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Integrative taxonomy reveals two sympatric species of the genus Eucriotettix Hebard, 1930 (Orthoptera: Tetrigidae)



Limno-terrestrial pygmy grasshoppers are mostly recognised using morphology but characters used are not always clear-cut for species identification. From a faunistic survey in Singapore, Eucriotettix were collected and two new species are identified and described here: E. guentheri sp. nov. and E. simulans sp. nov. This was done by iteratively incorporating morphological, molecular, morphometric, and ecological data. Mitochondrial COI trees, COI cluster analysis, microhabitat preference, and morphometric analyses provide strong enough indirect evidence that the specimens comprise a pair of species. A key to species of Eucriotettix from Singapore is also provided.


Scelimeninae, mitochondrial cytochrome c oxidase subunit I (COI), morphometric, microhabitat preference, taxonomy, new species, key, Singapore, Orthoptera

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Allegrucci, G., Massa, B., Trasatti, A. & Sbordoni, V. (2014) A taxonomic revision of western Eupholidoptera bush crickets (Orthoptera: Tettigoniidae): testing the discrimination power of DNA barcode. Systematic Entomology, 39 (1), 7–23.

Amédégnato, C. & Devriese, H. (2008) Global diversity of true and pygmy grasshoppers (Acridomorpha, Orthoptera) in freshwater. Hydrobiologia, 595 (1), 535–543.

Balian, E.V., Lévêque, C. & Segers, H. (2008) Freshwater Animal Diversity Assessment. Springer, Dordrecht, 656 pp.

Bensasson, D., Zhang, D.X. & Hewitt, G.M. (2000) Frequent assimilation of mitochondrial DNA by grasshopper nuclear genomes. Molecular Biology and Evolution, 17 (3), 406–415.

Bensasson, D., Zhang, D.X., Hartl, D.L. & Hewitt, G.M. (2001) Mitochondrial pseudogenes: evolution's misplaced witnesses. Trends in Ecology & Evolution, 16 (6), 314–321.

Blackith, R.E. (1992) Tetrigidae (Insecta: Orthoptera) of South-East Asia: Annotated catalogue with partial translated keys and bibliography. Ashford Co., JAPAGA, Rockbottom, 248 pp.

Blackith, R.E. & Blackith, R.M. (1987) Tridactylids and pygmy grasshoppers (Orthoptera) from Sulawesi, Indonesia. Tijschrift voor Entomologie, 130, 1–10.

Bonin, M.C., Srinivasan, M., Almany, G.R. & Jones, G.P. (2009) Interactive effects of interspecific competition and microhabitat on early post-settlement survival in a coral reef fish. Coral Reefs, 28 (1), 265–274.

Cigliano, M.M., Braun, H., Eades, D.C. & Otte, D. (2016) Orthoptera Species File Online. Version 5.0/5.0 Available from: (Accessed: 20 December 2016).

David, J.A.D.O., Zefa, E. & Fontanetti, C.S. (2003) Cryptic species of Gryllus in the light of bioacoustic (Orthoptera: Gryllidae). Neotropical Entomology, 32 (1), 75–80.

Deng, W.A., Zheng, Z.M., Li, X.D., Lin, M.P., Wei, S.Z., Yuan, B.D. & Lin, L.L. (2015) The groundhopper fauna (Orthoptera: Tetrigidae) of Shwanshan (Guangxi, China) with description of three new species. Zootaxa, 3925 (2), 151–178.

Deng, W.A., Zheng, Z.M., Yang, R.G., Guo, L.Y. & Zhou, P.N. (2012) Taxonomic Study of the Genus Eucriotettix Hebard (Orthoptera: Tetrigidea) from China. Neotropical entomology, 41 (6), 485–492.

Devriese, H. (1999) Revision des Xerophyllini d’Afrigue (Orthoptera, Tetrigidae). Belgian Journal of Entomology, 1 (1), 21–99.

Gellissen, G., Bradfield, J.Y., White, B.N. & Wyatt, G.R. (1983) Mitochondrial DNA sequences in the nuclear genome of a locust. Nature, 301 (5901), 631–634.

Günther, K. (1938) Revision der Acrydiinae (Orth.) II. Scelimenae spuriae. Stettiner Entomologische Zeitung, 99, 117–148, 161–230.

Günther, K. (1939) Revision der Acrydiinae (Orthoptera), III Sectio Amorphopi (Metrodorae Bol. 1887, auct.). Abhandlungen und Berichte aus den Staatlichen Museen für Tierkunde und Völkerkunde in Dresden, A20, Neue Folge, 1, 16–335.

Guo, H.F., Guan, B., Shi, F.M. & Zhou, Z.J. (2016) DNA Barcoding of genus Hexacentrus in China reveals cryptic diversity within Hexacentrus japonicus (Orthoptera, Tettigoniidae). ZooKeys, 596, 53.

Hancock, J.L. (1907) Orthoptera Fam. Acridiidae. Subfam. Tetriginae. In: Wytsman, P. (Ed.), Genera Insectorum, 48, 1–79.

Hebard, M. (1930) Acrydiinae (Orthoptera, Acrididae) of Southern India. Revue Suisse de Zoologie, 36 (19), 565–592.

Hebert, P.D., Cywinska, A. & Ball, S.L. (2003) Biological identifications through DNA barcodes. Proceedings of the Royal Society of London B: Biological Sciences, 270 (1512), 313–321.

Hebert, P.D., Penton, E.H., Burns, J.M., Janzen, D.H. & Hallwachs, W. (2004) Ten species in one: DNA barcoding reveals cryptic species in the neotropical skipper butterfly Astraptes fulgerator. Proceedings of the National Academy of Sciences of the United States of America, 101 (41), 14812–14817.

Hebert, P.D.N., Ratnasingham, S. & Dooh, R. (2004) Barcodes of Life. Available from: (accessed 10 March 2015)

Huang, J., Zhang, A., Mao, S. & Huang, Y. (2013) DNA barcoding and species boundary delimitation of selected species of Chinese Acridoidea (Orthoptera: Caelifera). PloS one, 8 (12), e82400.

Ichikawa, A. (1994) A revision of the family Tetrigidae (Orhoptera) of the Ryukyu Islands, Southern Japan, with descriptions of new species and subspecies (Part 1). Japanese Journal of Entomology, 62 (3), 457–474.

Ingrisch, S. (2001) Tetrigidae from Nepal in the Zoologische Staatssammlung München. Spixiana, 24 (2), 147–155.

Katoh, K. & Standley, D.M. (2013) MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Molecular biology and evolution, 30 (4), 772–780.

Kevan, D.K. M. (1966) Some Orthoptera-Caelifera from the Philippine, Bismarck and Solomon Islands, with a few interesting records from New Guinea and the Moluccas. Entomologiske Meddelelser, 34, 375–420.

Kim, T.W. & Pham, H.T. (2014) Checklist of Vietnamese Orthoptera (Saltatoria). Zootaxa, 3811 (1), 53–82.

Leray, M., Yang, J.Y., Meyer, C.P., Mills, S.C., Agudelo, N., Ranwez, V. Boehm, J.T. & Machida, R.J. (2013) A new versatile primer set targeting a short fragment of the mitochondrial COI region for metabarcoding metazoan diversity: application for characterizing coral reef fish gut contents. Frontiers in Zoology, 10 (1), 34.

Li, X.D., Deng, W.A., Zheng, Z.M. & Wei, S.Z. (2014) The tetrigid genus Eucriotettix Hebard (Orthoptera: Scelimeninae) from China with description of two new species. Journal of Natural History, 48 (33–34), 2083–2096.

Li, T.S., Zheng, Z.M. & Lu, W. (2000) A new species of Eucriotettix (Orthoptera: Scelimenidae) from Guangxi. Journal of Guangxi Agricultural and Biological Science, 19, 180–181. [in Chinese with English summary]

Liang, G.Q. & Zheng, Z.M. (1998) Fauna Sinica. Insecta. Vol. 12. Orthoptera Tetrigoidea. Science Press, Beijing, 278 pp. [in Chinese, with English summary]

Mahmood, K., Idris, A.B. & Salmah, Y. (2007) Tetrigidae (Orthoptera: Tetrigoidea) from Malaysia with the description of six new species. Acta Entomologica Sinica, 50 (12), 1272–1284.

Meier, R., Shiyang, K., Vaidya, G. & Ng, P.K. (2006) DNA barcoding and taxonomy in Diptera: a tale of high intraspecific variability and low identification success. Systematic biology, 55 (5), 715–728.

Meier, R., Wong, W., Srivathsan, A. & Foo, M. (2015) $1 DNA barcodes for reconstructing complex phenomes and finding rare species in specimen - rich samples. Cladistics, 32 (1), 100–110.

Mendelson, T.C. & Shaw, K.L. (2002) Genetic and behavioral components of the cryptic species boundary between Laupala cerasina and L. kohalensis (Orthoptera: Gryllidae). In: Genetics of Mate Choice: From Sexual Selection to Sexual Isolation. Springer, Dordrecht, pp. 301–310.

Moulton, M.J., Song, H. & Whiting, M.F. (2010) Assessing the effects of primer specificity on eliminating numt coamplification in DNA barcoding: a case study from Orthoptera (Arthropoda: Insecta). Molecular Ecology Resources, 10 (4), 615–627.

Ng, P.K.L. & Lim, K.K.P. (1992) The conservation status of the Nee Soon freshwater swamp forest of Singapore. Aquatic Conservation: Marine and Freshwater Ecosystems, 2 (3), 255–266.

Nylander, J.A.A. (2004) MrModeltest v2. Program distributed by the author. Evolutionary Biology Centre, Uppsala University. [software]

Otte, D. (1997) Orthoptera Species File 6. Tetrigoidea and Tridactyloidea (Orthoptera: Caelifera) and Addenda to OSF. Vols. 1-5. Orthopterists’ Society & Academy of Natural Sciences of Philadelphia, Philadelphia, 261 pp.

Padial, J.M., Miralles, A., De la Riva, I. & Vences, M. (2010) Review: The integrative future of taxonomy. Frontiers in Zoology, 7, 1–14.

Prosser, S.W., deWaard, J.R., Miller, S.E. & Hebert, P.D. (2015) DNA barcodes from century - old type specimens using next - generation sequencing. Molecular ecology resource, 16 (2), 487–497.

R Core Team (2016) R: a language and environment for statistical computing. Version 3.2. 2. Vienna: R Foundation for Statistical Computing. Available from: http://www. (accessed 10 April 2017)

Rehn, J.A.G. (1939) Studies in African Acrydiinae (Orthoptera, Acrididae) Part 2. New genera and species, and critical notes on previously-known forms. Proceedings of the Academy of Natural Sciences of Philadelphia, 90, 361–387.

Runkle, J.R. (1992) Guidelines and sample protocol for sampling forest gaps. Vol. 283. US Department of Agriculture, Forest Service, Pacific Northwest Research Station, Portland, 44 pp.

Schlick-Steiner, B.C., Steiner, F.M., Seifert, B., Stauffer, C., Christian, E. & Crozier, R.H. (2010) Integrative taxonomy: a multisource approach to exploring biodiversity. Annual Review of Entomology, 55, 421–438.

Shishodia, M.S. (1991) Taxonomy and zoogeography of the Tetrigidae (Orthoptera: Tetrigoidea) of North Eastern India. Records of the Zoological Survey of India. Occasional Paper, 140, 1–204.

Song, H., Buhay, J.E., Whiting, M.F. & Crandall, K.A. (2008) Many species in one: DNA barcoding overestimates the number of species when nuclear mitochondrial pseudogenes are coamplified. Proceedings of the National Academy of Sciences, 105 (36), 13486–13491.

Song, H., Moulton, M.J. & Whiting, M.F. (2014) Rampant nuclear insertion of mtDNA across diverse lineages within Orthoptera (Insecta). PloS one, 9 (10), e110508.

Song, H., Amédégnato, C., Cigliano, M.M., Desutter-Grandcolas, L., Heads, S.W., Huang, Y. & Whiting, M.F. (2015) 300 million years of diversification: elucidating the patterns of orthopteran evolution based on comprehensive taxon and gene sampling. Cladistics, 31 (6), 621–651.

Srivathsan, A. & Meier, R. (2012) On the inappropriate use of Kimura-2-parameter (K2P) divergences in the DNA-barcoding literature. Cladistics, 28 (2), 190–194.

Steinmann, H. (1970) Check-list of the Tetricidae (Orthoptera) of the Oriental faunal region. Acta Zoologica Academiae Scientiarum Hungaricae, 16 (1–2), 215–240.

Storozhenko, S. Yu. & Dawwrueng, P. (2015) New and little-known pygmy grasshoppers (Orthoptera: Tetrigidae) from Thailand. Zootaxa, 4052 (5), 527–554.

Swofford, D.L. (1990) PAUP: phylogenetic analysis using parsimony, version 3.0. Computer program distributed by Illinois Natural History Survey. [software]

Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. & Kumar, S. (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular biology and evolution, 28 (10), 2731–2739.

Tan, D.S.H., Ang, Y., Lim, G.S., Ismail, M.R., Meier, R. (2010) From ‘cryptic species’ to integrative taxonomy: an iterative process involving DNA sequences, morphology, and behaviour leads to the resurrection of Sepsis pyrrhosoma (Sepsidae: Diptera). Zoologica Scripta, 39, 51–61.

Tumbrinck, J. (2014) Taxonomic revision of the Cladonotinae (Orthoptera: Tetrigidae) from the islands of South-East Asia and from Australia, with general remarks to the classification and morphology of the Tetrigidae and descriptions of new genera and species from New Guinea and New Caledonia. In: Telnov, D. (Ed.), Biodiversity, Biogeography and Nature Conservation in Wallacea and New Guinea. Vol. II. the Entomological Society of Latvia, RHga, pp. 350–396, pls. 64–91.

Walker, T.J. (1964) Cryptic species among sound-producing ensiferan Orthoptera (Gryllidae and Tettigoniidae). Quarterly Review of Biology, 39 (4), 345–355.

Will-Wolf, S., Esseen, P.A. & Neitlich, P. (2002) Monitoring biodiversity and ecosystem function: forests. In: Monitoring with lichens – Monitoring lichens. Springer, Dordrecht, pp. 203–222.

Zheng, Z.M. (2005) Fauna of Tetrigoidea from Western China. Science Press, Beijing, 501 pp. [in Chinese with English summary]

Zheng, Z.M. & Ou, X.H. (2011) Four new species of Tetrigoidea (Orthoptera) from Western Area in Yunnan Province. Entomotaxonomia, 33 (3), 168–174. [in Chinese with English summary]

Zwickl, D.J. (2006) Genetic algorithm approaches for the phylogenetic analysis of large bioloigcal sequence datasets under the maximum likelihood criterion. Ph. D. dissertation, The University of Texas at Austin, Austin, Texas, 115 pp.



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