Research Article |
Corresponding author: Chao Jiang ( jiangchao0411@126.com ) Corresponding author: Lu-Qi Huang ( huangluqi01@126.com ) Academic editor: Marzio Zapparoli
© 2023 Tian-Yun Chen, Chao Jiang, Lu-Qi Huang.
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:
Chen T-Y, Jiang C, Huang L-Q (2023) A new species of Otostigmus (Chilopoda, Scolopendromorpha, Scolopendridae) from China, with remarks on the phylogenetic relationships of Otostigmus politus Karsch, 1881. ZooKeys 1168: 161-178. https://doi.org/10.3897/zookeys.1168.82750
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Through a combination of morphological and DNA data, a new scolopendrid centipede from southern and southwestern China was revealed: O. tricarinatus sp. nov. The species belong to the politus group but has three sharp tergal keels. Validation of phylogenetic status was performed through molecular analysis of the cytochrome c oxidase subunit I (COI), 16S rRNA, and 28S rRNA sequences from 16 Otostigmus species. Otostigmus tricarinatus sp. nov. was found to be two populations and varied in the number of spines on the ultimate prefemur, the sutures on a sternite, and a pore-free median longitudinal strip in the pore field. The Yunnan-Guizhou plateau population of O. tricarinatus sp. nov. was sister to the clade O. politus politus + O. politus yunnanensis + Guangxi population of O. tricarinatus sp. nov. with strong support from both BI (bayesian inference) and ML (maximum likelihood) analyses (PP = 1, BS = 97%).
Centipedes, key, phylogeny, politus species group, Yunnan-Guizhou plateau
The largest genus in the subfamily Otostigminae (Pocock, 1891), Otostigmus Porat, 1876, has approximately 110 recognised species. Otostigmus is classified into three subgenera: O. (Otostigmus), O. (Parotostigmus) Pocock, 1896, and O. (Dactylotergitius) Verhoeff, 1937 (
Eight species have been recorded in China (
In this study we describe a new species, Otostigmus tricarinatus sp. nov., which belongs to the politus group. The phylogenetic status of the new species was validated through the molecular analysis of cytochrome c oxidase subunit I (COI), 16S rRNA, and 28S rRNA sequences which were derived from 18 Otostigmus species.
Material was collected from different provinces in China (Table
No. | Species | Voucher | Locality | CO1 | 16S | 28S | References |
---|---|---|---|---|---|---|---|
1 | O. tricarinatus sp. nov. |
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Pingbian, Yunnan, China | OM791803 | OM793128 | – | This study |
2 | O. tricarinatus sp. nov. |
|
Maguan, Yunnan, China | OM791806 | OM793131 | OM793116 | This study |
3 | O. tricarinatus sp. nov. |
|
Libo, Guizhou, China | OM791804 | OM793129 | OM793114 | This study |
4 | O. tricarinatus sp. nov. |
|
Guiping, Guangxi, China | OM791805 | OM793130 | OM793115 | This study |
5 | O. tricarinatus sp. nov. |
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Guiping, Guangxi, China | OM791807 | OM793132 | OM793117 | This study |
6 | O. politus politus Karsch, 1881 |
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Tianjing, China | OM791808 | OM793133 | OM793118 | This study |
7 | O. scaber Porat, 1876 |
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Fuzhou, Fujian, China | OM791809 | OM793134 | OM793119 | This study |
8 | O. aculeatus Haase, 1887 |
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Xiamen, Fujian, China | OM791810 | OM793135 | OM793120 | This study |
9 | O. politus yunnanensis Lewis, 2003 |
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Pingbian, Yunnan, China | OM791812 | OM793137 | OM793122 | This study |
10 | O. politus yunnanensis Lewis, 2003 |
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Yingpan, Yunnan, China | OM791813 | OM793138 | OM793123 | This study |
11 | O. beroni Lewis, 2001 |
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Jilong, Xizang, China | OM791814 | OM793139 | OM793124 | This study |
12 | O. beroni Lewis, 2001 |
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Jilong, Xizang, China | OM791815 | OM793140 | OM793125 | This study |
13 | O. voprosus Schileyko, 1992 |
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Jingxiu, Guangxi, China | OM791816 | OM793141 | OM793126 | This study |
14 | O. voprosus Schileyko, 1992 |
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Xishuangbanna, Yunnan, China | OM791817 | OM793142 | OM793127 | This study |
15 | O. nudus Pocock, 1890 | CES091037 | Kerala, Periyar, India | JX531869 | JX531739 | JX531809 |
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16 | O. sulcipes Verhoeff, 1937 | CUMZ 00534 | Phra Cave, Ban Namyen, Myanmar | MF167805 | MF167738 | MF167872 |
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17 | O. sulcipes Verhoeff, 1937 | CUMZ 00533 | LaosChina borders, Luang Namtha, Laos | MF167804 | MF167737 | MF167871 |
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18 | O. sulcipes Verhoeff, 1937 | CUMZ 00532 | Chiang Mai, Khrai, Thailand | MF167803 | MF167736 | MF167870 |
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19 | O. spinosus Porat, 1876 | CUMZ 00553 | Khanom, Nakhon Si Thammarat, Thailand | MF167785 | MF167718 | MF167852 |
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20 | O. spinosus Porat, 1876 | CUMZ 00552 | Thung Khai Botanical Garden, Yantakhao, Trang, Thailand | MF167784 | MF167717 | MF167851 |
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21 | O. spinosus Porat, 1876 | CMUZ 00232 | Ban na ka som, Attapue, Laos | MF167783 | MF167716 | MF167850 |
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22 | O. spinicaudus Newport, 1844 | MCZ DNA104645 | Kasserine District, Tunisia | – | KF676472 | KF676370 |
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23 | O. scaber Porat, 1876 | CUMZ 00530 | Wat Ban Hu, Luang Phrabang, Laos | MF167802 | MF167735 | MF167869 |
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24 | O. scaber Porat, 1876 | CUMZ 00529 | Ban Sop Hun, Mueang Ngoi, Luang Phrabang, Laos | MF167801 | MF167734 | MF167868 |
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25 | O. rugulosus Porat, 1876 | CUMZ 00537 | Phu Phiang, Nan, Thailand | MF167787 | MF167720 | MF167854 |
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26 | O. rugulosus Porat, 1876 | CUMZ 00536 | Sai Yok, Kanchanaburi, Thailand | MF167786 | MF167719 | MF167853 |
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27 | O. rugulosus Porat, 1876 | CUMZ 00535 | Chum Ta Bong, Nakhon Sawan, Thailand | MF167782 | MF167715 | MF167849 |
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28 | O. ruficeps Pocock, 1890 | CES091342 | Kollam, Kerala, India | JX531901 | JX531771 | JX531822 |
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29 | O. politus politus Karsch, 1881 | MCZ DNA106768 | Dengfeng, Henan, China | KF676512 | KF676470 | KF676368 |
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30 | O. multidens Haase, 1887 | CUMZ 00527 | Pan Faen, Mae Taeng, Chiang Mai, Thailand | MF167795 | MF167728 | MF167862 |
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31 | O. multidens Haase, 188 | CUMZ 00526 | Taphome Stone Castle, Siem Reap, Cambodia | MF167793 | MF167726 | MF167860 |
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32 | O. multidens Haase, 188 | CUMZ 00523 | Gua I–Kan, Kelantan, Malaysia | MF167779 | MF167712 | MF167846 |
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33 | O. multidens Haase, 188 | MCZ DNA106502 | Central Province, Papua New Guinea | KF676511 | KF676469 | KF676367 |
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34 | O. astenus Kohlrausch,1878 | CUMZ 00521 | Ban Sop Laos, Huaphan, Laos | MF167800 | MF167733 | MF167867 |
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35 | O. astenus Kohlrausch, 1878 | CUMZ 00520 | Tham Mi Ka Ram, Mae Taeng, Chiang Mai, Thailand | MF167799 | MF167732 | MF167866 |
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36 | O. astenus Kohlrausch, 1878 | MCZ DNA102463 | Fiji/Vanuatu | HM453312 | HM453221 | HQ402532 |
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37 | O. angusticeps Pocock, 1898 | MCZ DNA106500 | Finisterre Mountains, Papua New Guinea | KF676509 | – | KF676365 |
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38 | O. aculeatus Haase, 1887 | CUMZ 00519 | Ban Pha Wong, Mueang Yommarat, Huaphan, Laos | MF167797 | MF167730 | MF167864 |
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39 | O. aculeatus Haase, 1887 | CUMZ 00518 | Wat Phra–Ong Thom, Siam Riep, Cambodia | MF167796 | MF167729 | MF167863 |
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40 | O. caraibicus Kraepelin, 1903 | MCZ DNA105633 | Puerto Rico | HQ402549 | HQ402498 | HQ402533 |
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41 | O. voprosus Schileyko, 1992 | IEBR–Chi 033 | Xuan Nha, Son La Province, Vietnam | MN861168 | – | – |
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42 | O. voprosus Schileyko, 1992 | IEBR–Chi 032 | Thuong Tien, Vietnam | MN861166 | – | – |
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43 | ‘O. amballae Chamberlin, 1913’ | IEBR–Chi 036 | Me Linh, Vietnam | MN861140 | – | – |
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44 | ‘O. amballae Chamberlin, 1913’ | IEBR–Chi 013 | Ta Xu, Vietnam | MN861139 | – | – |
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45 | O. angusticeps Pocock, 1898 | IZ–130684 | Finisterre Mountains, Papua New Guinea | KF676509 | – | KF676365 |
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46 | R. lewisi | Kurinjal, Kudremukh National Park, Chikkamagaluru district, India | MK273239 | MK273349 | MK273461 |
|
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47 | D. jangii | CES08915 | Kurinjal, Kudremukh National Park, Chikkamagaluru district, India | JX531843 | JX531713 | JX531789 |
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48 | R. lewisi | CES08920 | Tadoli, Kudremukh National Park, Chikkamagaluru district, India | MK273240 | MK273350 | MK273462 | Joshi and Edgecombe 2018 |
49 | D. jangii | CES08922 | Tadoli, Kudremukh National Park, Chikkamagaluru district, India | JX531845 | JX531715 | JX531791 |
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50 | A. crotalus | MCZ DNA100454 | Swaziland | AY288742 | AY288720 | HM453273 |
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51 | A. grandidieri | MCZ DNA106771 | Tanzania | KF676514 | KF676473 | KF676371 |
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The morphological terminology follows
Abbreviations: VL = ventrolateral, VM = ventromedial, M = medial, DM = dorsomedial, CS = corner spine, T = tergite, TT = tergites, SS = sternites, spm = specimen, coll. = collector, LBS = leg bearing segment.
Legs from each specimen were used to extract genomic DNA by using a Promega Wizard SV Genomic DNA Purification Kit (Promega, USA). Polymerase chain reaction (PCR) was used to amplify the cytochrome c oxidase subunit I (COI), mitochondrial ribosomal gene 16S, and nuclear ribosomal DNA 28S fragments. Table
Loci | Primers Sequence 5’– 3’ | Program | Refences |
---|---|---|---|
CO1 | LCO1490 GGTCAACAAATCATAAAGATATTGG | 5 min at 95 °C; 38 cycles of 20s at 95 °C, 20s at 45 °C and 1 min at 72 °C; 3 min at 72 °C |
|
Hcoutout GTAAATATATGRTGDGCTC | |||
CO1 | LCO1490 GGTCAACAAATCATAAAGATATTGG | 2 min at 94 °C; 35 cycles of 15s at 95 °C, 40s at 45–47 °C and 15s at 72 °C; 10 min at 72 °C |
|
Hco2198 TAAACTTCAGGGTGACCAAAAAATCA | |||
16S | 16Sar CGCCTGTTTATCAAAAACAT | 5 min at 95 °C; 35 cycles of 30s at 95 °C, 30s at 55 °C and 1 min at 72 °C; 3 min at 72 °C | Xiong and Kocher 1991 |
16Sb CTCCGGTTTGAACTCAGATC | |||
28S | Chilo28SF1 AGCCCAAGTCCCCCTGACC | 3 min at 95 °C; 35 cycles of 30s at 95 °C, 30s at 65 °C and 1 min at 72 °C; 3 min at 72 °C | This study |
Chilo28SR1 TATACTCAGGTCCGACGATCGATT | |||
28S | 28Sa GAC CCG TCT TGA AAC ACG GA | 2 min at 94 °C; 35 cycles of 15s at 95 °C, 40s at 52 °C and 15s at 72 °C; 10 min at 72 °C |
|
28Sb TCG GAA GGA ACC AGC TAC |
The genetic distance between Otostigmus species was calculated using the Kimura 2-parameter model in MEGA X (
Family Scolopendridae Leach, 1814
Subfamily Otostigminae Kraepelin, 1903
Genus Otostigmus Porat, 1876
Otostigmus carinatus Porat, 1876, by subsequent designation (
China: Beijing: Haidian District: Xiangshan Park, 1 spm,
China: Yunnan Province: Pingbian Miao Autonomous County: back mountain of Pingbian Memorial Park, 3 spms, CMMI20210315102–104, 22.9884°N, 103.6931°E, 1380 m asl., 15 March 2021, coll. Chao Jiang, Pingbian Railway Station, 2 spms,
Holotype.
Paratypes. 1 spm,
The name refers to the characteristics of the tergites. The tri- compounded with the Latin carinatus refers to the three sharp keels on the tergites.
Antennae with 17 articles, basal three glabrous dorsally, the apical article with a well-developed lateral depression. TT 3–20 with three longitudinal keels. SS 2–19(20) with paramedian sutures occupying anterior 20–100% of sternites, a median depression, and two posterolateral depressions. Coxopleural process with 1–3 apical spines and none or one lateral spine, pore-free median longitudinal strip in pore field from the posterior of sternite 21 to the end of coxopleural process. The ultimate leg prefemur typically with 0–7 spines, lacking corner spine.
Body length 26 mm. Antennae and anterior 1/2 of the cephalic plate, tergites and legs have blue colouration; posterior 1/2 of the cephalic plate, forcipule segment, and sternites have yellow colouration.
Antennae with 17 articles, 3 glabrous dorsally, 2.5 glabrous ventrally, apical article double the length of the penultimate, with a well-developed lateral depression (Fig.
Otostigmus tricarinatus sp. nov. (Guangxi population)
T1 lacks sutures and convex granules, T2 has sparse convex granules, TT3–21 has scattered convex granules. Tergites without paramedian sutures, TT3–20 with three sharp longitudinal keels, T21 with three incomplete longitudinal keels. Lateral margination at TT3–21. SS2–20 with paramedian sutures occupying anterior 80–100% of sternites (Fig.
The left coxopleural process has one apical and one lateral spine and the right one has two apical and one lateral spine (Fig.
Body length 16–50 mm (maximum in
Lateral margination at TT 2(3)-21. SS2–20 with paramedian sutures occupying anterior 20–100% of sternites. Coxopleural process with 1–3 apical spines, 0–1 lateral spine and lacking a dorsal spine. Ultimate leg prefemur with 0–7 spines: VL 0–1, M 0–3, VM 1–3, DM 1–2, lacking corner spine. Legs 1–3(4 or 5) typically with two tarsal spurs, subsequent legs to 20, with one tarsal spur.
The material here assigned to O. tricarinatus sp. nov. refers to two geographically separate groups: the Yunnan-Guizhou plateau population (Fig.
Otostigmus tricarinatus sp. nov. (Yunnan-Guizhou plateau population)
Fig.
We obtained sequences consisting of 658–812 bp COI, 408–423 bp 16S rRNA, and 368–886 bp28S rRNA. The average K2P genetic distance is 20.1% between Otostigmus species, and the range of the K2P distance between Otostigmus species is 17.2% (O. voprosus against O. astenus to 23.1% (O. politus yunnanensis against O. beroni) (Table
Mean K2P genetic distance between the politus group, aculeatus group and rugulosus group species based on COI sequences.
(1) | (2) | (3) | (4) | (5) | (6) | (7) | (8) | (9) | |
---|---|---|---|---|---|---|---|---|---|
O. angusticeps (1) | |||||||||
O. aculeatus (2) | 22.2% | ||||||||
O. astenus (3) | 18.3% | 22.9% | |||||||
O. beroni (4) | 21.8% | 23.9% | 17.7% | ||||||
O. politus politus (5) | 21.1% | 20.7% | 20.5% | 20.2% | |||||
O. politus yunnanensis (6) | 22.5% | 22.5% | 21.8% | 23.8% | 18.5% | ||||
O. rugulosus (7) | 22.7% | 20.1% | 20.1% | 21.9% | 20.0% | 20.6% | |||
O. spinosus (8) | 19.6% | 19.8% | 19.3% | 19.1% | 18.9% | 22.0% | 18.8% | ||
O. tricarinatus sp. nov. (9) | 22.5% | 22.4% | 20.5% | 21.8% | 19.3% | 21.1% | 21.7% | 19.0% | |
O. voprosus (10) | 20.8% | 22.9% | 17.4% | 17.5% | 22.1% | 21.9% | 22.4% | 20.0% | 21.5% |
Mean K2P genetic distance within the politus group, aculeatus group and rugulosus group species based on COI sequences.
Examined species | Mean distance | Standard error |
---|---|---|
O. angusticeps | – | – |
O. aculeatus | 18.2% | 1.5% |
O. astenus | 17.5% | 1.3% |
O. beroni | 0.2% | 0.1% |
O. politus politus | 8.4% | 1.3% |
O. politus yunnanensis | 16.9% | 1.7% |
O. rugulosus | 6.5% | 0.9% |
O. spinosus | 15.1% | 1.3% |
O. tricarinatus sp. nov. | 16.0% | 1.1% |
O. voprosus | 15.4% | 1.2% |
Sequences from the new species, as well as 47 other Otostigminae samples from different species groups, were aligned. Included in the alignment were two COI sequences from samples IEBR–Chi 013 and IEBR–Chi 036 from Vietnam, which were misidentified as “O. amballae Chamberlin, 1913” (
At last, the key for Otostigmus species in China is provided.
1 | The ultimate leg prefemur with two rows of ventrolateral spines | O. aculeatus |
– | The ultimate leg prefemur with one row of ventrolateral spines | 2 |
2 | Antennae with 19–22 articles | 3 |
– | Antennae with 17 or 18 articles | 5 |
3 | Tooth-plates with 3+3 teeth; tergites without keels | O. astenus |
– | Tooth-plates with 4+4 teeth; tergites with keels | 4 |
4 | TT5–20 with a median keel, coxopleural process curved in females | O. xizangensis |
– | TT3(5)–20 with 7–9 longitudinal keels, coxopleural without curved in females | O. scaber |
5 | Basal 3 antennal articles glabrous dorsally | 6 |
– | Basal 2–2.75 antennal articles glabrous dorsally | 10 |
6 | Tooth-plates with 3+3 teeth, coxopleural process with a dorsal spine | O. voprosus |
– | Tooth-plates with 4+4 teeth coxopleural process without a dorsal spine | 7 |
7 | Ultimate prefemur with 9–16 spines; with corner spine | 8 |
– | Ultimate prefemur with 0–7 spines; without corner spine | O. tricarinatus sp. nov. |
8 | Ultimate prefemur with 9–12 spines (3–4VL, 2–3VM, 2–3M, 1–1DM) | O. politus politus |
– | Ultimate prefemur with 15–16 spines (5VL, 4–5VM, 4M, 2DM) | O. politus yunnanensis |
9 | Coxopleural process with 1 apical spine | O. martensi |
– | Coxopleural process with 2 apical spines | 10 |
10 | Legs 1–4 or 5 with tibial spur, 2 tarsal spurs on legs 1–16(19) | O. beroni |
– | Legs 1–2 with tibial spur, the distribution of 2 tarsal spurs in legs without regularity | O. lewisi |
The presence of three well-developed longitudinal keels on TT3–20 is the main morphological character that distinguishes O. tricarinatus sp. nov. from other Otostigmus (Otostigmus) species. The keels on tergites are a widespread morphological characteristic in Otostigminae Kraepelin, 1903, and may be found in the genera/subgenera Alipes, Edentistoma, Otostigmus (Parotostigmus) and O. (Otostigmus). Otostigmus scaber, O. rugulosus Porat, 1876, and O. amballae generally have tergal longitudinal keels (
Comparison of four species with keels (1
Characteristics | O. scaber Porat, 1876 (1–3) | O. tricarinatus sp. nov. (5) | O. rugulosus Porat, 1876 (4) | O. amballae Chamberlin, 1913 (2) |
---|---|---|---|---|
Length(mm) | 14–69 | 16–50 | 32–43 | 38 |
Antennal articles | 20–21 | 17 | 19–21 | 17 |
Basal antennal articles glabrous | 2–2.5 | 3 dorsally, 2.5 ventrally | 2–2.25(2.4) | 2.5 |
Lateral depression on apical antennal article | absent | present | absent | present |
Teeth on coxosternal tooth-plates | 4–5 | 4 | 4 | 4 |
Teeth on the trochanteroprefemoral process | 2–4 | 2 | 2 | 2 |
Tergite paramedian sutures from | 5(6)–20 | without | 4(5) | Complete from tergite 3 |
Tergite longitudinal keels | 3(5)–20 with 5–7(9) longitudinal sutures | T3–20 with 3 longitudinal keels | low median ridge or keel from 3(5), low rounded lateral keels on posterior segments 7–17(10–14) | A low median keel from 3–20, paramedian sutures forming two lateral keels on each side from ca. 13 |
Tergites marginate starting from | 5–7 | 2 or 3 | 9(7) | 6 |
Sternal paramedian sutures/sulci | 3–19, incomplete sutures | 4–19 with 20–80% sutures on anterior part of sternites | 30–55% of mid and posterior sternites 55% on posterior sternites | 2–20 with low tubercles, 14–20 with round median depression |
Coxopleural process spines | AP2–3, LP1–3, DP1 | AP1–3, LP 0–1 | AP2, SAP1–3 LP1, DP1 | AP2, LAP1, DP1–2 |
pore-free median longitudinal strip in pore field | extending 50–75% length from the posterior of sternite 21 to the end of coxopleural process | from the posterior of sternite 21 to the end of coxopleural process | from the posterior of sternite 21 to the end of coxopleural process | from the posterior of sternite 21 to the end of coxopleural process |
Legs with tarsal spurs | 1–9(7, 8, 10, 11) with 2, the subsequent to 19 or 20 with one | 1–3(4, 5) with 2, the remainder with 1, 21 without | 1–11(up to 18) with 2, 20–21 without | Many legs missing. Legs 1 and 2 with two tarsal spines, legs 10 and 13–20 with one |
Spines of ultimate prefemur | VL3–4(5), VM2–3, DM2–3 | VL0–1, M0–3, VM0–3, DM0–2 | 3–4 rows of VL4, M0–3, M1–2, DM0–2 | VL3, M3, VM2, DM2 |
Corner spine | 1 | 0 | 1 | 1 |
Phylogenetic analysis revealed that O. tricarinatus sp. nov., O. politus politus, O. politus yunnanensis formed a clade constituting the politus group. Otostigmus politus politus and O. politus yunnanensis are reciprocally monophyletic and form a clade which is sister to O. tricarinatus sp. nov. (Yunnan-Guizhou plateau population). Furthermore, O. tricarinatus sp. nov. (Guangxi population) is a sister clade of O. tricarinatus sp. nov. (Yunnan-Guizhou plateau population), O. politus politus, and O. politus yunnanensis. However, O. politus politus, distributed in North China and Korea, is geographically isolated from other politus-group members. The records of O. politus politus from Vietnam with a long ultimate prefemur and spine arrangement were 5–6VL, 5 VM, and 3DM (
We thank Shi Mengxuan (Henan University, China) and Ji Quanyu (Hebei University, China) for contributing specimens to this study. We also sincerely thank Drs Arkady A. Schileyko (Moscow Lomonosov State University, Russia) and Gregory D. Edgecombe (Natural History Museum, UK) for their valuable comments to improve the manuscript.
The authors have declared that no competing interests exist.
No ethical statement was reported.
This work was supported by National Natural Science Foundation of China (82073972) and the Fundamental Research Funds for the Central public welfare research institutes (ZZXT2021011). Collections were made under the permit of the Fourth National Survey Chinese Material Medical Resources.
Tianyun Chen, performed the experiment; contributed significantly to analysis and manuscript preparation; Chao Jiang, performed the data analyses and wrote the manuscript;
Luqi Huang, helped perform the analysis with constructive discussions.
Tian-Yun Chen https://orcid.org/0000-0001-9761-0996
Chao Jiang https://orcid.org/0000-0003-1841-1169
All of the data that support the findings of this study are available in the main text or Supplementary Information.