Search bioRxiv⌕ Search

Biology subjects

Kozlov, A. M.

Publications and source records attributed to Kozlov, A. M..

13 recordsLinked to original sources

Adaptive RAxML-NG: Accelerating Phylogenetic inference under Maximum Likelihood using dataset difficulty

MotivationPhylogenetic inferences under the Maximum-Likelihood (ML) criterion deploy heuristic tree search strategies to explore the vast search space. Depending on the input dataset, searches from different starting trees might all converge to a single tree topology. Often, though, distinct searches infer multiple topologies with large log-likelihood score differences or yield topologically highly distinct, yet almost equally likely, trees. Recently, Haag et al. introduced an approach to quantify, and implemented machine learning methods to predict, the difficulty of an MSA with respect to phylogenetic inference. Easy MSAs exhibit a single likelihood peak on their likelihood surface, associated with a single tree topology to which most, if not all, independent searches rapidly converge. However, as difficulty increases, multiple locally optimal likelihood peaks emerge, yet from highly distinct topologies. ResultsTo this end, we introduce and implement an adaptive tree search heuristic in RAxML-NG, which modifies the thoroughness of the tree search strategy as a function of the predicted difficulty. Our adaptive strategy is based upon three observations. First, on easy datasets, searches converge rapidly and can hence be terminated at an earlier stage. Second, over-analyzing difficult datasets is hopeless and, thus, it suffices to quickly infer only one of the numerous almost equally likely topologies, to reduce overall execution time. Third, more extensive searches are justified and required on datasets with intermediate difficulty. While the likelihood surface exhibits multiple locally optimal peaks in this case, a small proportion of them is significantly better. Our experimental results for the adaptive heuristic on 9, 515 empirical and 5, 000 simulated datasets with varying difficulty exhibit substantial speedups, especially on easy and difficult datasets (53% of total MSAs), where we observe average speedups of more than 10x. Further, approximately 94% of the inferred trees using the adaptive strategy are statistically indistinguishable from the trees inferred under the standard strategy (RAxML-NG). AvailabilityGNU GPL at https://github.com/togkousa/raxml-ng/tree/adaptive. ContactAnastasis.Togkousidis@h-its.org Supplementary MaterialAvailable

bioinformatics↗

A representative Performance Assessment of Maximum Likelihood based Phylogenetic Inference Tools

SummaryThe evaluation of phylogenetic inference tools is commonly conducted on simulated and empirical sequence data alignments. An open question is how representative these alignments are with respect to those, commonly analyzed by users. Based upon the RAxMLGrove database, it is now possible to simulate DNA sequences based on more than 70, 000 representative RAxML and RAxML-NG tree inferences on empirical datasets conducted on the RAxML web servers. This allows to assess the phylogenetic tree inference accuracy of various inference tools based on realistic and representative simulated DNA alignments. We simulated 20, 000 MSAs based on representative datasets (in terms of signal strength) from RAxMLGrove, and used 5, 000 datasets from the TreeBASE database, to assess the inference accuracy of FastTree2, IQ-TREE2, and RAxML-NG. We find that on quantifiably difficult-to-analyze MSAs all of the analysed tools perform poorly, such that the quicker FastTree2, can constitute a viable alternative to infer trees. We also find, that there are substantial differences between accuracy results on simulated and empirical data, despite the fact that a substantial effort was undertaken to simulate sequences under as realistic as possible settings. ContactDimitri Hohler, dimitri.hoehler@h-its.org

bioinformatics↗

The Free Lunch is not over yet - Systematic Exploration of Numerical Thresholds in Phylogenetic Inference

Maximum Likelihood (ML) is a widely used phylogenetic inference model. ML implementations heavily rely on numerical optimization routines that use internal numerical thresholds to determine convergence. We systematically analyze the impact of these threshold settings on the log-likelihood and runtimes for ML tree inferences with RAxML-NG, IQ-TREE, and FastTree on empirical datasets. We provide empirical evidence that we can substantially accelerate tree inferences with RAxML-NG and IQ-TREE by changing the default values of two such numerical thresholds. At the same time, altering these settings does not significantly impact the quality of the inferred trees. We further show that increasing both thresholds accelerates the RAxML-NG bootstrap without influencing the resulting support values. For RAxML-NG, increasing the likelihood thresholds{epsilon} LnL and{epsilon} brlen to 10 and 103 respectively results in an average tree inference speedup of 1.9 {+/-} 0.6 on Data collection 1, 1.8 {+/-} 1.1 on Data collection 2, and 1.9 {+/-} 0.8 on Data collection 2 for the RAxML-NG bootstrap. Increasing the likelihood threshold{epsilon} LnL to 10 in IQ-TREE results in an average tree inference speedup of 1.3 {+/-} 0.4 on Data collection 1 and 1.3 {+/-} 0.9 on Data collection 2.

bioinformatics↗

NetRAX: Accurate and Fast Maximum Likelihood Phylogenetic Network Inference

Phylogenetic networks are used to represent non-treelike evolutionary scenarios. Current, actively developed approaches for phylogenetic network inference jointly account for non-treelike evolution and incomplete lineage sorting (ILS). Unfortunately, this induces a very high computational complexity. Hence, current tools can only analyze small data sets. We present NetRAX, a tool for maximum likelihood inference of phylogenetic networks in the absence of incomplete lineage sorting. Our tool leverages state-of-the-art methods for efficiently computing the phylogenetic likelihood function on trees, and extends them to phylogenetic networks via the notion of "displayed trees". NetRAX can infer maximum likelihood phylogenetic networks from partitioned multiple sequence alignments and returns the inferred networks in Extended Newick format. On simulated data, our results show a very low relative difference in BIC score and a near-zero unrooted softwired cluster distance to the true, simulated networks. With NetRAX, a network inference on a partitioned alignment with 8, 000 sites, 30 taxa, and 3 reticulations completes within a few minutes on a standard laptop. Our implementation is available under the GNU General Public License v3.0 at https://github.com/lutteropp/NetRAX.

bioinformatics↗

Exploring Parallel MPI Fault Tolerance Mechanisms for Phylogenetic Inference with RAxML-NG

Phylogenetic trees are now routinely inferred on large scale HPC systems with thousands of cores as the parallel scalability of phylogenetic inference tools has improved over the past years to cope with the molecular data avalanche. Thus, the parallel fault tolerance of phylogenetic inference tools has become a relevant challenge. To this end, we explore parallel fault tolerance mechanisms and algorithms, the software modifications required, and the performance penalties induced via enabling parallel fault tolerance by example of RAxML-NG, the successor of the widely used RAxML tool for maximum likelihood based phylogenetic tree inference. We find that the slowdown induced by the necessary additional recovery mechanisms in RAxML-NG is on average 2%. The overall slowdown by using these recovery mechanisms in conjunction with a fault tolerant MPI implementation amounts to 8% on average for large empirical datasets. Via failure simulations, we show that RAxML-NG can successfully recover from multiple simultaneous failures, subsequent failures, failures during recovery, and failures during checkpointing. Recoveries are automatic and transparent to the user. The modified fault tolerant RAxML-NG code is available under GNU GPL at https://github.com/lukashuebner/ft-raxml-ng Contact: lukas.huebner@{kit.edu,h-its.org};, alexey.kozlov@h-its.org, hespe@kit.edu, sanders@kit.edu, alexandros.stamatakis@hits.org Supplementary information: Supplementary data are available at bioR{chi}iv.

bioinformatics↗

Are fleas highly modified Mecoptera? Phylogenomic resolution of Antliophora (Insecta: Holometabola)

Insect orders have been defined and stable for decades, with few notable exceptions (e.g., Blattodea and Psocoptera). One of the few remaining questions of order-level monophyly is that of Mecoptera in respect to the phylogenetic placement of Siphonaptera (fleas). We used a large set of transcriptomic nucleotide sequence data representing 56 species and more than 3,000 single-copy genes to resolve the evolutionary history of Antliophora, including fleas (Siphonaptera), scorpionflies and relatives (Mecoptera), and true flies (Diptera). We find that fleas and mecopterans together are the sister group of flies. However, our data and/or analyses are unable to distinguish whether fleas are sister to a monophyletic Mecoptera, or whether they arose from within extant mecopteran families, rendering Mecoptera paraphyletic. We did not detect parameter bias in our dataset after applying a broad range of detection methods. Counter to a previous hypothesis that placed fleas within Mecoptera as the sister group to wingless boreids (snow fleas), we found a potential sister group relationship between fleas and the enigmatic family Nannochoristidae. Although we lack conclusive evidence, it seems possible that fleas represent the most-species rich group of modern mecopterans and that their parasitic lifestyle and morphological adaptations have simply made them unrecognizable in respect to their order-level classification.

evolutionary biology↗

CellPhy: accurate and fast probabilistic inference of single-cell phylogenies from scDNA-seq data

We introduce a maximum likelihood framework called CellPhy for inferring phylogenetic trees from single-cell DNA sequencing (scDNA-seq) data. CellPhy leverages a finite-site Markov genotype substitution model with 16 diploid states, akin to those typically used in statistical phylogenetics. It includes a dedicated error function for single cells that incorporates amplification/sequencing error and allelic dropout (ADO). Moreover, it can explicitly consider the uncertainty of the variant calling process by using genotype likelihoods as input. We implemented CellPhy in a widely used open-source phylogenetic inference package (RAxML-NG) that provides statistical confidence measurements on the estimated tree and scales particularly well on large scDNA-seq datasets with hundreds or thousands of cells. To benchmark CellPhy, we carried out 19,400 coalescent simulations of cell samples from exponentially-growing tumors for which the true phylogeny was known. We evolved single-cell diploid DNA genotypes along the simulated genealogies under different scenarios, including infinite- and finite-sites nucleotide mutation models, trinucleotide mutational signatures, sequencing, and amplification errors, allele dropouts, and cell doublets. Our simulations suggest that CellPhy is robust to amplification/sequencing errors and ADO and outperforms state-of-the-art methods under realistic scDNA-seq scenarios both in terms of accuracy and speed. Also, we sequenced 24 single-cell whole-genomes from a colorectal tumor. Together with three published scDNA-seq data sets, we analyzed these empirical data to illustrate how CellPhy can provide more reliable biological insights than most competing methods. CellPhy is freely available at https://github.com/amkozlov/cellphy.

bioinformatics↗

A Fast and Memory-Efficient Implementation of the Transfer Bootstrap

Recently, Lemoine et al. suggested the Transfer Bootstrap Expectation (TBE) branch support metric as an alternative to classical phylogenetic bootstrap support metric on taxon-rich datasets. However, the original TBE implementation in the booster tool is compute- and memory-intensive. Therefore, we developed a fast and memory-efficient TBE implementation. We improved upon the original algorithm described by Lemoine et al. by introducing multiple algorithmic and technical optimizations. On empirical as well as on random tree sets with varying taxon counts, our implementation is up to 480 times faster than booster. Furthermore, it only requires memory that is linear in the number of taxa, which leads to 10x - 40x memory savings compared to booster. Our implementation has been partially integrated into pll-modules and RAxML-NG and is available under the GNU Affero General Public License v3.0 at https://github.com/ddarriba/pll-modules and https://github.com/amkozlov/raxml-ng. The parallelized version that also computes additional TBE-related statistics is available in pll-modules and RAxML-NG forks at: https://github.com/lutteropp/pll-modules/tree/tbe and https://github.com/lutteropp/raxml-ng/tree/tbe.

bioinformatics↗

Long metabarcoding of the eukaryotic rDNA operon to phylogenetically and taxonomically resolve environmental diversity

High-throughput environmental DNA metabarcoding has revolutionized the analysis of microbial diversity, but this approach is generally restricted to amplicon sizes below 500 base pairs. These short regions contain limited phylogenetic signal, which makes it impractical to use environmental DNA in full phylogenetic inferences. However, new long-read sequencing technologies such as the Pacific Biosciences platform may provide sufficiently large sequence lengths to overcome the poor phylogenetic resolution of short amplicons. To test this idea, we amplified soil DNA and used PacBio Circular Consensus Sequencing (CCS) to obtain a ~4500 bp region of the eukaryotic rDNA operon spanning most of the small (18S) and large subunit (28S) ribosomal RNA genes. The CCS reads were first treated with a novel curation workflow that generated 650 high-quality OTUs containing the physically linked 18S and 28S regions of the long amplicons. In order to assign taxonomy to these OTUs, we developed a phylogeny-aware approach based on the 18S region that showed greater accuracy and sensitivity than similarity-based and phylogenetic placement-based methods using shorter reads. The taxonomically-annotated OTUs were then combined with available 18S and 28S reference sequences to infer a well-resolved phylogeny spanning all major groups of eukaryotes, allowing to accurately derive the evolutionary origin of environmental diversity. A total of 1019 sequences were included, of which a majority (58%) corresponded to the new long environmental CCS reads. Comparisons to the 18S-only region of our amplicons revealed that the combined 18S-28S genes globally increased the phylogenetic resolution, recovering specific groupings otherwise missing. The long-reads also allowed to directly investigate the relationships among environmental sequences themselves, which represents a key advantage over the placement of short reads on a reference phylogeny. Altogether, our results show that long amplicons can be treated in a full phylogenetic framework to provide greater taxonomic resolution and a robust evolutionary perspective to environmental DNA.

microbiology↗

ModelTest-NG: a new and scalable tool for the selection of DNA and protein evolutionary models

ModelTest-NG is a re-implementation from scratch of jModelTest and ProtTest, two popular tools for selecting the best-fit nucleotide and amino acid substitution models, respectively. ModelTest-NG is one to two orders of magnitude faster than jModelTest and ProtTest but equally accurate, and introduces several new features, such as ascertainment bias correction, mixture and FreeRate models, or the automatic processing of partitioned datasets. ModelTest-NG is available under a GNU GPL3 license at https://github.com/ddarriba/modeltest.

bioinformatics↗

Modelling structural constraints on protein evolution via side-chain conformational states

Few models of sequence evolution incorporate parameters describing protein structure, despite its high conservation, essential functional role and increasing availability. We present a structurally-aware empirical substitution model for amino acid sequence evolution in which proteins are expressed using an expanded alphabet that relays both amino acid identity and structural information. Each character specifies an amino acid as well a rotamer state: the discrete geometric pattern of permitted side-chain atomic positions. By assigning rotamer states in 251,194 protein structures and identifying 4,508,390 substitutions between closely related sequences, we generate a 55-state model that shows that the evolutionary properties of amino acids depend strongly upon side-chain geometry. The model performs as well as or better than traditional 20-state models for divergence time estimation, tree inference and ancestral state reconstruction. We conclude that the concomitant evolution of sequence and structure is a valuable source of phylogenetic information.

evolutionary biology↗

ParGenes: a tool for massively parallel model selection and phylogenetic tree inference on thousands of genes.

MotivationCoalescent- and reconciliation-based methods are now widely used to infer species phylogenies from genomic data. They typically use per-gene phylogenies as input, which requires conducting multiple individual tree inferences on a large set of multiple sequence alignments (MSAs). At present, no easy-to-use parallel tool for this task exists. Ad hoc scripts for this purpose do not only induce additional implementation overhead, but can also lead to poor resource utilization and long times-to-solution. We present ParGenes, a tool for simultaneously determining the best-fit model and inferring maximum likelihood (ML) phylogenies on thousands of independent MSAs using supercomputers.\n\nResultsParGenes executes common phylogenetic pipeline steps such as model-testing, ML inference(s), bootstrapping, and computation of branch support values via a single parallel program invocation. We evaluated ParGenes by inferring > 20, 000 phylogenetic gene trees with bootstrap support values from Ensembl Compara and VectorBase alignments in 28 hours on a cluster with 1024 nodes.\n\nAvailabilityGNU GPL at https://github.com/BenoitMorel/ParGenes.\n\nContactBenoit.Morel@h-its.org\n\nSupplementary informationSupplementary material is available at Bioinformatics online.

bioinformatics↗

EPA-ng: Massively Parallel Evolutionary Placement of Genetic Sequences

Next Generation Sequencing (NGS) technologies have led to a ubiquity of molecular sequence data. This data avalanche is particularly challenging in metagenetics, which focuses on taxonomic identification of sequences obtained from diverse microbial environments. To achieve this, phylogenetic placement methods determine how these sequences fit into an evolutionary context. Previous implementations of phylogenetic placement algorithms, such as the Evolutionary Placement Algorithm (EPA) included in RAxML, or O_SCPLOWPPLACERC_SCPLOW, are being increasingly used for this purpose. However, due to the steady progress in NGS technologies, the current implementations face substantial scalability limitations. Here we present EPA-O_SCPLOWNGC_SCPLOW, a complete reimplementation of the EPA that is substantially faster, offers a distributed memory parallelization, and integrates concepts from both, RAxML-EPA, and O_SCPLOWPPLACERC_SCPLOW. EPA-O_SCPLOWNGC_SCPLOW can be executed on standard shared memory, as well as on distributed memory systems (e.g., computing clusters). To demonstrate the scalability of EPA-O_SCPLOWNGC_SCPLOW we placed 1 billion metagenetic reads from the Tara Oceans Project onto a reference tree with 3,748 taxa in just under 7 hours, using 2,048 cores. Our performance assessment shows that EPA-O_SCPLOWNGC_SCPLOW outperforms RAxML-EPA and O_SCPLOWPPLACERC_SCPLOW by up to a factor of 30 in sequential execution mode, while attaining comparable parallel efficiency on shared memory systems. We further show that the distributed memory parallelization of EPA-O_SCPLOWNGC_SCPLOW scales well up to 3,520 cores. EPA-O_SCPLOWNGC_SCPLOW is available under the AGPLv3 license: https://github.com/Pbdas/epa-ng

bioinformatics↗