Wincladtree: Publication-quality tree-diagrams with TNT scripts.

This note describes the implementation and use of wincladtree, a TNT script to plot publication-quality tree-diagrams. This is intended to assist analysis of morphological datasets, where displaying the synapomorphies for the different groups in a compact "Hennigian" style is the norm.

Nothing to it: a reply to Wheeler's "much ado about nothing".

Wheeler (Cladistics 2023, 39, 475) recently suggested that the issues with inapplicable characters in phylogenetic analysis can be dealt with directly by treating observed absences of a feature not in a separate absence/presence character but as insertion/deletion events in a complex character that describes the feature in all its variation; and that this dynamic homology view can be achieved by imposing a sequence or linear order on a set of characters and by analysing the resulting sequence character using custom alphabet tree alignment algorithms. As Wheeler observed, this approach can lead to considering inappropriate character states (such as a head state and a foot state) homologous. We show that it is also sensitive to the specific ordering assumption used and that such different character orders can lead to a preference for different trees. We present a simple four-taxon dataset with observations of absence, but no inapplicable characters or other kinds of character dependence, for which the dynamic homology framework gives different results to classic algorithms for independent characters, including an optimal tree with biologically impossible reconstructions at inner nodes (every terminal has a head but the inner nodes are headless). We show how these issues can be solved by removing the character ordering assumption that the approach requires. Doing so, the dynamic homology framework reduces in general to Maddison's (Syst. Biol. 1993, 42, 576) well-known proposal to deal with inapplicability using step matrix analysis of complex characters. If in addition costs are interpreted in terms of homology, it reduces to Goloboff et al.'s (Cladistics 2021, 37, 596) step matrix implementation for maximization of homology as applied to inapplicable characters. However, if used with homogeneous costs, as Wheeler suggested, it reduces to unordered analysis of such complex characters, which is known to treat tails that may share many observed features as irrelevant for establishing kinship when they differ in just one feature, e.g. colour.

New TNT routines for parallel computing with MPI.

Phylogenetic inference, which involves time-consuming calculations, is a field where parallelization can speed up the resolution of many problems. TNT (a widely used program for phylogenetic analysis under parsimony) allows parallelization under the PVM system (Parallel Virtual Machine). However, as the basic aspects of the implementation remain unpublished, few studies have taken advantage of the parallelization routines of TNT. In addition, the PVM system is deprecated by many system administrators. One of the most common standards for high performance computing is now MPI (Message Passing Interface). To facilitate the use of the parallel analyses offered by TNT, this paper describes the basic aspects of the implementation, as well as a port of the parallelization interface of TNT into MPI. The use of the new routines is illustrated by reanalysis of seven significant datasets, either recent phylogenomic datasets with many characters (up to 2,509,064 characters) or datasets with large numbers of taxa (up to 13,921 taxa). Versions of TNT including the MPI functionality are available at: http://www.lillo.org.ar/phylogeny/tnt/.

TNT version 1.6, with a graphical interface for MacOS and Linux, including new routines in parallel.

A new graphical user interface (GUI) for the parsimony program TNT is presented that works under the Linux and Mac operating systems, as well as the Cygwin environment (which runs under Windows). The new interface is based on the GIMP Tool Kit, GTK (version 3). Formerly, only Windows versions of TNT had a GUI. The new interface improves upon the existing Windows GUI in several respects. These changes, together with several additions to the program since the publication of version 1.5, warrant a change in minor version, thus moving from version 1.5 to 1.6. Among the most notable improvements are the possibility to access graphical user dialogs by means of simple commands, to easily save trees in SVG format ("Scalable Vector Graphics") directly from any tree-diagram being displayed, and to manage analyses in parallel (using multiple processors, by means of the PVM system or "Parallel Virtual Machine"), as well as a generally more stable and consistent behaviour. As the binaries for the new version are compiled as native 64-bit applications, this removes the limitations for accessing large amounts of memory in the previous GUI Windows interface (which is a 32-bit application).

Farewell to the requirement for character independence: phylogenetic methods to incorporate different types of dependence between characters.

This paper discusses methods to take into account interactions between characters, in the context of parsimony analysis. These interactions can be in the form of some characters becoming inapplicable given certain states of other, primary characters; in the form of only certain states being allowed in some characters when a given state or set of states occurs for other characters; or in the form of transformation costs in some character being higher or lower when other characters have certain states or transformations between states. Character-state reconstructions and evaluation of trees under the assumption of independence may easily lead to ancestral assignments that violate elementary rules of biomechanics, well-established theories relating form and function or ideas about character co-variation. An obvious example is reconstructing an ancestral bird as wingless and flying at the same time; another is reconstructing a protein-coding gene as having a stop codon in some ancestors. If the characters are optimized independently, such chimeric ancestral reconstructions can occur even when no terminal displays the impossible combination of states. A set of conventions (implemented via new TNT commands and options) allows the definition of complex rules of interaction. By recoding groups of characters with proper step-matrix costs (and excluding impossible combinations from the set of permissible states), it is possible to find the ancestral reconstructions that maximize homology (and thus the degree to which similarities can be explained by common ancestry), within the constraints imposed by the rules specified by the user. We expect that considerations of biomechanics, functional morphology and natural history will be a source of many theories on possible character dependences, and that the present implementation will encourage users to take the possibility of character dependences into account in their phylogenetic analyses.

Quantification of congruence among gene trees with polytomies using overall success of resolution for phylogenomic coalescent analyses.

Gene-tree-inference error can cause species-tree-inference artefacts in summary phylogenomic coalescent analyses. Here we integrate two ways of accommodating these inference errors: collapsing arbitrarily or dubiously resolved gene-tree branches, and subsampling gene trees based on their pairwise congruence. We tested the effect of collapsing gene-tree branches with 0% approximate-likelihood-ratio-test (SH-like aLRT) support in likelihood analyses and strict consensus trees for parsimony, and then subsampled those partially resolved trees based on congruence measures that do not penalize polytomies. For this purpose we developed a new TNT script for congruence sorting (congsort), and used it to calculate topological incongruence for eight phylogenomic datasets using three distance measures: standard Robinson-Foulds (RF) distances; overall success of resolution (OSR), which is based on counting both matching and contradicting clades; and RF contradictions, which only counts contradictory clades. As expected, we found that gene-tree incongruence was often concentrated in clades that are arbitrarily or dubiously resolved and that there was greater congruence between the partially collapsed gene trees and the coalescent and concatenation topologies inferred from those genes. Coalescent branch lengths typically increased as the most incongruent gene trees were excluded, although branch supports typically did not. We investigated two successful and complementary approaches to prioritizing genes for investigation of alignment or homology errors. Coalescent-tree clades that contradicted concatenation-tree clades were generally less robust to gene-tree subsampling than congruent clades. Our preferred approach to collapsing likelihood gene-tree clades (0% SH-like aLRT support) and subsampling those trees (OSR) generally outperformed competing approaches for a large fungal dataset with respect to branch lengths, support and congruence. We recommend widespread application of this approach (and strict consensus trees for parsimony-based analyses) for improving quantification of gene-tree congruence/conflict, estimating coalescent branch lengths, testing robustness of coalescent analyses to gene-tree-estimation error, and improving topological robustness of summary coalescent analyses. This approach is quick and easy to implement, even for huge datasets.

Parsimony analysis of phylogenomic datasets (I): scripts and guidelines for using TNT (Tree Analysis using New Technology).

We discuss here the use of TNT (Tree Analysis using New Technology) for phylogenomic analysis. For such data, parsimony is a useful alternative to model-based analyses, which frequently utilize models that make unrealistic assumptions (e.g. low heterotachy), struggle with high levels of missing data, etc. Parsimony and model-based methods often yield trees with few topological differences, which can then be analyzed further in order to investigate whether these few topological differences are due to undesirable analysis artefacts. This is facilitated by the greater speed and computational efficiency of parsimony, which allow for a more in-depth analysis of datasets. We here briefly describe the computationally most efficient and versatile parsimony software, TNT, which can be used for phylogenetic and phylogenomic analyses. In particular, we describe and provide a series of scripts that are specifically designed for the analysis of phylogenomic datasets. This includes scripts for concatenation of gene data files in different formats, generation of plots and datasets with different levels of gene/taxon occupancy, calculation of different support measures and phylogenetic reconstruction based on concatenated matrices and single genes. The execution of the scripts is also demonstrated with video clips (https://www.youtube.com/channel/UCpIgK8sVH-yK0Bo3fK62IxA). Lastly, we describe the main commands and functions that enable efficient phylogenomic analyses in TNT.

Parsimony analysis of phylogenomic datasets (II): evaluation of PAUP*, MEGA and MPBoot.

This paper examines the implementation of parsimony methods in the programs PAUP*, MEGA and MPBoot, and compares them with TNT. PAUP* implements standard, well-tested algorithms, and flexible search strategies and options for handling trees; its main drawback is the lack of advanced search algorithms, which makes it difficult to find most parsimonious trees for large and complex datasets. In addition, branch-swapping can be much slower than in TNT for datasets with large numbers of taxa, although this is only occasionally a problem for phylogenomic datasets given that they typically have small numbers of taxa. The parsimony implementation of MEGA has major drawbacks. MEGA often fails to find parsimonious trees because it does not perform all possible branch swapping subtree pruning regrafting (SPR)/tree bisection-reconnection (TBR) rearrangements. It furthermore fails to properly handle ambiguity or multiple equally parsimonious trees, and it uses the same addition sequence for all bootstrap replicates. The latter yields values of group support that depend on the order in which taxa are listed in the dataset. In addition, tree searches are very slow and do not facilitate the exploration of different starting points (as random seed is fixed). MPBoot searches for optimal trees using the ratchet, but it is based on SPR instead of TBR (and only evaluates by default a subset of the SPR rearrangements). MPBoot approximates bootstrap frequencies by first finding a sample of trees and then selecting from those trees for every replicate, without performing a tree-search. The approximation is too rough in many cases, producing serious under- or overestimations of the correct support values and, for most kinds of datasets, slower estimations than can be obtained with TNT. In addition, bootstrapping with PAUP*, MEGA or MPBoot can attribute strong supports to groups that have no support at all under any meaningful concept of support, such as likelihood ratios or Bremer supports. In TNT, this problem is decreased by using the strict consensus tree to represent each replicate, or eliminated entirely by using different approximations of the Bremer support.

A phylogenetic C interpreter for TNT.

MOTIVATION: TNT (a widely used program for phylogenetic analysis) includes an interpreter for a scripting language, but that implementation is nonstandard and uses several conventions of its own. This article describes the implementation and basic usage of a C interpreter (with all the ISO essentials) now included in TNT. A phylogenetic library includes functions that can be used for manipulating trees and data, as well as other phylogeny-specific tasks. This greatly extends the capabilities of TNT. AVAILABILITY AND IMPLEMENTATION: Versions of TNT including the C interpreter for scripts can be downloaded from http://www.lillo.org.ar/phylogeny/tnt/.

Assessing topological congruence among concatenation-based phylogenomic approaches in empirical datasets.

Assessing the effect of methodological decisions on the resulting hypotheses is critical in phylogenetics. Recent studies have focused on evaluating how model selection, orthology definition and confounding factors affect phylogenomic results. Here, we compare the results of three concatenated phylogenetic methods (Maximum Likelihood, ML; Bayesian Inference, BI; Maximum Parsimony, MP) in 157 empirical phylogenomic datasets. The resulting trees were very similar, with 96.7% of all nodes shared between BI and ML (90.6% for ML-MP and 89.1% for BI-MP). Differing nodes were predominantly those of lower support. The main conclusions of most of the studies agreed for the three phylogenetic methods and the discordance involved nodes considered as recalcitrant problems in systematics. The differences between methods were proportionally larger in datasets that analyze the relationships at higher taxonomic levels (particularly phyla and kingdoms), and independent of the number of characters included in the datasets. Note: a spanish version of this article is available in the Supplementary material (Supplementary material online).

DUALCOR: a phylogenetic comparative method to evaluate phylogenetic correlation between a character and a non-evolving external variable.

A new phylogenetic comparative method called DUALCOR is presented to evaluate the evolutionary response of a character to non-evolving external factors, such as environmental variables. The method treats the character as a typical evolving feature of an organism that is reconstructed on a given tree, whereas the external factor is treated as unrelated to the phylogeny. DUALCOR first calculates the correlation/regression between the observed values of the character and the external factor; then it maps the character onto the phylogeny, shuffles the changes among branches, and re-evolves the character to yield new terminal values uncorrelated with the observed values of the external factor, allowing users to examine whether the observed degree of correlation can be attained at random. This is repeated n (say, 999) times, thereby using the dual nature of characters to construct a permutation test that is shown to satisfy requirements of Generalized Monte Carlo procedures. In addition, we provide an empirical example with a reverse test where the external variable (features determined largely by non-heritable factors) is the dependent variable.

Analysis of endemism of world arthropod distribution data supports biogeographic regions and many established subdivisions.

We analyzed 769 242 occurrence records for 115 424 species of terrestrial arthropods, from three biodiversity repositories (Global Biodiversity Information Facility (GBIF), Natural History Museum, London, and "Sistema de Informação Distribuído para Coleções Biológicas" (SpeciesLink)), to test the use of global-scale data points for quantitative assessments of areas of endemism. The data include Insecta (105,941 species), Arachnida (7984 species), Myriapoda (1229) and terrestrial crustaceans (270 Branchiopoda). The species were assigned to 14 543 higher taxonomic groups because such groups often characterize larger areas of endemism. Putative areas of endemism were visualized as sets of cells displaying unique groups of species without the assumption of hierarchical relationships. Yet, the use of 10° grid cells recovered many large areas broadly corresponding to biogeographic Regions (Nearctic, Neotropical, Panamanian, Palaearctic, Afrotropical, Australian, Oceanian and Oriental) albeit with the limits poorly defined. An analysis of 5° grids resulted in 306 sets included in the different biogeographic Realms: Afrotropical, Australian, Madagascan, Nearctic, Neotropical, Oceanian, Oriental, Palaearctic, Saharo-Arabian and Sino-Japanese. The Panamanian Realm comprises 89 partly overlapping sets, crossing the Nearctic and Neotropical boundaries. A total of 7338 species of Insecta were endemic to some areas (Sino-Japanese, Afrotropical, Panamanian, Palaearctic, among others), followed by Arachnida (412 spp) and 105 species in other clades ranked as "classes". Six sets were supported only by genera, except for Panamanian sets that were supported by genera and families. Many of the species in the dataset are included in IUCN red lists, but probably most of those have distributions more restricted than global areas of endemism; only 102 appear as endemic to some area (Neartic, Madagascan, Panamanian, Afrotropical, among others). The results show that data from global databases can be used to identify areas of endemism on a worldwide basis but-owing to their incompleteness-only at a relatively coarse level. At the level of resolution currently allowed by such databases, such global studies are only complementary to studies where areas are determined subjectively by systematists (instead of actual point records), or studies using point records in datasets for specific taxonomic groups curated and compiled by specialists.

A reconsideration of inapplicable characters, and an approximation with step-matrix recoding.

Evidence for phylogenetic analysis comes in the form of observed similarities, and trees are selected to minimize the number of similarities that cannot be accounted for by homology (homoplasies). Thus, the classical argument for parsimony directly links homoplasy with explanatory power. When characters are hierarchically related, a first character may represent a primary structure such as tail absence/presence and a secondary (subordinate) character may describe tail colour; this makes tail colour inapplicable when tail is absent. It has been proposed that such character hierarchies should be evaluated on the same logical basis as standard characters, maximizing the number of similarities accounted for by secondary homology, i.e. common ancestry. Previous evaluations of the homology of a given ancestral reconstruction contain the unintuitive quantity "subcharacters" (number of regions of applicability). Rather than counting subcharacters, this paper proposes an equivalent but more intuitive formulation, based on counting the number of changes into each separate state. In this formulation, x-transformations, the homoplasy for the reconstruction is simply the number of changes into the state beyond the first, summed over all states. There is thus no direct connection between homoplasy and number of steps, only between homoplasy and extra steps. The link between the two formulations is that, for any region of applicability of any character, a subcharacter can be interpreted as the change into the state that is plesiomorphic in that region. Although some authors have claimed that the equivalence between maximizing explanatory power and minimizing independent originations of similar features (i.e. the standard justification of parsimony) does not hold for inapplicable characters, evaluating homoplasy with x-transformations clearly connects the two sides of that equation. Furthermore, as the evaluation with x-transformations provides a direct count and a straightforward interpretation of homoplasy, it extends naturally into implied weighting, and sheds light on problems with additive, step-matrix or continuous characters. It also allows deriving transformation costs for recoding hierarchies as step-matrix characters (where recoded states correspond to permissible combinations of states in primary and secondary characters), so that homology of the original observations is properly measured. Those transformation costs set the cost of gaining the primary structure to the maximum difference between "present" states plus cost of loss, and difference between "present" states to the sum of user-defined transformation costs between secondary features. With such recoding, invoking multiple independent derivations of the structure and similar features will cost as many extra "steps" as the instances of similarities (in both original characters) that are not being homologized. The step-matrix recoding also can take into account nested dependences. We present a simple convention for naming characters, which TNT can use to automatically convert the original data into a step-matrix form and set the proper transformation costs. Finally, the basic elements for handling inapplicable characters in the context of maximum-likelihood inference are outlined, and some quantitative comparisons between different approaches to inapplicables are provided.

Morphological Data Sets Fit a Common Mechanism Much More Poorly than DNA Sequences and Call Into Question the Mkv Model.

The Mkv evolutionary model, based on minor modifications to models of molecular evolution, is being increasingly used to infer phylogenies from discrete morphological data, often producing different results from parsimony. The critical difference between Mkv and parsimony is the assumption of a "common mechanism" in the Mkv model, with branch lengths determining that probability of change for all characters increases or decreases at the same tree branches by the same exponential factor. We evaluate whether the assumption of a common mechanism applies to morphology, by testing the implicit prediction that branch lengths calculated from different subsets of characters will be significantly correlated. Our analysis shows that DNA (38 data sets tested) is often compatible with a common mechanism, but morphology (86 data sets tested) generally is not, showing very disparate branch lengths for different character partitions. The low levels of branch length correlation demonstrated for morphology (fitting models without a common mechanism) suggest that the Mkv model is too unrealistic and inadequate for the analysis of most morphological data sets. [Bayesian analysis; Mkv model; morphological data; phylogenetics.].

Likelihood approximations of implied weights parsimony can be selected over the Mk model by the Akaike information criterion.

A likelihood method that approximates the behaviour of implied weighting is described. This approach provides a likelihood perspective on several aspects of implied weighting, such as guidance for the choice of concavity values, a justification to use different concavities for different numbers of taxa, and a natural basis for extended implied weighting. In this approach, the number of free parameters in the estimation depends on C, the number of characters (in contrast to the standard Mk model, which estimates 2T-3 parameters for T taxa). Depending on the characteristics of the dataset, the likelihood obtained with this approach may in some cases be similar or superior to that of the Mk model, but with fewer parameters being adjusted. Because of that tradeoff, testing against the Mk model by means of the Akaike information criterion on a set of 182 morphological datasets indicated many cases (36) in which the likelihood approximation to implied weighting is the best method, from an information-theoretic point of view. Given that it is expected to produce (almost) the same results as this maximum-likelihood approximation, implied weighting can therefore be seen as a valid alternative to the Mk model often used for morphological datasets, on the basis of a criterion for model fit widely advocated by likelihoodists.

On defining a unique phylogenetic tree with homoplastic characters.

This paper discusses the problem of whether creating a matrix with all the character state combinations that have a fixed number of steps (or extra steps) on a given tree T, produces the same tree T when analyzed with maximum parsimony or maximum likelihood. Exhaustive enumeration of cases up to 20 taxa for binary characters, and up to 12 taxa for 4-state characters, shows that the same tree is recovered (as unique most likely or most parsimonious tree) as long as the number of extra steps is within 1/4 of the number of taxa. This dependence, 1/4 of the number of taxa, is discussed with a general argumentation, in terms of the spread of the character changes on the tree used to select character state distributions. The present finding allows creating matrices which have as much homoplasy as possible for the most parsimonious or likely tree to be predictable, and examination of these matrices with hill-climbing search algorithms provides additional evidence on the (lack of a) necessary relationship between homoplasy and the ability of search methods to find optimal trees.

Weighted parsimony outperforms other methods of phylogenetic inference under models appropriate for morphology.

One of the lasting controversies in phylogenetic inference is the degree to which specific evolutionary models should influence the choice of methods. Model-based approaches to phylogenetic inference (likelihood, Bayesian) are defended on the premise that without explicit statistical models there is no science, and parsimony is defended on the grounds that it provides the best rationalization of the data, while refraining from assigning specific probabilities to trees or character-state reconstructions. Authors who favour model-based approaches often focus on the statistical properties of the methods and models themselves, but this is of only limited use in deciding the best method for phylogenetic inference-such decision also requires considering the conditions of evolution that prevail in nature. Another approach is to compare the performance of parsimony and model-based methods in simulations, which traditionally have been used to defend the use of models of evolution for DNA sequences. Some recent papers, however, have promoted the use of model-based approaches to phylogenetic inference for discrete morphological data as well. These papers simulated data under models already known to be unfavourable to parsimony, and modelled morphological evolution as if it evolved just like DNA, with probabilities of change for all characters changing in concert along tree branches. The present paper discusses these issues, showing that under reasonable and less restrictive models of evolution for discrete characters, equally weighted parsimony performs as well or better than model-based methods, and that parsimony under implied weights clearly outperforms all other methods.

The spider tree of life: phylogeny of Araneae based on target-gene analyses from an extensive taxon sampling.

We present a phylogenetic analysis of spiders using a dataset of 932 spider species, representing 115 families (only the family Synaphridae is unrepresented), 700 known genera, and additional representatives of 26 unidentified or undescribed genera. Eleven genera of the orders Amblypygi, Palpigradi, Schizomida and Uropygi are included as outgroups. The dataset includes six markers from the mitochondrial (12S, 16S, COI) and nuclear (histone H3, 18S, 28S) genomes, and was analysed by multiple methods, including constrained analyses using a highly supported backbone tree from transcriptomic data. We recover most of the higher-level structure of the spider tree with good support, including Mesothelae, Opisthothelae, Mygalomorphae and Araneomorphae. Several of our analyses recover Hypochilidae and Filistatidae as sister groups, as suggested by previous transcriptomic analyses. The Synspermiata are robustly supported, and the families Trogloraptoridae and Caponiidae are found as sister to the Dysderoidea. Our results support the Lost Tracheae clade, including Pholcidae, Tetrablemmidae, Diguetidae, Plectreuridae and the family Pacullidae (restored status) separate from Tetrablemmidae. The Scytodoidea include Ochyroceratidae along with Sicariidae, Scytodidae, Drymusidae and Periegopidae; our results are inconclusive about the separation of these last two families. We did not recover monophyletic Austrochiloidea and Leptonetidae, but our data suggest that both groups are more closely related to the Cylindrical Gland Spigot clade rather than to Synspermiata. Palpimanoidea is not recovered by our analyses, but also not strongly contradicted. We find support for Entelegynae and Oecobioidea (Oecobiidae plus Hersiliidae), and ambiguous placement of cribellate orb-weavers, compatible with their non-monophyly. Nicodamoidea (Nicodamidae plus Megadictynidae) and Araneoidea composition and relationships are consistent with recent analyses. We did not obtain resolution for the titanoecoids (Titanoecidae and Phyxelididae), but the Retrolateral Tibial Apophysis clade is well supported. Penestomidae, and probably Homalonychidae, are part of Zodarioidea, although the latter family was set apart by recent transcriptomic analyses. Our data support a large group that we call the marronoid clade (including the families Amaurobiidae, Desidae, Dictynidae, Hahniidae, Stiphidiidae, Agelenidae and Toxopidae). The circumscription of most marronoid families is redefined here. Amaurobiidae include the Amaurobiinae and provisionally Macrobuninae. We transfer Malenellinae (Malenella, from Anyphaenidae), Chummidae (Chumma) (new syn.) and Tasmarubriinae (Tasmarubrius, Tasmabrochus and Teeatta, from Amphinectidae) to Macrobuninae. Cybaeidae are redefined to include Calymmaria, Cryphoeca, Ethobuella and Willisius (transferred from Hahniidae), and Blabomma and Yorima (transferred from Dictynidae). Cycloctenidae are redefined to include Orepukia (transferred from Agelenidae) and Pakeha and Paravoca (transferred from Amaurobiidae). Desidae are redefined to include five subfamilies: Amphinectinae, with Amphinecta, Mamoea, Maniho, Paramamoea and Rangitata (transferred from Amphinectidae); Ischaleinae, with Bakala and Manjala (transferred from Amaurobiidae) and Ischalea (transferred from Stiphidiidae); Metaltellinae, with Austmusia, Buyina, Calacadia, Cunnawarra, Jalkaraburra, Keera, Magua, Metaltella, Penaoola and Quemusia; Porteriinae (new rank), with Baiami, Cambridgea, Corasoides and Nanocambridgea (transferred from Stiphidiidae); and Desinae, with Desis, and provisionally Poaka (transferred from Amaurobiidae) and Barahna (transferred from Stiphidiidae). Argyroneta is transferred from Cybaeidae to Dictynidae. Cicurina is transferred from Dictynidae to Hahniidae. The genera Neoramia (from Agelenidae) and Aorangia, Marplesia and Neolana (from Amphinectidae) are transferred to Stiphidiidae. The family Toxopidae (restored status) includes two subfamilies: Myroinae, with Gasparia, Gohia, Hulua, Neomyro, Myro, Ommatauxesis and Otagoa (transferred from Desidae); and Toxopinae, with Midgee and Jamara, formerly Midgeeinae, new syn. (transferred from Amaurobiidae) and Hapona, Laestrygones, Lamina, Toxops and Toxopsoides (transferred from Desidae). We obtain a monophyletic Oval Calamistrum clade and Dionycha; Sparassidae, however, are not dionychans, but probably the sister group of those two clades. The composition of the Oval Calamistrum clade is confirmed (including Zoropsidae, Udubidae, Ctenidae, Oxyopidae, Senoculidae, Pisauridae, Trechaleidae, Lycosidae, Psechridae and Thomisidae), affirming previous findings on the uncertain relationships of the "ctenids" Ancylometes and Cupiennius, although a core group of Ctenidae are well supported. Our data were ambiguous as to the monophyly of Oxyopidae. In Dionycha, we found a first split of core Prodidomidae, excluding the Australian Molycriinae, which fall distantly from core prodidomids, among gnaphosoids. The rest of the dionychans form two main groups, Dionycha part A and part B. The former includes much of the Oblique Median Tapetum clade (Trochanteriidae, Gnaphosidae, Gallieniellidae, Phrurolithidae, Trachelidae, Gnaphosidae, Ammoxenidae, Lamponidae and the Molycriinae), and also Anyphaenidae and Clubionidae. Orthobula is transferred from Phrurolithidae to Trachelidae. Our data did not allow for complete resolution for the gnaphosoid families. Dionycha part B includes the families Salticidae, Eutichuridae, Miturgidae, Philodromidae, Viridasiidae, Selenopidae, Corinnidae and Xenoctenidae (new fam., including Xenoctenus, Paravulsor and Odo, transferred from Miturgidae, as well as Incasoctenus from Ctenidae). We confirm the inclusion of Zora (formerly Zoridae) within Miturgidae.

TNT version 1.5, including a full implementation of phylogenetic morphometrics.

Version 1.5 of the computer program TNT completely integrates landmark data into phylogenetic analysis. Landmark data consist of coordinates (in two or three dimensions) for the terminal taxa; TNT reconstructs shapes for the internal nodes such that the difference between ancestor and descendant shapes for all tree branches sums up to a minimum; this sum is used as tree score. Landmark data can be analysed alone or in combination with standard characters; all the applicable commands and options in TNT can be used transparently after reading a landmark data set. The program continues implementing all the types of analyses in former versions, including discrete and continuous characters (which can now be read at any scale, and automatically rescaled by TNT). Using algorithms described in this paper, searches for landmark data can be made tens to hundreds of times faster than it was possible before (from T to 3T times faster, where T is the number of taxa), thus making phylogenetic analysis of landmarks feasible even on standard personal computers.

Problems with supertrees based on the subtree prune-and-regraft distance, with comments on majority rule supertrees.

This paper examines a recent proposal to calculate supertrees by minimizing the sum of subtree prune-and-regraft distances to the input trees. The supertrees thus calculated may display groups present in a minority of the input trees but contradicted by the majority, or groups that are not supported by any input tree or combination of input trees. The proponents of the method themselves stated that these are serious problems of "matrix representation with parsimony", but they can in fact occur in their own method. The majority rule supertrees, being explicitly clade-based, cannot have these problems, and seem much more suited to retrieving common clades from a set of trees with different taxon sets. However, it is dubious that so-called majority rule supertrees can always be interpreted as displaying those clades present (or compatible with) with a majority of the trees. The majority rule consensus is always a median tree, in terms of the Robinson-Foulds distances (i.e. it minimizes the sum of Robinson-Foulds distances to the input trees). In contrast, majority rule supertrees may not be median-different, contradictory trees may minimize Robinson-Foulds distances, while their strict consensus does not. If being "majority" results from being median in Robinson-Foulds distances, this means that in the supertree setting a "majority" is ambiguously defined, sometimes achievable only by mutually contradictory trees.