title: An assembly and alignment-free method of phylogeny reconstruction from next-generation sequencing data.
abstract: BACKGROUND
  Next-generation sequencing technologies are rapidly generating whole-genome datasets for an increasing number of organisms. However, phylogenetic reconstruction of genomic data remains difficult because de novo assembly for non-model genomes and multi-genome alignment are challenging.
  
  
  RESULTS
  To greatly simplify the analysis, we present an Assembly and Alignment-Free (AAF) method ( https://sourceforge.net/projects/aaf-phylogeny ) that constructs phylogenies directly from unassembled genome sequence data, bypassing both genome assembly and alignment. Using mathematical calculations, models of sequence evolution, and simulated sequencing of published genomes, we address both evolutionary and sampling issues caused by direct reconstruction, including homoplasy, sequencing errors, and incomplete sequencing coverage. From these results, we calculate the statistical properties of the pairwise distances between genomes, allowing us to optimize parameter selection and perform bootstrapping. As a test case with real data, we successfully reconstructed the phylogeny of 12 mammals using raw sequencing reads. We also applied AAF to 21 tropical tree genome datasets with low coverage to demonstrate its effectiveness on non-model organisms.
  
  
  CONCLUSIONS
  Our AAF method opens up phylogenomics for species without an appropriate reference genome or high sequence coverage, and rapidly creates a phylogenetic framework for further analysis of genome structure and diversity among non-model organisms.
authors.affiliation: Key Laboratory of Tropical Forest Ecology, Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Mengla, Yunnan, 666303, China. hfan22@wisc.edu.|University of Chinese Academy of Sciences, Beijing, 100049, China. hfan22@wisc.edu.|Department of Zoology, University of Wisconsin-Madison, Madison, WI, 53706, USA. hfan22@wisc.edu.
authors.forename: Huan
authors.initials: H
authors.lastname: Fan
authors.affiliation: Department of Zoology, University of Wisconsin-Madison, Madison, WI, 53706, USA. arives@wisc.edu.
authors.forename: Anthony R
authors.initials: AR
authors.lastname: Ives
authors.affiliation: Institut des Sciences de la Forêt Tempérée, Université du Québec en Outaouais, 58 rue Principale, Ripon, QC, J0V 1V0, Canada. yann.surget-groba@uqo.ca.
authors.forename: Yann
authors.initials: Y
authors.lastname: Surget-Groba
authors.affiliation: Key Laboratory of Tropical Forest Ecology, Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Mengla, Yunnan, 666303, China. chuck.cannon@ttu.edu.|Department of Biological Sciences, Texas Tech University, Lubbock, TX, 79410, USA. chuck.cannon@ttu.edu.
authors.forename: Charles H
authors.initials: CH
authors.lastname: Cannon
mesh_terms: D000465:Algorithms; D000818:Animals; D001483:Base Sequence; D016678:Genome; D023281:Genomics / Q000379:methods; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D008322:Mammals / Q000235:genetics; D010802:Phylogeny