Neotropical diversification: the effects of a complex history on diversity within the poison frog genus Dendrobates

Aim We study the Neotropical poison frogs of the genus Dendrobates Wagler, 1830 in order to clarify their phylogenetic relationships and biogeographical history. The genus Dendrobates is an excellent taxon for examining patterns of Neotropical diversification as the four major species groups appear to correspond roughly to distinct geographical regions: (1) trans‐Andean, (2) Andean foreland, (3) Brazilian Shield and (4) Guianan Shield/Central America. In order to test the agreement of five of the most prominent hypotheses of Amazonian diversification, phylogenetic patterns were examined for agreement with patterns predicted by these hypotheses. Location Central and South America Methods The phylogenetic relationships of the genus Dendrobates were examined from novel and existing (GenBank) sequences of four mitochondrial loci totalling c. 1400 bp from 40 specimens of 22 different species using maximum parsimony and Bayesian methods. Results were compared with traditional taxonomic arrangements by means of SH tests. Phylogenetic relationships and genetic distances were used to test the adequacy of various diversification hypotheses. Results Phylogenetic analyses support the restructuring of two species groups of Dendrobates and the creation of a new species group. Statistical tests of the traditional taxonomic arrangement indicate a significantly bad fit to the molecular data. This restructuring has important implications for the understanding of the historical biogeography of Dendrobates. Biogeographical patterns within this genus suggest that a complex interaction of biotic and abiotic factors since the Eocene have produced the diversity observed today. Main conclusions The current classification of Dendrobates into discrete species groups does not accurately reflect evolutionary history. Data presented here strongly support a monophyletic Brazilian Shield lineage whose members have previously been split among the quinquevittatus and tinctorius groups. Furthermore, previous attempts at elucidating the historical biogeography of this genus were compromised by incomplete sampling and conclusions drawn from a paraphyletic ingroup. Our findings demonstrate a role for numerous hypotheses of diversification (e.g. river, refuge, disturbance–vicariance) in the history of Dendrobates, supporting previous warnings about the dangers of over‐simplification in the study of Neotropical diversification.     

A new scolenaspidid (Osteostraci, Vertebrata) from the Lower Devonian of Podolia, Ukraine

Anew osteostracan genus and species,Victoraspis longicornualis n. gen., n. sp., is described based on material from Rakovets’, present day Ukraine. This new taxon shares characters with the two generaStensiopelta Denison, 1951 andZychaspis Janvier, 1985. A phylogenetic analysis supports the position ofVictoraspis as the sister group to a monophyleticStensiopelta, while the interrelationships of the various species ofZychaspis are poorly resolved. A morphometric analysis is carried out in an attempt to further resolve the taxonomic affinity. This analysis groups all examinedZychaspis species closely together, and further supports the establishment ofVictoraspis as separate genus.      

Phylogenetic analyses of Tolpis Adans. (Asteraceae) reveal patterns of adaptive radiation,multiple colonization and interspecific hybridization

The plant genus Tolpis (Asteraceae) has been the subject of several investigations on the evolution of oceanic island plants. Its insular species were utilized in studies of artificial hybrid fertility, testing the validity of Baker’s law, the application of DNA barcodes, and the phylogenetic utility of inter‐simple sequence repeat markers. Despite this considerable interest in Tolpis, little is known about its phylogenetic history. Past investigations were unable to resolve most of the interspecific relationships, especially within the Canary Islands, where the genus is particularly diverse. Incomplete taxon sampling, the use of ambiguous outgroups and the limited utility of slowly evolving chloroplast DNA markers precluded detailed reconstructions. The present investigation presents a comprehensive molecular phylogeny of Tolpis. By utilizing highly variable nuclear DNA markers and a comprehensive taxon set, we have resolved the majority of interspecific relationships in the genus. Evaluations of competing tree topologies and ancestral area reconstructions complemented the analyses. Our results highlight the presence of three dominant mechanisms of island plant evolution—island colonization, adaptive radiation and interspecific hybridization—in Tolpis: (i) the extant distribution of the genus is the result of two independent colonization pathways, (ii) Tolpis has colonized at least one archipelago multiple times, (iii) the present insular diversity is the product of adaptive radiation, (iv) potential hybridization was detected between species now inhabiting different islands and archipelagoes, indicating sympatric historical distributions, and (v) several undescribed species await taxonomic recognition.     

Phylogenetic resolution and systematics of the Asian tree frogs,Rhacophorus (Rhacophoridae,Amphibia)

Li, J.T., Li, Y., Murphy, R.W., Rao, D.‐Q. & Zhang, Y.‐P. (2012). Phylogenetic resolution and systematics of the Asian tree frogs, Rhacophorus (Rhacophoridae, Amphibia). —Zoologica Scripta, 41, 557–570. The treefrog genus Rhacophorus, a large genus with 80 species, has a wide range, occurring eastward from India to China, Japan, South‐east Asia, the Greater Sunda Islands and the Philippines. The phylogenetic relationships and taxonomic recognition of many species are very controversial. To stabilize the taxonomy, the phylogenetic relationships among about 52 species are investigated from 96 samples using mtDNA sequence data. Matrilineal relationships based on maximum likelihood and Bayesian inference methods resolve three well‐supported lineages (A, B and C), although the phylogenetic relationships among three lineages remain ambiguous. Analyses support recognition of two previously assigned subgenera, Leptomantis and Rhacophorus, and these correspond to lineages A and B, respectively. Given that we have three strongly supported lineages, that these lineages are morphologically distinct, and the constrained geographic distributions of these groups, we recognize each lineage as a taxon. Subgenus Leptomantis includes species mainly from Malaysia, Indonesia and the Philippines. Subgenus Rhacophorus contains a mix of species occurring in India, Indochina and southern China. Lineage C accommodates species distributed mostly in East Asia, including Japan and China. Based on genetic and morphological data from type localities, the taxonomic recognition of some species needs to be reconsidered. Rhacophorus pingbianensis and Polypedates spinus are considered as junior synonyms of Rhacophorus duboisi. Specimens of Rhacophorus rhodopus from Vietnam and Hainan, China likely represent an undescribed, cryptic species.     

A taxonomic revision of Encyclia (Orchidaceae: Laeliinae) in Costa Rica

A taxonomic revision of Encyclia (Orchidaceae) in Costa Rica is presented. The taxonomic history of the genus and its phylogenetic position are discussed. Characters of vegetative and floral morphology are described and their taxonomic significance is discussed. The genus is treated as comprising nine species in the country and a key to species is provided. Each taxon is described on the basis of Costa Rican material, illustrated in a composite plate, and its distribution within the country is assessed. Distribution maps for all the taxa are given. Overall distribution, derivation of name, synonymy, notes on species ecology and diagnostic features are presented for each taxon. The names Encyclia tonduziana and Epidendrum mooreanum are typified. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 168 , 395–448.     

Survivorship of Cyclops abyssorum tatricus (Cyclopoida, Copepoda) and Boeckella gracilipes (Calanoida, Copepoda) under ambient levels of solar UVB radiation in two high-mountain lakes

We performed in situ experiments during the summer of 1995 and 1996 to assess the potential effect of solar ultraviolet B (UVB) radiation (290-320 nm) on the survival of Cyclops abyssorum tatricus Kozminski and Boeckella gracilipes Daday. These species are numerically dominant within the crustacean zooplankton living in two high-mountain lakes, one located in the Austrian Alps [Gossenkollesee (GKS), 2417 m above sea level, maximum depth 9.9 m] and another in the Chilean Andes (Laguna Negra, 2700 m above sea level, maximum depth 320 m). The copepods were incubated in quartz tubes (1 1) or in quartz tubes wrapped with Mylar D® to exclude most of the UVB radiation. The organisms were exposed at 0.5 m depth for 10-72 h on cloudless days. Both lakes were very transparent to UVB and 10% of the surface radiation at the nominal wavelength of 305 nm was still present at 9.6 m in GKS and at 12.8 in Laguna Negra. These species migrate vertically and have a maximum daytime distribution close to the bottom (C.abyssorum tatricus) or below 15 m depth (B.gracilipes). Both species were red, but the carotenoid concentration was higher in C.abyssorum tatricus than in B.gracilipes (6.5 and 2.3 g mg^-1 dry weight, respectively). UV-absorbing compounds with a maximum absorption at 334 nm were also detected. Cyclops abyssorum tatricus was highly resistant to UVB radiation and no significant lethal effect was observed. Boeckella gracilipes had a mortality 5 times higher in the treatment receiving full sunlight than in the Mylar treatment (3.2%) only when exposed for 70 h. The resistance of B.gracilipes was higher than that reported in the literature for the same species, suggesting the existence of intraspecific differences in UV sensitivity.      

Zooplankton of Lake Titicaca on the Bolivian side

Summary The zooplankton, especially the Cladocera and the Copepoda, of Lake Titicaca on the Bolivian side is reported on the basis of a collection made in April 1961. The zooplankton at that time was sparse, consisting only of seven species. Boeckella titicacae Harding is the dominant copepod and B. occidentalis Marsh is also abundant, as in the previous records obtained by the Percy Sladen Expedition in 1937. The Cladocera are poorly represented in the plankton by Ceriodaphnia quadrangula (O. F. Müller), Bosmina cf. hagmanni Stingelin and a few Daphnia pulex Leydig. In a pond near the village of Belen, close to the east coast of the large lake were found 14 species of zooplankters, most of which were living among aquatic vegetation and did not occur in Lake Titicaca. Some taxonomic and zoogeographic remarks are given on some species.Kyoto UniversitySakarazuka-Hondori 2–18, Toyonaka-shi near Osaka, Japan     

Boeckella antiqua n. sp. (Copepoda,Calanoida, Centropagidae) from Patagonia

Boeckella antiqua n. sp. from samples collected in an ephemeral pond on the Patagonian plateau is described and illustrated. Diagnostic features of B. antiqua are almost exclusively related to the male fifth pair of legs, females being almost indistinguishable from those of the closely related B. poppei. The two-segmented, spine-bearing left endopod of the male fifth leg suggests that it may be the basal species in the genus.     

A new record and an evaluation of the phylogenetic relationships of Abrothrix olivaceus markhami (Rodentia: Sigmodontinae)

Recent phylogenetic studies of the subfamily of cricetid rodents (Sigmodntinae) have validated the taxonomic classification at the tribal level of the Andean Clade. It is possible that some endemic species from Patagonian South America are part of this new tribe, but previous studies have not evaluated this hypothesis due to the difficulty of obtaining samples. In this study, we evaluate the phylogenetic relationships of Akodon markhami (Pine, 1973), a species endemic to the Chilean Patagonia, using individuals recently captured at the type locality of this taxon. Our results indicate that this form of Akodon corresponds to a subspecies of Abrothrix olivaceus, and should be incorporated into the Andean Clade as a geographic race of this widely distributed species on the South American continent. Based on a molecular clock calibration, we dated the origin of this geographic race during the last glacial cycles of the Quaternary, as the result of a vicariant process.     

Interspecific Aggression and Resource Monopolization of the Invasive Ant Anoplolepis gracilipes in Malaysian Borneo

Invasions by introduced ant species can be ecologically destructive and affect a wide range of taxa, particularly native ants. Invasive ant species often numerically dominate ant communities and outperform native ant species in effective resource acquisition. Here, we describe interactions between the invasive ant Anoplolepis gracilipes (Smith) and resident ant species in disturbed habitats in NE Borneo. We measured interference competition abilities of A. gracilipes by performing arena bioassays between two A. gracilipes colonies and seven local ant species, and measured its effective resource competition at baits within supercolonies and at supercolony boundaries. Furthermore, we compared ant species diversity and composition at baits among (A) core areas of A. gracilipes supercolonies, (B) supercolony boundaries and (C) outside supercolonies. Anoplolepis gracilipes was behaviorally dominant over most ant species except Oecophylla smaragdina. Within supercolonies, A. gracilipes discovered all food baits first, and monopolized the vast majority throughout the course of the experiment. At supercolony boundaries, A. gracilipes discovered baits later than resident ant species, but subsequently monopolized half of the baits. Furthermore, the activity and diversity of the ant community within A. gracilipes supercolonies was lower than at its boundaries and outside supercolonies, and the ant communities differed significantly between infested and noninfested areas. Our study supports the hypothesis that successful establishment of A. gracilipes in anthropogenically disturbed habitats may negatively affect resident ant communities through high levels of direct interspecific aggression and almost complete monopolization of resources within high‐density supercolonies.     

Revision and phylogeny of the bee genus Paratetrapedia Moure,with description of a new genus from the Andean Cordillera (Hymenoptera,Apidae, Tapinotaspidini)

The bee genus Paratetrapedia represents a commonly collected group of bees and is especially diverse in forested areas of the Neotropics. Its taxonomy has remained poorly understood because of a lack of modern revisionary work and numerous species described as Tetrapedia whose type specimens have not been re‐examined in recent times. Here, a comprehensive study was carried out to review the taxonomy of the genus Paratetrapedia and to investigate cladistically the relationships amongst its species. Eighteen new species of Paratetrapedia are described, giving a total of 32 species in the genus. A phylogenetic analysis of the species of Paratetrapedia was carried out using 61 morphological characters for 41 terminal taxa. The phylogenetic results confirmed the monophyly of Paratetrapedia and allowed the recognition of five species groups: the lugubris, moesta, bicolor, lineata, and flavipennis groups. Nasutopedia gen. nov. , recognized as the sister group of Paratetrapedia and with its distribution restricted to the western forested portions of the Andean highlands, is proposed based on distinct morphology, its placement in the phylogenetic tree, and biogeographical patterns. Species of Paratetrapedia are especially diverse in the Amazon Forest; the eastern Brazilian Atlantic Forest contains four endemic species, and one species is endemic to the Cerrado of central Brazil. Paratetrapedia shows a biogeographical pattern similar to other Neotropical groups of bees and birds, with wide distribution and high diversity in lowland forests and whose sister taxon occurs on highlands of north‐western portions of the Andean cordillera. Identification keys for males and females of all species are provided, as well as distribution maps and illustrations of general external morphology and genitalia. © 2011 The Linnean Society of London, Zoological Journal of the Linnean Society, 2011, 162 , 351–442.     

Phylogenetic structure of Neotropical annual fish of the genus Cynopoecilus (Cyprinodontiformes: Rivulidae), with an assessment of taxonomic implications

The definition of species boundaries constitutes an important challenge in biodiversity studies. Cynopoecilus Regan, 1912 encompasses several endangered species of annual fish, occurring in temporary ponds in a restricted area of Southern Brazil and Uruguay. Divergences about the taxonomic status of Cynopoecilus species highlight the importance of species delimitation studies. Therefore, we address here the phylogenetic structure of Cynopoecilus, while assessing its taxonomic implications. For this, fragments of the mitochondrial COI and nuclear RAG1 genes were characterized and analyzed for a set of 275 and 280 specimens, respectively. DNA barcoding and phylogenetic analyses detected subdivision of these specimens in 8–10 clusters, which comprise the six previously described species, and suggest one invalid taxon and at least 3–5 putative new species. The phylogenetic structure also suggests that the Jacuí River and the Patos Lagoon historically acted as effective barriers to gene flow between populations, although some isolated dispersal events across these water bodies could be evidenced, especially for C. melanotaenia Regan, 1912. In general, the results highlight the need of independent conservation strategies within the distribution area of each of the endemic allopatric killifish clusters, while questioning several taxonomic boundaries and distribution data.     

Patterns and timing of diversification in a tropical montane butterfly genus,Lymanopoda (Nymphalidae,Satyrinae)

Species distributions are a product of contemporary and historical forces. Using phylogenetic and geographic data, we explore the timing of and barriers to the diversification of the Andean butterfly genus Lymanopoda (Nymphalidae, Satyrinae). Clade and species level diversification is coincident with Andean orogeny and Pleistocene glaciation cycles. Lymanopoda has primarily diversified within elevational bands, radiating horizontally throughout the Andes with occasional speciation across elevational boundaries, often associated with ecotones. Narrow elevational ranges and infrequent speciation into adjacent elevational strata suggest that expansion across elevational gradients is relatively difficult. These results are similar to those found in studies of other Andean taxa.     

Integrative taxonomy reveals cryptic Amazonian species of Pristimantis (Anura: Strabomantidae)

Hypotheses on the taxonomic status of two Bolivian Pristimantis with taxonomic problems are assessed by an integrative taxonomic approach that integrates three independent lines of evidence: external morphology, prezygotic reproductive barriers (advertisement calls) and reciprocal monophyly (phylogenetic analyses of partial 16S mtDNA sequences). Central Andean Bolivian populations previously assigned to either P. peruvianus or P. dundeei, and lowland Amazonian populations from southern Peru and northern Bolivia previously considered P. peruvianus do not correspond to these species. Indeed, multivariate analyses of qualitative and quantitative morphological and bioacoustic characters, and phylogenetic analyses support the hypothesis that they represent different, previously unknown, cryptic lineages. They are herein described as new species. The former is a sibling species of P. fenestratus that inhabits the Amazonian and semideciduous forests of the Andean foothills in central Bolivia. The latter is sibling to the Andean species P. danae and is parapatric to it in the Amazonian lowland forests and adjacent foothills of northern Bolivia, southern Peru and adjacent Brazil. Most species of Neotropical frogs, and especially Pristimantis, have been described by using external qualitative morphological characters only. An extended integrative taxonomic approach, as exemplified herein, may lead to the discovery of many other cryptic and sibling lineages that would increase the species numbers of tropical areas. © 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 155 , 97–122.     

BRASILONEMA ANGUSTATUM SP. NOV. (NOSTOCALES), A NEW FILAMENTOUS CYANOBACTERIAL SPECIES FROM THE HAWAIIAN ISLANDS1

A new filamentous cyanobacterial species of the genus Brasilonema was isolated from the island of Oahu, Hawaii. The taxon is distinguished from the seven other species in the genus by attenuation of trichomes, and is here described as Brasilonema angustatum sp. nov. It possesses the cytoplasmic kerotimization and reddish‐brown coloration of several species in the genus. Phylogenetic analysis based on 16S rRNA gene sequence shows B. angustatum within a highly supported clade containing all sequenced Brasilonema species. We compared the secondary structure of the 16S–23S ITS regions for B. octagenarum and B. angustatum. The structurally conservative D1–D1′ and V3 helices show similar motifs between the two taxa, but differ structurally and in sequence, providing additional justification for erection of the new species. The Box‐B helix has identical secondary structure. The existence of tapering in a Brasilonema species is unique in this genus, and requires modification of the current concept of the genus Brasilonema, which was described as being unattenuated. Our phylogenetic evidence supports the hypothesis that tapering has developed repeatedly in separate cyanobacterial lineages and lacks the taxonomic significance once assumed by early workers.     

Molecular Phylogeny and Biogeography of Petaurista Inferred from the Cytochrome b Gene,with Implications for the Taxonomic Status of P. caniceps,P. marica and P. sybilla

The polymorphic genus Petaurista includes a group of diverse species and subspecies that are adapted for gliding and arboreal life. This morphological diversity has resulted in taxonomic discrepancies, and molecular phylogenetic studies have been limited by taxon sampling. To clarify this controversial taxonomy, we used the cytochrome b gene to reconstruct the phylogeny to obtain a more accurate picture of the evolutionary relationships, species differentiation and divergence pattern of Petaurista. The results revealed a significant inconsistency between taxonomic designations, phylogeny and genetic distances. When 6 recognized species were included, species delimitation revealed 15 putative species, a finding that warrants a comprehensive morphological diagnosis and a re-assessment of the species status. The validity of P. caniceps and P. marica was discussed. An estimation of the molecular divergence time demonstrated that the diversification and speciation of Petaurista began during the later Miocene and may have been affected by the uplifting of the Qinghai-Tibet plateau and subsequent climate change.     

Molecular phylogeny and species delimitation of Stachyuraceae: Advocating a herbarium specimen‐based phylogenomic approach in resolving species boundaries

Species concept and delimitation are fundamental to taxonomic and evolutionary studies. Both inadequate informative sites in the molecular data and limited taxon sampling have often led to poor phylogenetic resolution and incorrect species delineation. Recently, the whole chloroplast genome sequences from extensive herbarium specimen samples have been shown to be effective to amend the problem. Stachyuraceae are a small family consisting of only one genus Stachyurus of six to 16 species. However, species delimitation in Stachyurus has been highly controversial because of few and generally unstable morphological characters used for classification. In this study, we sampled 69 individuals of seven species (each with at least three individuals) covering the entire taxonomic diversity, geographic range, and morphological variation of Stachyurus from herbarium specimens for genome‐wide plastid gene sequencing to address species delineation in the genus. We obtained high‐quality DNAs from specimens using a recently developed DNA reconstruction technique. We first assembled four whole chloroplast genome sequences. Based on the chloroplast genome and one nuclear ribosomal DNA sequence of Stachyurus, we designed primers for multiplex polymerase chain reaction and high throughput sequencing of 44 plastid loci for species of Stachyurus. Data of these chloroplast DNA and nuclear ribosomal DNA internal transcribed spacer sequences were used for phylogenetic analyses. The phylogenetic results showed that the Japanese species Stachyurus praecox Siebold & Zucc. was sister to the rest in mainland China, which indicated a typical Sino‐Japanese distribution pattern. Based on diagnostic morphological characters, distinct distributional range, and monophyly of each clade, we redefined seven species for Stachyurus following an integrative species concept, and revised the taxonomy of the family based on previous reports and specimens, in particular the type specimens. Furthermore, our divergence time estimation results suggested that Stachyuraceae split from its sister group Crossosomataceae from the New World at ca. 54.29 Mya, but extant species of Stachyuraceae started their diversification only recently at ca. 6.85 Mya. Diversification time of Stachyurus in mainland China was estimated to be ca. 4.45 Mya. This research has provided an example of using the herbarium specimen‐based phylogenomic approach in resolving species boundaries in a taxonomically difficult genus.     

Feeding of Boeckella gracilipes (Copepoda, Calanoida) on ciliates and phytoflagellates in an ultraoligotrophic Andean lake

The calanoid copepod, Boeckella gracilipes, is the dominantcrustacean zooplankton in South Andean deep ultra-oligotrophiclakes. Combining field and experimental data we explored thefeeding of the copepod and its access to the mixotrophic ciliate,Ophrydium naumanni, in Lake Moreno Oeste (Patagonia, Argentina).Phytoplankton was dominated by nanoflagellates throughout thewater column. Ophrydium naumanni, which accumulates much ofthe chlorophyll a, as do copepodites and adults of B. gracilipes,has a deep distribution during the day, with maximal abundancesaround 30 m depth. Mouth-part morphology analysis of B. gracilipesindicated that the copepod has an omnivorous diet. Laboratoryexperiments showed that B. gracilipes could access O. naumannionly when it is offered as a single food item. However, whennatural phytoplankton and ciliate assemblages (including O.naumanni)are offered, B. gracilipes did not eat Ophrydium and preyedon the oligotrich, Strombidium viride, and phytoflagellateslike Chrysochromulina parva. The range of ingested sizes wasbroad (3.9–33 µm of equivalent spherical diameter)but all selected particles were motile ones with distinctivemovements, which would enhance the copepod particle detection.     

Relationships within the Munnopsidae (Crustacea, Isopoda, Asellota) based on three genes

The Munnopsidae are a diverse group of asellote isopods that are an important component of deep‐sea fauna. Morphologically‐based phylogenetic inference attempts have proven to be of limited use due to the ecological and morphological diversity within the clade. Monophyly of the family is well‐established but relationships within the group remain unresolved. This project is the first molecularly‐based effort focused specifically on resolving phylogenetic relationships within the Munnopsidae. Partial 28S and COI and complete 18S genes were sequenced for 28 asellotes, 15 additional taxa were included from which only one or two of the three target sequences could be obtained, and 18S sequences for five additional taxa were available from GenBank. Sequences were analysed both as individual genes and in combination using Bayesian and maximum parsimony approaches. Each gene provided a phylogenetic signal that could be identified in the combined analyses, with 18S analyses providing the most resolution of phylogenetic relationships. The available representatives of subfamilies Munnopsinae and Ilyarachninae were monophyletic, as was the genus Munneurycope. Relationships within the subfamily Munnopsinae were well‐resolved by thorough taxon sampling, several new species were placed, and the need for taxonomic revision of Munnopsis/Munnopsoides was supported. These analyses supported putative Eurycope paraphyly and emphasized the need for careful revision of this highly variable genus. Tytthocope was sister to Munnopsurus. Syneurycope was suggested as the sister group to the ilyarachnines. Combined analyses provided increased support for clades suggested in at least two individual gene analyses and for clades not strongly contradicted by individual analyses. Further work is required to fully resolve the munnopsid phylogeny and should consist of increased taxon sampling for the complete 18S sequence and possibly identification of at least one slowly evolving, nuclear protein‐coding gene to resolve the basal polytomy and enable placement of the root.