Abstract
The family Perlidae is a diverse group of stoneflies frequently found in Mid-Cretaceous Kachin amber from northern Myanmar. However, no detailed descriptions of genuine perlid larvae from amber specimens have been reported. In this study, the first well-preserved genuine stonefly larva from Kachin amber is described and classified within the Perlidae. Morphological analysis reveals its closest affinity to larvae of the extant genus Caroperla Kohno, 1946 (subfamily Acroneuriinae, tribe Kiotinini), yet highlights unique traits suggesting affiliation with an undescribed lineage. Phylogenetic and morphospace analyses based on larval morphology consistently recover this fossil in an intermediate phylogenetic position, raising questions about the monophyly of subfamilies and tribes within Perlidae. Notably, the specimen exhibits functional adaptations such as predatory mouthparts and chloride cell-bearing gills identical to modern Perlidae, indicating morphological stasis since the mid-Cretaceous. This discovery not only underscores the critical role of amber in preserving aquatic immature stages but also provides novel insights into the evolutionary trajectories and ecological resilience of perlid larvae in ancient freshwater ecosystems.
References
- Asiah, W.N.W., Salmah, M.C. & Sivec, I. (2009) Description of Etrocorema belumensis sp. n. from Royal Belum State Park, Perak, Malaysia. Illiesia, 5 (17), 182–187.
- Cao, T.K.T. & Bae, Y.J. (2007) New species of Acroneuria (Plecoptera: Perlidae: Acroneuriinae) from tropical Southeast Asia. Journal of the Kansas Entomological Society, 80 (3), 192–204. https://doi.org/10.2317/0022-8567(2007)80[192:NSOAPP]2.0.CO;2
- Cao, T.K.T. & Bae, Y.J. (2013) A rare tropical stonefly Brahmana flavomarginata (Plecoptera: Perlidae: Acroneuriinae) from Vietnam. Journal of Animal Systematics, Evolution and Diversity, 29 (2), 179–183. https://doi.org/10.5635/ASED.2013.29.2.179
- Chen, Z.T. (2022a) The larval morphology of a new mid-Cretaceous stonefly and its systematic position in Plecoptera. Arthropod Systematics & Phylogeny, 80, 423–438. https://doi.org/10.3897/asp.80.e82549
- Chen, Z.T. (2022b) Comparative larval morphology of two stoneflies of Perlodidae (Insecta: Plecoptera). Zoologischer Anzeiger, 299, 115–127. https://doi.org/10.1016/j.jcz.2022.06.001
- Chen, Z.T. (2024a) The structure of a gilled stonefly larva from the mid-Cretaceous Kachin amber. Arthropod Structure & Development, 82, 101380. https://doi.org/10.1016/j.asd.2024.101380
- Chen, Z.T. (2024b) Comparative larval morphology and DNA barcodes of Kamimuria petasus Chen and Neoperla tuberculata Wu (Plecoptera: Perlidae). Zoologischer Anzeiger, 309, 75–89. https://doi.org/10.1016/j.jcz.2024.02.003
- Cruz, I.N.B.D., Nuneza, O.M. & Lin, C.P. (2016) Description of a new Oriental stonefly species, Phanoperla constanspina (Plecoptera: Perlidae) from Mindanao, Philippines and association of life stages using DNA barcoding. Zootaxa, 4193 (1), 102–116. https://doi.org/10.11646/zootaxa.4193.1.4
- DeWalt, R.E. & Kondratieff, B.C. (2019) Plecoptera. In: Merritt, R.W., Cummins, K.W. Berg, M.B. (Eds.), An Introduction to the Aquatic Insects of North America. 5th Edition. Kendall Hunt Publishing Company, Dubuque, pp. 429–520.
- DeWalt, R.E., Hopkins, H., Neu-Becker, U. & Stueber, G. (2025) Plecoptera Species File Online. Version 5.0/5.0. Available from: https://plecoptera.speciesfile.org/ (accessed 7 April 2025)
- Dorvillé, L.F.M. (1997) Caracterização Taxonômica e Biologia de Uma Espécie de Kempnyia Klapálek (Plecoptera, Perlidae) em um Riacho da Floresta da Tijuca, Rio de Janeiro (RJ). UFRJ, Rio de Janeiro, 120 pp. https://doi.org/10.4257/oeco.1998.0501.05
- Dorvillé, L.F., Nessimian, J.L. & Sanseverino, A.M. (2000) First record of symphoresy between nymphs of the stonefly Kempnyia tijucana, and chironomid larvae, Nanocladius (Plecopteracoluthus) sp., in the Neotropics. Studies on Neotropical Fauna and Environment, 35 (2), 109–114. https://doi.org/10.1076/0165-0521(200008)35:2;1-9;FT109
- Goloboff, P.A., Farris, J.S. & Nixon, K.C. (2008) TNT, a free program for phylogenetic analysis. Cladistics, 24, 774–786. https://doi.org/10.1111/j.1096-0031.2008.00217.x
- Guindon, S., Dufayard, J.F., Lefort, V., Anisimova, M., Hordijk, W. & Gascuel, O. (2010) New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Systematic Biology, 59, 307–321. https://doi.org/10.1093/sysbio/syq010
- Hamada, N. & Couceiro, S.R.M. (2003) An illustrated key to nymphs of Perlidae (Insecta, Plecoptera) genera in Central Amazonia, Brazil. Revista Brasileira de Entomologia, 47, 477–480. https://doi.org/10.1590/S0085-56262003000300020
- Hammer, O., Harper, D.A. & Ryan, P.D. (2001) PAST: paleontological statistics software package for education and data analysis. Palaeontologia Electronica, 4, 1–9.
- Heckman, C.W. (2003) Encyclopedia of South American Aquatic Insects: Plecoptera: Illustrated Keys to Known Families, Genera, and Species in South America. Springer, Dordrecht, 329 pp. https://doi.org/10.1007/978-94-017-1423-5
- Illies, J. (1966) Katalog der rezenten Plecoptera. Das Tierreich, 82, 1–632.
- Kawai, T. (1968) Stoneflies (Plecoptera) from Thailand and India with descriptions of one new genus and two new species. Oriental Insects, 2 (2), 107–139. https://doi.org/10.1080/00305316.1968.10433877
- Kohno, M. (1944) On Niponiella limbatella Klapálek: with special reference to nymph. Japanese Journal of Entomology, 16 (3–4), 87–91.
- Kohno, M. (1946) Notes on two new genera of Plecoptera from Japan. Collecting and Breeding, 8 (3), 57–65.
- Lamine, S., Lounaci, A., Reding, J.P.G. & Vincon, G. (2019) Marthamea bayae, a new species of stonefly from Algeria (Plecoptera: Perlidae). Zootaxa, 4603 (2), 311–326. https://doi.org/10.11646/zootaxa.4603.2.5
- Letsch, H., Simon, S., Frandsen, P.B., Liu, S., Machida, R., Mayer, C., Misof, B., Niehuis, O., Zhou, X. & Wipfler, B. (2021) Combining molecular datasets with strongly heterogeneous taxon coverage enlightens the peculiar biogeographic history of stoneflies (Insecta: Plecoptera). Systematic Entomology, 46 (4), 952–967. https://doi.org/10.1111/syen.12505
- Minh, B.Q., Nguyen, M.A. & von Haeseler, A. (2013) Ultrafast approximation for phylogenetic bootstrap. Molecular Biology and Evolution, 30, 1188–1195. https://doi.org/10.1093/molbev/mst024
- Mo, R.R., Wang, G.Q., Li, W.H. & Muranyi, D. (2020) Review of the Oriental genus Neoperlops Banks, 1939 (Plecoptera: Perlidae). Zootaxa, 4763 (3), 405–418. https://doi.org/10.11646/zootaxa.4763.3.5
- Nguyen, L.T., Schmidt, H.A., von Haeseler, A. & Minh, B.Q. (2015) IQ-TREE: a fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies. Molecular Biology and Evolution, 32, 268–274. https://doi.org/10.1093/molbev/msu300
- Nixon, K.C. (2002) WinClada. Version 1.00.08. Computer software and documentation distributed by the Author, Ithaca, New York.
- Ribeiro, J.M.F. & Gorayeb, I.S. (2015) Description of immatures associated with adults of two species of Macrogynoplax Enderlein and one species of Enderleina Jewett (Plecoptera: Perlidae) of the Brazilian Amazon. Zootaxa, 3948 (1), 109–124. https://doi.org/10.11646/zootaxa.3948.1.7
- Rippel, M.L.S., Novaes, M.C. & Krolow, T.K. (2019) First records of Kempnyia and Macrogynoplax (Plecoptera: Perlidae) from Tocantins State, Brazil with description of the immatures and the adult female. Zootaxa, 4700 (4), 471–478. https://doi.org/10.11646/zootaxa.4700.4.5
- Ronquist, F., Teslenko, M., Mark, P.V.D., Ayres, D.L., Darling, A., Höhna, S., Larget, B., Liu, L., Suchard, M.A. & Huelsenbeck, J.P. (2012) MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology, 61, 539–542. https://doi.org/10.1093/sysbio/sys029
- Sánchez, K.N.C., Aguirre, Y.P., González, T.A.R. & Vega, J.A.B. (2019) The Plecoptera of Panama. IV. Description of the last nymphal instar for two species of Anacroneuria Klapálek (Plecoptera: Perlidae) from the Caldera River, Chiriquí, Panamá. Zootaxa, 4711 (3), 446–458. https://doi.org/10.11646/zootaxa.4711.3.2
- Shepard, W.D. & Stewart, K.W. (1983) Comparative study of nymphal gills in North American stonefly (Plecoptera) genera and a new, proposed paradigm of Plecoptera gill evolution. Miscellaneous Publications of the Entomological Society of America, 13, 1–57. https://doi.org/10.4182/JSYY2711.13-55.1
- Shi, G., Grimaldi, D.A., Harlow, G.E., Wang, J., Wang, J., Wang, M., Lei, W., Li, Q. & Li, X. (2012) Age constraint on Burmese amber based on U-Pb dating of zircons. Cretaceous Research, 37, 155–163. https://doi.org/10.1016/j.cretres.2012.03.014
- Sinitshenkova, N.D. (1987) Istoricheskoe razvitie vesnyanok [Historical development of stoneflies.]. Trudy Paleontologicheskogo Instituta Akedemii Nauk SSSR, 221, 1–144.
- Sivec, I. (1997) Helenoperla malickyi, a new genus of European Perlidae (Plecoptera) from Greece. In: Landolt, P. & Satori, M. (Eds.), Ephemeroptera & Plecoptera: Biology-Ecology-Systematics. MTL, Fribourg, pp. 205–208.
- Sivec, I. & Stark, B.P. (2010) Caroperla longiseta (Plecoptera: Perlidae), a new stonefly species from Thailand. Illiesia, 6 (7), 58–61.
- Sivec, I., Stark, B.P. & Uchida, S. (1988) Synopsis of the world genera of Perlinae (Plecoptera: Perlidae). Scopolia, 16, 1–66.
- South, E.J., Skinner, R.K., DeWalt, R.E., Kondratieff, B.C., Johnson, K.P., Davis, M.A., Lee, J.J. & Durfee, R.S. (2021) Phylogenomics of the North American Plecoptera. Systematic Entomology, 46 (1), 287–305. https://doi.org/10.1111/syen.12462
- Sroka, P., Staniczek, A.H. & Kondratieff, B.C. (2018) “Rolling” stoneflies (Insecta: Plecoptera) from mid-Cretaceous Burmese amber. PeerJ, 6, e5354. https://doi.org/10.7717/peerj.5354
- Stark, B.P. (1989) The genus Enderleina (Plecoptera: Perlidae). Aquatic Insects, 11 (3), 153–160. https://doi.org/10.1080/01650428909361364
- Stark, B.P. (2001) A synopsis of neotropical Perlidae (Plecoptera). In: Dominguez, E. (Ed.), Trends in Research in Ephemeroptera and Plecoptera. Kluwer Academic Plenum Publisher, New York, New York, pp. 405–422. https://doi.org/10.1007/978-1-4615-1257-8_48
- Stark, B.P. & Gaufin, A.R. (1976) The Nearctic genera of Perlidae (Plecoptera). Entomological Society of America, 10, 1–80. https://doi.org/10.4182/KWHH7779.10-1.1
- Stark, B.P. & Zwick, P. (1989) New species of Macrogynoplax from Venezuela and Surinam (Plecoptera: Perlidae). Aquatic Insects, 11 (4), 247–255. https://doi.org/10.1080/01650428909361378
- Stewart, K.W. & Stark, B.P. (2002) Nymphs of North American Stonefly Genera (Plecoptera). 2nd Edition. The Caddis Press, Columbus, Ohio, 510 pp.
- Terry, M.D. (2003) Phylogeny of the Polyneopterous Insects with Emphasis on Plecoptera: Molecular and Morphological Evidence. Brigham Young University, Provo, 118 pp.
- Teslenko, V.A. & Zhiltzova, L.A. (2009) Key to the Stoneflies (Insecta, Plecoptera) of Russia and Adjacent Countries. Imagines and Nymphs. Russian Academy of Sciences, Vladivostok, 381 pp.
- Uchida, S. (1990) A Revision of the Japanese Perlidae (Insecta: Plecoptera), with Special Reference to Their Phylogeny. Tokyo Metropolitan University, Tokyo, 228 pp.
- Walters, D.M., Ford, M.A. & Zuellig, R.E. (2017) A digital reference collection for aquatic macroinvertebrates of North America. Freshwater Science, 36 (4), 693–697. https://doi.org/10.1086/694539
- Wu, C.F. (1934) A homonym of a plecopterous genus. Annals of the Entomological Society of America, 27, 256. https://doi.org/10.1093/aesa/27.2.256
- Yu, T., Kelly, R., Mu, L., Ross, A., Kennedy, J., Broly, P., Xia, F., Zhang, H., Wang, B. & Dilcher, D. (2019) An ammonite trapped in Burmese amber. Proceedings of the National Academy of Sciences, 116, 11345–11350. https://doi.org/10.1073/pnas.1821292116
- Zwick, P. (1973) Insecta: Plecoptera Phylogenetisches System und Katalog. Das Tierreich, 94, 1–465.
- Zwick, P. (1980) Plecoptera (Steinfliegen). Handbuch der Zoologie, 4, 1–115.
- Zwick, P. (2000) Phylogenetic System and Zoogeography of the Plecoptera. Annual Review of Entomology, 45, 709–746. https://doi.org/10.1146/annurev.ento.45.1.709
- Zwick, P. (2004) Key to the West Palaearctic genera of stoneflies (Plecoptera) in the larval stage. Limnologica, 34 (4), 315–348. https://doi.org/10.1016/S0075-9511(04)80004-5
