Research Article |
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Corresponding author: Huifeng Zhao ( zhaohf@lfnu.edu.cn ) Academic editor: Samuel James
© 2026 Saiyu Qiao, Jiangni Li, Yufeng Zhang, Nonillon M. Aspe, Huifeng Zhao.
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:
Qiao S, Li J, Zhang Y, Aspe NM, Zhao H (2026) Establishment of the Drawida gisti species complex and description of a new species from northeastern China (Oligochaeta, Moniligastridae). ZooKeys 1293: 327-346. https://doi.org/10.3897/zookeys.1293.204408
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The family Moniligastridae Claus, 1880 remains taxonomically poorly resolved, and its actual species diversity is likely severely underestimated. Despite its ecological significance as an East Asian endemic species, Drawida gisti Michaelsen, 1931 has been studied almost exclusively from non-taxonomic perspectives. Consequently, its species boundaries remain poorly understood. Here, we formally establish the Drawida gisti species complex (DiSC), comprising eight morphologically and geographically related species. We employed an integrative taxonomic approach combining morphology and molecular data to examine specimens of DiSC from northeastern China, leading to the description of a new species, Drawida xuqini sp. nov. The new species is characterized by a colorless body in alcohol, 3–5 gizzards, M-shaped prostates, a very short and uncoiled vas deferens, and thick penes with a swollen base and blunt, rounded apex. Phylogenetic analyses based on 13 protein-coding genes of the mitochondrial genome of Drawida employing two molecular species delimitation methods—Assemble Species by Automatic Partitioning (ASAP), and Bayesian Phylogenetics and Phylogeography (BPP)—recovered D. xuqini sp. nov. as a monophyletic lineage, and COI genetic distances (19.4–22.0% between D. xuqini sp. nov. and D. gisti gisti) confirmed the species status of D. xuqini sp. nov. Additionally, we provide an updated redescription of D. gisti gisti based on newly examined specimens from multiple localities. Our findings highlight considerable external morphological plasticity in D. gisti gisti, which may likely be environmentally induced, and underscore the necessity of integrative approaches for accurate species delimitation in this group.
Annelida, mitogenome, new species, species group
The family Moniligastridae Claus, 1880 comprises five genera and 193 species. Among them, Drawida Michaelsen, 1900 is the largest genus with 163 species and subspecies (
Drawida gisti is distributed in eastern China (
We examined other morphologically similar species recorded from the distributional range of D. gisti gisti. These taxa were only reviewed based on published literature, as type specimens are presumed lost: Drawida cheni Gates, 1935 (body length 122–177 mm, diameter 4 mm) is distributed in Jiangsu; D. linhaiensis Chen, 1933 (37–58 mm, 3–4 mm) in Zhejiang, D. sinica Chen, 1933 (50–95 mm, 2.2–3.2 mm) in Jiangxi (
Key morphological characteristics of the Drawida gisti species complex (NA: not available).
| Characters | D. xuqini sp. nov. | D. gisti gisti | D. anchingiana | D. gisti nanchangiana | D. cheni | D. linhaiensis | D. sinica | D. sulcata | D. syringa |
|---|---|---|---|---|---|---|---|---|---|
| Color | light gray | Red-brown | light blue-green | light gray, without pigment | light blue | Light blue-gray | light green | brown | dusky flesh |
| Length | 48–87 | 48–66 | 56–80 | 55–98 | 122–177 | 37–58 | 50–95 | 60–86 | 50–100 |
| Width | 4.5–7.0 | 5.0–7.0 | 3.0–5.0 | 2.2–3.0 | 4.0 | 3.0–4.0 | 2.2–3.2 | 9.0–12.5 | 2.5–3.0 |
| Genital markings | VII–XI, unpaired, irregular | absent or present VIII–XII | VII–XI, unpaired | VII–XIII, XIV, unpaired, numerous | 7–10, irregular | 8–10, irregular | 8–11 | absent | absent or present in VII–IX |
| Clitellum | X–XIII; Pale pink | IX/X–XIII/XIV | X–XIII | X–XIII | ½IX–XIV,½XV | X–XIII, pale | X–XIII, pale white | NA | X–XII |
| Penis | 10/11, large, rounded at tip, large at base | 10/11, long, large at base gradually tapering to sharp | 10/11, small | 10/11, small, pointed at tip, large at base | 10/11, broad triangular at base | 10/11, large, transversely ovoid, rounded at tip | 10/11, small, rounded or transversely flat at tip, large at base 0.18 mm | Conical | 10/11 thick at base tapering at tip |
| Form of male pores | small pore on penis | longitudinal slit on penis | small pore on penis | paired | at tip of penis | 10/11 | small, at tip or slightly lateral of penis | 10/11, oval or eye-shaped | NA |
| Female pore | 11/12 | 11/12 | 11/12, transverse slit-like | paired | 12; in ab line | 11/12; in b line | 11/12; in b line | 11/12, Small crack-shaped | NA |
| Spermathecae ducts | medium, coiled | medium, coiled | medium, less coiled | coiled | long, coiled | slender, coiled | long, coiled | long, coiled | coiled |
| Spermathecae Ampulla | VIII | VIII | VII | VIII | VIII | VIII | VIII | VIII | VIII |
| Spermathecae Atrium | large, elliptical in 7/8 | large, comma-shaped in 8 | short, cylindrical in 7/8 | thumb-like and small in 8 | NA | in 7/8 | zigzag, long tube in 7/8 | cylindrical and slightly flattened | thick and short, stick-shaped |
| Ovaries | ovary chambers present, ovaries in XI–XII | ovary chambers present, ovaries in X | ovary chambers present, ovaries in XI | ovary chambers present, ovaries in XI | NA | ovary chambers present, ovaries in XI | ovary chambers present, ovaries in XI | ovary chambers present, ovaries in XI | NA |
| Ovisacs | XIII–XXI | X–XIII/XVI/XVIII | NA | XII–XIIII | XII, XIV, or XV | XVI–XX | X/XI/XII–XVI or XXII | NA | Extend to XXV |
| Gizzard segments and position | 3–5; IX–XVI, XII/XIII–XVIII or XI–XVII | 3; IX/X–XV/XVII/XVIII | 2 or 3; XI–XIII | 3; XII–XV | 3; XIII–XV | 3; XIII–XVI | 3 or 4; XI–XIII | 3, XI/XII–XIV | 2–5 |
| Vas deferens | short, uncoiled | extremely long, two coils | long, coiled | long, coiled | short, twisted | multi-coiled | extremely long, two coils | short coiled | NA |
| Prostate | M-shaped, white, 4.705–11.614 mm | Shape | slender, cylindrical, maximum 2 mm | Slender, short, club-shaped, 2.0–2.5 mm | multi-curved, 10–12 mm | large, long | long | smooth, long, 5.0–6.8 mm | short and broad, with thick glands on exterior |
Here, we provide an updated redescription of D. gisti gisti based on the newly examined specimens. Moreover, we employ an integrative taxonomic approach to examine and analyze earthworm specimens belonging to the DiSC collected from northeastern China, leading to the delimitation of a new species, Drawida xuqini sp. nov. Mitochondrial genomes and three nuclear markers were sequenced and analyzed for D. gisti gisti and D. xuqini sp. nov.
Specimens of the new species were collected from Weiyuan Park, Shanhaiguan District, Qinhuangdao Prefecture, Hebei Province, China. Specimens of D. gisti gisti were collected from Mentougou district in Beijing, Rizhao and Heze prefectures in Shandong Province, and Chengde Prefecture in Hebei Province, China (Table
List of Drawida species, incorporated into analysis (hyphen indicate not available).
| Specimen ID | Species | Location | Accession Number | ||||
|---|---|---|---|---|---|---|---|
| COI | 28S | AKAP17 | FLAD1 | Mitogenome | |||
| DLS_35 | D. gisti gisti | China: Beijing: Dongling Mountain | PX369606 | PZ484227 | PZ495527 | PZ495535 | PX369606 |
| L668_09_01 | D. gisti gisti | China: Hebei: Wuling Mountain | PZ462180 | PZ484228 | PZ495528 | PZ495536 | PZ462180 |
| SDRZ_03 | D. gisti gisti | China: Shandong: Rizhao | PZ462182 | PZ484230 | PZ495530 | PZ495537 | PZ462182 |
| SDHZ04_01 | D. gisti gisti | China: Shandong: Heze | PZ462181 | PZ484229 | PZ495529 | PZ495534 | PZ462181 |
| BJMTG01_II_3 | D. gisti gisti | China: Beijing: Mentougou | PZ462178 | – | – | – | PZ462178 |
| 345R_01 | D. xuqini sp. nov. | China: Hebei: Qinhuangdao | – | PZ484221 | – | – | PZ168347 |
| 345R_33 | D. xuqini sp. nov. | China: Hebei: Qinhuangdao | PZ486062 | PZ484222 | – | – | – |
| 345R_34 | D. xuqini sp. nov. | China: Hebei: Qinhuangdao | PZ486063 | PZ484223 | – | – | – |
| 345R_38 | D. xuqini sp. nov. | China: Hebei: Qinhuangdao | – | PZ484225 | PZ495524 | PZ495533 | PZ462175 |
| 345R_39 | D. xuqini sp. nov. | China: Hebei: Qinhuangdao | PZ486064 | – | – | – | – |
| 345R_40 | D. xuqini sp. nov. | China: Hebei: Qinhuangdao | – | PZ484224 | PZ495525 | PZ495532 | PZ462176 |
| 345R_42 | D. xuqini sp. nov. | China: Hebei: Qinhuangdao | – | PZ484226 | PZ495526 | PZ495531 | PZ462177 |
| HNLN_GR_I1_02 | D. henanensis | China: Henan: Luonling | PQ288546 | – | – | – | PQ278996 |
| HNLN_GR_I2_23 | D. henanensis | China: Henan: Luonling | PQ288549 | – | – | – | PQ278997 |
| HNLN_GR_I2_34 | D. sinensis | China: Henan: Luonling | PQ288559 | – | – | – | PQ278998 |
| HNLN_GR_I2_40 | D. sinensis | China: Henan: Luonling | PQ288561 | – | – | – | PQ278999 |
| HNLN_GR_I3_01 | D. japonica s. s. | China: Henan: Luonling | PQ288577 | – | – | – | PQ279000 |
| 360R02 | D. japonica s. s. | – | PZ168348 | – | – | – | PZ168348 |
| CQWX11_45 | D. wuxiensis | China: Chongqing: Wuxi | PX552887 | – | – | – | PX369605 |
| CQWX05_37 | D. wuxiensis | China: Chongqing: Wuxi | PX552881 | – | – | – | PZ210301 |
| CNWN02_01 | D. xinjiangensis | China: Xinjiang: Shuanghe | PX978833 | – | – | – | PZ099866 |
| nxGXJ1_1_1 | D. xinjiangensis | China: Ningxia: Xiji | PX978843 | – | – | – | PZ210303 |
| MN539609 | D. japonica s. l. | China: Jiangsu: Nanjing | – | – | – | – | MN539609 |
| – | E. irregularis | – | – | – | – | – | PZ168348 |
| – | M. anisodiverticulus | – | – | – | – | – | PV591013 |
External and internal characters were examined under a ZEISS stereomicroscope (ZEN v. 3.3 Pro). Measurements (to 0.1 mm) and dissections followed
Genomic DNA was extracted from posterior tissues (TIANGEN kit). Sanger sequencing targeted four markers: mitochondrial COI, nuclear 28S ribosomal RNA gene, 28S; A-kinase anchor protein 17A, AKAP17; flavin adenine dinucleotide synthetase 1, FLAD1 primers in Table
| Marker | Primer | Sequence (5’–3’) | Round | Source |
|---|---|---|---|---|
| COI | LCO1490 | GGTCAACAAATCATAAAGATATTGG | ||
| HCO2198 | TAAACTTCAGGGTGACCAAAAAATCA | |||
| 28S | 28sF1 | GAGTACGTGAAACCGTCTAG | ||
| 28SR1 | CGTTTCGTCCCCAAGGCCTC | |||
| AKAP17 | AKAP17-F1 | AAYTGGGARGTNATGGARAA | Round1 | |
| AKAP17-R1 | TCYTTRAACATNARYTTCAT | |||
| AKAP17-F2 | AARATGATHAARCCNGAYCARTT | Round2 | ||
| AKAP17-R2 | GCYTTNACRAANCCCATRTAYTC | |||
| FLAD1 | FLAD1-F1 | GGNCCNACNCAYGAYGAYAT | Round1 | |
| FLAD1-R1 | TTNGGRTGNGTRTTYTCCAT | |||
| FLAD1-F2 | TGYAARGCNTTYTTYGGNACNGA | Round2 | ||
| FLAD1-R2 | TTNACNCKCATRAAYTCNGGCCA |
Phylogenetic analyses were performed on a concatenated dataset of 13 protein-coding genes (PCGs) extracted from mitogenomes by a script of gbseqextractor_v2.py (
Species boundaries were assessed using Assemble Species by Automatic Partitioning (ASAP,
Family Moniligastridae Claus, 1880
Drawida gisti:
Drawida gisti f. typica:
Drawida gisti:
Drawida gisti:
Drawida gisti gisti:
• Five clitellates (SDRZ_03, SDRZ_04, L668_09_01, DLS_35, SDHZ4_01). • SDRZ_03, SDRZ_04: Rizhao, Shandong, China, Oct-2025, collected by Jiangni Li; • L668_09_01: Xinglong, Hebei, China, Jul-2025, collected by Huifeng Zhao; • DLS_35: Dongling Mountain Beijing, China, Jul-2025; • SDHZ4_01: Heze, Shandong, China, Oct-2025, collected by Yanmeng Bi. All specimens were preserved in the Hebei Key Laboratory of Animal Diversity, Langfang Normal University, Hebei, China (C-HLU).
Length 48–66 mm, diameter 5.0–7.0 mm. Segments 135–194. The setae begin on II and are closely paired; aa slightly less than bc. Setae clearly visible. Prostomium prolobous. Dorsal pores absent. The clitellar coloration variable, extending across segments IX/X–XIII/XIV. A pair of spermathecal pores are broad transverse slits on 7/8, at the middle of the cd, with the anterior and posterior margins swollen so that the pores appear elevated. The female pores are very minute, narrow transverse slits on 11/12, the slits in ab, external invisible. On 10/11 there is a larger transverse slit, in which a pair of penes is situated in bc, slightly nearer to b than to c. The true male pore is much smaller and opens at the ventral end of the penis as a longitudinal slit, about half the length of the penis. The penis of the examined specimens is 2.827 mm in length.
Drawida gisti gisti (SDRZ_03). A. Ventral view of the prostomium; B. Dorsal view of clitellum region; C, K. Ventral view of male pores, spermathecal pores, seta, and genital markings; D. Lateral view of the spermathecal pores, penis, and clitellum; E. Dorsal view of gizzards, ovisacs, and ovary chamber; F, J. Dorsal view of right testis sac, with vas deferens and prostate gland; G. Dorsal view of left testis sac and penial chamber; H, I. Dorsal view of right spermathecae. Scale bars: 1.0 mm (A, B); 0.5 mm (C–K).
Hearts from V to IX. Septa 5/6, 6/7, 7/8 and 8/9 thickened. Three gizzards in IX/X–XV/XVII/XVIII, occupying seven or nine segments. Testis sacs in two pairs, kidney-shaped, the concave side directed ventrally; in IX–X, or IX, or IX–XI, unconstricted by 9/10, but enveloped by fine membranes and fixed in the ventral body cavity. From the middle of each testis sac near the ventral median line a slender vas deferens emerges, first coiled into a tight bunch of loops on the anterior face of 9/10, then into another tight bunch on the posterior face; the anterior bunch smaller. The vas deferens finally opens on the ventral body wall of segment X. Prostate glands are in two parts: glandular portion and the prostatic duct. The glandular portion is covered with an outer layer of glandular tissue; when the outer layer is removed, a firm, slender, glossy central body is visible (Fig.
The eastern China (except northeastern China).
Drawida gisti gisti, D. anqingiana, and D. gisti nanchangiana share several morphological similarities: all three species possess three gizzards, and their prostates terminate in a slender duct connecting to the penial chamber, and ovarian chambers surrounded by septa, and spermathecal ampulla are relatively small with coiled ducts. The differences among the three are as follows: the prostate of D. gisti nanchangiana measures 2.0–2.5 mm in length, and D. gisti gisti is 7.6 mm, and reaching a maximum of 2.0 mm of D. anchingiana; the spermathecal atrium of D. gisti gisti is a larger (2.4 mm in length), that of D. anchingiana is a smaller, short-cylindrical to globose structure reaching a maximum of 2.0 mm, and that of D. g. nanchangiana is thumb-shaped and small. Prostate morphology differs substantially among D. gisti gisti and the other species of the complex. In D. gisti gisti, D. cheni, D. linhaiensis, D. sinica, and D. sulcata, the prostates are cylindrical, relatively long, but curved in various configurations. In D. syringa, however, the prostates are shorter and possess an additional outer glandular layer; comparative measurements are given in Table
Moreover, there is variation in penis length within D. gisti gisti. SDRZ_03 and SDRZ_04 possess long, well-developed penises, which is consistent with D. gisti.The penes of the remaining specimens are relatively small and lamellate, presumably due to morphological variation induced by differences in habitat environment. Additionally, these specimens exhibit some color variation, yet their internal morphology is highly uniform.
Holotype: • One clitellate (mature) specimen (345R_39) (87 mm in total length, dissected), Weiyuan Park, Shanhaiguan District, Qinhuangdao Prefecture, Hebei, China, 40.0139°N, 119.7485°E, 21-Aug-2019, collected by Yufeng Zhang. Paratypes: • Five clitellates (345R_33, 345R_34, 345R_37, 345R_38, 345R_40), data the same as the holotype. All specimens were preserved in C-HLU.
Length 48–87 mm, diameter 4.5–7.0 mm. Segments 160–181. Body light gray in 95% ethanol. Prostomium prolobous, dorsal pores absent. Clitellum pale pink, on segments X–XIII. Gizzards 3–5 in number. Vas deferens very short and uncoiled, a key character distinguishing it from most congeners in the DiSC. Prostates distinctly M-shaped, having a very slender terminal prostatic duct. Penes thick, with swollen base and blunt rounded apex; male pores minute, opens on the ventral face of the penis. Ovaries enclosed in ovarian chambers in segments XI–XII. Spermathecae situated in segment VIII, with large ampulla and small elliptical atrium.
Drawida xuqini sp. nov., holotype (345R_39). A. Ventral view of the prostomium; B. Dorsal view of clitellum region; C, K. Ventral view of male pores, spermathecal pores and genital markings; D. Lateral view of the spermathecal pores, penis and clitellum; E. Dorsal view of ovisacs, ovaries and ovary chamber; F. Dorsal view of gizzards; G, J. Dorsal view of right testis sac, with vas deferens and prostate gland; H, I. Dorsal view of right spermathecae. Scale bars: 1.0 mm (A–D, K); 0.5 mm (E–J).
Holotype: length 87 mm, diameter 7.0 mm. Segments 164. The setae begin on II and are closely paired; ab:cd:aa:bc = 1:1ab:10ab:10ab. Prostomium prolobous. Dorsal pores absent or invisible. Body color brown, head greyish blue, clitellum pale pink. Clitellum X–XIII. The spermathecal pores are narrow transverse slits on 7/8, at bc, close to c than b. The female pores are very minute, narrow transverse slits on 11/12, external invisible. On 10/11 there is a larger transverse slit, in which a pair of penes are situated in bc, slightly nearer to b than to c. The true male pore is much smaller and opens on the ventral face of the penis but is inconspicuous, no longitudinal or transverse slit observed. The left penis is 0.929 mm in length; the right is 1.066 mm in length. The penes of this new species are rather thick, swollen at the base and gradually tapering and the apex blunt resembling a triangular cone. Genital markings present, irregular and three in number, on segment VIII. The genital markings are mainly concentrated around the spermathecal pores.
Holotype: hearts from V to IX. Septa 5/6, 6/7, 7/8 and 8/9 thickened. The gizzards three to five, in IX–XVI, XII/XIII–XVIII or XI–XVII, occupying six, seven or eight segments. The testis sacs two pairs, 2.362 × 3.545 mm in size, elliptical; in IX–X, or IX/X, or IX–XI, occupying one to two segments, occasionally three, unconstricted by 9/10. A very short, uncoiled vas deferens arises from the margin at two-thirds of the testis sac, finally opening on the ventral body wall of segment X, 2.977 mm in length. One pair of prostates are M-shaped, 11.614 mm in length, only the glandular portion enveloped by granulations (external glandular layer), and when this outer layer is removed a smooth central body is visible. The prostate width gradually narrows, 0.828 mm at the apex and 0.302 mm at the distal end. On the inner side of the prostate near the ventral median line there are one or two accessory glands. The penial chambers in 10/11. A very short prostatic duct arises from the apex of the prostate, 0.437 mm in length, lying close to the opening of the vas deferens. The ovaries are on each side of the gizzard, in XI–XII, inverted U-shaped; the septa form ovarian chambers, and when these are removed the white ovaries are visible. The ovisacs appear as brown sacs covering the gizzard, extending to segment XXI. The spermathecae are in VIII; the ampulla large, milky white, 1.497 × 1.694 mm in size; the atrium small, elliptical, pale yellow, 1.853 × 0.859 mm in size, situated on the septum 7/8. The duct tightly coiled, gradually widening from the end of the ampulla and finally ending in the middle of the atrium; the anterior portion 0.129 mm in width, the posterior portion 0.173 mm in width, the duct about 10.0 mm in length.
In the paratypes, the genital markings are irregular, unpaired, and scattered on segments VII–XI. The number and position of gizzards vary as follows: specimen 345R_34 has three gizzards in segments 13–18; 345R_33 has five gizzards in segments 9–16; 345R_38 has four gizzards in segments 9–16; 345R_37 has three gizzards in segments 12–18; and 345R_40 has three gizzards in segments 11–17. Thus, the gizzard number ranges from three to five, and the segmental distribution ranges from nine to 18 among the paratypes. This variation is considered intraspecific, as is commonly observed in the genus Drawida. The testis sacs are two pairs, elliptical, measuring 2.362 × 3.545 mm, and occur in IX–X, or at IX/X, or in IX–XI, occupying one or two segments (occasionally three), and are not constricted by septum 9/10. The vas deferens ranges from 2.208 to 3.113 mm in length. The prostates are one pair, M-shaped, 4.705–11.614 mm long. The spermathecal ampulla is large, milky white, about 1.43 × 1.47 mm; the atrium is small, elliptical, pale yellow, about 1.585 × 0.859 mm. The ovisacs appear as brown sacs covering the gizzard and extend to segments XVI–XX.
The specific epithet xuqini is a patronymic genitive noun formed, in honor of Mr Qin Xu, the first author of “Terrestrial Earthworms (Opisthopora: Oligochaeta) of China”.
Known only from the type locality (Qinhuangdao, Hebei).
Drawida xuqini sp. nov. is readily distinguished from other members of the D. gisti species complex by a combination of clear morphological characters. The penis is thick, with a swollen base and a blunt, rounded apex, whereas in D. gisti gisti it is slender, elongated, and tapers to a sharp tip. The new species possesses 3–5 gizzards located in segments 9–18 (with intraspecific variation), while D. gisti gisti and most other species consistently have three gizzards. The vas deferens is very short and entirely uncoiled, a unique character among the complex (vs long and coiled in D. gisti gisti, D. anchingiana, D. g. nanchangiana, D. linhaiensis, D. sinica, and D. syringa; short but coiled in D. sulcata and D. cheni). The spermathecal atrium is elliptical and large in the new species (comma-shaped in D. gisti gisti), and the prostate is M-shaped without a slender terminal duct. The body color in alcohol is light gray (vs reddish brown in D. gisti gisti).
The mitogenomes of D. gisti gisti and D. xuqini sp. nov. are 14,647–14,648 bp and 14,668–14,750 bp in length, respectively, and all of them contain the typical mitochondrial 37 genes: 13 protein-coding genes (PCGs), two ribosomal RNA genes (rRNAs), and 22 transfer RNA genes (tRNAs). (Fig.
A, B. Mitogenomes of Drawida gisti gisti and D. xuqini sp. nov.; C. Phylogenetic relationship of D. xuqini sp. nov. and other species in DjSC and DiSC using 13 coding genes of mitogenomes. Numbers near branches indicate the maximum likelihood bootstrap support/Bayesian posterior probabilities. Values with a bootstrap value below 50 and a posterior probability lower than 80 have been omitted. Orange tips showed the new species.
Phylogenetic analysis based on the concatenated 13 PCGs (Fig.
Three methods—ASAP, GMYC, and BPP—were employed to delimit molecular operational taxonomic units (MOTUs) (Fig.
BPP analysis of the four-locus dataset consistently distinguished D. gisti gisti and D. xuqini sp. nov. as two distinct species congruent with morphological evidence. The K2P distances based on COI further supported the genetic integrity of the new species: intraspecific divergence ranged from 0 to 0.5%, whereas distances between D. xuqini sp. nov. and D. gisti gisti fell between 19.4% and 22.0% (Table
The COI genetic distance of K2P of groups of Drawida (values are percentages; intraspecific genetic distances are given in parentheses).
| Species | 1 | 2 | 3 | 4 | 5 | |
|---|---|---|---|---|---|---|
| 1 | D. xuqini sp. nov. | (0–0.5) | ||||
| 2 | D. gisti gisti | 19.4–22.0 | (0.3–5.7) | |||
| 3 | D. henanensis | 19.9–23.5 | 22.6–27.1 | (0.–5.7) | ||
| 4 | D. japonica s. s. | 23.3–24.3 | 22.3–23.4 | 19.2–22.8 | (0–0.2) | |
| 5 | D. sinensis | 22.3–23.8 | 20.8–22.0 | 20.2–22.5 | 17.6–22.2 | (13.3) |
Morphological examinations reveal considerable external plasticity in Drawida gisti gisti, with body color and size varying markedly among specimens from different localities. This external plasticity contrasts with the relatively conserved internal characters, which show only minor variations across specimens. Such disparity complicates species delimitation in Drawida, underscoring the necessity of integrative taxonomic approaches (
Notably, specimens with divergent penial morphology from Rizhao populations showed K2P genetic distances of 4.8–5.7% from typical D. gisti gisti populations, and species-delimitation analyses using ASAP and GMYC (based on COI) all recovered them as distinct evolutionary lineages. However, it should be noted that GMYC tends to separate biological species into several OTUs due to excessive lineage splitting (
The discovery of D. xuqini sp. nov. at Shanhaiguan in northeastern China hints at an expansion of distribution in DiSC. The Shanhaiguan population may represent a peripheral isolate that diverged through geographic isolation and local adaptation under different climatic regimes. Conversely, given the wide distribution of allied taxa across eastern China, additional cryptic species may await discovery in the southern portion of the range. Both hypotheses remain to be tested through denser sampling across the northern and southern provinces of China.
We thanked the anonymous referee for his improvement of the manuscript, and International Joint Research Center for Invertebrate Biology in Hebei Province.
The authors have declared that no competing interests exist.
No ethical statement was reported.
The authors accept full responsibility for the content of the manuscript, including the disclosure of any use of AI.
No AI tools were used in the preparation of this manuscript.
This study is funded by Science & Technology Fundamental Resources Investigation Program (Grant No. 2025FY100800 and 2024FY100400) and the Hebei Natural Science Foundation (Grant No. C2025408018).
Conceptualization, Qiao S., Li J., Zhang Y. and Zhao H.; specimen identification, Qiao S. and Zhao H.; methodology and experiments, Qiao S. and Zhao H.; data analysis, Qiao S. and Zhao H.; writing-original draft prepara-tion, Qiao S. and Zhao H.; writing-review and editing, Qiao S., Li J., Zhang Y. and Zhao H.
All of the data that support the findings of this study are available in the main text or Supplementary Information.
GMYC species delimition results
Data type: pdf
Explanation note: fig. S1. GMYC species delimition results based on COI (A) and 28S (B).
The result of molecular species delimitaiton
Data type: xlsx
Explanation note: table SS1. The result of molecular species delimitaiton of ASAP on COI and 28S in Drawida gisti gisti and D. xuqini sp. nov.
BPP related files
Data type: rar