Research Article |
Corresponding author: Piyatida Pimvichai ( piyatida_pimvichai@yahoo.com ) Academic editor: Luiz Felipe Iniesta
© 2022 Piyatida Pimvichai, Henrik Enghoff, Somsak Panha, Thierry Backeljau.
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:
Pimvichai P, Enghoff H, Panha S, Backeljau T (2022) A new genus of Pseudospirobolellidae (Diplopoda, Spirobolida) from limestone karst areas in Thailand, with descriptions of three new species. Zoosystematics and Evolution 98(2): 313-326. https://doi.org/10.3897/zse.98.90032
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A new genus of the millipede family Pseudospirobolellidae, Siliquobolellus gen. nov., is described from limestone mountains in Thailand, based on three new species, viz. Siliquobolellus amicusdraconis gen. et sp. nov. from Uthaithani Province, Siliquobolellus constrictus gen. et sp. nov. from Prachuap Khiri Khan Province and Siliquobolellus prasankokae gen. et sp. nov. (type species) from Lampang Province. The descriptions are based on gonopod morphology and mitochondrial DNA data (COI barcodes). The COI barcodes grouped the three new species in a well-supported Siliquobolellus gen. nov. clade. The mean interspecific COI sequence divergence among the three new species was 12% (range: 8–15%). The mean intergeneric COI sequence divergence between Siliquobolellus gen. nov., Coxobolellus Pimvichai, Enghoff, Panha & Backeljau, 2020, and Pseudospirobolellus Carl, 1912 was 19% (range: 14–23%). Three conspicuous gonopodal synapomorphies differentiate Siliquobolellus gen. nov. from other pseudospirobollellid genera: (1) the telopodital part of the posterior gonopod forms a deep concavity, (2) the telopodite of the anterior gonopod is directed distad and does not reach the tip of the coxal part of the anterior gonopod, and (3) the tip of the anterior gonopod coxa is narrowed, curving mesad. As such, the monophyly of the new genus is well supported by both morphological and mitochondrial DNA data. A distribution map and an identification key to the species are provided.
COI, gonopod, monophyly, synapomorphy
Hitherto, the small SE Asian spirobolidan millipede family Pseudospirobolellidae comprised < 20 species in 2 or 3 genera, viz. Pseudospirobolellus Carl, 1912, Coxobolellus Pimvichai, Enghoff, Panha & Backeljau, 2020, and Benoitolus Mauriès, 1980 (
In this context, the present study uses gonopod morphology and mitochondrial DNA sequence data to describe three new species in a new genus of Pseudospirobolellidae from limestone karst areas in Thailand.
Live specimens were hand-collected and were partly preserved in 70% ethanol for morphological study and partly placed in a freezer at –20 °C for DNA analysis.
This research was conducted under the approval of the Animal Care and Use regulations (numbers U1-07304-2560 and IACUC-MSU-037/2019) of the Thai government.
Gonopods were photographed with a digital camera. Samples for scanning electron microscopy (SEM: Hitachi TM4000Plus) were air-dried directly from alcohol and sputter-coated for 60 s with gold (Hitachi: MC1000). Scanning electron micrographs were taken at the Central Lab of Mahasarakham University. Drawings were made using a stereomicroscope and photographs. Voucher specimens were deposited in the collections of CUMZ.
Total genomic DNA was extracted from legs of single specimens of Siliquobolellus amicusdraconis gen. et sp. nov. (Hub Pa Tard, Uthaithani Province; CUMZ-D00149), Siliquobolellus constrictus gen. et sp. nov. (Ban Yang Chum, Prachuap Khiri Khan Province; CUMZ-D00150) and Siliquobolellus prasankokae gen. et sp. nov. (Pha Thai, Lampang Province; CUMZ-D00148) using the NucleoSpin Tissue kit (Macherey-Nagel, Düren, Germany) following the manufacturer’s instructions. PCR amplifications and sequencing of the standard mitochondrial COI DNA barcoding fragment (
Specimens from which the COI gene fragment was sequenced. CUMZ, Museum of Zoology, Chulalongkorn University, Bangkok, Thailand; NHMD, Natural History Museum of Denmark; NHMW, Naturhistorisches Museum, Vienna, Austria; NHM, The Natural History Museum, London, United Kingdom. Names of countries are in capitals. Abbreviations after species names refer to the isolate of each sequence. GenBank accession numbers are indicated for each species.
Voucher code | Locality | COI | |
---|---|---|---|
Genus Siliquobolellus gen. nov. | |||
S. amicusdraconis gen. et sp. nov. | CUMZ- D00149 | Hub Pa Tard, Lan-Sak, Uthaithani, THAILAND | OP174621 |
S. constrictus gen. et sp. nov. | CUMZ- D00150 | Ban Yang Chum, Kui Buri, Prachuap Khiri Khan, THAILAND | OP174622 |
S. prasankokae gen. et sp. nov. | CUMZ- D00148 | Pha Thai, Ngao, Lampang, THAILAND | OP174623 |
Genus Apeuthes | |||
A. maculatus Amc | NHMW-Inv. No.2395 | South Annam, VIETNAM | MF187404 |
A. maculatus Am26 | NHMD-621697 | Nha Trang, Bao Dai Villas Hotel, in garden, VIETNAM | MZ567159 |
A. fimbriatus BMP | CUMZ-D00144 | Bach Ma Peak, Da Nang, VIETNAM | MZ567160 |
A. longeligulatus TPP | CUMZ-D00140 | Tham Phet Po Thong, Klong Hard, Sa Kaeo, THAILAND | MZ567161 |
A. pollex SMR | CUMZ-D00141 | Sra Morakot, Klongthom, Krabi, THAILAND | MZ567162 |
A. pollex SML | CUMZ-D00142 | Koh 8, Similan islands, Phang-Nga, THAILAND | MZ567163 |
A. pollex WTS | CUMZ-D00143 | Tham Sue Temple, Muang, Krabi, THAILAND | MZ567164 |
?A. spininavis ABB | CUMZ-D00145 | Air Banun, Perak, MALAYSIA | MZ567165 |
Genus Atopochetus | |||
A. anaticeps SVL | CUMZ- D00091 | Srivilai temple, Chalermprakiet, Saraburi, THAILAND | MF187405 |
A. dollfusii DOL | NHM | Cochinchina, VIETNAM | MF187412 |
A. helix SPT | CUMZ- D00094 | Suan Pa Thong Pha Phum, Kanchanaburi, Thailand | MF187416 |
A. moulmeinensis TAK | CUMZ- D00095 | Km 87, Tha Song Yang, Tak, THAILAND | MF187417 |
A. setiferus HPT | CUMZ- D00097 | Hub Pa Tard, Lan-Sak, Uthaithani, THAILAND | MF187419 |
A. spinimargo Ton27 | NHMD- 00047013 | Koh Yo, Songkhla, THAILAND | MF187423 |
A. truncatus SML | CUMZ- D00101 | Koh 8, Similan islands, Phang-Nga, THAILAND | MF187424 |
A. uncinatus KMR | CUMZ- D00102 | Khao Mar Rong, Bangsapan, Prachuapkhirikhan, THAILAND | MF187425 |
A. weseneri Tos29 | NHMD-00047003 | Supar Royal Beach Hotel, Khanom, Nakhonsrithammarat, THAILAND | MF187431 |
Genus Aulacobolus | |||
A. uncopygus Auc | NHMW-Inv. No.2375 | Nilgiris, South India, INDIA | MF187433 |
Genus Benoitolus | |||
B. birgitae BBG | NHMD 621687 | Chiang Dao, Chiang Mai, THAILAND | MT328992 |
Genus Coxobolellus | |||
C. albiceps Stpw | CUMZ-D00121 | Tham Pha Tub, Muang District, Nan Province, THAILAND (green individual) | MT328994 |
C. albiceps Stpl | CUMZ-D00122 | Tham Pha Tub, Muang District, Nan Province, THAILAND (small, brown individual) | MT328993 |
C. albiceps TPB | CUMZ-D00123 | Wat Tham Bampen Bun, Pan District, Chiang-Rai Province, THAILAND | MT328996 |
C. albiceps Stvd | CUMZ-D00124 | Tham Wang Daeng, Noen Maprang District, Phitsanulok Province, THAILAND | MT328995 |
C. compactogonus SKR | CUMZ-D00134 | Sakaerat Environmental Research Station, Wang Nam Khiao District, Nakhon Ratchasima Province, THAILAND | MT328998 |
C. compactogonus KLC | CUMZ-D00135 | Khao Look Chang, Pak Chong District, Nakhon Ratchasima Province, THAILAND | MT328997 |
C. fuscus HKK | CUMZ-D00133 | Kroeng Krawia waterfall, Sangkhla Buri District, Kanchanaburi Province, THAILAND | MT328999 |
C. nodosus SPW | CUMZ-D00126 | Chao Por Phawo Shrine, Mae Sot District, Tak Province, THAILAND | MT329000 |
C. serratus KKL | CUMZ-D00132 | Khao Kalok, Pran Buri District, Prachuap Khiri Khan Province, THAILAND | MT329001 |
C. simplex TNP | CUMZ-D00136 | Tham Pha Pha Ngam, Mae Prik District, Lampang Province, THAILAND | MT329002 |
C. tenebris KWP | CUMZ-D00119 | Wat Khao Wong Phrohm-majan, Ban Rai District, Uthai Thani Province, THAILAND | MT329003 |
C. tenebris TPL | CUMZ-D00120 | Wat Tham Phrom Lok Khao Yai, Sai Yok District, Kanchanaburi Province, THAILAND | MT329004 |
C. tigris TKP | CUMZ-D00130 | Wat Tham Khao Plu, Pathio District, Chumphon Province, THAILAND | MT329005 |
C. tigris TYE | CUMZ-D00131 | Tham Yai I, Pathio District, Chumphon Province, THAILAND | MT329006 |
C. transversalis Stpg | CUMZ-D00125 | Tham Pha Tub, Muang District, Nan Province, THAILAND | MT329007 |
C. valvatus TCD | CUMZ-D00127 | Wat Tham Chiang Dao, Chiang Dao District, Chiang-Mai Province, THAILAND | MT329009 |
C. valvatus BRC | CUMZ-D00128 | Tham Borichinda, Chom Thong District, Chiang-Mai Province, THAILAND | MT329008 |
C. valvatus TST | CUMZ-D00129 | Tham Sam Ta, Muang District, Mae Hong Son Province, THAILAND | MT329010 |
Genus Leptogoniulus | |||
L. sorornus BTN | CUMZ- D00109 | Botanical Garden, Penang, MALAYSIA | MF187434 |
Genus Litostrophus | |||
L. chamaeleon PPT | CUMZ- D00111 | Phu Pha terb, Mukdahan, THAILAND | MF187436 |
L. saraburensis PKS | CUMZ- D00113 | Phukhae Botanical Garden, Saraburi, THAILAND | MF187438 |
L. segregatus Ls19 | NHMD 621686 | Koh Kut, Trad, THAILAND | MF187440 |
Genus Macrurobolus | |||
M. macrurus INT | CUMZ- D00147 | Wat Tham Inthanin, Mae Sot District, Tak Province, THAILAND | MZ905519 |
Genus Madabolus | |||
M. maximus Mm4 | NHMD-00047007 | de Toliara Province, Parc National de Bermaraha, South Bank of Manambolo River, Near Tombeau Vazimba, MADAGASCAR | MF187441 |
Genus Narceus | |||
N. annularis | NC_003343.1 | ||
Genus Parabolus | |||
P. dimorphus Pd34 | NHMD-00047004 | Dar es Salaam, TANZANIA | MF187442 |
Genus Paraspirobolus | |||
P. lucifugus | AB608779.1 | ||
Genus Pelmatojulus | |||
P. tigrinus Pt2 | NHMD-00047008 | Southern part of the Comoé N.P., 30 km north of Kakpin, CÔTE d’IVOIRE | MF187443 |
P. togoensis Pto6 | NHMD-00047006 | Biakpa, GHANA | MF187444 |
Genus Pseudospirobolellus | |||
Pseudospirobolellus avernus GPG | CUMZ-D00117 | Gua Pulai, Gua Musang, Kelantan, MALAYSIA | MT329011 |
Pseudospirobolellus sp. KCS | CUMZ-D00118 | Koh Chuang, Sattahip, Chonburi, THAILAND | MT329012 |
Genus Rhinocricus | |||
R. parcus Rp49 | NHMD-00047009 | Puerto Rico, USA | MF187449 |
Genus Trachelomegalus | |||
T. sp. Tr54 | NHMD-00047012 | Borneo Sabah, MALAYSIA | MF187445 |
Genus Trigoniulus | |||
T. corallinus Tco15 | NHMD-00047010 | Vientiane, LAOS | MF187446 |
Outgroup | |||
Genus Anurostreptus | |||
A. barthelemyae Tlb | CUMZ-D00003 | Thale-Ban N.P., Khuan-Don, Satun, THAILAND | KC519469 |
Genus Chonecambala | |||
C. crassicauda Ttp | CUMZ-D00001 | Ton-Tong waterfall, Pua, Nan, THAILAND | KC519467 |
Genus Thyropygus | |||
T. allevatus Bb | CUMZ-D00013 | BangBan, Ayutthaya, THAILAND | KC519479 |
The COI data included 56 specimens, representing 18 genera and 46 nominal species of ingroup taxa (Table
CodonCode Aligner (ver. 4.0.4, CodonCode Corporation) was used to assemble the forward and reverse sequences and to check for errors and ambiguities. All sequences were checked with the Basic Local Alignment Search Tool (BLAST) provided by NCBI and compared with reference sequences in GenBank. All sequences were aligned using MUSCLE (ver. 3.6, see http://www.drive5.com/muscle;
Phylogenetic trees were constructed using maximum likelihood (ML) and Bayesian inference (BI). The shape parameter of the gamma distribution, based on 16 rate categories, was estimated using maximum-likelihood analysis. ML trees were inferred with RAxML (ver. 8.2.12, see http://www.phylo.org/index.php/tools/raxmlhpc2_tgb.html;
BI trees were run for 2 million generations (heating parameter: 0.02), sampling every 1000 generations. Convergences were confirmed by verifying that the standard deviations of split frequencies were below 0.01. Then the first 1000 trees were discarded as burn-in, so that the final consensus tree was built from the last 3002 trees. Support for nodes was assessed by posterior probabilities.
For ML we consider branches with bootstrap values (BV) of ≥ 70% to be well supported (
The uncorrected p-distances between the sequences ranged from 0.00 to 0.26 (Suppl. material
The mean intergeneric sequence divergences between Siliquobolellus gen. nov. and various pachybolid genera were: 0.20 (range: 0.18–0.23) for Apeuthes Attems, 1938, 0.23 (range: 0.20–0.26) for Atopochetus Attems, 1953, 0.21 (range: 0.18–0.22) for Litostrophus Chamberlin, 1921, and 0.20 (range: 0.19–0.22) for Pelmatojulus De Saussure, 1860.
The mean intergeneric sequence divergences between Siliquobolellus gen. nov. and the other Spirobolida families were: 0.22 (range: 0.21–0.24) for Narceus annularis (Rafinesque, 1820) (Spirobolidae) and 0.24 (range: 0.23–0.26) for Paraspirobolus lucifugus (Gervais, 1836) (Spirobolellidae).
Both the BI and ML trees (Fig.
Phylogenetic relationships of Siliquobolellus gen. nov. and several other spirobolidan millipede taxa based on maximum likelihood analysis (ML) and Bayesian inference (BI) of a 660 bp COI gene fragment. Numbers at nodes indicate branch support based on bootstrapping (ML) / posterior probabilities (BI). Scale bar = 0.3 substitutions/site. # indicates branches with < 50% ML bootstrap support or < 0.95 posterior probability, - indicates non-supported branches. The colored areas mark the families Pseudospirobolellidae (minus Benoitolus) (purple) and Pachybolidae (plus Benoitolus) (yellow).
Class Diplopoda de Blainville in Gervais, 1844
Order Spirobolida Bollman, 1893
Suborder Spirobolidea Bollman, 1893
Family Pseudospirobolellidae Brölemann, 1913
From Latin siliqua = pod, and referring to the podlike shape of the gonopod telopodite.
Siliquobolellus prasankokae gen. et sp. nov.
Siliquobolellus amicusdraconis gen. et sp. nov. and Siliquobolellus constrictus gen. et sp. nov.
Siliquobolellus species are characterised by (1) the unique shape of the telopodital part of their posterior gonopod, which forms a deep concavity. In contrast, the telopodital part of the posterior gonopod is extremely slender and sickle-shaped in Pseudospirobolellus, does not form a deep concavity in Coxobolellus, or is slender with a broad apical canopy in Benoitolus; (2) the telopodite of anterior gonopod (at) simple, directed distad, not reaching tip of coxal part of the anterior gonopod (cx). In contrast, the telopodite of anterior gonopod (at) is overreaching coxa in the other three genera; and (3) tip of anterior gonopod coxa narrowed, curving mesad. In contrast the tip of the anterior gonopod coxa directed distad in Coxobolellus, or is rounded in Pseudospirobolellus and Benoitolus.
Head capsule smooth. Occipital furrow extending down between, but not beyond eyes; clypeal furrow reaching level of antennal sockets. Area below antennal sockets and eyes impressed, forming part of antennal furrow. Incisura lateralis (IL) open (Fig.
External morphology of Siliquobolellus gen. nov. A–C. S. amicusdraconis gen. et sp. nov. A. Head, lateral view; B. Head, ventral view; C. Posterior end, lateral view; D. Posterior end, lateral view (S. prasankokae gen. et sp. nov.); E. Posterior end, lateral view (S. constrictus gen. et sp. nov.); F–I. S. prasankokae gen. et sp. nov.; F. Posterior end, ventral view; G, H. Male leg, latero-ventral view; I. Body rings, dorsal colour pattern; J. S. constrictus gen. et sp. nov., body rings, dorsal colour pattern. Av = anal valves; Gst = gnathochilarial stipes; IL = incisura lateralis Me = mentum; Mst = Mandibular stipes; Pre = preanal ring; Sub = subanal scale.
Collum smooth, with a marginal furrow along lateral part of anterior margin; lateral lobes narrowly rounded, extending as far ventrad as the ventral margin of body ring 2.
Body rings 2–3 ventrally concave, hence with distinct ventrolateral “corners”. Body rings very smooth, parallel-sided in dorsal view. Prozona smooth. ‘Tergo-pleural’ suture visible on pro- and mesozona; mesozona ventrally with fine oblique striae, dorsally punctate; metazona ventrally with fine longitudinal striae, otherwise smooth. “Pleural” parts of rings with fine oblique striae. Sterna transversely striate. Ozopores from ring 6, situated in mesozona, ~1/2 pore diameter in front of metazona.
Telson smooth; preanal ring (Pre) with slightly recurved dorsal profile, with short process protruding to vertical tangent to anal valves or slightly beyond (Fig.
Legs (Fig.
Male sexual characters. Apical part of coxa and entire ventral surface of prefemur from third to the last body rings with large ventral soft pad. Body ring 7 entirely fused ventrally, no trace of a suture. Tip of anterior gonopods visible when the animal is stretched out (not when it is rolled up).
Anterior gonopods (Figs
Siliquobolellus amicusdraconis gen. et sp. nov., holotype, gonopods (specimen from Hub Pa Tard, CUMZ-D00149-1). A. Anterior gonopod, anterior view; B. Anterior gonopod, posterior view, arrow indicates a thumb-like process; C. Right posterior gonopod, lateral view; D. Anterior gonopod, anterior view; E. Anterior gonopod, posterior view; F. Right posterior gonopod, lateral view; G. SEM, Right posterior gonopod, latero-mesal view; H. SEM, Mesal part of right posterior gonopod, lateral view; I. SEM, Right posterior gonopod, latero-mesal view; J. SEM, Tip of right posterior gonopod, latero-mesal view, arrow indicates a canopy; K. SEM, lateral part of pt, latero-mesal view; L. SEM, left female vulva, posterior mesal view. at = anterior gonopod telopodite; cx = coxa; oeg = opening of efferent groove; op = operculum of vulva; pcx = coxal part of the posterior gonopod telopodite; pt = telopodital part of the posterior gonopod telopodite; st = sternal process.
Posterior gonopods (Figs
Female vulvae (Figs
Siliquobolellus constrictus gen. et sp. nov., holotype, gonopods (specimen from Ban Yang Chum, CUMZ-D00150-1). A. Anterior gonopod, anterior view; B. Anterior gonopod, posterior view, arrow indicates a thumb-like process, with longitudinal ridge; C. Right posterior gonopod, lateral view; D. Anterior gonopod, anterior view; E. Anterior gonopod, posterior view; F. Right posterior gonopod, lateral view; G. SEM, Left posterior gonopod, lateral view; H. SEM, Left posterior gonopod, latero-mesal view; I. SEM, tip of left posterior gonopod, latero-mesal view; J. SEM, Mesal part of left posterior gonopod, lateral view; K. SEM, left female vulva, posterior mesal view. at = anterior gonopod telopodite; cx = coxa; oeg = opening of efferent groove; op = operculum of vulva; pcx = coxal part of the posterior gonopod telopodite; pt = telopodital part of the posterior gonopod telopodite; st = sternal process.
Siliquobolellus prasankokae gen. et sp. nov., holotype, gonopods (specimen from Pha Thai, CUMZ-D00148-1). A. Anterior gonopod, anterior view; B. Anterior gonopod, posterior view, arrow indicates a a thumb-like process with strong ridge distomesally; C. Right posterior gonopod, lateral view, arrow indicates a short, sharp protrusion; D. Anterior gonopod, anterior view; E. Anterior gonopod, posterior view; F. Right posterior gonopod, lateral view; G. SEM, Right posterior gonopod, lateral view, arrow indicates a short, sharp protrusion; H. SEM, Mesal part of right posterior gonopod, lateral view; I. SEM, Right posterior gonopod, mesal view, arrow indicates a canopy; J. SEM, Tip of right posterior gonopod, mesal view; K. SEM, left female vulva, posterior mesal view. at = anterior gonopod telopodite; cx = coxa; oeg = opening of efferent groove; op = operculum of vulva; pcx = coxal part of the posterior gonopod telopodite; pt = telopodital part of the posterior gonopod telopodite; st = sternal process.
Species descriptions
Holotype. 1 male (CUMZ-D00149-1), Thailand, Uthaithani Province, Lan-Sak District, Hub Pa Tard; 15°22'37.13"N, 99°37'49.98"E; 119 m a.s.l.; 26 July 2020; P. Pimvichai, T. Backeljau and P. Prasankok leg.
Paratypes. 3 males (CUMZ-D00149-3), 3 females (CUMZ-D00149-2); same data as holotype.
The species epithet means “friend of the dragon” and refers to the type locality, which is shared with the “shocking pink dragon millipede”, Desmoxytes purpurosea Enghoff, Sutcharit & Panha, 2007.
Differing from other species in the genus by having the tip of anterior gonopod crossing over with tip of opposite side, the two together delimiting a drop-shaped “window”, whereas in the other two species the tips of anterior gonopod are separated from each other; externally differing by its bright orange color on the anterior and posterior ends. In contrast, S. constrictus gen. et sp. nov. has a row of triangular dark brown spots middorsally on its body rings, while S. prasankokae gen. et sp. nov. has a row of rectangular dark brown spots middorsally on the body rings.
Adult males with 41 or 42 podous rings, 1 apodous ring. Length ~3 cm, diameter 2.7–2.9 mm. Adult females with 41–43 podous rings, 1–3 apodous rings. Length ~3 cm, diameter 2.8–3.1 mm.
Colour. Living animal mainly dark brown. Head, dorsal part of the first four body rings and telson orange, antenna light brown, middorsal metazona orange (Fig.
Anterior gonopods (Fig.
Posterior gonopods (Fig.
Female vulvae (Fig.
The GenBank accession number of the COI barcode of the paratype is OP174621 (voucher code CUMZ-D00149).
Found under leaf litter and on rocks.
Known only from the type locality in Uthaithani Province, Thailand (Fig.
Holotype. 1 male (CUMZ-D00150-1), Thailand, Prachuap Khiri Khan Province, Kui Buri District, Ban Yang Chum; 12°04'19.64"N, 99°42'57.63"E; 149 m a.s.l.; 7 August 2014; C. Sutcharit leg.
Paratypes. 1 male (CUMZ-D00150-2), 2 females (CUMZ-D00150-3); same data as holotype.
The species epithet is a Latin adjective derived from the verb constringere = “to tighten” and refers to the constricted anterior gonopod coxa.
Differing from other species in the genus by having the anterior gonopod coxae constricted at middle of lateral margin, while the other two species have no constriction at middle of lateral margin of anterior gonopod coxae; externally differing by having a middorsal row of triangular dark brown spots, with dark brown bands on both lateral sides running parallel to the dorsal one.
Adult males with 41 or 42 podous rings, 1 apodous ring. Length ~3 cm, diameter 3.6–3.7 mm. Adult females with 42 or 43 podous rings, 1 apodous ring. Length ~3 cm, diameter 4.1–4.5 mm.
Colour. After 8 years in alcohol overall beige with 3 stripes from head to telson: a mid-dorsal row of triangular dark brown spots, with dark brown bands on both lateral side running parallel to the dorsal one (Fig.
Anterior gonopods (Fig.
Posterior gonopods (Fig.
Female vulvae (Fig.
The GenBank accession number of the COI barcode of the paratype is OP174622 (voucher code CUMZ-D00150).
Found under leaf litter.
Holotype. 1 male (CUMZ-D00148-1), Thailand, Lampang Province, Ngao District, Pha Thai; 18°36'01.05"N, 99°54'01.00"E; 350 m a.s.l.; 28 July 2020; P. Pimvichai, P. Prasankok and S. Saratan leg.
Paratypes. 6 females (CUMZ-D00148-2); same data as holotype.
The species honors Associate Professor Dr. Pongpun Prasankok, biologist and devoted millipede collector.
Differing from other species in the genus by having the posterior gonopod apically with a short, sharp protrusion; externally differing by having a row of rectangular dark brown spots middorsally on the body rings.
Adult male with 41 podous rings, 1 apodous ring. Length ~3 cm, diameter ~3 mm. Adult females with 40 or 41 podous rings, 1 apodous ring. Length ~3 cm, diameter 3.2–3.6 mm.
Color of living animal: collum and telson beige; antenna and leg light brown; dorsal part of body rings yellowish brown, with a row of rectangular dark brown spots middorsally on the body rings; head and lateral part of body rings dark brown (Fig.
Anterior gonopods (Fig.
Posterior gonopods (Fig.
Female vulvae (Fig.
The GenBank accession number of the COI barcode of the paratype is OP174623 (voucher code CUMZ-D00148).
Found under leaf litter and inside a rotten log.
Known only from the type locality in Lampang Province, Thailand (Fig.
1 | Telopodital part of the posterior gonopod forming a deep concavity; telopodite of anterior gonopod not reaching tip of coxal part of anterior gonopod; tip of anterior gonopod coxa narrowed, curving mesad | Siliquobolellus gen. nov. |
– | Telopodital part of the posterior gonopod simple, not forming a deep concavity; telopodite of anterior gonopod protruding above the tip of coxal part of anterior gonopod | 2 |
2 | The coxae of the 3rd male leg-pair with extremely large, protruding processes | Coxobolellus Pimvichai, Enghoff, Panha & Backeljau, 2020 |
– | The coxae of the 3rd male leg-pair without large processes | 3 |
3 | The posterior gonopod extremely slender, sickle-shaped | Pseudospirobolellus Carl, 1912 |
– | The posterior gonopod slender, apically with broad canopy | Benoitolus Mauriès, 1980 |
1 | Tip of anterior gonopod coxa crossing over with tip of opposite side (Fig. |
S. amicusdraconis gen. et sp. nov. |
– | Tip of anterior gonopod coxa separate from that of the opposite side | 2 |
2 | Posterior gonopod telopodite apically with a short, sharp protrusion (Fig. |
S. prasankokae gen. et sp. nov. |
– | Posterior gonopod telopodite apically without a protrusion, anterior gonopod coxae constricted at middle of lateral margin (Fig. |
S. constrictus gen et sp. nov. |
Morphologically, the three new species clearly belong to the Pseudospirobolellidae because they share the unique characters of the family viz., (1) prefemora of male legs with large ventral soft pads, (2) posterior gonopod simple and independent of each other, (3) small and slender species, (4) anterior gonopods sternum slightly fused with coxae or present only by a small inward pleat, and (5) tracheal apodeme is articulated at the base of the coxae (
While the new pseudospirobolellid genus seems to be well-founded, it does show a remarkably high mean interspecific COI sequence divergence of 12% (range: 8–15%). Yet, this appears to be a common phenomenon in millipedes, with reported mean interspecific COI divergences between congeneric species of, for example, 11% (range: 6–15%) in Coxobolellus (
The new genus and the three new species constitute an important expansion of the family Pseudospirobolellidae, but at the same time further expose the problem of the highly divergent position of Benoitolus birgitae within Pachybolidae, instead of Pseudospirobolellidae (
This research was funded by the Thailand Science Research and Innovation (TSRI) together with Mahasarakham University as a TRF Research Career Development Grant (2019–2022; RSA6280051) (to P. Pimvichai). Additional funding was provided by the Royal Belgian Institute of Natural Sciences. We thank Sathit Saratan (Sirindhorn Museum) for assistance in collecting material. We are indebted to Thita Krutchuen (College of Fine Arts, Bunditpatanasilpa Institute) for the excellent drawings. We thank Thomas Wesener (Zoological Research Museum Alexander Koenig, Bonn, Germany) for the critical comments and helpful discussions, which have substantially improved our manuscript.
Table S1
Data type: excel file
Explanation note: Estimates of COI sequence divergences within and among Siliquobolellus gen. nov. species and related taxa expressed as uncorrected p-distances (rounded off to two decimals).