Map showing locations of Pristimantis gretathunbergae sp. nov. and Pristimantis cruentus in Panama. Numbers in the map correspond to localities mentioned in methods. Internal divisions in the map correspond to provinces in black letters.

Phylogenetic tree of Pristimantis spp. based on mtDNA genes 16S and COI performed by a Shimodaira-Hasegawa approximate likelihood ratio test (SH-aLRT test). Numbers on nodes indicate estimated SH-aLRT support/bootstrap support with SH-aLRT values ≥ 80% are considered reliable for a clade (Guindon et al. 2010). The tree is drawn to scale, with branch lengths measured in the number of substitutions per site. Colored bars and G-numbers represent the groups generated by ABGD analysis (see results of phylogenetic analysis and Suppl. material 1: Table S1 for more details).

Map of the Multiple Correspondence Analysis (MCA) of P. gretathunbergae sp. nov. (red dots) and P. cruentus (black dots): Number labels for individual frog with lines pointing to specimen location on the map. Following correlation ratio (Dimension 1/Dimension 2) resulted from the MCA: iris coloration 0.937/0.001; iris reticulation 0.933/0.637; upper eyelid tubercle 0.751/0.331; upper lip coloration 0.735/0.326; groin coloration 0.852/0; ventral coloration 0.810/0.001. The qualitative scoring of the variables and its species-specific expression is explained in the methods.

Coloration in life of specimens of Pristimantis gretathunbergae sp. nov. and P. cruentus from eastern Panama A holotype male (MHCH 3082), Cerro Chucantí B paratype female (SMF97520), Cerro Chucantí C left, paratype female (MHCH 3081), right P. cruentus female (MHCH3034) D female from Cerro Chucantí, not collected E female (MHCH3115) La Javillosa F female, Cerro La Javillosa, Ambroya, Maje Mountain Range (SMF97517) G female (MHCH3079), Rio Tuquesa. Colored lines point to some diagnostic characters as follow: red: blackish iris; yellow: single spine-like tubercle; turquoise: light-colored upper lip; pink: cream, yellow to red groin (red groin also shown in Suppl. material 2: Fig. S10).

Preserved holotype of Pristimantis gretathunbergae sp. nov. (MHCH 3082) A left hand in ventral view B left foot in ventral view C dorsal view D ventral view E ventral view of head F lateral view of head. Scale bars: 10 mm.

Habitat, mating, and parental care in females of Pristimantis gretathunbergae sp. nov. from Cerro Chucantí A Habitat on Cerro Chucantí at ca. 1300 m a.s.l. B understory bromeliad with a P. gretathunbergae sp. nov. in situ (blue line) and zoomed in on inset (MHCH 3115) C amplectant pair on axillary part of bromeliad leaf (not collected) D same female after amplexus guarding eggs E female of P. gretathunbergae taking care of its eggs with a male P. cruentus species holding on the female in reverse position (not collected) F female with eggs about to hatch, note the transparency of the egg membrane (not collected).