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RESEARCH ARTICLE

Integrative species delimitation reveals fine-scale allopatric speciation in a good-flying insect: a case study on Cylindera pseudocylindriformis complex (Coleoptera, Cicindelidae)

Ming-Hsun Chou https://orcid.org/0000-0002-7426-0257 A * , I-Hsuan Chu B , Daniel Lau B and Jen-Pan Huang https://orcid.org/0000-0002-9329-8867 A *
+ Author Affiliations
- Author Affiliations

A Biodiversity Research Center, Academia Sinica, Taipei, Taiwan.

B Department of Entomology, National Chung Hsing University, Taichung, Taiwan.


Handling Editor: Bruno de Medeiros

Invertebrate Systematics 36(10) 910-925 https://doi.org/10.1071/IS22011
Submitted: 14 February 2022  Accepted: 2 August 2022   Published: 6 October 2022

© 2022 The Author(s) (or their employer(s)). Published by CSIRO Publishing.

Abstract

Alpha taxonomy is fundamental for many biological fields. Delineation of the species boundary, however, can be challenging in a species complex, where different species share a similar morphology and diagnostic characters may not be available. In this context, integrative approaches that incorporate molecular and morphological data sets, and account for speciation history can be helpful to alpha taxonomy. Different approaches to species delimitation based on different assumptions are complementary and by integrating the results from multiple approaches we can generate a more reliable and objective taxonomic decision. In this study, we applied three molecular approaches to species delimitation and inferred the demographic history based on an isolation with migration model to test a morphologically based taxonomic hypothesis for the Cylindera pseudocylindriformis complex. We discuss the association between genetic divergence and microhabitat specialisation, and further corroborate that C. subtilis sp. nov. is a valid new species by integrating the results from model-based species delimitation and the genealogical divergence index. We argue that genetic endemism can occur at a small geographic scale, even in a winged insect like tiger beetles. Our results also indicated that there may still be undocumented species diversity of Taiwanese Cylindera remaining to be discovered.

ZooBank LSID: urn:lsid:zoobank.org:pub:9DEC1432-365C-4872-8D06-73B95F30624F

Keywords: alpha taxonomy, cryptic diversity, dispersal, GDI, genealogical divergence index, microhabitat, Taiwan, tiger beetle, unified species concept.


References

Andriamampianina, L, Kremen, C, Vane-Wright, D, Lees, D, and Razafimahatratra, V (2000). Taxic richness patterns and conservation of Madagascar tiger beetles (Coleoptera: Cicindelidae). Journal of Insect Conservation 4, 109–128.
Taxic richness patterns and conservation of Madagascar tiger beetles (Coleoptera: Cicindelidae).Crossref | GoogleScholarGoogle Scholar |

Ballesteros, JA, and Hormiga, G (2018). Species delimitation of the North American orchard-spider Leucauge venusta (Walckenaer, 1841) (Araneae, Tetragnathidae). Molecular Phylogenetics and Evolution 121, 183–197.
Species delimitation of the North American orchard-spider Leucauge venusta (Walckenaer, 1841) (Araneae, Tetragnathidae).Crossref | GoogleScholarGoogle Scholar |

Barley, AJ, Brown, JM, and Thomson, RC (2018). Impact of model violations on the inference of species boundaries under the multispecies coalescent. Systematic Biology 67, 269–284.
Impact of model violations on the inference of species boundaries under the multispecies coalescent.Crossref | GoogleScholarGoogle Scholar |

Benham, PM, and Cheviron, ZA (2019). Divergent mitochondrial lineages arose within a large, panmictic population of the Savannah sparrow (Passerculus sandwichensis). Molecular Ecology 28, 1765–1783.
Divergent mitochondrial lineages arose within a large, panmictic population of the Savannah sparrow (Passerculus sandwichensis).Crossref | GoogleScholarGoogle Scholar |

Bickford, D, Lohman, DJ, Sodhi, NS, Ng, PK, Meier, R, Winker, K, Ingram, KK, and Das, I (2007). Cryptic species as a window on diversity and conservation. Trends in Ecology & Evolution 22, 148–155.
Cryptic species as a window on diversity and conservation.Crossref | GoogleScholarGoogle Scholar |

Bouckaert, R, Vaughan, TG, Barido-Sottani, J, Duchêne, S, Fourment, M, Gavryushkina, A, Heled, J, Jones, G, Kühnert, D, De Maio, N, Matschiner, M, Mendes, FK, Müller, NF, Ogilvie, HA, Du Plessis, L, Popinga, A, Rambaut, A, Rasmussen, D, Siveroni, I, Suchard, MA, Wu, CH, Xie, D, Zhang, C, Stadler, T, and Drummond, AJ (2019). BEAST 2.5: an advanced software platform for Bayesian evolutionary analysis. PLoS Computational Biology 15, e1006650.
BEAST 2.5: an advanced software platform for Bayesian evolutionary analysis.Crossref | GoogleScholarGoogle Scholar |

Brown, DM, Brenneman, RA, Koepfli, KP, Pollinger, JP, Milá, B, Georgiadis, NJ, Louis Jr, EE, Grether, GF, Jacobs, DK, and Wayne, RK (2007). Extensive population genetic structure in the giraffe. BMC Biology 5, 57.
Extensive population genetic structure in the giraffe.Crossref | GoogleScholarGoogle Scholar |

Camargo, A, Morando, M, Avila, LJ, and Sites Jr, JW (2012). Species delimitation with ABC and other coalescent‐based methods: A test of accuracy with simulations and an empirical example with lizards of the Liolaemus darwinii complex (Squamata: Liolaemidae). Evolution: International Journal of Organic Evolution 66, 2834–2849.
Species delimitation with ABC and other coalescent‐based methods: A test of accuracy with simulations and an empirical example with lizards of the Liolaemus darwinii complex (Squamata: Liolaemidae).Crossref | GoogleScholarGoogle Scholar |

Cardoso, A, Serrano, A, and Vogler, AP (2009). Morphological and molecular variation in tiger beetles of the Cicindela hybrida complex: is an ‘integrative taxonomy’possible? Molecular Ecology 18, 648–664.
Morphological and molecular variation in tiger beetles of the Cicindela hybrida complex: is an ‘integrative taxonomy’possible?Crossref | GoogleScholarGoogle Scholar |

Carroll, SS, and Pearson, DL (1998). Spatial modeling of butterfly species richness using tiger beetles (Cicindelidae) as a bioindicator taxon. Ecological Applications 8, 531–543.
Spatial modeling of butterfly species richness using tiger beetles (Cicindelidae) as a bioindicator taxon.Crossref | GoogleScholarGoogle Scholar |

Carstens, BC, Pelletier, TA, Reid, NM, and Satler, JD (2013). How to fail at species delimitation. Molecular Ecology 22, 4369–4383.
How to fail at species delimitation.Crossref | GoogleScholarGoogle Scholar |

Cassola, F (2002). Studies on tiger beetles. CXXX. On four presumed “Prothyma” species from China and the Oriental region (Coleoptera: Cicindelidae). Zeitschrift der Arbeitsgemeinschaft Österreichischer Entomologen 54, 87–94.

Cassola, F, and Pearson, DL (2000). Global patterns of tiger beetle species richness (Coleoptera: Cicindelidae): their use in conservation planning. Biological Conservation 95, 197–208.
Global patterns of tiger beetle species richness (Coleoptera: Cicindelidae): their use in conservation planning.Crossref | GoogleScholarGoogle Scholar |

Chambers, EA, and Hillis, DM (2020). The multispecies coalescent over-splits species in the case of geographically widespread taxa. Systematic Biology 69, 184–193.
The multispecies coalescent over-splits species in the case of geographically widespread taxa.Crossref | GoogleScholarGoogle Scholar |

Chan, KO, Hutter, CR, Wood Jr, PL, Grismer, LL, Das, I, and Brown, RM (2020). Gene flow creates a mirage of cryptic species in a Southeast Asian spotted stream frog complex. Molecular Ecology 29, 3970–3987.
Gene flow creates a mirage of cryptic species in a Southeast Asian spotted stream frog complex.Crossref | GoogleScholarGoogle Scholar |

Chou, M-H, and Yeh, W-B (2019). Delineation of two new, highly similar species of Taiwanese Cylindera tiger beetles (Coleoptera, Carabidae, Cicindelinae) based on morphological and molecular evidence. ZooKeys 875, 31–62.
Delineation of two new, highly similar species of Taiwanese Cylindera tiger beetles (Coleoptera, Carabidae, Cicindelinae) based on morphological and molecular evidence.Crossref | GoogleScholarGoogle Scholar |

Chou, MH, Tseng, WZ, Sang, YD, Morgan, B, De Vivo, M, Kuan, YH, Wang, LJ, Chen, WY, and Huang, JP (2021). Incipient speciation and its impact on taxonomic decision: a case study using a sky island sister-species pair of stag beetles (Lucanidae: Lucanus). Biological Journal of the Linnean Society 134, 745–759.
Incipient speciation and its impact on taxonomic decision: a case study using a sky island sister-species pair of stag beetles (Lucanidae: Lucanus).Crossref | GoogleScholarGoogle Scholar |

Cicero, C, Mason, NA, Jiménez, RA, Wait, DR, Wang-Claypool, CY, and Bowie, RCK (2021). Integrative taxonomy and geographic sampling underlie successful species delimitation. Ornithology 138, ukab009.
Integrative taxonomy and geographic sampling underlie successful species delimitation.Crossref | GoogleScholarGoogle Scholar |

Cook, LG, Edwards, RD, Crisp, MD, and Hardy, NB (2010). Need morphology always be required for new species descriptions. Invertebrate Systematics 24, 322–326.
Need morphology always be required for new species descriptions.Crossref | GoogleScholarGoogle Scholar |

Darriba, D, Taboada, GL, Doallo, R, and Posada, D (2012). jModelTest 2: more models, new heuristics and parallel computing. Nature Methods 9, 772.
jModelTest 2: more models, new heuristics and parallel computing.Crossref | GoogleScholarGoogle Scholar |

Dasmahapatra, KK, Elias, M, Hill, RI, Hoffman, JI, and Mallet, J (2010). Mitochondrial DNA barcoding detects some species that are real, and some that are not. Molecular Ecology Resources 10, 264–273.
Mitochondrial DNA barcoding detects some species that are real, and some that are not.Crossref | GoogleScholarGoogle Scholar |

Dayrat, B (2005). Towards integrative taxonomy. Biological Journal of the Linnean Society 85, 407–415.
Towards integrative taxonomy.Crossref | GoogleScholarGoogle Scholar |

De Queiroz, K (2007). Species concepts and species delimitation. Systematic Biology 56, 879–886.
Species concepts and species delimitation.Crossref | GoogleScholarGoogle Scholar |

Després, L (2019). One, two or more species? Mitonuclear discordance and species delimitation. Molecular Ecology 28, 3845–3847.
One, two or more species? Mitonuclear discordance and species delimitation.Crossref | GoogleScholarGoogle Scholar |

Diniz-Filho, JAF, Soares, TN, Lima, JS, Dobrovolski, R, Landeiro, VL, de Campos Telles, MP, Rangel, TF, and Bini, LM (2013). Mantel test in population genetics. Genetics and Molecular Biology 36, 475–485.
Mantel test in population genetics.Crossref | GoogleScholarGoogle Scholar |

Dong, F, Hung, CM, Li, XL, Gao, JY, Zhang, Q, Wu, F, Lei, FM, Li, SH, and Yang, XJ (2017). Ice age unfrozen: severe effect of the last interglacial, not glacial, climate change on East Asian avifauna. BMC Evolutionary Biology 17, 244.
Ice age unfrozen: severe effect of the last interglacial, not glacial, climate change on East Asian avifauna.Crossref | GoogleScholarGoogle Scholar |

Dupuis, JR, Roe, AD, and Sperling, FAH (2012). Multi‐locus species delimitation in closely related animals and fungi: one marker is not enough. Molecular Ecology 21, 4422–4436.
Multi‐locus species delimitation in closely related animals and fungi: one marker is not enough.Crossref | GoogleScholarGoogle Scholar |

Duran, DP, Herrmann, DP, Roman, SJ, Gwiazdowski, RA, Drummond, JA, Hood, GR, and Egan, SP (2019). Cryptic diversity in the North American Dromochorus tiger beetles (Coleoptera: Carabidae: Cicindelinae): a congruence-based method for species discovery. Zoological Journal of the Linnean Society 186, 250–285.
Cryptic diversity in the North American Dromochorus tiger beetles (Coleoptera: Carabidae: Cicindelinae): a congruence-based method for species discovery.Crossref | GoogleScholarGoogle Scholar |

Eberle, J, Bazzato, E, Fabrizi, S, Rossini, M, Colomba, M, Cillo, D, Uliana, M, Sparacio, I, Sabatinelli, G, Warnock, RCM, Carpaneto, G, and Ahrens, D (2019). Sex-biased dispersal obscures species boundaries in integrative species delimitation approaches. Systematic Biology 68, 441–459.
Sex-biased dispersal obscures species boundaries in integrative species delimitation approaches.Crossref | GoogleScholarGoogle Scholar |

Ence, DD, and Carstens, BC (2011). SpedeSTEM: a rapid and accurate method for species delimitation. Molecular Ecology Resources 11, 473–480.
SpedeSTEM: a rapid and accurate method for species delimitation.Crossref | GoogleScholarGoogle Scholar |

Fišer, C, Robinson, CT, and Malard, F (2018). Cryptic species as a window into the paradigm shift of the species concept. Molecular Ecology 27, 613–635.
Cryptic species as a window into the paradigm shift of the species concept.Crossref | GoogleScholarGoogle Scholar |

Flouri, T, Jiao, X, Rannala, B, and Yang, Z (2018). Species tree inference with BPP using genomic sequences and the multispecies coalescent. Molecular Biology and Evolution 35, 2585–2593.
Species tree inference with BPP using genomic sequences and the multispecies coalescent.Crossref | GoogleScholarGoogle Scholar |

Folmer, O, Black, M, Hoeh, W, Lutz, R, and Vrijenhoek, R (1994). DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology 3, 294–299.

Fujita, MK, Leaché, AD, Burbrink, FT, McGuire, JA, and Moritz, C (2012). Coalescent-based species delimitation in an integrative taxonomy. Trends in Ecology & Evolution 27, 480–488.
Coalescent-based species delimitation in an integrative taxonomy.Crossref | GoogleScholarGoogle Scholar |

Giska, I, Sechi, P, and Babik, W (2015). Deeply divergent sympatric mitochondrial lineages of the earthworm Lumbricus rubellus are not reproductively isolated. BMC Evolutionary Biology 15, 217.
Deeply divergent sympatric mitochondrial lineages of the earthworm Lumbricus rubellus are not reproductively isolated.Crossref | GoogleScholarGoogle Scholar |

Hall, TA (1999). BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41, 95–98.

Hebert, PD, Ratnasingham, S, and de Waard, JR (2003a). Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species. Proceedings of the Royal Society of London – B. Biological Sciences 270, S96–S99.
Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species.Crossref | GoogleScholarGoogle Scholar |

Hebert, PD, Cywinska, A, and Ball, SL (2003b). Biological identifications through DNA barcodes. Proceedings of the Royal Society of London – B. Biological Sciences 270, 313–321.
Biological identifications through DNA barcodes.Crossref | GoogleScholarGoogle Scholar |

Hebert, PDN, Penton, EH, Burns, JM, Janzen, DH, and 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 101, 14812–14817.
Ten species in one: DNA barcoding reveals cryptic species in the neotropical skipper butterfly Astraptes fulgerator.Crossref | GoogleScholarGoogle Scholar |

Hey, J (2010). The divergence of chimpanzee species and subspecies as revealed in multipopulation isolation-with-migration analyses. Molecular Biology and Evolution 27, 921–933.
The divergence of chimpanzee species and subspecies as revealed in multipopulation isolation-with-migration analyses.Crossref | GoogleScholarGoogle Scholar |

Hillis, DM (2019). Species delimitation in herpetology. Journal of Herpetology 53, 3–12.
Species delimitation in herpetology.Crossref | GoogleScholarGoogle Scholar |

Hinojosa, JC, Dapporto, L, Brockmann, E, Dincă, V, Tikhonov, V, Grishin, N, Lukhtanov, VA, and Vila, R (2021). Overlooked cryptic diversity in Muschampia (Lepidoptera: Hesperiidae) adds two species to the European butterfly fauna. Zoological Journal of the Linnean Society 193, 847–859.
Overlooked cryptic diversity in Muschampia (Lepidoptera: Hesperiidae) adds two species to the European butterfly fauna.Crossref | GoogleScholarGoogle Scholar |

Hogner, S, Laskemoen, T, Lifjeld, JT, Porkert, J, Kleven, O, Albayrak, T, Kabasakal, B, and Johnsen, A (2012). Deep sympatric mitochondrial divergence without reproductive isolation in the common redstart Phoenicurus phoenicurus. Ecology and Evolution 2, 2974–2988.
Deep sympatric mitochondrial divergence without reproductive isolation in the common redstart Phoenicurus phoenicurus.Crossref | GoogleScholarGoogle Scholar |

Horn, W (1913). 50 neue Cicindelinae. Archiv für Naturgeschichte 79, 1–33.

Hu, GL, Gao, K, Wang, J-S, Hebert, PDN, and Hua, B-Z (2019). Molecular phylogeny and species delimitation of the genus Dicerapanorpa (Mecoptera: Panorpidae). Zoological Journal of the Linnean Society 187, 1173–1195.
Molecular phylogeny and species delimitation of the genus Dicerapanorpa (Mecoptera: Panorpidae).Crossref | GoogleScholarGoogle Scholar |

Huang, J-P (2018). What have been and what can be delimited as species using molecular data under the multi-species coalescent model? A case study using Hercules beetles (Dynastes; Dynastidae). Insect Systematics and Diversity 2, 3.
What have been and what can be delimited as species using molecular data under the multi-species coalescent model? A case study using Hercules beetles (Dynastes; Dynastidae).Crossref | GoogleScholarGoogle Scholar |

Huang, J-P (2020). Is population subdivision different from speciation? From phylogeography to species delimitation. Ecology and Evolution 10, 6890–6896.
Is population subdivision different from speciation? From phylogeography to species delimitation.Crossref | GoogleScholarGoogle Scholar |

Huang, J-P (2021). The genealogical divergence index across a speciation continuum in Hercules beetles. Insect Systematics and Diversity 5, 1.
The genealogical divergence index across a speciation continuum in Hercules beetles.Crossref | GoogleScholarGoogle Scholar |

Huang, J-P, and Knowles, LL (2016). The species versus subspecies conundrum: quantitative delimitation from integrating multiple data types within a single Bayesian approach in Hercules beetles. Systematic Biology 65, 685–699.
The species versus subspecies conundrum: quantitative delimitation from integrating multiple data types within a single Bayesian approach in Hercules beetles.Crossref | GoogleScholarGoogle Scholar |

Huang, J, Bennett, J, Flouri, T, Leaché, AD, and Yang, Z (2022). Phase resolution of heterozygous sites in diploid genomes is important to phylogenomic analysis under the multispecies coalescent model. Systematic Biology 71, 334–352.
Phase resolution of heterozygous sites in diploid genomes is important to phylogenomic analysis under the multispecies coalescent model.Crossref | GoogleScholarGoogle Scholar |

Jackson, ND, Carstens, BC, Morales, AE, and O’Meara, BC (2017). Species delimitation with gene flow. Systematic Biology 66, 799–812.
Species delimitation with gene flow.Crossref | GoogleScholarGoogle Scholar |

Jaskuła, R (2011). How unique is the tiger beetle fauna (Coleoptera Cicindelidae) of the Balkan Peninsula? ZooKeys 100, 487–502.
How unique is the tiger beetle fauna (Coleoptera Cicindelidae) of the Balkan Peninsula?Crossref | GoogleScholarGoogle Scholar |

Jaskuła, R (2015). The Maghreb – one more important biodiversity hot spot for tiger beetle fauna (Coleoptera, Carabidae, Cicindelinae) in the Mediterranean region. ZooKeys 482, 35–53.
The Maghreb – one more important biodiversity hot spot for tiger beetle fauna (Coleoptera, Carabidae, Cicindelinae) in the Mediterranean region.Crossref | GoogleScholarGoogle Scholar |

Jaskuła, R, and Płóciennik, M (2020). Water is needed to exist: Habitat preferences of tiger beetles (Coleoptera: Cicindelidae) in a desert country. Insects 11, 809.
Water is needed to exist: Habitat preferences of tiger beetles (Coleoptera: Cicindelidae) in a desert country.Crossref | GoogleScholarGoogle Scholar |

Jaskuła, R, Rewicz, T, Płóciennik, M, and Grabowski, M (2016). Pleistocene phylogeography and cryptic diversity of a tiger beetle, Calomera littoralis, in North-Eastern Mediterranean and Pontic regions inferred from mitochondrial COI gene sequences. PeerJ 4, e2128.
Pleistocene phylogeography and cryptic diversity of a tiger beetle, Calomera littoralis, in North-Eastern Mediterranean and Pontic regions inferred from mitochondrial COI gene sequences.Crossref | GoogleScholarGoogle Scholar |

Jaskuła, R, Płóciennik, M, and Schwerk, A (2019). From climate zone to microhabitat—environmental factors affecting the coastal distribution of tiger beetles (Coleoptera: Cicindelidae) in the south-eastern European biodiversity hotspot. PeerJ 7, e6676.
From climate zone to microhabitat—environmental factors affecting the coastal distribution of tiger beetles (Coleoptera: Cicindelidae) in the south-eastern European biodiversity hotspot.Crossref | GoogleScholarGoogle Scholar |

Jones, G (2017). Algorithmic improvements to species delimitation and phylogeny estimation under the multispecies coalescent. Journal of Mathematical Biology 74, 447–467.
Algorithmic improvements to species delimitation and phylogeny estimation under the multispecies coalescent.Crossref | GoogleScholarGoogle Scholar |

Jones, G, Aydin, Z, and Oxelman, B (2015). DISSECT: an assignment-free Bayesian discovery method for species delimitation under the multispecies coalescent. Bioinformatics 31, 991–998.
DISSECT: an assignment-free Bayesian discovery method for species delimitation under the multispecies coalescent.Crossref | GoogleScholarGoogle Scholar |

Jörger, KM, and Schrödl, M (2013). How to describe a cryptic species? Practical challenges of molecular taxonomy. Frontiers in Zoology 10, 59.
How to describe a cryptic species? Practical challenges of molecular taxonomy.Crossref | GoogleScholarGoogle Scholar |

Korshunova, T, Picton, B, Furfaro, G, Mariottini, P, Pontes, M, Prkić, J, Fletcher, K, Malmberg, K, Lundin, K, and Martynov, A (2019). Multilevel fine-scale diversity challenges the ‘cryptic species’ concept. Scientific Reports 9, 6732.
Multilevel fine-scale diversity challenges the ‘cryptic species’ concept.Crossref | GoogleScholarGoogle Scholar |

Kuchta, SR, Brown, AD, and Highton, R (2018). Disintegrating over space and time: Paraphyly and species delimitation in the Wehrle’s salamander complex. Zoologica Scripta 47, 285–299.
Disintegrating over space and time: Paraphyly and species delimitation in the Wehrle’s salamander complex.Crossref | GoogleScholarGoogle Scholar |

Leaché, AD, Zhu, T, Rannala, B, and Yang, Z (2019). The spectre of too many species. Systematic Biology 68, 168–181.
The spectre of too many species.Crossref | GoogleScholarGoogle Scholar |

Lee, KH, Shaner, PJL, Lin, YP, and Lin, SM (2016). Geographic variation in advertisement calls of a Microhylid frog – testing the role of drift and ecology. Ecology and Evolution 6, 3289–3298.
Geographic variation in advertisement calls of a Microhylid frog – testing the role of drift and ecology.Crossref | GoogleScholarGoogle Scholar |

Lin, TJ (2017). A new species of the tiger beetle (Coleoptera: Cicindelidae) from Taiwan. Journal of Experience Forest. National Taiwan University 22, 177–185.

Lin, TJ, and Acciavatti, RE (2019). Description of Cylindera (Cylindera) triangulata, sp. nov., and Its Comparison with Cylindera (Cylindera) cylindriformis (Horn, 1912) (Coleoptera: Cicindelidae) in Taiwan. Journal of Experience Forest. National Taiwan University 33, 295–310.

Lin, JS, Yeh, W-B, and Wang, CL (2003). Molecular identification of multiplex-PCR and PCR-RFLP for the quarantine pest, Frankliniella occidentalis (Pergande). Formosan Entomologist 23, 353–366.
Molecular identification of multiplex-PCR and PCR-RFLP for the quarantine pest, Frankliniella occidentalis (Pergande).Crossref | GoogleScholarGoogle Scholar |

Lin, HD, Chen, YR, and Lin, SM (2012). Strict consistency between genetic and topographic landscapes of the brown tree frog (Buergeria robusta) in Taiwan. Molecular Phylogenetics and Evolution 62, 251–262.
Strict consistency between genetic and topographic landscapes of the brown tree frog (Buergeria robusta) in Taiwan.Crossref | GoogleScholarGoogle Scholar |

López‐López, A, Hudson, P, and Galián, J (2012). The blackburni/murchisona species complex in Australian Pseudotetracha (Coleoptera: Carabidae: Cicindelinae: Megacephalini): evaluating molecular and karyological evidence. Journal of Zoological Systematics and Evolutionary Research 50, 177–183.
The blackburni/murchisona species complex in Australian Pseudotetracha (Coleoptera: Carabidae: Cicindelinae: Megacephalini): evaluating molecular and karyological evidence.Crossref | GoogleScholarGoogle Scholar |

López-López, A, Hudson, P, and Galián, J (2016). Islands in the desert: species delimitation and evolutionary history of Pseudotetracha tiger beetles (Coleoptera: Cicindelidae: Megacephalini) from Australian salt lakes. Molecular Phylogenetics and Evolution 101, 279–285.
Islands in the desert: species delimitation and evolutionary history of Pseudotetracha tiger beetles (Coleoptera: Cicindelidae: Megacephalini) from Australian salt lakes.Crossref | GoogleScholarGoogle Scholar |

Makhov, IA, Gorodilova, YYU, and Lukhtanov, VA (2021). Sympatric occurrence of deeply diverged mitochondrial DNA lineages in Siberian geometrid moths (Lepidoptera: Geometridae): cryptic speciation, mitochondrial introgression, secondary admixture or effect of Wolbachia? Biological Journal of the Linnean Society 134, 342–365.
Sympatric occurrence of deeply diverged mitochondrial DNA lineages in Siberian geometrid moths (Lepidoptera: Geometridae): cryptic speciation, mitochondrial introgression, secondary admixture or effect of Wolbachia?Crossref | GoogleScholarGoogle Scholar |

Mason, NA, Fletcher, NK, Gill, BA, Funk, WC, and Zamudio, KR (2020). Coalescent-based species delimitation is sensitive to geographic sampling and isolation by distance. Systematics and Biodiversity 18, 269–280.
Coalescent-based species delimitation is sensitive to geographic sampling and isolation by distance.Crossref | GoogleScholarGoogle Scholar |

Morgan, B, and Huang, JP (2021). Isolation by geographical distance after release from Pleistocene refugia explains genetic and phenotypic variation in Xylotrupes siamensis (Coleoptera: Scarabaeidae). Zoological Journal of the Linnean Society 192, 117–129.
Isolation by geographical distance after release from Pleistocene refugia explains genetic and phenotypic variation in Xylotrupes siamensis (Coleoptera: Scarabaeidae).Crossref | GoogleScholarGoogle Scholar |

Newton, LG, Starrett, J, Hendrixson, BE, Derkarabetian, S, and Bond, JE (2020). Integrative species delimitation reveals cryptic diversity in the southern Appalachian Antrodiaetus unicolor (Araneae: Antrodiaetidae) species complex. Molecular Ecology 29, 2269–2287.
Integrative species delimitation reveals cryptic diversity in the southern Appalachian Antrodiaetus unicolor (Araneae: Antrodiaetidae) species complex.Crossref | GoogleScholarGoogle Scholar |

Osozawa, S, Fukuda, H, Kwon, HY, and Wakabayashi, J (2016). Quaternary vicariance of tiger beetle, Cicindela chinensis, in Ryukyu, Japan, Taiwan and Korea–China. Entomological Research 46, 122–127.
Quaternary vicariance of tiger beetle, Cicindela chinensis, in Ryukyu, Japan, Taiwan and Korea–China.Crossref | GoogleScholarGoogle Scholar |

Pante, E, Schoelinck, C, and Puillandre, N (2015). From integrative taxonomy to species description: one step beyond. Systematic Biology 64, 152–160.
From integrative taxonomy to species description: one step beyond.Crossref | GoogleScholarGoogle Scholar |

Patterson, DJ, Cooper, J, Kirk, PM, Pyle, RL, and Remsen, DP (2010). Names are key to the big new biology. Trends in Ecology & Evolution 25, 686–691.
Names are key to the big new biology.Crossref | GoogleScholarGoogle Scholar |

Pazhenkova, EA, and Lukhtanov, VA (2019). Nuclear genes (but not mitochondrial DNA barcodes) reveal real species: evidence from the Brenthis fritillary butterflies (Lepidoptera, Nymphalidae). Journal of Zoological Systematics and Evolutionary Research 57, 298–313.
Nuclear genes (but not mitochondrial DNA barcodes) reveal real species: evidence from the Brenthis fritillary butterflies (Lepidoptera, Nymphalidae).Crossref | GoogleScholarGoogle Scholar |

Pearson, DL (1988). Biology of tiger beetles. Annual Review of Entomology 33, 123–147.
Biology of tiger beetles.Crossref | GoogleScholarGoogle Scholar |

Pearson, DL, and Carroll, SS (1998). Global patterns of species richness: spatial models for conservation planning using bioindicator and precipitation data. Conservation Biology 12, 809–821.

Pearson, DL, and Ghorpade, K (1989). Geographical distribution and ecological history of tiger beetles (Coleoptera: Cicindelidae) of the Indian subcontinent. Journal of Biogeography 16, 333–344.
Geographical distribution and ecological history of tiger beetles (Coleoptera: Cicindelidae) of the Indian subcontinent.Crossref | GoogleScholarGoogle Scholar |

Pinto, BJ, Colli, GR, Higham, TE, Russell, AP, Scantlebury, DP, Vitt, LJ, and Gamble, T (2019). Population genetic structure and species delimitation of a widespread, Neotropical dwarf gecko. Molecular Phylogenetics and Evolution 133, 54–66.
Population genetic structure and species delimitation of a widespread, Neotropical dwarf gecko.Crossref | GoogleScholarGoogle Scholar |

Rambaut, A, Drummond, AJ, Xie, D, Baele, G, and Suchard, MA (2018). Posterior summarization in Bayesian phylogenetics using Tracer 1.7. Systematic Biology 67, 901–904.
Posterior summarization in Bayesian phylogenetics using Tracer 1.7.Crossref | GoogleScholarGoogle Scholar |

Ronquist, F, Teslenko, M, van der Mark, P, Ayres, DL, Darling, A, Höhna, S, Larget, B, Liu, L, Suchard, MA, and Huelsenbeck, JP (2012). MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology 61, 539–542.
MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space.Crossref | GoogleScholarGoogle Scholar |

Rowson, B, Anderson, R, Turner, JA, and Symondson, WOC (2014). The slugs of Britain and Ireland: undetected and undescribed species increase a well-studied, economically important fauna by more than 20%. PLoS One 9, e91907.
The slugs of Britain and Ireland: undetected and undescribed species increase a well-studied, economically important fauna by more than 20%.Crossref | GoogleScholarGoogle Scholar |

Satoh, A, and Hori, M (2005). Microhabitat segregation in larvae of six species of coastal tiger beetles in Japan. Ecological Research 20, 143–149.
Microhabitat segregation in larvae of six species of coastal tiger beetles in Japan.Crossref | GoogleScholarGoogle Scholar |

Satoh, A, Sota, T, Uéda, T, Enokido, Y, Paik, JC, and Hori, M (2004). Evolutionary history of coastal tiger beetles in Japan based on a comparative phylogeography of four species. Molecular Ecology 13, 3057–3069.
Evolutionary history of coastal tiger beetles in Japan based on a comparative phylogeography of four species.Crossref | GoogleScholarGoogle Scholar |

Satoh, A, Uéda, T, Ichion, E, and Hori, M (2006). Distribution and habitat of three species of riparian tiger beetle in the Tedori River System of Japan. Environmental Entomology 35, 320–325.
Distribution and habitat of three species of riparian tiger beetle in the Tedori River System of Japan.Crossref | GoogleScholarGoogle Scholar |

Schlesinger, MD, and Novak, PG (2011). Status and conservation of an imperiled tiger beetle fauna in New York State, USA. Journal of Insect Conservation 15, 839–852.
Status and conservation of an imperiled tiger beetle fauna in New York State, USA.Crossref | GoogleScholarGoogle Scholar |

Schlick-Steiner, BC, Seifert, B, Stauffer, C, Christian, E, Crozier, RH, and Steiner, FM (2007). Without morphology, cryptic species stay in taxonomic crypsis following discovery. Trends in Ecology & Evolution 22, 391–392.
Without morphology, cryptic species stay in taxonomic crypsis following discovery.Crossref | GoogleScholarGoogle Scholar |

Schlick-Steiner, BC, Steiner, FM, Seifert, B, Stauffer, C, Christian, E, and Crozier, RH (2010). Integrative taxonomy: a multisource approach to exploring biodiversity. Annual Review of Entomology 55, 421–438.
Integrative taxonomy: a multisource approach to exploring biodiversity.Crossref | GoogleScholarGoogle Scholar |

Smith, ML, and Carstens, BC (2020). Process‐based species delimitation leads to identification of more biologically relevant species. Evolution 74, 216–229.
Process‐based species delimitation leads to identification of more biologically relevant species.Crossref | GoogleScholarGoogle Scholar |

Sota, T, Liang, H, Enokido, Y, and Hori, M (2011). Phylogeny and divergence time of island tiger beetles of the genus Cylindera (Coleoptera: Cicindelidae) in East Asia. Biological Journal of the Linnean Society 102, 715–727.
Phylogeny and divergence time of island tiger beetles of the genus Cylindera (Coleoptera: Cicindelidae) in East Asia.Crossref | GoogleScholarGoogle Scholar |

Sukumaran, J, and Knowles, LL (2017). Multispecies coalescent delimits structure, not species. Proceedings of the National Academy of Sciences USA 114, 1607–1612.
Multispecies coalescent delimits structure, not species.Crossref | GoogleScholarGoogle Scholar |

Tsai, CL, Kubota, K, Pham, HT, and Yeh, WB (2021). Ancestral Haplotype Retention and Population Expansion Determine the Complicated Population Genetic Structure of the Hilly Lineage of Neolucanus swinhoei Complex (Coleoptera, Lucanidae) on the Subtropical Taiwan Island. Insects 12, 227.
Ancestral Haplotype Retention and Population Expansion Determine the Complicated Population Genetic Structure of the Hilly Lineage of Neolucanus swinhoei Complex (Coleoptera, Lucanidae) on the Subtropical Taiwan Island.Crossref | GoogleScholarGoogle Scholar |

Tseng, SP, Wang, CJ, Li, SH, and Lin, SM (2015). Within-island speciation with an exceptional case of distinct separation between two sibling lizard species divided by a narrow stream. Molecular Phylogenetics and Evolution 90, 164–175.
Within-island speciation with an exceptional case of distinct separation between two sibling lizard species divided by a narrow stream.Crossref | GoogleScholarGoogle Scholar |

Tsuji, K, Hori, M, Phyu, MH, Liang, H, and Sota, T (2016). Colorful patterns indicate common ancestry in diverged tiger beetle taxa: Molecular phylogeny, biogeography, and evolution of elytral coloration of the genus Cicindela subgenus Sophiodela and its allies. Molecular Phylogenetics and Evolution 95, 1–10.
Colorful patterns indicate common ancestry in diverged tiger beetle taxa: Molecular phylogeny, biogeography, and evolution of elytral coloration of the genus Cicindela subgenus Sophiodela and its allies.Crossref | GoogleScholarGoogle Scholar |

Vitecek, S, Kučinić, M, Previšić, A, Živić, I, Stojanović, K, Keresztes, L, Bálint, M, Hoppeler, F, Waringer, J, Graf, W, and Pauls, SU (2017). Integrative taxonomy by molecular species delimitation: multi-locus data corroborate a new species of Balkan Drusinae micro-endemics. BMC Evolutionary Biology 17, 129.
Integrative taxonomy by molecular species delimitation: multi-locus data corroborate a new species of Balkan Drusinae micro-endemics.Crossref | GoogleScholarGoogle Scholar |

Webb, WC, Marzluff, JM, and Omland, KE (2011). Random interbreeding between cryptic lineages of the Common Raven: evidence for speciation in reverse. Molecular Ecology 20, 2390–2402.
Random interbreeding between cryptic lineages of the Common Raven: evidence for speciation in reverse.Crossref | GoogleScholarGoogle Scholar |

Wiens, JJ, and Graham, CH (2005). Niche conservatism: Integrating evolution, ecology, and conservation biology. Annual Review of Ecology, Evolution, and Systematics 36, 519–539.
Niche conservatism: Integrating evolution, ecology, and conservation biology.Crossref | GoogleScholarGoogle Scholar |

Wild, AL, and Maddison, DR (2008). Evaluating nuclear protein-coding genes for phylogenetic utility in beetles. Molecular Phylogenetics and Evolution 48, 877–891.
Evaluating nuclear protein-coding genes for phylogenetic utility in beetles.Crossref | GoogleScholarGoogle Scholar |

Yang, Z (2015). The BPP program for species tree estimation and species delimitation. Current Zoology 61, 854–865.
The BPP program for species tree estimation and species delimitation.Crossref | GoogleScholarGoogle Scholar |

Yang, Z, and Rannala, B (2010). Bayesian species delimitation using multilocus sequence data. Proceedings of the National Academy of Sciences 107, 9264–9269.
Bayesian species delimitation using multilocus sequence data.Crossref | GoogleScholarGoogle Scholar |

Yang, L, Kong, H, Huang, JP, and Kang, M (2019). Different species or genetically divergent populations? Integrative species delimitation of the Primulina hochiensis complex from isolated karst habitats. Molecular Phylogenetics and Evolution 132, 219–231.
Different species or genetically divergent populations? Integrative species delimitation of the Primulina hochiensis complex from isolated karst habitats.Crossref | GoogleScholarGoogle Scholar |

Yeates, DK, Seago, A, Nelson, L, Cameron, SL, Joseph, L, and Trueman, JWH (2011). Integrative taxonomy, or iterative taxonomy? Systematic Entomology 36, 209–217.
Integrative taxonomy, or iterative taxonomy?Crossref | GoogleScholarGoogle Scholar |

Zhang, J, Kapli, P, Pavlidis, P, and Stamatakis, A (2013). A general species delimitation method with applications to phylogenetic placements. Bioinformatics 29, 2869–2876.
A general species delimitation method with applications to phylogenetic placements.Crossref | GoogleScholarGoogle Scholar |