Bioacoustics Reveal Hidden Diversity in Frogs: Two New Species of the Genus Limnonectes from Myanmar (Amphibia, Anura, Dicroglossidae)
Abstract
:1. Introduction
2. Materials and Methods
3. Results and Conclusions
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Appendix A. Comparative Specimens Examined
Appendix B
Species | GenBank Accession No. | Voucher |
---|---|---|
Fejervarya limnocharis | AF206466 | USNM 520407 |
Limnonectes bagoensis | MZ578030 | HML-030 |
Limnonectes bagoensis | MZ578031 | HML-033 |
Limnonectes bagoensis | MZ578010 | CAS 213272 |
Limnonectes bagoensis | MZ578025 | GK-6742/SMF 104090 |
Limnonectes bagoensis | MZ578026 | GK-6743/SMF 104091 |
Limnonectes bagoensis | MZ578027 | GK-7091/SMF 106038 |
Limnonectes bagoensis | MZ578028 | GK-7093/SMF 106039 |
Limnonectes bagoensis | MZ578029 | GK-7142/SMF 106040 |
Limnonectes bagoyoma | MZ578045 | GK-7110/SMF 106034 |
Limnonectes bagoyoma | MZ578046 | GK-7111/SMF 106035 |
Limnonectes bagoyoma | MZ578047 | GK-7136/SMF 106036 |
Limnonectes bagoyoma | MZ578050 | HML-032 |
Limnonectes bannaensis | KR827879 | #2004.0342 |
Limnonectes coffeatus | KY768796 | NCSM 77785 |
Limnonectes coffeatus | KY768797 | NCSM 77786 |
Limnonectes dabanus | MK688608 | MVZ 258240 |
Limnonectes dabanus | MK688609 | MVZ 258244 |
Limnonectes doriae | GU934330 | CAS 208425 |
Limnonectes doriae | MG935863 | USNM 587327 |
Limnonectes doriae | MG935864 | USNM 587096 |
Limnonectes doriae | MG935865 | CAS 248173 |
Limnonectes doriae | MG935866 | USNM 587326 |
Limnonectes doriae | MG935867 | USNM 587306 |
Limnonectes doriae | MG935868 | USNM 587303 |
Limnonectes doriae | MG935871 | USNM 586919 |
Limnonectes doriae | MG935877 | USNM 586913 |
Limnonectes doriae | MK688573 | CAS 211632 |
Limnonectes doriae | MK688574 | CAS 229581 |
Limnonectes doriae | MK688575 | CAS 229714 |
Limnonectes doriae | MK688576 | FMNH 268506 |
Limnonectes doriae | MK688577 | FMNH 268509 |
Limnonectes doriae | MK688578 | FMNH 270111 |
Limnonectes doriae | MK688579 | FMNH 270113 |
Limnonectes doriae | MW567806 | ZMKU 01521 |
Limnonectes doriae | MW567807 | ZMKU 01522 |
Limnonectes doriae | MW567808 | ZMKU AM 01523 |
Limnonectes doriae | MW567809 | ZMKU AM 01524 |
Limnonectes doriae | MW567810 | ZMKU AM 01525 |
Limnonectes doriae | MW567811 | ZMKU AM 01526 |
Limnonectes doriae | MW567812 | ZMKU AM 01527 |
Limnonectes doriae | MW567813 | ZMKU AM 01528 |
Limnonectes doriae | MW567814 | ZMKU AM 01530 |
Limnonectes doriae | MW567815 | ZMKU AM 01534 |
Limnonectes doriae | MW567816 | ZMKU AM 01543 |
Limnonectes doriae | MW567817 | ZMKU AM 01545 |
Limnonectes doriae | MW567818 | ZMKU AM 01547 |
Limnonectes doriae | MW567819 | ZMKU AM 01552 |
Limnonectes doriae | MZ578008 | CAS 211632 |
Limnonectes doriae | MZ578009 | CAS 211633 |
Limnonectes doriae | MZ578011 | CAS 229581 |
Limnonectes doriae | MZ578012 | CAS 229769 |
Limnonectes doriae | MZ578013 | CAS 232304 |
Limnonectes doriae | MZ578014 | CAS 245709 |
Limnonectes doriae | MZ578015 | CAS 247238 |
Limnonectes doriae | MZ578016 | CAS 247239 |
Limnonectes doriae | MZ578017 | CAS 247792 |
Limnonectes doriae | MZ578018 | CAS 247851 |
Limnonectes doriae | MZ578019 | CAS 247908 |
Limnonectes doriae | MZ578020 | GK-6679 |
Limnonectes doriae | MZ578021 | GK-6682 |
Limnonectes doriae | MZ578022 | GK-6683 |
Limnonectes doriae | MZ578023 | GK-6684 |
Limnonectes doriae | MZ578024 | GK-6685 |
Limnonectes fujianensis | AF315152 | not given |
Limnonectes gyldenstolpei | KR827890 | #0155Y |
Limnonectes gyldenstolpei | KR827891 | #0175Y |
Limnonectes hascheana | AY880449 | MNHN 1997.5355 |
Limnonectes hascheanus | GU934337 | FMNH 270118 |
Limnonectes hascheanus | GU934338 | FMNH 270119 |
Limnonectes hascheanus | GU934349 | LSUHC 6777 |
Limnonectes hascheanus | GU934356 | CAS 229588 |
Limnonectes hascheanus | GU934366 | THNHM 4488 |
Limnonectes hascheanus | GU934368 | THNHM 4490 |
Limnonectes hascheanus | GU934369 | THNHM 4491 |
Limnonectes hascheanus | MZ578033 | CAS 247245 |
Limnonectes hascheanus | MZ578034 | CAS 247853 |
Limnonectes heinrichi | AY313747 | A167137 |
Limnonectes kohchangae | KR827893 | #2003.0317 |
Limnonectes kohchangae | KY768809 | NCSM 79546 |
Limnonectes lauhachindai | KP939077 | ZMKU AM 01109 |
Limnonectes lauhachindai | MK688587 | FMNH 266153 |
Limnonectes limborgi | AB981417 | KUHE 15614 |
Limnonectes limborgi | DQ347343 | isolate 1218 |
Limnonectes limborgi | GU934334 | FMNH 271338 |
Limnonectes limborgi | GU934335 | MVZ 258223 |
Limnonectes limborgi | GU934344 | FMNH 262817 |
Limnonectes limborgi | GU934345 | FMNH 262818 |
Limnonectes limborgi | GU934353 | CAS 228513 |
Limnonectes limborgi | GU934354 | CAS 228514 |
Limnonectes limborgi | GU934355 | CAS 228516 |
Limnonectes limborgi | GU934357 | CAS 229833 |
Limnonectes limborgi | GU934358 | CAS 230378 |
Limnonectes limborgi | GU934359 | CAS 232167 |
Limnonectes limborgi | GU934360 | CAS 232168 |
Limnonectes limborgi | GU934361 | CAS 232174 |
Limnonectes limborgi | GU934362 | CAS 232189 |
Limnonectes limborgi | GU934363 | CAS 232267 |
Limnonectes limborgi | GU934364 | CAS 232274 |
Limnonectes limborgi | GU934365 | CAS 232725 |
Limnonectes limborgi | MG935880 | USNM 586920 |
Limnonectes limborgi | MG935881 | USNM 587305 |
Limnonectes limborgi | MG935882 | USNM 587100 |
Limnonectes limborgi | MZ578032 | CAS 221424 |
Limnonectes limborgi | MZ578035 | CAS 221462 |
Limnonectes limborgi | MZ578036 | CAS 225209 |
Limnonectes limborgi | MZ578037 | CAS 228510 |
Limnonectes limborgi | MZ578038 | CAS 228511 |
Limnonectes limborgi | MZ578039 | CAS 228512 |
Limnonectes limborgi | MZ578040 | CAS 228515 |
Limnonectes limborgi | MZ578041 | CAS 228519 |
Limnonectes limborgi | MZ578042 | CAS 240382 |
Limnonectes limborgi | MZ578043 | CAS 240383 |
Limnonectes limborgi | MZ578044 | CAS 247950 |
Limnonectes limborgi | MZ578048 | GK-7210 |
Limnonectes limborgi | MZ578049 | GK-7216 |
Limnonectes limborgi | MZ578051 | Kovac-3a |
Limnonectes limborgi | MZ578052 | Kovac-6 |
Limnonectes limborgi | MZ578053 | PT-1597 |
Limnonectes limborgi | MZ578054 | PT-0237 |
Limnonectes longchuanensis | KU599868 | KIZYPX 33448 |
Limnonectes macrocephalus | AY313716 | TNHC 61913 |
Limnonectes macrodon | KX055957 | not given |
Limnonectes macrognathus | MK688588 | FMNH 268503 |
Limnonectes macrognathus | MK688589 | FMNH 268505 |
Limnonectes macrognathus | MK688590 | FMNH 270114 |
Limnonectes macrognathus | MK688591 | FMNH 270115 |
Limnonectes plicatellus | KJ720982 | LSUHC 6582 |
Limnonectes plicatellus | KJ720983 | LSUHC 4001 |
Limnonectes pseudodoriae | MW567820 | ZMKU AM 01553 |
Limnonectes pseudodoriae | MW567821 | ZMKU AM 01555 |
Limnonectes pseudodoriae | MW567822 | ZMKU AM 01563 |
Limnonectes pseudodoriae | MW567823 | ZMKU AM 01565 |
Limnonectes pseudodoriae | MW567824 | ZMKU AM 01567 |
Limnonectes pseudodoriae | MW567825 | ZMKU AM 01577 |
Limnonectes pseudodoriae | MW567826 | ZMKU AM 01579 |
Limnonectes pseudodoriae | MW567827 | ZMKU AM 01580 |
Limnonectes pseudodoriae | MW567828 | ZMKU AM 01582 |
Limnonectes pseudodoriae | MW567829 | ZMKU AM 01583 |
Limnonectes pseudodoriae | MW567830 | ZMKU AM 01585 |
Limnonectes savan | MK688602 | NCSM 84943 |
Limnonectes savan | MK688603 | NUOL 00061 |
Limnonectes taylori | KU599866 | KIZ 022399 |
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Clade_3 | Clade_1 | |
---|---|---|
Clade_3 | ||
Clade_1 | 2.4% | |
Clade_2 | 2.5% | 1.9% |
Taxon ID | Call Duration (sec) | Dom. Freq (Hz) | Gap Duration (sec) | Notes Per Call | Notes/s | Freq 5% (Hz) | Freq 95% (Hz) | |
---|---|---|---|---|---|---|---|---|
SMF 104084_0170 (n = 12 calls) ZMKU 1552 (n = 3 calls) | L. doriae, Clade 3, near Mawlamyine L. doriae, Clade 3, Thailand, Mae Hong Son Province | 1.25–2.24 (1.74 ± 0.32) 6.75–7.71 (7.16 ± 0.40) | 775–1033 (947 ± 80.6) 840–1055 (902 ± 51.7) | 2.29–8.43 (4.39 ± 1.94) 32.2 | 15–26 (20.6 ± 3.6) 79–90 (83.7 ± 4.64) | 11.6–12.1 (11.8 ± 0.15) 11.7–11.7 (11.7 ± 0.02) | 302–517 (398 ± 68) 345–431 (367 ± 19) | 1292–1507 (1385 ± 52) 1335–1486 (1414 ± 53) |
SMF 104090_0185 (n = 10 calls) | L. cf. doriae, Clade 1, Bago | 2.09–3.27 (2.72 ± 0.32) | 861–991 (945 ± 30.6) | 5.11–10.82 (7.42 ± 2.44) | 5–9 (7.1 ± 1.0) | 4.2–4.6 (4.4 ± 0.14) | 431–495 (458 ± 19) | 1271–1335 (1302 ± 19) |
SMF 106039_418 (n = 5 calls) | L. cf. doriae, Clade 1, Bago Yoma | 2.62–3.32 (2.99 ± 0.24) | 732–818 (772 ± 21.2) | 49.25–72.67 (59.64 ± 9.74) | 7–8 (7.2 ± 0.4) | 3.6–4.1 (3.9 ± 0.19) | 345–388 (367 ± 22) | 1163–1206 (1201 ± 13) |
L. bagoensis ♂ 4 ♀ 4 | L. doriae ♂ 3 ♀ 3 | L. bagoyoma ♂ 2 ♀ 2 | L. limborgi ♂ 1 ♀ 2 | ||
---|---|---|---|---|---|
SVL | males | 32.40–48.50 (42.25 ± 7.387) | 47.40–54.50 (50.83 ± 3.556) | 29.3–30.8 (30.05 ± 1.06) | 31.20 |
females | 29.80–39.70 (35.73 ± 4.631) | 37.50–42.10 (36.57 ± 2.335) | 23.2–26.8 (25.00 ± 2.55) | 26.9–27.4 (27.15 ± 0.35) | |
SHL/SVL | males | 0.529–0.586 (0.553 ± 0.024) | 0.534–0.560 (0.549 ± 0.013) | 0.471–0.515 (0.493 ± 0.031) | 0.526 |
females | 0.554–0.580 (0.566 ± 0.010) | 0.560–0.568 (0.563 ± 0.004) | 0.522–0.522 (0.522 ± 0.0) | 0.494–0.533 (0.513 ± 0.027) | |
FL/SVL | males | 0.459–0.509 (0.488 ± 0.021) | 0.449–0.490 (0.465 ± 0.021) | 0.393–0.468 (0.430 ± 0.053) | 0.490 |
females | 0.487–0.501 (0.495 ± 0.007) | 0.496–0.517 (0.505 ± 0.010) | 0.470–0.487 (0.480 ± 0.009) | 0.446–0.493 (0.469 ± 0.033) | |
HL/SVL | males | 0.370–0.440 (0.412 ± 0.030) | 0.430–0.470 (0.453 ± 0.020) | 0.390–0.400 (0.395 ± 0.007) | 0.350 |
females | 0.360–0.400 (0.375 ± 0.019) | 0.370–0.380 (0.373 ± 0.005) | 0.330–0.340 (0.335 ± 0.007) | 0.310–0.370 (0.340 ± 0.042) | |
HW / SVL | males | 0.330–0.400 (0.380 ± 0.033) | 0.390–0.450 (0.420 ± 0.030) | 0.880–1.030 (0.955 ± 0.106) | 0.350 |
females | 0.320–0.350 (0.332 ± 0.015) | 0.340–0.350 (0.343 ± 0.005) | 0.880–0.910 (0.895 ± 0.021) | 0.300–0.330 (0.315 ± 0.021) | |
HW/HL | males | 0.900–0.940 (0.920 ± 0.023) | 0.910–0.960 (0.930 ± 0.026) | 0.880–1.03 (0.955 ± 0.106) | 1.020 |
females | 0.800–0.940 (0.892 ± 0.063) | 0.890–0.950 (0.916 ± 0.030) | 0.880–0.910 (0.895 ± 0.021) | 0.890–0.960 (0.925 ± 0.049) | |
IOD/SVL | males | 0.102–0.120 (0.114 ± 0.008) | 0.118–0.134 (0.126 ± 0.008) | 0.104–0.109 (0.106 ± 0.003) | 0.119 |
females | 0.095–0.101 (0.097 ± 0.002) | 0.093–0.105 (0.099 ± 0.006) | 0.101–0.108 (0.104 ± 0.004) | 0.104–0.109 (0.106 ± 0.003) | |
TYD / SVL | males | 0.680–0.860 (0.075 ± 0.007) | 0.089–0.109 (0.101 ± 0.010) | 0.071–0.075 (0.073 ± 0.002) | 0.064 |
females | 0.058–0.064 (0.060 ± 0.002) | 0.056–0.061 (0.058 ± 0.002) | 0.073–0.075 (0.074 ± 0.001) | 0.077–0.082 (0.079 ± 0.003) | |
EYD/SVL | males | 0.099–0.114 (0.103 ± 0.007) | 0.094–0.105 (0.098 ± 0.005) | 0.104–0.119 (0.111 ± 0.010) | 0.131 |
females | 0.101–0.117 (0.107 ± 0.008) | 0.096–0.115 (0.105 ± 0.009) | 0.091–0.127 (0.109 ± 0.025) | 0.126–0.128 (0.127 ± 0.001) | |
NED/SVL | males | 0.090–0.103 (0.095 ± 0.005) | 0.093–0.105 (0.097 ± 0.006) | 0.065–0.075 (0.070 ± 0.007) | 0.071 |
females | 0.081–0.093 (0.089 ± 0.005) | 0.087–0.096 (0.092 ± 0.004) | 0.075–0.095 (0.085 ± 0.014) | 0.078–0.080 (0.079 ± 0.001) | |
TYD/EYD | males | 0.649–0.854 (0.734 ± 0.089) | 0.910–0.960 (0.930 ± 0.026) | 0.629–0.688 (0.658 ± 0.041) | 0.488 |
females | 0.500–0.641 (0.574 ± 0.057) | 0.890–0.950 (0.916 ± 0.030) | 0.588–0.810 (0.699 ± 0.157) | 0.600–0.647 (0.623 ± 0.033) |
Cl_4 | Cl_8 | Cl_3 | Hasch. | Cl_1 | Cl_7 | Cl_5 | Cl_6 | Cl_2 | |
---|---|---|---|---|---|---|---|---|---|
Cl_4 | |||||||||
Cl_8 | 3.2% | ||||||||
Cl_3 | 1.8% | 3.1% | |||||||
hasch. | 5.5% | 5.6% | 5.3% | ||||||
Cl_1 | 4.8% | 4.7% | 4.7% | 5.6% | |||||
Cl_7 | 2.2% | 3.0% | 2.1% | 4.7% | 4.5% | ||||
Cl_5 | 3.4% | 3.7% | 3.4% | 5.7% | 4.8% | 3.1% | |||
Cl_6 | 3.0% | 3.5% | 2.8% | 5.4% | 4.9% | 2.8% | 3.7% | ||
Cl_2 | 4.3% | 4.5% | 4.0% | 5.7% | 4.2% | 4.4% | 4.5% | 4.4% | |
Cl_9 | 4.3% | 4.4% | 3.8% | 4.3% | 3.5% | 3.5% | 3.9% | 3.9% | 2.9% |
Taxon ID | Call Duration (sec) | Dom. Freq (Hz) | Gap Duration (sec) | Number of Notes Per Call | Note Duration (sec) | Freq 5% (Hz) | Freq 95% (Hz) | |
---|---|---|---|---|---|---|---|---|
GK-7209_0446 (n = 5 calls) | L. limborgi, Clade 7, Leiktho | 1.13–1.58 (1.23 ± 0.18) | 1335–1528 (1417 ± 29.2) | 15.8–56.5 (33.3 ± 16.28) | 3–4 (3.2 ± 0.40) | 0.241–0.329 (0.274 ± 0.027) | 904–1033 (986 ± 36.7) | 1895–1981 (1951 ± 20.0) |
GK-7209_0447 (n = 4 calls) | L. limborgi, Clade 7, Leiktho | 1.57–2.87 (2.48 ± 0.53) | 1550–1637 (1589 ± 24.1) | 27.4–37.0 (32.2 ± 3.94) | 3–5 (4.5 ± 0.87) | 0.293–0.419 (0.336 ± 0.039) | 1098–1163 (1120 ± 18.9) | 2046–2132 (2099 ± 24.8) |
GK-7110_0438 (n = 7 calls) | L. cf. limborgi, Clade 6, Bago Yoma | 0.222–0.249 (0.235 ± 0.009) | 1141–1464 (1307 ± 94.0) | 21.5–64.6 (32.3 ± 3.3) | 1 | 0.222–0.249 (0.235 ± 0.009) | 753–797 (781 ± 15.0) | 1830–1938 (1889 ± 45.6) |
GK-7110_0439 (n = 9 calls) | L. cf. limborgi, Clade 6, Bago Yoma | 0.218–0.252 (0.231 ± 0.010) | 1335–1615 (1428 ± 78.6) | 11.3–21.3 (16.1 ± 3.3) | 1 | 0.218–0.252 (0.231 ± 0.010) | 818–861 (846 ± 14.3) | 1960–2046 (2007 ± 30.1) |
SMF 104090 | SMF 106034 | |
---|---|---|
Raw read pairs (Illumina) | 302,829,670 | 278,081,641 |
Read length | 150 | 150 |
Total bases | 90,848,901,000 | 83,424,492,300 |
Survived Reads (%) | 93.5 | 92.3 |
Heterozygosity (%) | 0.78 | 0.78 |
Assembled Nuclear Genome | 1,941,736,683 | 2,099,953,831 |
Number of Scaffolds | 647,264 | 691,036 |
Scaffold N50 | 4270 | 4236 |
Largest Scaffold | 42,771 | 42,830 |
Smallest Scaffold | 500 | 500 |
GC% | 34.74 | 42.54 |
Circular Mito-Genome | 17,480 bp | 17,604 bp |
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Köhler, G.; Zwitzers, B.; Than, N.L.; Gupta, D.K.; Janke, A.; Pauls, S.U.; Thammachoti, P. Bioacoustics Reveal Hidden Diversity in Frogs: Two New Species of the Genus Limnonectes from Myanmar (Amphibia, Anura, Dicroglossidae). Diversity 2021, 13, 399. https://doi.org/10.3390/d13090399
Köhler G, Zwitzers B, Than NL, Gupta DK, Janke A, Pauls SU, Thammachoti P. Bioacoustics Reveal Hidden Diversity in Frogs: Two New Species of the Genus Limnonectes from Myanmar (Amphibia, Anura, Dicroglossidae). Diversity. 2021; 13(9):399. https://doi.org/10.3390/d13090399
Chicago/Turabian StyleKöhler, Gunther, Britta Zwitzers, Ni Lar Than, Deepak Kumar Gupta, Axel Janke, Steffen U. Pauls, and Panupong Thammachoti. 2021. "Bioacoustics Reveal Hidden Diversity in Frogs: Two New Species of the Genus Limnonectes from Myanmar (Amphibia, Anura, Dicroglossidae)" Diversity 13, no. 9: 399. https://doi.org/10.3390/d13090399