr. Lepidoglossa Peru, van der Werff et al. 15697 (NY); Ecuador, Pitman & Romero 353 (NY) ... E. terrestre A. Rojas (E. sp. nov. cf. terrestre of Rouhan et al. 2004) Pachyglossa Costa Rica, Moraga & Rojas 509 (NY) 2F E. tomentosum (Bory ex Willd.) H. Christ Lepidoglossa Mauritius, Rouhan 178 (NY) 9G E. tonduzii H. Christ Lepidoglossa Costa Rica, Herrera 3511 (NY) ... E. tripartitum (Hook. & Grev.) Mickel Squamipedia Ecuador, Øllgaard 99199 (AAU) 5K E. trivittatum (Sodiro) H. Christ Lepidoglossa Ecuador, Øllgaard & Balslev 8272 (AAU) ... E. truncatum Rosenst. Polytrichia Bolivia, Williams 1031 (NY) ... E. unduaviense Rosenst. Pachyglossa Bolivia, Tate 345 (NY) ... E. vestitum (Schltdl. & Cham.) T. Moore Lepidoglossa Mexico, Mickel & Leonard 4705 (NY) 11K E. vieillardii (Mett.) T. Moore Pachyglossa New Caledonia, Munzinger 1276 (P) 3D E. villosum (Sw.) J. Sm. Setosa Jamaica, Yuncker 17696 (NY) ... E. wardiae Mickel Setosa Peru, Foster et al. 10890 (NY) ... E. wawrae (Luerss.) C. Chr. Platyglossa Hawaii, Forbes 350 (NY) 5A E. webbii T. Moore Setosa Colombia, Lehmann BT760 (NY) ... E. welwitschii (Baker) C. Chr. Lepidoglossa Tanzania, Taylor 9099 (P) 10G E. wrightii (Mett.) T. Moore Squamipedia Cuba, Ekman 3882 (NY) ... E. yatesii (Sodiro) H. Christ Lepidoglossa Ecuador, Øllgaard & Balslev 8411 (AAU) ... E. yungense de la Sota Lepidoglossa Bolivia, Kessler et al. 6444 (NY) ... E. yunnanense (Baker) C. Chr. Platyglossa China, Henry 13110 (NY) ... E. zebrinum Mickel Setosa Peru, Foster 10891 (NY) ...

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 Atehortu´a (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.

927 928 INTERNATIONAL JOURNAL OF PLANT SCIENCES

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) Characters Characters Characters Species 12345 Species 12345 Species 12345 Outgroups: E. eximium 1--01 E. papillosum 1--01 Athyrium filix-femina 00011 E. flaccidum 01000 E. peltatum 00001 Bolbitis auriculata 00001 E. forsythii-majoris 00010 E. petiolatum 00011 Lomariopsis marginata 1--01 E. fournierianum 01*00 E. phanerophlebium 01000 Rumohra adiantiformis 00011 E. furfuraceum 00011 E. pillosellioides 00011 Elaphoglossum: E. gayanum 00011 E. pilosius 00000 E. achroalepis 01011 E. glabellum 00011 E. poolii 00011 E. acrostichoides 00*11 E. glaucum 00011 E. prestonii 00001 E. aemulum 00011 E. grayumii 01000 E. pringlei 00011 E. affine 00011 E. guatamalense 00011 E. productum 00011 E. amygdalifolium 01011 E. herminieri 00111 E. proximum 00011 E. angulatum 00011 E. heterolepis 00001 E. pteropus 01000 E. apodum 00011 E. hoffmannii 01000 E. pygmaeum 1--01 E. asterolepis 00011 E. hornei 00*10 E. randii 00001 E. aubertii 1--01 E. huacsaro 00001 E. rapense 01010 E. auricomum 00011 E. humbertii 00001 E. richardii 00011 E. auripilum 00011 E. hybridum 00011 E. rimbachii 00011 E. avaratraense 00011 E. hybridum var. vulcanii 00011 E. rufidulum 00011 E. backhousianum 00011 E. killipii 00011 E. russelliae 01100 E. biolleyi 01000 E. lanatum 00001 E. samoense 01000 E. boryanum var. eutecnum 01100 E. lancifolium 00011 E. sartorii 01000 E. burchellii 00001 E. aff. latifolium 00011 E. scolopendriforme 00001 E. cardenasii 01100 E. latifolium 01000 E. setigerum 1--01 E. cardiophyllum 00011 E. lepervanchei 01000 E. sieberi 00011 E. caricifolium 00011 E. leucolepis 00010 E. siliquoides 00010 E. ciliatum 00001 E. lingua 00011 E. smithii 01000 E. cismense 01000 E. luridum 01100 E. spatulatum 00011 E. conspersum 00011 E. macropodium 00011 E. splendens 00011 E. coriaceum 00011 E. malgassicum 00011 E. squamatum 01000 E. costaricense 01000 E. marojejyense 00010 E. squamipes 00001 E. coursii 00011 E. marquisearum 01*00 E. stipitatum 00001 E. crinitum 00011 E. metallicum 01000 E. subsessile 01000 E. croatii 01100 E. micropogon 00011 E. succisaefolium 00011 E. cuspidatum 00111 E. minutissimum 00011 E. tectum 00011 E. davidsei 01000 E. mitorrhizum 00011 E. terrestre 01000 E. decaryanum 01010 E. muscosum 1--11 E. tomentosum 00011 E. deckenii 00011 E. nigrescens 00001 E. tovii 00011 E. decoratum 00011 E. oblanceolatum 1--01 E. tripartitum 00001 E. doanense 00001 E. orbignyanum 00011 E. vestitum 00011 E. dussii 00011 E. ovalilimbatum 00011 E. vieillardii 00001 E. edwallii 00011 E. ovatum 00011 E. wawrae 00111 E. erinaceum 00011 E. paleaceum 00011 E. welwitschii 00011 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 aster- isk indicates a polymorphism.

Literature Cited

Atehortu´ a L 1983 Taxonomic study of the Elaphoglossum apodum Fe´e ALA 1845 Histoire des acrostiche´es me´moires fougeres. Vol 2. complex (Elaphoglossaceae). PhD diss. City University of New York. Berger-Levrault, Strasbourg. Christ KHH 1899 Monographie des Genus Elaphoglossum. Neue Holttum RE 1978 Lomariopsis group. Flora Malesiana Ser II Denkschr Allg Schweiz Ges Gesammten Naturwiss 36:1–159. Pteridophyta 1:255–330. MORAN ET AL.—SPORES OF ELAPHOGLOSSUM 929

Korall P, WA Taylor 2006. Megaspore morphology in the Selaginella- Palmer DD 2003 Hawai’i’s ferns and fern allies. University of ceae in a phylogenetic context: a study of the megaspore surface and Hawai’i Press, Honolulu. wall structure using scanning electron microscopy. Grana 45:22–60. Rodrı´guez R 1995 Pteridophyta. Pages 119–351 in C Marticorena, R Kramer KU 1990 Lomariopsidaceae. Pages 164–170 in KU Kramer, Rodrı´guez, eds. Flora de Chile. Universidad de Concepcio´ n. PS Green, eds. The families and genera of vascular plants. Vol 1. Rouhan G, J-Y Dubuisson, F Rakotondrainibe, TJ Motley, JT Mickel, Pteridophytes and gymnosperms. Springer, New York. J-N Labat, RC Moran 2004 Molecular phylogeny of the fern Lorence DH, G Rouhan 2004 A revision of the Mascarene species genus Elaphoglossum (Elaphoglossaceae) based on chloroplast non- of Elaphoglossum (Elaphoglossaceae). Ann Mo Bot Gard 91:536– coding DNA sequences: contributions of species from the Indian 565. Ocean area. Mol Phylogenet Evol 33:745–763. Maddison DR, WP Maddison 2000 MacClade 4. Sinauer, Sunder- Skog JE, JT Mickel, RC Moran, M Volovsek, EA Zimmer 2004 land, MA. Molecular studies of representatives species in the fern genus Mickel JT 1980 Relationships of the dissected elaphoglossoid ferns. Elaphoglossum (Dryopteridaceae) based on cpDNA sequences Brittonia 32:109–117. rbcL, trnL-F, and rps4-trnS. Int J Plant Sci 165:1063–1075. ——— 1985 The proliferous species of Elaphoglossum (Elaphoglos- Smith AR, KM Pryer, E Schuettpelz, P Korall, H Schneider, PG saceae) and their relatives. Brittonia 37:261–278. Wolf 2006 A classiﬁcation for extant ferns. Taxon 55:705–731. Mickel JT, L Atehortu´ a 1980 Subdivision of the genus Elaphoglos- Tryon AF, B Lugardon 1991 Spores of the Pteridophyta: surface, wall sum. Am Fern J 70:47–68. structure,anddiversitybasedonelectronmicroscopestudies.Springer, Moran RC, JT Mickel 2004 Three new Neotropical species of New York. Elaphoglossum (Elaphoglossaceae) with subulate scales. Brittonia Tryon RM, AF Tryon 1982 Ferns and allied plants with special 56:200–204. reference to tropical America. Springer, Berlin.