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Tempo and mode of evolution of Oryzomyine rodents (Rodentia, Cricetidae, Sigmodontinae): a phylogenomic approach


Percequillo, Alexandre et al. (2021), Tempo and mode of evolution of Oryzomyine rodents (Rodentia, Cricetidae, Sigmodontinae): a phylogenomic approach, Dryad, Dataset,


The tribe Oryzomyini is an impressive group of rodents, comprising 30 extant genera and an estimated 147 species. Recent remarkable advances in the understanding of the diversity, taxonomy and systematics of the tribe have mostly derived from analyses of single or few genetic markers. However, the evolutionary history and biogeography of Oryzomyini, its origin and diversification across the Neotropics, remain unrevealed. Here we use a multi-locus dataset (over 400 loci) obtained through anchored phylogenomics to provide a genome-wide phylogenetic hypothesis for Oryzomyini and to investigate the tempo and mode of its evolution. Species tree and supermatrix analyses produced topologies with strong support for most branches, with all genera confirmed as monophyletic, a result that previous studies failed to obtain. Our analyses also corroborated the monophyly and phylogenetic relationship of three main clades of Oryzomyini (B, C and D). The origin of the tribe is estimated to be in the Miocene (8.93–5.38 million years ago). The cladogenetic events leading to the four main clades occurred during the late Miocene and early Pliocene and most speciation events in the Pleistocene. Geographic range estimates suggested an east of Andes origin for the ancestor of oryzomyines, most likely in the Boreal Brazilian region, which includes the north bank of Rio Amazonas and the Guiana Shield. Oryzomyini rodents are an autochthonous South America radiation, that colonized areas and dominions of this continent mainly by dispersal events. The evolutionary history of the tribe is deeply associated with the Andean cordillera and the landscape history of Amazon basin.


We selected 25 genera and 50 species of the tribe Oryzomyini, representing a fairly comprehensive sampling of generic diversity that includes 83% of the extant genera, and an evenly distributed sampling of species, with about 34% of known species included. We also included 10 species of other Sigmodontinae as outgroups (Table S1, Supplementary Material), representing five tribes, Akodontini, Phyllotini, Sigmodontini, Thomasomyini and Wiedomyini, as well as two incertae sedis lineages.

We improved the vertebrate Anchored Hybrid Enrichment (AHE) target loci of Lemmon et al (2012) for optimal use in mammals. We first identified the genomic coordinates in the human genome (hg19) corresponding to the coordinates of the extended anchor regions of Gallus gallus (galGal4) obtained by Prum et al. (2015) using the UCSC liftover tool ( The corresponding genomic sequences were then extracted and aligned using MAFFT v7.023b to that of the regions used by Prum et al (2015) for probe design. After inspecting the alignments and masking any misaligned regions in Geneious R9 (Biomatters Ltd.), 120 bp probes were tiled uniformly across the human sequences at 1.5x density: conserved regions targeted for AHE enrichment were identified using alignments across divergent species. Once the alignments were constructed/improved and the conserved regions were identified, probe sequences of length of 120bp were generated by sliding a window across the human sequence from the alignment. A new probe was generated starting every 90bp in the sequences.

Sequence data were generated and analyzed at the Florida State University Center for Anchored Phylogenomics ( Extracted DNA was quantified using Qubit and run on gels to assess initial DNA quality. Approximately 500ng of input DNA was sheared to 200-500bp fragments using a Covaris ultrasonicator. Indexing Illumina library preparations were performed on a Beckman Coulter FxP liquid-handling robot. Libraries were then pooled in groups of 16 samples and enriched using an enrichment kit produced by Agilent technologies that contained the probes described above. Enriched libraries were pooled for sequence on two HiSeq 2500 lanes, using a paired-end 150bp protocol with 8bp (single) indexing. Sequencing was performed at the Translational Laboratory in the College of Medicine at Florida State University.

Reads passing the Illumina CASAVA high-chastity filter were demultiplexed using the 8-bp indexes (with no mismatches tolerated). Overlapping read pairs were merged using the Bayesian approach developed by Rokita et al (2012). Adapters were removed and sequencing errors in overlapping regions were corrected during the process. Reads were assembled using the quasi-de novo approach described by Hamilton et al (2016), with Homo sapiens serving as the reference during assembly. Consensus sequences were called from assembly clusters containing at least 100 reads. Orthology among consensus sequences for each locus was determined using a neighbor-joining approach that utilized alignment free pairwise distances computed using the % of shared kmers (short sequences), with one sequence per individual being allowed in each orthologous group. Putative orthologous sets containing more than 50% of the individuals were utilized downstream as independent loci. Sequences were aligned using MAFFT v7.023b.

Prum, R.O., Berv, J.S., Dornburg, A., Field, D.J., Townsend, J.P., Lemmon, E.M., Lemmon, A.R., 2015. A comprehensive phylogeny of birds (Aves) using targeted next-generation DNA sequencing. Nature 526, 569–573.

Lemmon, A.R., Emme, S.A., Lemmon, E.M., 2012. Anchored hybrid enrichment for massively high-throughput phylogenomics. Syst. Biol. 61, 727–744.

Hamilton, C.A., Lemmon, A.R., Lemmon, E.M., Bond, J.E., 2016. Expanding anchored hybrid enrichment to resolve both deep and shallow relationships within the spider tree of life. BMC Evol. Biol. 16, 1–20.

Usage Notes

We are uploading three folders: 

Alignments (with four folders: Astral, BioGeoBears, Divergence times and RAxML and IQ-Tree)


Final_Raw reads_oryzomyini (with five folders: Clade B, Clade C, Clade D, Scolomys and Outgroup; each folder contains the raw reads, by specimen included in the analysis)


Fundação de Amparo à Pesquisa do Estado de São Paulo, Award: 09/16009-1