Bioacoustics of Gracixalus lumarius

Ecology. The holotype was found underwater in a tree hole with conspecific eggs on the wall of the phyptotelm ( Fig 7B ). All other specimens were found on arboreal vegetation, away from streams or ponds in montane evergreen or bamboo forest (eg. Fig 7A ). The advertisement call and larvae of the new species are unknown. At present, the species is only known from montane bamboo and evergreen forest (> 1800 m ) on Mount Ngoc Linh and adjacent peaks, with less than 20 km between the farthest localities. The distribution of the species is unknown, but is likely to be restricted to high-elevation forest on Mount Ngoc Linh and adjacent peaks. Comparisons. Gracixalus lumarius sp. nov. is distinguished from all other species of Gracixalus , and all other medium-sized ( 35–45 mm adult SVL) rhacophorids with brownish dorsum in Indochina by a combination of the following characters: dorsum brown diurnally and yellowish brown nocturnally; venter pink; tympanum and supratympanic fold indistinct; vomerine teeth absent; iris dark gold with a dense, relatively uniformly distributed network of black reticulations; dorsum with distinctive, white conical asperities in males; and eggs deposited as a tightly spaced array of nonpendent eggs on the wall of a phyptotelm. Within the genus Gracixalus , Gracixalus lumarius sp. nov. is distinguished from Clade I of Rowley et al . (2011) , G. gracilipes (Bourret) , G. quangi Rowley, Dau, Nguyen , Cao & Nguyen, G. quyeti (Nguyen, Hendrix, Böhme, Vu & Ziegler) , and G. supercornutus (Orlov, Ho, & Nguyen) by having a dorsum brown diurnally and yellow nocturnally and a pink venter (versus greenish dorsum and white or yellow venter), an indistinct (versus distinct) tympanum and supratympanic fold, a dark gold iris with a dense, relatively uniformly distributed network of black reticulations (versus bright copper or olive-gold iris with limited, non-uniform black reticulations), distinctive white, conical asperities on the dorsum (versus dorsum mostly smooth or slightly tuberculate) in males, and eggs deposited on wall of a phyptotelm (versus on vegetation above pools for G. gracilipes , G. quangi , and G. supercornutus , Rowley pers. obs.; unknown in G. quyeti ). G. lumarius sp. nov. also has a larger body size ( 38.9– 41.6 mm in three adult males) compared to G. gracilipes , G. quangi , and G. supercornutus (all with male SVL < 25 mm ). Gracixalus lumarius sp. nov. is distinguished from G. carinensis (Boulenger) by having a pink (versus white) venter, an indistinct (versus distinct) tympanum and supratympanic fold, dark gold iris with a dense, relatively uniformly distributed network of black reticulations (versus bright gold iris with limited, non-uniform black reticulations), dorsum with distinctive, white conical asperities (versus dorsum smooth or granulated with small warts on head and anterior part of back only) in males. From G. jinxiuensis (Hu) , the new species is distinguished by having a larger body size ( 38.9–41.6 mm in three adult males versus 23.5 mm in adult male holotype ), indistinct (versus distinct) tympanum and supratympanic fold, dark gold iris with a dense, relatively uniformly distributed network of black reticulations (versus bright gold iris with limited, non-uniform black reticulations), and distinctive, white conical asperities (versus low, scattered tubercles on dorsum) in males. From G. medogensis (Ye & Hu) , the new species is distinguished by having a larger body size ( 38.9–41.6 mm in three adult males compared to 26.5 mm in one adult male), pink (versus light grey or whitish) venter, indistinct (versus distinct) tympanum and supratympanic fold, and distinctive, white conical asperities (versus smooth) in males. From the recently described G. nonggangensis Mo, Zhang, Luo & Chen and G. waza Nguyen, Le, Pham, Nguyen, Bonkowski & Ziegler , the new species differs by having a larger body size ( 38.9–41.6 mm in three adult males versus < 36 mm in adult males), a pink venter (versus white venter with brown spots), indistinct (versus distinct) tympanum and supratympanic fold, dark gold iris with a dense, relatively uniformly distributed network of black reticulations (versus olive iris with limited, non-uniform brown reticulations), and dorsum with distinctive, white conical asperities (versus dorsum smooth or with some small tubercles) in males. The new species differs from Kurixalus ananjevae (Matsui & Orlov) , which is morphologically and likely molecularly similar to Gracixalus Clade II ( Rowley et al . 2011 ), by having a larger body size ( 38.9–41.6 mm in three adult males compared to 32.4 mm in one adult male), pink (versus greyish white) venter, indistinct (versus distinct) tympanum and supratympanic fold, and dorsum with distinctive, white conical asperities (versus dorsum with irregularly distributed, small tubercles) in males. Gracixalus lumarius sp. nov. also differs from all other medium-sized ( 35–45 mm adult SVL) rhacophorids from Indochina. From Kurixalus banaensis (Bourret) , the new species is distinguished by having a pink (versus white) venter, indistinct (versus distinct) tympanum and supratympanic fold, dorsum with distinctive, white conical asperities (versus dorsum granular) in males, no dermal fringing along limbs (versus marked fringing along forearm and tarsus). From Kurixalus odontotarsus (Ye & Fei) , the new species differs by having a pink venter (versus white venter with black or grey marbling), indistinct (versus distinct) tympanum and supratympanic fold, vomerine teeth absent (versus present), and dorsum with distinctive, white conical asperities (versus dorsum rugose) in males. From Polypedates leucomystax (Gravenhorst) and P. megacephalus Hallowell , the new species differs by having a pink (versus white) venter, indistinct (versus distinct) tympanum and supratympanic fold, vomerine teeth absent (versus present), dark gold iris with a dense, relatively uniformly distributed network of black reticulations (versus bright gold iris lacking distinct black reticulations), dorsum with distinctive, white conical asperities (versus dorsum smooth) in males and eggs deposited as a tightly spaced array of nonpendent eggs on the wall of a phyptotelm (versus eggs in foam nest). From Rhacophorus bipunctatus Ahl the new species differs by having a pink (versus yellow) venter, indistinct (versus distinct) supratympanic fold, vomerine teeth absent (versus present), dark gold iris with a dense, relatively uniformly distributed network of black reticulations (versus bright gold iris with limited, non-uniform black reticulations), dorsum with distinctive, white conical asperities (versus dorsum smooth) in males and eggs deposited as a tightly spaced array of nonpendent eggs on the wall of a phyptotelm (versus eggs in foam nest). From R. calcaneus Smith , the new species differs by having a brown dorsum diurnally and yellow nocturnally (versus green or deep reddish-brown dorsum), pink (versus yellow) venter, indistinct (versus distinct) tympanum and supratympanic fold, vomerine teeth absent (versus present), dark gold iris with a dense, relatively uniformly distributed network of black reticulations (versus iris with a bright red periphery), and dorsum with distinctive, white conical asperities (versus dorsum smooth) in males. From R. orlovi Ziegler & Köhler , the new species differs by having a brown dorsum diurnally and yellow nocturnally (versus a reddish brown dorsum with dark brown markings), pink (versus white to light brown) venter, indistinct (versus distinct) tympanum and supratympanic fold, vomerine teeth absent (versus present), dark gold iris with a dense, relatively uniformly distributed network of black reticulations (versus bright gold iris with limited, non-uniform black reticulations), dorsum with distinctive, white conical asperities (versus dorsum smooth) in males and eggs deposited as a tightly spaced array of nonpendent eggs on the wall of a phyptotelm (versus eggs in foam nest). From R. rhodopus Liu and Hu the new species differs by having a pink (versus yellowish) venter, indistinct (versus distinct) tympanum and supratympanic fold, vomerine teeth absent (versus present), dark gold iris with a dense, relatively uniformly distributed network of black reticulations (versus bright gold iris with limited, non-uniform black reticulations), dorsum with distinctive, white conical asperities (versus dorsum smooth) in males and eggs deposited as a tightly spaced array of nonpendent eggs on the wall of a phyptotelm (versus eggs in foam nest). From R. spelaeus Orlov, Gnophanxay, Phimminith, & Phomphoumy , the new species differs by having a pink venter (versus light gray venter covered by dark specks), indistinct (versus distinct) tympanum and supratympanic fold, vomerine teeth absent (versus present), dark gold iris with a dense, relatively uniformly distributed network of black reticulations (versus bright gold or yellowish cream iris with limited, non-uniform black reticulations), and dorsum with distinctive, white conical asperities (versus dorsum smooth) in males. From R. robertingeri Orlov, Poyarkov, Vassilieva, Ananjeva, Nguyen, Nguyen, & Geissler , Gracixalus lumarius sp. nov. differs by having a pink (versus white, grey or cream) venter, indistinct (versus distinct) tympanum and supratympanic fold, vomerine teeth absent (versus present), dark gold iris with a dense, relatively uniformly distributed network of black reticulations (versus bright gold or yellowish cream iris with limited, non-uniform black reticulations), dorsum with distinctive, white conical asperities (versus dorsum smooth) in males and eggs deposited as a tightly spaced array of nonpendent eggs on the wall of a phyptotelm (versus eggs in foam nest). Lastly, from Theloderma ryabovi Orlov, Dutta, Ghate, & Kent , the new species differs by having a brown dorsum diurnally and yellow nocturnally (versus light beige with lilac patterns and black spots), pink (versus dark grey) venter, dark gold iris with a dense, relatively uniformly distributed network of black reticulations (versus dark brown iris), and dorsum with distinctive, white conical asperities (versus dorsum with only small dorsal asperities) in males. Molecular data. Our preliminary molecular analysis supports our placement of G. lumarius sp. nov. in the genus Gracixalus (1.00 Bayesian posterior probability; Fig. 8 ). G. lumarius sp. nov. falls within “Clade I” of Gracixalus ( Rowley et al . 2011 ) , which also includes the type specimen of the genus, G. gracilipes (1.00 Bayesian posterior probability). Tree toplogy was near-identical when we excluded pairwise comparisons for gapped sites. Molecular differentiation among G. lumarius sp. nov . and all other Gracixalus species for which comparable molecular data is available, is high, with 13.3–15.91% uncorrected sequence divergence at the 16S rRNA gene fragment examined. This degree of pairwise divergence in the 16S rRNA gene in frogs is strongly indicative of differentiation at the species level ( Vences et al . 2005 ). Our preliminary molecular analysis recovers the same relationships within the genus Gracixalus as Rowley et al . 2011 (eg. ‘Clade I’ and ‘Clade II’), but more extensive molecular analysis will be required to resolve the evolutionary relationships within the genus and among the family Rhacophoridae .