Lankesteriana International Journal on Orchidology ISSN: 1409-3871 [email protected] Universidad de Costa Rica Costa Rica Karremans, Adam P. LANKESTERIANA, A NEW GENUS IN THE PLEUROTHALLIDINAE (ORCHIDACEAE) Lankesteriana International Journal on Orchidology, vol. 13, núm. 3, enero, 2013, pp. 319- 332 Universidad de Costa Rica Cartago, Costa Rica Available in: http://www.redalyc.org/articulo.oa?id=44339826010 How to cite Complete issue Scientific Information System More information about this article Network of Scientific Journals from Latin America, the Caribbean, Spain and Portugal Journal's homepage in redalyc.org Non-profit academic project, developed under the open access initiative LANKESTERIANA 13(3): 319—332. 2014. I N V I T E D P A P E R* LANKESTERIANA, A NEW GENUS IN THE PLEUROTHALLIDINAE (ORCHIDACEAE) ADAM P. KARREMANS Lankester Botanical Garden, University of Costa Rica, P.O. Box 302-7050 Cartago, Costa Rica. Naturalis Biodiversity Center - NHN Universiteit Leiden, The Netherlands [email protected] ABSTRACT. We estimated phylogenetic relationships within Anathallis and related genera using Bayesian analyses of nrITS sequence data. The genus is biphyletic in the molecular trees. A novel generic concept, Lankesteriana, is proposed for the species Anathallis barbulata and 19 close relatives. The genus is more closely related to some species of Trichosalpinx and Zootrophion than to Anathallis s.s. Species previously transferred from Pleurothallis subgen. Acuminatia sect. Acuminatae to Anathallis, are here transferred to Stelis, to which they are related phylogenetically. A few additional transfers to Anathallis are made. Lankesteriana is described and characterized, and the necessary taxonomic transfers are made. KEY WORDS: Anathallis, Lankesteriana, Specklinia, Stelis, phylogenetics, systematics Introduction. The most recent reorganization of the However, Pridgeon’s data set included only species generic classification of the Pleurothallidinae proposed of Pleurothallis subgen. Acuminatia sect. Alatae Luer by Pridgeon and Chase (2001) was largely based on and did not include representatives of sect. Acuminatae the results of the molecular phylogenetic studies of the Lindl. had been initially analyzed. Karremans (2010) subtribe (Pridgeon et al. 2001). The initial analyses noted that species belonging to sect. Acuminatae were were made on a representative set of species and their not related to those of sect. Alatae, but instead were results were extrapolated to the whole subtribe by found embedded within Stelis Sw. (sensu Pridgeon correlation with the classification previously proposed 2005), and suggested that, based on morphology, the by Luer (1986), based on morphological similarities. same would be true for all other species in the section. The circumscription of each genus was discussed and The studies by Chiron et al. (2012) and Karremans et refined by Pridgeon (2005). al. (2013a) confirmed that additional species of the sect. Subsequent molecular studies have shown that Acuminatae belonged in Stelis. The first set of authors several of the genera of Pleurothallidinae still require even proposed a new combination for Anathallis rubens a modified circumscription in order to comply with (Lindl.) Pridgeon & M.W.Chase in Stelis, but neglected the monophyly criterion. Anathallis Barb.Rodr. is to transfer all other species of the section. no exception. In the phylogenetic trees of Pridgeon Luer (2006) later segregated species of Pleurothallis et al. (2001), species of Pleurothallis R.Br. subgen. subgen. Specklinia (Lindl.) Garay sect. Muscosae Lindl. Acuminatia Luer (Luer 1999), including the type into Panmorphia Luer resulting in a genus of 73 highly species of genus Anathallis, formed a clade together heterogeneous species with “Specklinia-like habit and with species of Pleurothallis subgen. Specklinia sect. Anathallis-like flowers”. Luer later decided that the Muscosae Lindl. The clade was found sister to a clade variation within Panmophia graded into the concept of which includes Trichosalpinx Luer and Lepanthes Anathallis, and he reduced his Panmophia as a synonym Sw., among others, and a broad concept of genus of the latter (Luer 2009). Analyses of molecular data by Anathallis was re-established (Prodgeon & Chase Stenzel (2004) demonstrated that species of Panmorphia 2001; Pridgeon 2005). (including the type) were embedded within Anathallis. * This paper was prepared in the framework of the celebration of Lankester Botanical Garden’s 40th anniversary. 320 LANKESTERIANA This conclusion was confirmed by Chiron et al. (2012), taken using a DFC295 Leica digital microscope color who included a broad representation of Anathallis camera with Leica FireCam version 3.4.1 software. species in their analyses. Scanning electron microscope (SEM) micrographs One Anathallis species, the broadly distributed were taken from flowers fixed in FAA (formalin 10%, and highly variable Anathallis barbulata (Lindl.) glacial acetic acid 5%, water 35%, ethanol 50%). Pridgeon & Chase, was shown to be distinct from all The floral samples were then dehydrated through a the other species (Chiron et al. 2012). It is probably series of ethanol steps and subjected to critical-point- the most well known species of the group here drying using liquid CO2. Dried samples were mounted discussed. In Luer’s subgeneric classification of and sputter-coated with gold and observed with a genus Pleurothallis R.Br., A. barbulata and a few JEOL JSM-5300 scanning electron microscope at an close relatives were placed in Pleurothallis subgen. accelerating voltage of 10kV. Specklinia sect. Muscosae Lindl. (Luer 1986). Later Phylogenetic analysis —. The data matrix included on, they were transferred to Anathallis by Pridgeon 56 individuals (Table 1), 18 of which were produced and Chase (2001) and Panmorphia by Luer (2006). in this study. The remaining data were obtained from We present nrITS analyses showing that most species GenBank (Pridgeon et al. 2001, Chiron et al. 2012, of Panmorphia, including the type species, Anathallis Karremans et al. 2013a). Plants were obtained from sertularioides (Sw.) Pridgeon & Chase, are embedded living collections at Lankester Botanical Garden within Anathallis. Our data also show that Anathallis in Costa Rica, the Hortus Botanicus in Leiden, and barbulata and a few sister species are not closely related private collections. Vouchers were deposited in to other Anathallis and require generic recognition to spirit collections at JBL and L. Fresh leaf and flower maintain monophyly. cuttings of approximately 1 cm2 were dried with silica Most of these Specklinia-like species of Anathallis gel. Samples (20 mg) were pulverized and extraction have also been treated as species of Specklinia Lindl. performed following the DNEasy procedure (Qiagen). at some point or another. A more extensive molecular The nuclear ribosomal internal transcribed spacer phylogenetic analysis of Specklinia (Karremans et (nrITS) region was amplified using the methods and al. unpublished), excludes the species here treated as primers for sequencing and amplification described Anathallis (Pupulin et al. 2012, Bogarín et al. 2013, by Sun et al. (1994), and Sanger sequencing was Karremans et al. 2013b), requiring the circumscription done commercially by Macrogen on a 96-capillary of those genera in the present manuscript. It 3730xl DNA Analyzer automated sequencer (Applied becomes necessary as well to propose the systematic Biosystems, Inc.) using standard dye-terminator modifications required in order to attain monophyly chemistry (Macrogen, Inc.). within Anathallis, Specklinia, and Stelis and to propose The Staden et al. (2003) package was used for a segregated generic concept for the A. barbulata and editing of the sequences. Contigs were exported as its close relatives. .fas files and opened in Mesquite v2.72 (Maddison & Maddison 2007), where they were checked for Material and Methods. This study was conducted at base calling errors, the matrix was aligned manually. Jardín Botánico Lankester (JBL) of the Universidad de The ends of each data set were trimmed to eliminate Costa Rica and Naturalis Biodiversity Center - Leiden possible erroneous data, and gaps were regarded University, between October 2011 and October 2013. as missing data (filled with Ns). The data matrix is Living material was studied at Lankester Botanical deposited in the Dryad Digital Repository (Heneghan Garden and the Hortus Botanicus in Leiden, while dried et al. 2011). Echinosepala aspasicensis was used as and spirit material was deposited at CR, JBL-spirit and the outgroup, as it was found to be one of the most L-spirit. Taxon names mostly follow Pridgeon (2005). distantly related of all included species (Pridgeon et Photography —. Color illustrations of complete al. 2001). The trees were produced with an analysis flowers were made using a Nikon D5100 digital camera, of the nrITS dataset of 43 sequences using BEAST while photographs of the columns and pollinaria were v1.6.0. (Drummond & Rambaut 2007). Parameters LANKESTERIANA 13(3), January 2014. © Universidad de Costa Rica, 2014. KARREMANS — Lankesteriana, new genus 321 were set to preset, except for substitution model Discussion . The DNA based evidence obtained here GTR with 10 categories, clock model uncorrelated supports the results of Chiron et al. (2012), showing lognormal, tree prior Yule process, and number of that Anathallis is non-monophyletic. The addition generations 20,000,000. The resulting trees were of other