Research Article |
Corresponding author: Yan-Li Che ( shirleyche2000@126.com ) Academic editor: Fred Legendre
© 2022 Wei Han, Lu Qiu, Jing Zhu, Zong-Qing Wang, Yan-Li Che.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Han W, Qiu L, Zhu J, Wang Z-Q, Che Y-L (2022) Exploring the diversity of Eupolyphaga Chopard, 1929 (Blattodea, Corydioidea): species delimitation based on morphology and molecular analysis. ZooKeys 1120: 67-94. https://doi.org/10.3897/zookeys.1120.87483
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Eupolyphaga Chopard, 1929 is a cockroach genus mainly endemic to China. In this study, the species diversity of this genus is further explored through morphology and molecular analysis. Four species are described new to science: Eupolyphaga miracidia Qiu, sp. nov., Eupolyphaga udenostyla Qiu, sp. nov., Eupolyphaga reducta Qiu, sp. nov., and Eupolyphaga simila Qiu, sp. nov. New knowledge on some known species is added, including new distribution records and characteristics of females. Forty-five COI sequences were newly sequenced and a molecular species delimitation analysis was performed using ABGD method. Eighteen molecular operational taxonomic units were obtained by ABGD analysis, which are nearly consistent with the results of morphological delimitation.
ABGD, Corydiinae, DNA barcoding, new species, Polyphagini
Eupolyphaga Chopard, 1929 is the largest genus of Corydioidea in China, including 20 species and two subspecies (
Species delimitation of the genus has been based on external morphological characters, male genitalia and some oothecae characters (
Automatic Barcode Gap Discovery (ABGD) (
We recently obtained more Eupolyphaga material from various collecting sites in China. In this study, we use morphological characteristics combined with ABGD to delimit species. Four new species are established. Some descriptions of females and oothecae of known species, and some new distributional data are added to complement species descriptions and facilitate future identifications.
Specimens, including the voucher specimens (Table
Species | Abbreviation | GenBank ID | Collecting information | Remark |
---|---|---|---|---|
E. sinensis | EupoSineLN | OP215845 | Tongnai Village, Fuxin, Liaoning; 31 July 2016; Lu-Yu Wang | nymph |
EupoSineBJ | OP215846 | Mt. Xishan, Beijing; 28 April 2015; Bing-Qiang Wang | male | |
EupoSineWH | OP215847 | Sushansi, Wuhan, Hubei; 16 August 2019; Chen-Liang Li | male | |
E. hanae | EupoHanaSM | OP215848 | Mt. Simianshan, Jiangjin, Chongqing; 31 August 2018; Lu Qiu | male |
EupoHanaDG | OP215849 | Daguan Town, Dujiangyan, Sichuan; 19 May 2015; Lu Qiu, Jing-Fei Han | male | |
EupoHanaSN1 | OP215850 | Shehong, Suining, Sichuan; 8 May 2016; Lei Wang | male | |
EupoHanaSN2 | OP215851 | ibid | female | |
EupoHanaJY | OP215852 | Mt. Jinyunshan, Beibei, Chongqing; 22 August 2018; Lu Qiu | male | |
EupoHanaGL | OP215853 | Jianzhu Township, Gulin, Sichuan; 1 February 2019; Lu Qiu | female | |
E. hupingensis | EupoHupiSM | OP215854 | Hupingshan Town, Shimen, Hunan; 21 May 2016; Hao Xu | male |
EupoHupiJZ | OP215855 | Jingzhai, Lu,an, Anhui; 2 August 2018; Yu-chen Zheng | male | |
E. robusta | EupoRobu1 | OP215856 | Maoxian, Sichuan; 6 August 2019; Lu Qiu, Wei Han, Huan-Yu Ren | male, orange abdominal type |
EupoRobu2 | OP215857 | ibid | female | |
EupoRobu3 | OP215858 | Miancu Village, Maoxian, Sichuan; 7 August 2019; Zong-Qing Wang | male, orange abdominal type | |
EupoRobu4 | OP215859 | Miancu Village, Maoxian, Sichuan; 7 August 2019; Wei Han, Huanyu Ren | female | |
EupoRobu5 | OP215860 | Wenchuan, Sichuan; 7 August 2019; Wei Han, Huanyu Ren | male, orange abdominal type | |
EupoRobu6 | OP215861 | Wenchuan, Sichuan; 7 August 2019; Zongqing Wang | female | |
EupoRobu8 | OP215862 | Wenchuan, Sichuan; 7 August 2019; Wei Han, Huan-Yu Ren | male, black abdominal type | |
EupoRobu7 | OP215863 | Miansi Town, Wenchuan, Sichuan; 29 May 2020; Jian-Yue Qiu | male, black abdominal type | |
E. yunnanensis | EupoYunnCB | OP215864 | Zayü, Tibet; 1 August 2014; Weiwei Zhang | female |
EupoYunnCY | OP215865 | Zayü, Tibet; 14 August 2015; Lu Qiu | male | |
EupoYunnBO | OP215866 | Bomê, Tibet; 11 July 2016; Jian-Yue Qiu, Hao Xu | male | |
EupoYunnTM2 | OP215867 | Tongmai Town, Bomê, Tibet; 13 August 2017; HaoXu, Jian-Yue Qiu | male | |
EupoYunnTM3 | OP215868 | Tongmai Town, Bomê, Tibet; 11 August 2017; Hao Xu, Jian-Yue Qiu | male | |
EupoYunnTM1 | OP215869 | Tongmai Town, Bomê, Tibet; 12 August 2017; Jian-Yue Qiu, Hao Xu | male | |
E. fengi | EupoFeng1 | OP215870 | Mt. Zixishan, Chuxiong, Yunnan; 7 July 2012; Dong Wang | male |
EupoFeng2 | OP215871 | ibid | male | |
E. dongi | EupoDong1 | OP215872 | Mt. Gaoligongshan, Baoshan, Yunnan; 13 April 2017; Zhi-Wei Dong | male |
EupoDong2 | OP215873 | Mt. Gaoligongshan, Baoshan, Yunnan; June 2020; Lu Qiu, Jin-Lin Liu | nymph | |
E. wooi | EupoWooi | OP215874 | Mt. Ailaoshan, Xinping, Yunnan; 11 May 2016; Lu Qiu | female |
E. xuorum | EupoXuor1 | OP215875 | Caoke Township, Shimian, Sichuan; 25 August 2016; Hao Xu, Jian-Yue Qiu | male |
EupoXuor2 | OP215876 | ibid | male | |
E. daweishana | EupoDawe | OP215877 | Mt. Daweishan, Pinbian, Yunnan; 16 May 2016; Lu Qiu | nymph |
E. miracidia sp. nov. | EupoMira | OP215878 | Maqiao Town, Xiangyang, Hubei; 13 July 2017; Lu Qiu | male |
E. nigrinotum | EupoNigr1 | OP215879 | Mt. Jizushan, Bingchuan, Yunnan; 20 February 2016; Hao Xu, Jian-Yue Qiu | male |
EupoNigr3 | OP215880 | Mt. Jizushan, Bingchuan, Yunnan; 20 February 2016; Hao Xu, Jian-Yue Qiu | female | |
EupoNigr2 | OP215881 | Mt. Jizushan, Bingchuan, Yunnan; 7 June 2019, local | male | |
E. pilosa | EupoPilo | OP215882 | Pantiange Township, Weixi, Yunnan; 21 August 2015; Lu Qiu | male |
E. simila sp. nov. | EupoSimi1 | OP215883 | Miyaluo Town, Lixian, Sichuan; 6 October 2019; Lu Qiu, Hao Xu, Zhi-Teng Chen | female |
EupoSimi2 | OP215884 | ibid | nymph | |
EupoSimi3 | OP215885 | ibid | male | |
E. reducta sp. nov. | EupoRedu | OP215886 | Wadi Township, Maoxian, Sichuan; 3 October 2019; Hao Xu, Zhi-Teng Chen, Lu Qiu | nymph |
E. udenostyla sp. nov. | EupoUden1 | OP215887 | Keku Township, Wenchuan, Sichuan; 7 August 2019; Huan-Yu Ren, Wei Han | female |
EupoUden2 | OP215888 | ibid | male | |
EupoUden3 | OP215889 | ibid | male | |
Outgroup | ||||
Periplaneta americana | PeriAmer | HM386405 | / | |
Eucorydia dasytoides | EucuDasy | LC480880 | / | |
Diploptera punctata | DiplPunc | MF479156 | / |
The morphological terminology follows
The hind legs or thoracic muscle were used for DNA extraction, and the other body parts were stored in 95% ethanol as voucher specimens. The extraction procedure was performed according to the Hipure Tissue DNA Mini Kit (Magen Bio-tech, Guangzhou) and the extracted total DNA was stored in a -20 °C environment. Fragments of COI were amplified using PCR; primers used for the amplifications are F: 5’-GGTCAACAAATCATAAAGATATTGG-3’ and R: 5’-TAAACTTCAGGGTGACCAAAAAATCA-3’ (
In total, 48 COI sequences were used for analysis (45 sequences representing Eupolyphaga species and three sequences representing outgroups downloaded from GenBank). All 45 newly acquired sequences are deposited in GenBank (https://www.ncbi.nlm.nih.gov/nuccore) with accession numbers OP215845 to OP215889 (Table
We also conducted an ABGD analysis to confront its results to our morphologically defined species and thus further refine our species delimitation within the genus Eupolyphaga. ABGD analysis was performed using a web interface (https://bioinfo.mnhn.fr/abi/public/abgd/abgdweb.html); the default parameters were used except for the relative gap width set at 1.0 and using the Jukes-Cantor (JC69) model.
Combining male and female external morphology, male genitalia, and some oothecae features, we identified 16 morphospecies (including four new species) of Eupolyphaga among 266 samples examined from China (Fig.
In this study, we acquired 45 COI sequences of Eupolyphaga representing 16 morphospecies of Eupolyphaga. The maximum likelihood tree shows that, for each morphological species, all samples (including the different locality samples, different sex samples, and adult and nymphal samples) are monophyletic, although most of the nodes do not have high bootstrap values (Fig.
Eighteen molecular operational taxonomic units (MOTUs) were estimated by ABGD for the 45 samples (Fig.
Holotype : China · male; Hubei Prov., Xiangyang City, Maqiao Town, roadside of Ganxigou; 480 m–600 m; 31°46.99'N, 110°55.06'E; 13 July 2017; Lu Qiu leg.; SWU-B-CC-010001.
Paratypes : China · 2 males & 2 females, same collection data as holotype; SWU-B-CC-010002 to SWU-B-CC-010005.
This species can be easily distinguished from other species by its small body size. It resembles E. hupingensis by its dark coloration and dense maculae on tegmina, but it can be distinguished from the latter by its small-sized body in both sexes (12.1–12.5 mm excluding tegmina and wings in males, 22.3–24.1 mm in females; Fig.
Eupolyphaga miracidia Qiu, sp. nov. A male holotype, dorsal view B male holotype, ventral view C female paratype, dorsal view D female paratype, ventral view E pronotum, male holotype, dorsal view F head, male holotype, ventral view G pronotum, female paratype, dorsal view H head, female paratype, ventral view I supra-anal plate, male holotype, ventral view J subgenital plate, male holotype, ventral view K genitalia, male holotype, dorsal view L right phallomere, holotype, right-ventral view M the serration of oothecae N oothecae, lateral view O–R habitats of E. miracidia Qiu, sp. nov., in Maqiao Town, Xiangyang City, Hubei Province S living male T living female. Scale bars: 1.0 cm (A–D); 0.1 cm (E–L, N); 0.02 cm (M). Photographs O–T by LQ.
Male holotype. Measurements (mm). Overall length: 26.9; body length: 12.1; body width (tegmina not included): 9.8; tegmen length × width: 22.3 × 9.5; pronotum length × width: 7.4 × 4.4.
Coloration. Body blackish brown, covered with brown setae (Fig.
Head
: Subrounded, almost hidden under pronotum. Interocular space moderate, about half of the distance between ocelli. Ocelli large, ocelli ridge protruded and curved, two dimples symmetrically situated below the ridge. Two yellowish brown semicircular pits each situated next to the inner side of antennal sockets. Clypeus distinct, labrum almost quadrate, posterior margin concave in the middle (Fig.
Male paratypes. Similar to the holotype, no distinct differences.
Female paratypes. Body length: 22.3–24.1 mm, body width: 16.2–16.9 mm. Body uniformly dark reddish brown (Fig.
Nymph. Similar to the female, body darkish brown.
Ootheca. Reddish brown. The longitudinal lines distinct. Serrations on the keel large and curved, apex of serrations slightly truncated. Space between the serrations of the curved portion moderate (Fig.
Found in dry earth under a cliff of the roadside (Fig.
The species epithet miracidia is derived from Greek mirakos referring to it's small size.
Holotype : China · male; Sichuan Prov., Aba Prefecture, Wenchuan County, Keku Township; 1555 m; 31°30.93'N, 103°34.27'E, 5 May 2019; Lu Qiu leg.; SWU-B-CC-010006.
Paratypes : China · 3 males & 1 female, same collection data as holotype; SWU-B-CC-010007 to 010010 · 3 males, same collection data as holotype, but 7 August 2019; Huan-Yu Ren, Wei Han leg; SWU-B-CC-010011 to 010013 · 2 males & 4 females, Sichuan Prov., Wenchuan County, mountains behind the 5·12 Wenchuan Earthquake Memorial Museum; July–August 2019; Qi Li leg.; SWU-B-CC-010014 to 010019.
This species is remarkable for the absence of styli and the short tegmina and strongly reduced anal fields of hind wings in male, which can easily distinguish the males of E. udenostyla from all other species. The serrations of ootheca are distinctly reduced.
Male holotype: Body stout. Measurements (mm). Overall length: 23.5; body length: 19.7; body width (tegmina not included): 12.1; pronotum length × width: 17.1 × 9.4; tegmina length × width: 17.6 × 8.4.
Coloration. Body almost black, covered with black setae (Fig.
Eupolyphaga udenostyla Qiu, sp. nov. A male holotype, dorsal view B male holotype, ventral view C female paratype, dorsal view D female paratype, ventral view E male head, ventral view F tegmen, ventral view G hind wing, ventral view H supra-anal plate, ventral view I subgenital plate, ventral view J left portion of subgenital plate, dorsal view K genitalia, dorsal view L right phallomere, right-ventral view. Scale bars: 1.0 cm (A–D); 0.2 cm E; 0.5 cm (F, G); 0.1 cm (H, I, K, L); 0.02 cm (J). Black arrows indicate the strongly reduced anal field and absence of styli.
Head
: Ovoid, longer than wide. Vertex concealed under pronotum. Eyes small, slightly reduced. Interocular space wide but narrower than the distances between antennal sockets and between ocelli. Ocelli moderate, ocellar ridge distinct, with a row of setae on the upper edge (Fig.
A–D Characteristics and habitats of Eupolyphaga species from Minjiang River Basin A head of female E. robusta, ventral view B head of female Eupolyphaga udenostyla Qiu, sp. nov., ventral view C supra-anal plate of female E. robusta, dorsal view D supra-anal plate of female E. udenostyla Qiu, sp. nov., dorsal view E ootheca of E. udenostyla Qiu, sp. nov., lateral view F same, close-up view to show the serration G a living male E. udenostyla Qiu, sp. nov. H a living female E. udenostyla Qiu, sp. nov. I, J habitats of E. udenostyla Qiu, sp. nov., in Keku Township, Wenchuan County, Sichuan Province. Scale bars: 0.2 cm (E); 0.1 cm (F). Photographs G–J by LQ. White arrows indicate the ocelli.
Male paratypes: Similar to the holotype, no significant variation.
Female paratypes: Body length: 23.5–24.7 mm, body width: 17.5–18.1 mm. Body uniformly dark brown (Fig.
Nymph. Similar to the female, body uniformly black or dark brown.
Ootheca. Reddish brown. Serrations of keel small, blunt. Surface with elevated longitudinal lines, the width of ridge wider near the serrations (Fig.
This new species is distributed in the Minjiang River Basin (Wenchuan County) (Fig.
uden- (Greek) + styla (Greek) indicate that the males of the new species have no styli.
The absence of styli in males makes this new species morphologically remarkable in Eupolyphaga. Males of the species also have distinctly shortened tegmina and wings, and extremely reduced anal areas on hind wings. The ootheca has reduced serrations. The type locality of the new species is close to the one of Eupolyphaga robusta, they are sympatric, distributed in Minjiang River Basin. Males and ootheca of this new species are easily distinguished from E. robusta, but females may be confused because of their similar black and large body. However, females of this new species have reduced ocelli (Fig.
Holotype : China · male, Sichuan Prov., Maoxian County, Wadi Township, Shahuzhai Village; 1938 m; 31°53.20'N, 103°28.62'E; 3 October 2019; Hao Xu, Zhi-Teng Chen, Lu Qiu leg (the holotype was reared from nymph by LQ).; SWU-B-CC-010020.
1 male (an incompletely eclosed individual, unsuitable to be designated as a type specimen); SWU-B-CC-010021 · 1 female nymph, same collection data as holotype; SWU-B-CC-010022.
This species is easily distinguished from its congeners by the reduced and stout tegmina and hind wings in male. In comparison with other species also possessing reduced tegmina and hind wings in males (e.g., E. everestiana reni and E. udenostyla Qiu, sp. nov.), this new species is obviously larger (24.7 mm without tegmina and wings in length), the tegmina with sparse macula only on distal half; while E. everestiana reni is small in size (18.0–18.5 mm without tegmina and hind wings in length) and with evenly distributed maculae on tegmina. E. udenostyla Qiu, sp. nov. (19.7 mm without tegmina and wings in length) is an almost unicolored black species with no styli in males.
Male holotype: Body short and broad (Fig.
Eupolyphaga reducta Qiu, sp. nov., male holotype A habitus, dorsal view B habitus, ventral view C pronotum, male holotype, dorsal view D head, male holotype, ventral view E supra-anal plate, ventral view F subgenital plate, ventral view G genitalia, dorsal view H right phallomere, right-ventral view I–J the living holotype after moulting K habitat of E. reducta Qiu, sp. nov., in Shahuzhai Village, Maoxian County, Sichuan Province L Dr. Zhi-Teng Chen and Dr. Hao Xu collecting this species. Scale bars: 1.0 cm (A, B); 0.2 cm (C, D); 0.1 cm (E–H). Photographs I–L by LQ.
Coloration. Body yellowish brown. Pronotum brown, anterior margin yellow (Fig.
Head
: Interocular space narrow, obviously narrower than the distance between antennal sockets, and slightly narrower than the distance between ocelli. Ocelli large, ocellar ridge short, straight, covered with yellowish brown setae. Clypeus small (Fig.
Female and ootheca: Unknown.
Nymph: Similar to the male in color, yellowish brown.
This new species is distributed in the Heishui River Basin (Maoxian County to Heishui County). Nymphs of this new species live beneath the withered grass at the edge of a soil slope (Hao Xu, Lu Qiu, pers. obs.; Fig.
The species epithet is from the Latin reductus indicating its reduced tegmina and wings.
Holotype : China · male; Sichuan Prov., Lixian County, Miyaluo Town, Siboguo Village; 2944 m; 31°41.58'N, 102°44.80'E; 6 October 2019; Hao Xu, Lu Qiu, Zhi-Teng Chen leg.; SWU-B-CC-010023.
Paratypes : 2 males, same collection data as holotype; SWU-B-CC-010024 to 010025.
2 nymphs & 4 oothecae, same collection data as holotype; SWU-B-CC-010026 to 010027.
The male of this new species resembles Eupolyphaga yunnanensis in external morphology. However, the male of E. simila Qiu sp. nov. has a darker abdomen (Fig.
Eupolyphaga simila Qiu, sp. nov., male holotype A habitus, dorsal view B habitus, ventral view C pronotum, dorsal view D head, ventral view E supra-anal plate, ventral view F subgenital plate (right stylus missing), ventral view G genitalia, dorsal view H right phallomere, right-ventral view. Scale bars: 1.0 cm (A, B); 0.2 cm (C, D); 0.1 cm (E–H).
Eupolyphaga simila Qiu, sp. nov. A ootheca, lateral view B serrations of ootheca C, D habitats of Eupolyphaga simila, Siboguo Village, Lixian County, Sichuan Province E, F the living holotype after emergence G the ventral side of a nymph H the ventral side of a female. Scale bars: 0.1 cm (A); 0.01 cm (B). Photographs C–H by LQ.
Male holotype: Measurements (mm). Overall length: 28.9; body length: 16.2; body width (tegmina not included): 9.4; pronotum length × width: 8.3 × 5.1; tegmina length × width: 26.3 × 9.0.
Coloration. Body fulvous (Fig.
Head
. Round, longer than wide. Interocular space narrower than the distances between antennal sockets and ocelli. Ocelli bulged, ocellar ridge pubescent (Fig.
Male paratype: Body length 29.1 mm including tegmina and hind wings, no significant differences from the holotype.
Nymph. Yellowish brown, dorsal surface black-brown to pale yellow-brown, with many black maculae. Vertex with a black strip between eyes, the space between ocelli with some black spots. Hind-clypeus black. Legs darkish brown to light yellowish brown, coxa with some black maculae. Abdomen yellow, with dark margins (Fig.
Ootheca. Light reddish brown. Surface with longitudinal lines. Serrations of the keel small and bluntly rounded. Respiratory canals absent (Fig.
This new species can be found in dry soil under cliffs along the roadside (Fig.
The species epithet is from similis (Latin) indicating this species is similar to Eupolyphaga yunnanensis.
Due to the outbreak of COVID-19, all living females of the new species were uncared-for in the lab during the research time. Thus, no female specimens survived for further study, but LQ has taken a photograph of a female in the wild (Fig.
Polyphaga sinensis Walker, 1868: 14.
Homoeogamia sinensis
Saussure, 1869: 282;
Heterogamia sinensis:
Heterogamia dohrniana
Saussure, 1893: 309;
Polyphaga limbata Kirby, 1903: 379.
Eupolyphaga sinensis:
“North China”.
China · 2 males; Hubei Prov., Xiangyang City, Xianshan Mountain; July 2020; Mao Ye leg; SWU-B-CC-010028 to 010029 · 1 male; Hubei Prov., Wuhan City, Huangling District, Sushan Temple; 16 August 2019; Chen-Liang Li leg.; SWU-B-CC-010030.
China (Beijing, Hebei, Henan, Inner Mongolia, Liaoning, Jilin, Tianjin, Shaanxi, Shanxi, Ningxia, Shandong, Jiangsu, Anhui, Hubei (new record), Hunan, Chongqing, Guizhou, Yunnan).
This species is newly recorded from Hubei Province. This is the most widely distributed Eupolyphaga species in China, but in general it is more common in Northern China than in the south. The distribution of this species in south China needs to be further surveyed in the future. The ventral abdomen of this species in males is usually uniformly yellowish white, while a specimen from Wuhan City has dark brown markings on the legs and abdomen (Fig.
Eupolyphaga sinensis, male from Sushan Temple, Wuhan City, Hubei Province A habitus, dorsal view B habitus, ventral view C a living male D head, ventral view E pronotum, dorsal view F legs and abdomen (showing dark brown markings), ventral view G supra-anal plate, ventral view H subgenital plate, ventral view I genitalia, dorsal view J right phallomere, right-ventral view. Scale bars: 1.0 cm (A, B); 0.2 cm (D–J). Photographs C by Chen-Liang Li.
Eupolyphaga hanae
Qiu, Che & Wang., 2018: 16;
“Sichuan Prov., Chengdu City, Dujiangyan Prefectural-Level City, Daguan Town, Puzhao Temple; 770 m.”
China · 2 males, 1 female, 10 nymphs, 4 oothecae; Sichuan Prov., Luzhou City, Gulin County, Jianzhu Township, Daheidong Scenic Area, Xiazhai; 899 m; 28°03.50'N, 105°34.70'E, 1 February 2019, Lu Qiu leg.; SWU-B-CC-010031 to 010043 · 1 male, Sichuan Prov., Aba Prefecture, Wenchuan County, Yingxiu Town; 1000 m; 17 September 1983; Xue-Zhong Zhang leg.; SWU-B-CC-010044 · 2 males, Chongqing City, Jiangjin District, Simianshan Mountain, Dawopu; 31 August 2018; Lu Qiu leg.; SWU-B-CC-010045 to 010046 · 1 male, Chongqing City, Jiangjin District, Simianshan Mountain; 22 October 2006, Wei-Wei Zhang leg.; SWU-B-CC-010047 · 1 male, Chongqing City, Jiangjin District, Simianshan Mountain; August 2018; De-Yao Zhou leg.; SWU-B-CC-010048.
This species is mainly distributed in Sichuan, Chongqing. We obtained COI sequences of this species from various localities (Simianshan Mountain and Jinyunshan Mountain in Chongqing, Shehong, Dujiangyan and Gulin in Sichuan). Genetic distances ranged from 2.02–6.12% between samples from these localities (Suppl. material
Eupolyphaga robusta
Qiu, Che & Wang., 2018: 19;
“Sichuan Prov., Aba Prefecture, Wenchuan County; 1100 m.”
China · 2 males, 1 female, 1 nymph, 6 oothecae; Sichuan Prov., Aba Prefecture, Maoxian County, Nanxin Town, Miancu Village; 1518 m; 31°35.73'N, 103°44.97'E; 7 August 2019; Zong-Qing Wang, Lu Qiu, Wei Han, Huan-Yu Ren leg.; SWU-B-CC-010049 to 010052 · 1 nymph, 1 ootheca; Sichuan Prov., Aba Prefecture, Maoxian County, Nanxin Town, Miancu Village; 1503 m; 31°35.77'N, 103°45.00'E; 2 October 2019; Lu Qiu, Hao Xu, Zhi-Teng Chen leg.; SWU-B-CC-010053 · 1 male, 1 female, 10 oothecae; Sichuan Prov., Aba Prefecture, Maoxian County, Xiaomiao Mountain; 1650 m; 31°40.85'N, 103°51.33'E; 6 August 2019; Lu Qiu, Wei Han, Huan-Yu Ren leg.; SWU-B-CC-010054 to 010055 · 1 male, 1 female, 1 nymph, 4 oothecae; Sichuan Prov., Aba Prefecture, 013 Township road near Wenchuan County; 1473 m; 31°28.63'N, 103°35.90'E; Zong-Qing Wang, Lu Qiu, Wei Han, Huan-Yu Ren leg.; SWU-B-CC-010056 to 010057 · 2 males, 6 females, 2 oothecae; Sichuan Prov., Aba Prefecture, Wenchuan County, Miansi Town; 29 March 2020; Jian-Yue Qiu leg.; SWU-B-CC-010058 to 010065.
Male: Tegmina almost black, with a few white maculae, some individuals have more and larger maculae that are randomly distributed on tegmina. Abdomen orange or black, if black, the terminal two segments dimly yellowish.
Female: Body length 29.1–32.5 mm, body width 20.5–22.5 mm. Body uniformly black (Fig.
Eupolyphaga robusta from Minjiang River basin A male, dorsal view B male, ventral view C female, dorsal view D female, ventral view E–G habitats of Eupolyphaga robusta, Maoxian County, Sichuan Province H the variation of the abdominal coloration in males I ootheca, lateral view J serrations of ootheca. Scale bars: 1.0 cm (A–D); 0.1 cm (I); 0.01 cm (J). Photographs E–H by LQ.
Nymph: Similar to female, blackish brown.
Ootheca: Yellowish brown. Surface with shallow longitudinal lines. Serrations on the keel large and curved, compactly arranged. Respiratory canals indistinct (Fig.
This species is distributed in the Minjiang River basin of Sichuan, which is a typical arid river valley. It can be found in multiple habitats, such as in soil, under abandoned bricks or rocks, or inside dead wood.
This species was originally described based on a single male specimen collected in 1983. The holotype is dark brown, the last two segments of abdomen are yellowish white, and its vertex has a small yellowish spot. During two expeditions in Minjiang River basin in 2019, we obtained numerous specimens of this species. We found that the fresh samples of the males are black rather than dark brown. The maculae on tegmina show some variation, most male individuals are black, with sparse whitish maculae, but there are individuals with larger maculae. We also found that most of the newly collected individuals differ from the holotype by the coloration of abdomen. Most males have a uniformly orange abdomen, while the specimens from Miansi Town have the same abdominal color as the holotype, i.e., the apical two segments of abdomen are yellowish, the remaining portions are black. Among the newly collected male specimens, no individual was found with a yellow spot at vertex, which indicates the yellow spot at vertex in the holotype may be a variation or aberration.
We sequenced the COI marker from eight samples and found that the genetic distances between samples from Miansi Town (male with black abdomen) and samples from other localities (males with orange abdomens) are somewhat distant (3.29–3.57%), while the genetic distances between samples with orange abdomen are much lower (0.15–0.47%) (Suppl. material
Eupolyphaga hupingensis
Qiu, Che & Wang., 2018: 18;
“China: Hunan: Daling Village, Hupingshan Town, Shimen County”, 444 m.
China · 1 male, 2 females, 1 ootheca; Hunan Prov., Zhangjiajie City, Sangzhi County, Badagongshan Nature Reserve, Tianpingshan Mountain; 1400 m; 29°46.99'N, 110°05.44'E; 30–31 July 2019; Hao Xu, Jian-Yue Qiu leg.; SWU-B-CC-010066 to 010068 · 1 male; Anhui Prov., Jinzhai County, Mazongling; 580 m; 2 August 2018, Yu-Chen Zheng leg.; SWU-B-CC-010069.
Female: Body length 32.3–41.0 mm, body width 21.5–23.7 mm. Body uniformly dark reddish brown. Antennal sockets, ocelli, ante-clypeus, and labrum white. Ocelli large. Distance between ocelli almost equal to interocular space, and slightly narrower than distance between antennal sockets. Arolia and pulvilli absent. Posterior margin of supra-anal plate protruded, medially with a large incision (Fig.
Eupolyphaga hupingensis from Mt. Badagongshan, Hunan Province (A–D) and Eupolyphaga dongi from Mt. Gaoligongshan, Baoshan City, Yunnan Province (E–H). A female, dorsal view B female, ventral view C ootheca, lateral view D serrations of ootheca E female, dorsal view F female, ventral view G ootheca, lateral view H serrations of ootheca. Scale bars: 1.0 cm (A, B, E, F); 0.2 cm (C, G); 0.05 cm (D, H).
Nymph. Similar to female, blackish brown.
Ootheca. Reddish brown. Longitudinal lines distinct and evenly arranged (Fig.
China: Hunan, Anhui (new record).
The individual from Jinzhai County (North Anhui) is almost morphologically identical to other specimens of E. hupingensis, only the color of its clypeus and labrum, abdomen and legs are lighter than those of holotype (Fig.
Eupolyphaga hupingensis, male from Mazongling, Jinzhai County, Anhui Province A habitus, dorsal view B habitus, ventral view C head, ventral view D tegmen, ventral view E supra-anal plate, ventral view F subgenital plate, ventral view G genitalia, dorsal view H right phallomere, right-ventral view. Scale bars: 1.0 cm (A, B, D); 0.2 cm (C, G, H); 0.1 cm (E, F).
Eupolyphaga dongi
Qiu, Che & Wang, 2018: 52;
“Yunnan Prov., Baoshan City, Longyang District, Nankang Village; 1980 m.”
China · 1 male; Yunnan Prov., Baoshan City, Mt. Gaoligongshan Nature Reserve, Baihualing; 13 April 2017; Zhi-Wei Dong leg.; SWU-B-CC-010070 · 2 females, 1 nymph, 8 oothecae; Yunnan Prov., Baoshan City, Mt. Gaoligongshan Nature Reserve, Baihualing; 20–21 June 2020; Lu Qiu, Jin-Lin Liu leg.; SWU-B-CC-010071 to 010073.
Female: Body length 20.1–21.2 mm, body width 14.6–14.9 mm. Body dark brown, with small fuzzy markings (Fig.
Nymphs: Similar to female.
Ootheca: Reddish brown. Surface with obvious longitudinal lines (Fig.
This species can be found in dry soil under huge rocks in the forest of Baihualing.
In recent years, ABGD has been successfully applied in the species delimitation of some Blattodea genera (e.g., Cryptocercus,
Samples of E. sinensis from three different localities (Beijing, Liaoning, and Wuhan) were divided into two MOTUs. Compared to the genetic distance between individuals from Beijing and Liaoning (4.76%), the individual from Wuhan was significantly more genetically distant from the first two (8.81% and 9.54%, respectively) (Suppl. material
Not all species have as unusually large intraspecific genetic distances as E. sinensis and E. hupingensis. For E. hanae, E. yunnanensis, and E. robusta, samples from different geographic populations were not significantly different morphologically, and their genetic distances (E. hanae, 2.02%–6.12%; E. yunnanensis, 0.30%–2.01%; E. robusta, 0.15%–3.57%, Suppl. material
The genus Eupolyphaga is widely distributed in China, especially in the mountainous regions of Western China. It is still under-researched due to its secretive habits, which make it not easily detected in the natural environment. The low dispersal ability and the habit of inhabiting only in specific micro-habitats may lead to their long-term geographical isolation (
We sincerely thank Yu-Chen Zheng (China Agricultural University), Wei-Wei Zhang, Dong Wang (Chongqing), Bing-Qiang Wang (Beijing), De-Yao Zhou (Shanghai), Hao Xu, Jian-Yue Qiu (Mianyang Normal University), Zhi-Teng Chen (Jiangsu University of Science and Technology), Mao Ye (Xiangyang, Hubei), Chen-Liang Li (Wuhan, Hubei), Lu-Yu Wang, Jin-Lin Liu, Huan-Yu Ren (SWU), Qi Li (Chengdu, Sichuan), and Lei Wang (Suining, Sichuan) for their efforts in specimen collecting. We thank Dr. John Richard Schrock for proofreading the draft. We also cordially thank Dr. Frédéric Legendre, Dr. Tobias Kovacs, and an anonymous reviewer for their numerous comments and revisionary suggestions. This study is supported by the National Natural Sciences Foundation of China (Nos. 32070468, 31872271).
Table S1
Data type: Genetic distance.
Explanation note: Genetic distances between COI gene of 45 Eupolyphaga species.