Theloderma pyaukkya is recorded for the first time in China based on a specimen collected from western Yunnan. Morphologically, the specimen shows good agreement with the original description of T. pyaukkya, and phylogenetically is clustered with the type specimens and holotype of T. pyaukkya from Kachin State (northern Myanmar) with strong support. The taxonomic status of T. pyaukkya from Chin State (western Myanmar) needs further examination. In addition, Theloderma moloch is also recorded in Yunnan for the first time. This brings the number of Theloderma species recorded in Yunnan, China, to seven, namely, T. albopunctatum, T. baibungense, T. bicolor, T. gordoni, T. moloch, T. pyaukkya, and T. rhododiscus.
Theloderma Tschudi, a genus of the family Rhacophoridae, is distributed widely throughout Southeast Asia, China, India, and Sri Lanka, and presently includes 26 recognized species (Frost, 2020). To date, eight species have been recorded in China, namely, T. albopunctatum (Liu and Hu), T. baibungense (Jiang, Fei, and Huang), T. bicolor (Bourret), T. corticale (Boulenger), T. gordoni Taylor, T. lateriticum Bain, Nguyen, and Doan, T. moloch (Annandale), and T. rhododiscus (Liu and Hu) (AmphibiaChina, 2020; Chen et al., 2019; Hou et al., 2017; Qi et al., 2018). Of these eight species, five occur in Yunnan, i.e., T. albopunctatum, T. baibungense, T. bicolor, T. gordoni, and T. rhododiscus (Qi et al., 2018).
Theloderma pyaukkya Dever, a species highly similar in appearance and size to T. albopunctatum and T. asperum (Dever, 2017), was previously known only from Kachin State in northern Myanmar and Chin State in western Myanmar (Dever, 2017). Western Yunnan borders northern Myanmar (Figure 1A) and previous studies have shown these regions to be similar in amphibian fauna (e.g., Hui et al., 2019; Yu et al., 2018, 2019). Thus, it is expected that T. pyaukkya could also be found in western Yunnan, where only one species of Theloderma (T. baibungense) has been reported (Hou et al., 2017).
Figure 1. Collection sites, habitat, phylogeny, and photos of Theloderma pyaukkya (GXNU YU000116), T. moloch (GXNU YU000115), and T. albopunctatum
During recent field surveys in western Yunnan, China, we collected a Theloderma specimen similar to members of the T. asperum complex (T. asperum, T. albopunctatum, T. baibungense, and T. pyaukkya) in appearance due to its splattered brown-and-white dorsal coloration. Molecular comparison indicated this individual to be T. pyaukkya. Herein, we describe this new record for China in detail.
Field surveys were conducted in Yingjiang County, Yunnan Province, China (Figure 1A). Specimens were collected and euthanized with ethyl acetate and then fixed in 80% ethanol for storage after taking photographs. Liver tissue samples were preserved in 99% ethanol for molecular analysis. The specimens were deposited in Guangxi Normal University (GXNU YU000115 and GXNU YU000116).
Total genomic DNA was extracted from the liver tissues. Tissue samples were digested using proteinase K, and subsequently purified following standard phenol/chloroform isolation and ethanol precipitation. Fragments encoding partial 16S rRNA (16S) and cytochrome c oxidase subunit I (COI) genes were amplified and sequenced (see Supplementary Methods). All new sequences were deposited in GenBank under accession Nos. MT509809, MT509810, and MT522176 (Supplementary Table S1). Phylogenetic relationships among Theloderma species were inferred using Bayesian inference (Supplementary Methods). Uncorrected pairwise distances between species were calculated in MEGA 7 (Kumar et al., 2016).
Measurements were taken with a digital caliper to the nearest 0.1 mm. Morphological terminology followed Fei et al. (2009) (Supplementary Methods).
The obtained sequence alignments for the 16S and COI genes were 496 bp and 681 bp long, respectively. As shown in previous study (Dever, 2017), T. pyaukkya consists of two lineages, one containing the holotype and paratype from the type locality, Kachin State (clade I), and one containing the paratypes from Chin State (clade II). GXNU YU000116 clustered with clade I with strong support (Figures 1C and 1D), whereas clade II was sister to T. baibungense with strong support (Figure 1D). The genetic distances between GXNU YU000116 and T. pyaukkya in clade I were 1.2%–2.1% for 16S and 0.6%–1.4% for COI, whereas clade I differed from clade II by 5.8%–6.2% for 16S and 10.1%–10.4% for COI (Table 1) and from T. baibungense by 5.7%–6.4% for 16S. The distance between T. baibungense and clade II was 4.0%–4.6% for 16S, slightly higher than the conventional threshold of species-level divergence in other groups of Anura (3.0% of divergence in the 16S rRNA gene; Vences et al., 2005a, 2005b; Vieites et al., 2009). Therefore, we considered GXNU YU000116 to belong to T. pyaukkya (type locality: Kachin State).
Species Clade Voucher 1 2 3 4 5 6 7 T. pyaukkya I 1. YU000166 0.6 1.4 10.1 10.1 N/A N/A I 2. CAS 236133 1.2 1.3 10.4 10.4 N/A N/A I 3. CAS 226113 2.1 1.0 10.3 10.3 N/A N/A II 4. CAS 234869 6.2 6.0 6.0 0 NA N/A II 5. CAS 234857 6.2 5.8 5.8 0.2 N/A N/A T. baibungense 6. YPX37270 5.7 6.1 5.9 4.0 4.2 N/A 7. YPX31940 6.0 6.4 6.2 4.3 4.6 0 N/A: Not available.
Table 1. Divergence (P-distance; %) between and within Theloderma pyaukkya and T. baibungense estimated from 16S (lower triangle) and COI (upper triangle) sequences
GXNU YU000115 was clustered with T. moloch from Tibet with strong support and short branches (Figure 1D).
First record of Theloderma pyaukkya Dever, 2017 (Anura: Rhacophoridae) in China, with range extension of Theloderma moloch (Annandale, 1912) to Yunnan
- Received Date: 2020-04-15
- Accepted Date: 2020-06-18
- Available Online: 2020-07-21
|Citation:||Li-Na Du, Shuo Liu, Mian Hou, Guo-Hua Yu. First record of Theloderma pyaukkya Dever, 2017 (Anura: Rhacophoridae) in China, with range extension of Theloderma moloch (Annandale, 1912) to Yunnan. Zoological Research. doi: 10.24272/j.issn.2095-8137.2020.083|