A comprehensive molecular phylogeny of the genus Sylvirana (Anura: Ranidae) highlights unrecognized diversity, revised classification and historical biogeography

The genus Sylvirana includes 12 species widely distributed in South China and Southeast Asia. The phylogenetic relationships and species diversity for Sylvirana and allied genera remain unresolved and controversial due to insufficient data and incomplete taxon sampling. Using a combined dataset of mitochondrial genes (16S and COI) and 101 nuclear genes obtained through the amplicon sequence capture approach, we generated the most comprehensive phylogenetic analysis for the genus Sylvirana to date, inferring diversity, phylogenetic relationships, and historical biogeography with unprecedented levels of taxon and geographic sampling. Our results conservatively reveal six undescribed species, mostly distributed in peninsular Indochina. Phylogenetic analyses strongly support the non-monophyly of Sylvirana with respect to Pterorana. Additionally, phylogenetic results place Sylvirana guentheri and Pelophylax lateralis into genus Humerana, supporting the inclusion of Hylarana latouchii, Papura..., Library preparation, bait preparation, hybridization, and sequencing were conducted at Sun Yat-sen University. The protocol followed that of Zhang et al. (2019). In brief, we mixed 20 ng of genomic DNA of each of the 38 samples to make a DNA pool. The pooled DNA was used as template to amplify 101 nuclear protein-coding (NPC) markers using the primers and protocol developed by Shen et al. (2013). The 101 PCR products were then mixed together and purified. The purified PCR mixture was subjected to end-repair and A-tailing and then ligated with a biotinylated adapter. The biotinylated amplicons were subsequently immobilized on Dynabeads MyOne streptavidin magnetic beads (Life Technologies) to obtain bait-coated beads.
For each hybridization reaction, 500 ng of pooled libraries and 2.5 μl of bait-coated beads (containing 100 ng of biotinylated amplicons) were used. A touch-down hybridization program was adopted. After denaturation, the hybridization started from 65°C, decreased by 5°C ever..., , # A comprehensive molecular phylogeny of the genus Sylvirana (Anura: Ranidae) highlights unrecognized diversity, revised classification and historical biogeography

[https://doi.org/10.5061/dryad.c59zw3rjj](https://doi.org/10.5061/dryad.c59zw3rjj)

## Description of the data and file structure

To further resolve the phylogenetic relationship of genus Sylvirana, we obtained nuclear sequence capture data from 101 loci for a subset of 12 species, six allied species (H. latouchii, H. bannanica, H. attigua, Pterorana khare, Pelophylax lateralis, and Papurana milleti), and five candidate species. To better understand the temporal evolution of the genus Sylvirana, 15 species were selected as outgroup taxa. In total, 38 species and candidate species were sequenced for nuclear loci.

### Files and variables

#### File: Nu\_Hylarana2.fas

**Description:** 101 nuclear sequence data for a subset of 12 species, six allied species (*H.* *latouchii*, *H. bannanica*, *H. attigua*, *Pterorana khare*, *...