Cryptogamie, Bryologie, 2017, 38 (3): 253-263 © 2017 Adac. Tous droits réservés Lepidolejeunea grandiocellata sp. nov. (Lejeuneaceae, Porellales), a new leafy liverwort from the West Indies based on morphological and molecular evidence Alfons SCHÄFER-VERWIMP a*, Julia BECHTELER b, Huub VAN MELICK c, Matthew A. M. RENNER d & Jochen HEINRICHS b aMittlere bSystematic Letten 11, 88634 Herdwangen-Schönach, Germany Botany and Mycology, GeoBio-Center, Ludwig Maximilian University, Menzinger Str. 67, 80638 Munich, Germany cMerellaan dRoyal 13, 5552 BZ Valkenswaard, The Netherlands Botanic Gardens and Domain Trust, Mrs Macquaries Road, Sydney, NSW 2000, Australia Abstract – Lepidolejeunea grandiocellata, sp. nov. is described for two specimens from the Blue Mountains of Jamaica. This delicate species is characterized by ocelli in basal parts of leaves, underleaves, bracteoles and bracts often being conspicuously larger than the surrounding cells, whereas ocelli in central and upper parts of these structures are more or less the same size as surrounding cells. Further defining characters include sharply 5-keeled perianths without inflated horns, and orbicular underleaves with a sharp, v-shaped notch. In a molecular phylogeny based on the nrITS region and the cp DNA markers rbcL and trnL-F, L. grandiocellata is placed sister to a clade including accessions of L. delessertii from Réunion and accessions of L. cuspidata, L. eluta, and L. sullivantii from the Neotropics [L. subg. Perilejeunea]. Based on the phylogeny and the characters of the ocelli, L. grandiocellata is placed in a new subgenus Caribeolejeunea. Bryophyte / classification / integrative taxonomy / Jamaica / Jungermanniopsida / ocelli INTRODUCTION Recent taxonomic research on liverworts provided evidence that documentation of extant species is still incomplete. On one hand, integrative approaches have identified numerous morphologically cryptic or near cryptic species that were not accepted based solely on morphology (Bakalin & Vilnet, 2014; Buczkowska et al., 2015; Renner et al., 2017). On the other hand, morphologically well-circumscribed species are still being discovered through revision of herbarium material or fieldwork (Potemkin et al., 2015; Bastos et al., 2016; Szabó & Pócs, 2016). * Corresponding author: moos.alfons@kabelbw.de doi/10.7872/cryb/v38.iss3.2017.253 254 A. Schäfer-Verwimp et al. In 2013, ASV and HvM collected bryophytes in Jamaica and have so far identified some 200 liverwort species (Schäfer-Verwimp & van Melick, 2016). However, among their collections were several species that could not yet be identified or were new to science, including distinctive plants belonging to the pantropical genus Lepidolejeunea R.M.Schust. Lepidolejeunea is well characterized by gynoecial innovations with a pycnolejeuneoid leaf arrangement, pyriform perianths with 5 smooth to crenulate keels, presence of scattered ocelli in leaves, underleaves, bracts, bracteoles, and perianths, lack of well-developed oil bodies in cells other than ocelli, and a marginal hyaline papilla at the proximal base of the first tooth of the leaf lobule. Lepidolejeunea was monographed by Piippo (1986) who accepted 12 species based on morphological evidence. A recent integrative study (Heinrichs et al., 2015) led to the recognition of 14 species and corroborated the importance of ocelli characters for the classification of Lepidolejeunea species. Ocelli – modified cells containing only a single large oil body – are of prime importance for the classification of Lejeuneaceae yet are difficult to study in older herbarium specimens because the oil bodies usually have disappeared (He & Piippo, 1999; Dong et al., 2013). Fortunately, the Lepidolejeunea specimens were investigated only a few weeks after collecting, and the ocelli were still in nearly perfect condition. Here we describe this plant as Lepidolejeunea grandiocellata sp. nov. based on morphological evidence and the outcome of a molecular phylogenetic study. MATERIALS AND METHODS Herbarium material of Lepidolejeunea grandiocellata sp. nov. was investigated using an Olympus CH2 microscope and its morphology was documented using a drawing tube (Figs 1-17). Morphological distinctness was confirmed by comparing L. grandiocellata to Lepidolejeunea specimens from the herbarium Schäfer-Verwimp and using the Lepidolejeunea treatments of Piippo (1986) and Heinrichs et al. (2015). Gametophytical tissue was isolated from the holotype of Lepidolejeunea grandiocellata. Total genomic DNA was extracted, and PCR and sequencing reactions were performed as described in Bechteler et al. (2016a). The chloroplast rbcL gene and trnL-trnF region, as well as the nuclear ribosomal ITS region (ITS15.8S-ITS2) were amplified, and the newly obtained sequences were compared with GenBank sequences using the nucleotide BLAST search (https://blast.ncbi.nlm.nih. gov/Blast.cgi?PAGE_TYPE=BlastSearch; Altschul et al., 1990). In a second step the sequences were aligned with published sequences downloaded from GenBank (http://www.ncbi.nlm.nih.gov/genbank/). Representatives of the genera Metalejeunea Grolle, Otolejeunea Grolle & Tixier, Rectolejeunea A.Evans and Vitalianthus R.M.Schust. & Giancotti were chosen as outgroup based on the phylogenetic hypotheses of Bechteler et al. (2016a, b) (Appendix 1). Sequences were aligned manually in PhyDE v.0.9971 (http://www.phyde.de/index.html) and ambiguous positions were excluded for phylogenetic analyses. Phylogenetic analyses The best fit models of evolution determined under the Akaike Information Criterion (AIC; Akaike, 1973) in jModelTest2 (Darriba et al., 2012) were TIM3+I+G Lepidolejeunea grandiocellata sp. nov. (Lejeuneaceae, Porellales) 255 for rbcL, TPM1uf+I for trnL-trnF, and TrN+I+G for ITS. RAxML 8.2.4 (Stamatakis, 2014) was used for maximum likelihood (ML) inference. Clades with bootstrap percentage values (BP) of 70-94 % were regarded as moderately supported and those with BP ≥ 95 % as strongly supported (Erixon et al., 2003). Since no incongruence among the single marker datasets was evident, they were concatenated. For the combined analysis, the dataset was partitioned by marker and a GTR+G model was used, following the recommendation given by Stamatakis (2016). Multiparametric bootstrapping using the autoMRE function (Pattengale et al., 2010) and ten thorough ML searches were executed. MrBayes 3.2.6 (Ronquist & Huelsenbeck, 2003) was used for Bayesian inference (BI) of the partitioned dataset. The GTR+I+G model as suggested by jModelTest and two Metropolis-coupled Markov Chain Monte Carlo (MCMC) analyses, including three heated chains and one cold chain running for 10 million generations, sampling every 1,000 generations were employed. An average standard deviation of split frequencies below 0.01 indicated a sufficiently long run and TRACER 1.6 (http://tree.bio.ed.ac.uk/software/tracer/) was used to check for convergence and stationarity. Burn-in was set to the first 25 % of sampled trees and the remainder was summarized with TreeAnnotator 1.8.2 (Drummond et al., 2012). FigTree 1.4.2 (http://tree.bio.ed.ac.uk/software/figtree/) was used to visualize the resulting maximum clade credibility (MCC) tree. Bayesian Posterior Probability (PP) values ≥ 0.95 were regarded as significant (Larget & Simon, 1999). RESULTS Taxonomy Lepidolejeunea subg. Caribeolejeunea Schäf.-Verw., Bechteler, van Melick, M.A.M.Renner & Heinrichs subg. nov. Type – Lepidolejeunea grandiocellata Schäf.-Verw., Bechteler, van Melick, M.A.M.Renner & Heinrichs This subgenus includes Lepidolejeunea species with ventral merophytes two cells wide, a short underleaf insertion comprising two inferior cells and two enlarged basal cells, and ocelli in basal parts of leaves, underleaves, bracteoles and bracts being often conspicuously larger than the surrounding cells whereas the ocelli in central and upper parts of these structures are more or less the same size as surrounding cells. Lepidolejeunea grandiocellata Schäf.-Verw., Bechteler, van Melick, M.A.M.Renner & Heinrichs, sp. nov. Figs 1-17 Type – JAMAICA: St. Andrew, Blue Mountains, military road from Newcastle to Catherines Peak, secondary forest near summit, on bark of young hardwood tree, 1525 m, WGS84: 18°04,7’ N, 76°42,2’ W, 4. Dez. 2013, leg. A. Schäfer-Verwimp 35071 & H. van Melick (Holotype: JE, isotypes: M, FR). Paratype collection from the same site, leg. A. Schäfer-Verwimp 35058/A (JE). Plants delicate, shoots up to 8 mm long and 0.9-1.1 mm wide, three year old herbarium material light green, greyish green or greyish, sparsely to irregularly pinnately branched, branches 1-1.5(-2) mm long, slightly to strongly adhering to substrate. Stem (60-)65-75(-80) µm in diameter, in cross section with 7 cortical cell 256 A. Schäfer-Verwimp et al. Figs 1-17. Lepidolejeunea grandiocellata. 1. Upper portion of shoot with perianth, ventral view. 2. Cross section of stem. 3-5. Leaf lobules. 6-7. Underleaves. 8. Leaf cells above leaf base with two ocelli. 9. Female bracteole. 10. Female bract. 11. Upper part of female bracteole. 12. Upper part of female bract lobe. 13. Perianth. 14. Upper part of perianth keel. 15-16. Two leaf lobes with lobules. 17. Part of dorsal leaf lobe (all from holotype). Scales: 400 µm for 1, 9-10, 13, 17; 100 µm for 2-7, 11-12, 14, 17; 60 µm for 8; 270 µm for 15-16. Lepidolejeunea grandiocellata sp. nov. (Lejeuneaceae, Porellales) 257 rows and 9-12 medullary cells, cortical cells up to 22 × 30 µm, medullary cells up to 7 × 14 µm; stem cells on ventral side of stem in top view subquadrate to rectangular to irregularly polygonal, rarely wider than long, 32-36 × 40-65 µm. Leaves imbricate, spreading widely to slightly obliquely upward, insertion nearly straight, the lobe (480-)490-520(-550) µm long, 400-420(-440) µm wide, asymmetrically broadly ovate (Figs 15-16), slightly apiculate, slightly incurved at apex, dorsal margin strongly arched, ventral margin nearly straight with keel of lobule slightly convex and junction of lobule and free margin of lobe concave, margin irregularly crenulatedenticulate (Fig. 17). Leaf lobe cells (sub-)quadrate to irregularly polygonal in outline, irregular in size, walls thin, trigones small (Fig. 8), intermediate thickenings absent, cuticle smooth. Apical marginal leaf cells 16-20 × 18-24 µm, central cells from 20 × 24 µm up to 26 × 30 µm, basal cells from 20 × 24 µm and 16 × 30 µm up to 40 × 45 µm; oil bodies seen only in ocelli. Ocelli present in all examined parts of gametophyte except stem, ca. 15-25(-30) per leaf lobe, scattered, in distal part of leaf lobe more or less of the same size as surrounding cells, in proximal half of leaf lobe often conspicuously larger than surrounding cells, the largest ones up to 40 × 45 µm, consisting of numerous small oil droplets of different size and filling almost the whole cell lumen, chloroplasts absent. Leaf lobules ovate, reaching 0.33-0.4 of length of lobes, basal portion covered by underleaves (Fig. 1), inflated, oriented at an angle of 70°-90° to stem, attached to stem by 3-(4)-5 elongated cells; lobule cells ± isodiametric to subrectangular to polygonal, similar in size and shape as in leaf lobes; 3-6(-7) scattered ocelli present per lobule, those along and near lobule insertion line conspicuously larger (up to 40 × 40 µm) than surrounding cells, others ± of the same size as surrounding cells (Figs 3-5); free lobule margin entire, 6-8 cells long, slightly incurved, apex semicircular, consisting of (4-)5-6 cells (incl. apical tooth); keel slightly to moderately arched, apical tooth falcate, 10-12 × (26-)3035 µm, hyaline papilla conspicuously smaller than apical tooth, proximal between first lobule tooth and first cell of free margin, ca. 7 × 15 µm; stylus not seen. Ventral merophytes two cells wide; underleaves (Figs 1, 6-7) contiguous to imbricate, 3-4.5 × stem width, rounded, often slightly wider than long, (200-)220280 µm wide, 200-250(-280) µm long (subgynoecial underleaves somewhat larger), bases cuneate, lobed to 0.5 of their length, lobes acute, sinus v-shaped, margins entire or slightly and irregularly crenulate-denticulate, line of insertion straight, attached to stem by two inferior central cells and two enlarged elliptical cells (Figs 6-7), underleaf cells mostly isodiametric to short rectangular, variable in size, from 15 × 16 µm up to 38 × 40 µm (incl. ocelli); ocelli scattered, 7-8(-10), those near underleaf base usually conspicuously larger than surrounding cells, others more or less of the same size as surrounding cells. Rhizoid initial disc small, rhizoids not seen. Dioicous. Androecia usually intercalary on short or long branches, bracts in 3-7 pairs, imbricate, saccate, mostly smaller than ordinary leaves, lobules slightly to considerably smaller than lobes. Bracteoles present throughout the androecium, similar to underleaves but sometimes less deeply incised. Scattered ocelli present in bracts and bracteoles. Gynoecia on main shoots with one or two Pycnolejeunea type innovations; bracts longer than vegetative leaves, their lobes 720-750 µm long, 420450 µm wide, asymmetrically ovate-oblong, apiculate, margin irregularly crenulatedenticulate; lobules extending 0.7-0.75 of length of lobes, subrectangular to ± rhombic; apex apiculate, margin irregularly crenulate-denticulate, keel about 1/3 of length of bract lobe, occasionally narrowly winged. Bracteole connate with bracts on both sides, ovate-oblong, narrowed towards base, ca. 600 µm long and 315 µm wide, apex incised, sinus sharp and narrow, lobe acute-acuminate, ending in 1-2(-3) 258 A. Schäfer-Verwimp et al. superposed cells, margin slightly and irregularly crenulate-denticulate. Bracts and bracteoles with 10-20 scattered ocelli, those near base often conspicuously larger than surrounding cells, others ± of same size as surrounding cells; cells and ocelli of bracts and bracteoles similar to those of leaf lobes. Perianth ± as long as bracts, 675-750 µm long, 520-540 µm wide, pyriform, not inflated distally, sharply 5-keeled, upper portions of keels irregularly denticulate; beak small, often inconspicuous, 1(-2) cells high; perianth cells resembling those of bracts, bracteoles and leaves, scattered ocelli present. Mature sporophyte not observed (only a single immature one observed inside of a perianth). Vegetative reproduction not observed. Molecular Phylogeny The ML phylogram (Fig. 18) largely resembles the MCC tree of the Bayesian analysis. All species with multiple accessions were monophyletic, with strong BP and significant PP. An Asian clade (BP = 100, PP = 1.0) with specimens of L. integristipula (J.B.Jack. & Steph.) R.M.Schust. and L. bidentula (J.B.Jack & Steph.) R.M.Schust. [L. subg. Lepidolejeunea] is placed sister to a clade (BP = 94, PP = 1.0) containing a Neotropical lineage (BP = 91, PP = 1.0) with accessions of L. cordifissa, L. auriculata Schäf.-Verw. & Heinrichs, L. grossepapulosa (Steph.) Piippo and L. involuta (Gottsche) Grolle [L. subg. Kingiolejeunea (H.Rob.) R.M.Schust.], and a lineage (BP = 100, PP = 1.0) with the Jamaican L. (subg. Caribeolejeunea) grandiocellata sp. nov. in a sister relationship to anAfromadagascanNeotropical clade (BP = 94, PP = 1.0) with accessions of L. delessertii (Nees & Mont.) Grolle, L. sullivantii (Gottsche) M.E.Reiner, L. eluta (Nees) R.M.Schust. and L. cuspidata (Gottsche) Heinrichs & Schäf.-Verw. [L. subg. Perilejeunea (Kachroo & R.M.Schust.) Schust.] DISCUSSION Distinction of Lepidolejeunea (subg. Caribeolejeunea) grandiocellata Lepidolejeunea grandiocellata was initially recognized by its unusual morphology, particularly the ocelli larger than the surrounding leaf cells in the proximal half of the leaf and lobule. Lepidolejeunea grandiocellata was also well separated from the other Lepidolejeunea species included in the molecular phylogeny (Fig. 18), indicating congruence of morphology-based taxonomy and molecular data. Based on morphology, L. grandiocellata can be recognized by the following combination of characters: (1) delicate plants being less than 1 cm long and ca. 1 mm wide, (2) underleaves orbicular, being 3-4.5 × the stem width and having a sharp, v-shaped notch of ½ underleaf length, (3) perianths sharply 5-keeled and without inflated horns, and (4) ocelli of variable size of which those in basal parts of leaves, underleaves, bracteoles and bracts are conspicuously larger than surrounding cells and thus recognisable in older herbarium material. The latter character separates L. grandiocellata from nearly all other Lepidolejeunea taxa. Similar ocelli have been reported for only a single Lepidolejeunea taxon, the Australasian L. (subg. Lepidolejeunea) novae-caledoniae (Piippo) R.L.Zhu & Frank Müll. (Piippo, 1986, as L. bidentula var. novae-caledoniae; Shu et al., 2016) but its ocelli are almost all larger than the surrounding cells. Further differences include the deviant underleaf insertion and the more robust stature of the gametophytes. Lepidolejeunea grandiocellata sp. nov. (Lejeuneaceae, Porellales) 259 Fig 18. Most likely phylogram including Lepidolejeunea grandiocellata resulting from maximum likelihood analysis of the concatenated rbcL, trnL-trnF, nrITS dataset. Bootstrap percentage values ≥ 50 are displayed. Stars indicate Bayesian Posterior Probability values ≥ 0.95. Subgeneric classification Lepidolejeunea grandiocellata is placed sister to a clade with representatives of L. subg. Perilejeunea in the circumscription of Heinrichs et al. (2015). This subgenus includes species with ocelli being in general slightly smaller than the 260 A. Schäfer-Verwimp et al. surrounding leaf cells or partly as large as the surrounding cells. The very large basal leaf ocelli of L. grandiocellata and the considerable variation in ocellus size do not correspond to this circumscription. Accordingly, we place L. grandiocellata in a new subgenus Caribeolejeunea. Distribution and Ecology Currently, Lepidolejeunea grandiocellata is known only from the type locality in the Blue Mountains of Jamaica. The plants were growing on bark of two young hardwood trees in an open secondary forest just below the summit of Catherines Peak, at 1525 m. Söderström et al. (2011) consider knowledge of the Caribbean, and especially the Jamaican, liverwort flora too poor to reliably comment about range-restricted or endemic species. Local endemics are rare in Lepidolejeunea (Piippo, 1986), however, plants with the morphology of L. grandiocellata have neither been observed by Piippo (1986) nor Heinrichs et al. (2015), and were also not present in other collections made by SV in Jamaica. The plants of the Lepidolejeunea grandiocellata-paratype grew intermingled with Rectolejeunea emarginuliflora (Gottsche ex Schiffn.) A.Evans. Further liverworts at the type locality include Cheilolejeunea unciloba (Lindenb.) Malombe, Cheilolejeunea adnata (Kunze) Grolle, Cryptolophocolea martiana (Nees) L.Söderstr., Crand.-Stotl. & Stotler, Calypogeia peruviana Nees & Mont., Telaranea nematodes (Gottsche ex Aust.) M.A.Howe, Radula pallens (Sw.) Nees & Mont., Riccardia fucoidea (Sw.) Schiffn., Bazzania gracilis (Hampe & Gottsche) Steph., Trichocolea elliottii Steph., Mnioloma cyclostipum (Spruce) R.M.Schust., and Odontoschisma longiflorum (Taylor) Trevis. Mosses at the type locality were Leskeodon auratus (Müll. Hal.) 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Heidelberg Institute for Theoretical Studies. Available from: http://sco.h-its.org/exelixis/web/software/raxml/#documentation (accessed 20 July 2016) SZABÓ A. & PÓCS T., 2016 — New or little known epiphyllous liverworts, XX Cololejeunea nosykombae A.Szabó & Pócs sp. nov. from Madagascar. Journal of bryology 38: 302-307. APPENDIx 1 Taxa, voucher information, and GenBank accession numbers for specimens of Lepidolejeunea and outgroup species used in this study, ‘xx’ indicates missing sequence. Species, voucher, rbcL, trnL-trnF, nrITS OUTGROUP: Metalejeunea crassitexta (J.B.Jack & Steph.) Pócs, Fiji, Pócs & Pócs 03303/R (EGR), KT626918, KT626934, KT626899; M. cucullata (Reinw., Blume & Nees) Grolle, Fiji, Pócs & Pócs 03305/Q (EGR), KT626913, KT626929, KT626894; Otolejeunea moniliata Grolle, Madagascar (I), 9448/L (EGR), KT626923, KT626937, KT626904; O. moniliata, Madagascar (II), Pócs et al. 90113/EA (EGR), KT626922, KT626936, KT626903; Rectolejeunea flagelliformis A.Evans, Panama, Schäfer-Verwimp & Verwimp 34286 (JE), KT626924, KT626938, KT626905; R. versifolia (Schiffn.) L.Söderstr. & A.Hagborg, Guadeloupe, SchäferVerwimp & Verwimp 22245/A (GOET), DQ983724, DQ987444, DQ987342; Vitalianthus aphanellus (Spruce) Bechteler et al., Brazil, Yano & Zartman 32771 (SP), KT626920, xx, KT626900; V. bischlerianus (K.C. Pôrto & Grolle) R.M.Schust. & Giancotti, Brazil, Schäfer-Verwimp & Verwimp 9505 (M), KT626921, xx, KT626901; – INGROUP: Lepidolejeunea auriculata Schäf.-Verw. & Heinrichs, Colombia, Wolf 1595 (GOET), KP635310, xx, KP635281; L. auriculata Ecuador, Schäfer-Verwimp & Preussing 23265 (JE), KP635309, KP635335, KP635280; L. bidentula (J.B.Jack & Steph.) R.M.Schust., China, Koponen et al. 51525 (H) AY125936, AY144476, AY125340; L. bidentula, Thailand, Schäfer-Verwimp & Verwimp 16266 (JE), xx, KP635355, KP635303; L. cordifissa (Taylor) M.E.Reiner, Costa Rica, Schäfer-Verwimp & Holz SV/H-0284/A (JE), KP635306, KP635332, KP635277; L. cordifissa, Ecuador (I), Schäfer-Verwimp & Nebel 31716/B (JE), KP635305, KP635331, KP635276; L. cordifissa, Ecuador (II), Schäfer-Verwimp & Nebel 33219 (M), KP635307, KP635333, KP635278; L. cuspidata (Gottsche) Lepidolejeunea grandiocellata sp. nov. (Lejeuneaceae, Porellales) 263 Heinrichs & Schäf.-Verw., Brazil (I), Schäfer-Verwimp & Verwimp 12659/B (JE), KP635318, KP635343, KP635289; L. cuspidata, Brazil (II), Schäfer-Verwimp & Verwimp 14363 (M), KP635319, KP635344, KP635290; L. cuspidata, Costa Rica, Dauphin & Morales s.n. (GOET), xx, KP635347, KP635294; L. cuspidata, Dominica, Schäfer-Verwimp & Verwimp 17905/B (JE), KP635324, KP635349, KP635296; L. cuspidata, Ecuador (I), Schäfer-Verwimp & Nebel 32123/C (JE), KP635320, KP635345, KP635291; L. cuspidata, Ecuador (II), Gradstein & al. 10054 (GOET), KP635322, xx, KP635293; L. cuspidata, Guadeloupe, SchäferVerwimp & Verwimp 22193 (M), KP635323, KP635348, KP635295; L. cuspidata, Panama, Schäfer-Verwimp & Verwimp 30835 (JE), KP635321, KP635346, KP635292; L. delessertii (Nees & Mont.) Grolle, Réunion (I), Schäfer-Verwimp & Verwimp 19985/A (JE), KP635316, KP635341, KP635287; L. delessertii, Réunion (II), Schäfer-Verwimp & Verwimp 20276/A (JE), KP635317, KP635342, KP635288; L. delessertii, Réunion (III), Gyarmati et al. 9651/CB (JE), KP635315, KP635340, KP635286; L. delessertii, Réunion (IV), Schäfer-Verwimp & Verwimp 20355/B (M), KF039819, KF039853, KF039781; L. eluta (Nees) R.M.Schust., Bolivia (I), Gradstein 9937 (GOET), KP635329, KP635354, KP635302; L. eluta, Bolivia (II), Acebey & Krömer 855 (GOET), KP635328, KP635353, KP635301; L. eluta, Bolivia (III), Drehwald 4833 (GOET), DQ983696, DQ987379, DQ987257; L. eluta, Bolivia (IV), Churchill & Vásquez 21800 (GOET), AY548066, DQ238579, DQ987266; L. eluta, Ecuador, Schäfer-Verwimp & Nebel 33212/B (JE), KP635327, xx, KP635300; L. grandiocellata, Jamaica, Schäfer-Verwimp 35071 (JE), KY550264, KY550265, KY550263; L. grossepapulosa (Steph.) Piippo, Colombia, Gradstein & Varon 11065 (GOET), KP635308, KP635334, KP635279; L. integristipula (J.B.Jack. & Steph.) R.M.Schust., Fiji Isls., Pócs & Pócs 03307/AC (GOET), DQ983697, DQ987417, DQ987313; L. involuta (Gottsche) Grolle, Colombia, Gradstein 8626 (GOET), xx, KP635351, KP635298; L. involuta, Costa Rica, Gradstein & Dauphin 9347 (GOET), KP635311, KP635336, KP635282; L. involuta, Dominica, SchäferVerwimp & Verwimp 17855 (JE), KP635314, KP635339, KP635285; L. involuta, Guadeloupe (I), Schäfer-Verwimp 22178 (M), KP635312, KP635337, KP635283; L. involuta, Guadeloupe (II), Schäfer-Verwimp & Verwimp 22242 (JE), KP635326, KP635352, KP635299; L. involuta, Guadeloupe (III), Schäfer-Verwimp & Verwimp 22309 (M), KP635313, KP635338, KP635284; L. sullivantii (Gottsche) M.E.Reiner, Ecuador, Schäfer-Verwimp & Preussing 23373 (JE), KP635325, KP635350, KP635297.