Research Article |
Corresponding author: Yi-Tong Ма ( mayitong@ntu.edu.cn ) Academic editor: Louis Deharveng
© 2023 Mei-Dong Jing, Yin-Huan Ding, Yi-Tong Ма.
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:
Jing M-D, Ding Y-H, Ма Y-T (2023) Description of three new species of Callyntrura (Japonphysa) (Collembola, Entomobryidae) from China with the aid of DNA barcoding. ZooKeys 1187: 237-260. https://doi.org/10.3897/zookeys.1187.109608
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Callyntrura(s.l.) Börner, 1906 is the largest genus of the subfamily Salininae and contains 11 subgenera and 98 species from all over the world (mainly Asia), with eight species recorded from China. In the present paper, three new species of Callyntrura(s.l.) are described from China: C. (Japonphysa) xinjianensis sp. nov.; C. (J.) tongguensis sp. nov. and C. (J.) raoi sp. nov. Their differences in colour pattern, chaetotaxy and other characters are slight, however distances of COI mtDNA support their validation as three new distinct species. A key to the Chinese Callyntrura(s.l.) is provided.
Chaetotaxy, DNA sequence, identification key, Salininae, subgenus, taxonomy, Yoshii
The genus Callyntrura (s.l.) Börner, 1906 was previously considered a member of the family Paronellidae, but now belongs to the family Entomobryidae (
Callyntrura(s.l.) specimens are medium-sized and their colour pattern plays a key role in its classification. So far, 97 species of Callyntrura have been described from Southeast and South Asia and one species from Africa and the descriptions of most species were quite simple (
Specimens were collected with an aspirator and stored in 99% alcohol. They were mounted on glass slides in Marc André II solution, and were studied with a Leica DM2500 phase contrast microscope. Photographs were taken under a Leica DFC300 FX digital camera which mounted on the microscope and enhanced with Photoshop CS2 (Adobe Inc.). SEM photographs were taken under a ZEISS Gemini SEM 300 after the specimens were coated with a Leica EM ACE600. Type specimens are deposited in School of Life Sciences, Nantong University, Jiangsu, China.
The nomenclature of the dorsal macrochaetotaxy of the head and interocular chaetae are described following
DNA was extracted from one specimen per species by using an Ezup Column Animal Genomic DNA Purification Kit (Sangon Biotech, Shanghai, China) following the manufacturer’s standard protocols. Amplification of a 658 bp fragment of the mitochondrial COI gene was carried out using a Prime Thermal Cycler (TECHNE, Bibby Scientific Limited, Stone, Staffordshire, UK) in 25 μl volumes using Premix Taq polymerase system (Takara Bio, Otsu, Shiga, Japan). The primers and PCR progams followed
DNA sequences were assembled using Sequencher 4.5 (Gene Codes Corp), and then deposited in GenBank. Sequences were aligned by ClustalW implemented in MEGA 6 (
Ant. antennal segment(s);
Th. thoracic segment(s);
Abd. abdominal segment(s);
mac macrochaeta(e);
mes mesochaeta(e);
ms specialised microchaeta(e);
sens specialized ordinary chaeta(e).
The distribution in China of the species described in present paper is shown in Fig.
Class Collembola Lubbock, 1873
Order Entomobryomorpha Börner, 1913
Family Entomobryidae Tömösvary, 1882
Moderate size, usually 2–3 mm; antennae four segmented and without apical bulb; eyes 8+8; frontal spines on head 4+4; scales present on body; dens smooth; mucro almost square and with more than three teeth.
Holotype. ♀ on slide, China, Jiangxi Province, Nanchang City, Xinjian District, Meiling Town, Jiuxi Village, 12-XI-2020, 28°47'56"N, 115°45'11"E, 168 m asl, sample number 1243. Paratypes. 3♀♀ on slides, same data as holotype; 6♀♀ on slides, China, Jiangxi Province, Nanchang City, Xinjian District, Meiling Town, Shizifeng Park, 12-XI-2020, 28°49'26"N, 115°43'06"E, 100 m asl, sample number 1242. All collected by Y-T Ma.
Size. Body length up to 2.75 mm.
Coloration. Ground colour pale yellow; eye patches dark blue; brown stripe present from head to Abd. III laterally; middle part of Abd. II–III with brown pigment; medial and posterior margin of Abd. IV with pair of irregular brown patches, respectively; brown pigment scattered on basal Ant. I and distal Ant. IV, legs, anterior part of ventral tube and distal dentes (Figs
Head. Antenna not annulated and 1.37–1.57 times length of body. Ratio of Ant. I–IV as 1.00/0.85–1.00/0.49–0.72/1.37–2.00. Distal part of Ant. IV with many sensory chaetae and normal ciliate chaetae, without apical bulb (Figs
Callyntrura xinjianensis sp. nov. 7 SEM photomicrograph of apex of Ant. IV 8 apex of Ant. IV (dorsal view) 9 photomicrograph of distal Ant. III (arrow showing rod-like chaeta of Ant. III organ, ventral view) 10 distal Ant. III (ventral view) 11 SEM photomicrograph of basal Ant. I 12 basal Ant. I (dorsal view) 13 basal Ant. I (ventral view) 14 SEM photomicrograph of anterior part of dorsal head 15 dorsal head (right side) 16 SEM photomicrograph of frontal spines of head 17 frontal spine 18 SEM photomicrograph of prelabrum and labrum 19 prelabrum and labrum. Scale bars: 20 μm.
Callyntrura xinjianensis sp. nov. 20 photomicrograph of maxillary palp and outer lobe (arrow showing basal chaeta, right side) 21 maxillary palp and outer lobe (right side) 22 SEM photomicrograph of labial palp E (right side) 23 labial palp E (right side) 24 photomicrograph of labial base (left side) 25 labial base (left side). Scale bars: 20 μm.
Thorax. Tergal ms formula on Th. II–Abd. V as 1, 0/1, 0, 1, 0, 0, sens as 1, 1/0, 2, 2, 31–44, 3 (Figs
Callyntrura xinjianensis sp. nov. 26 chaetotaxy of Th. II–III (right side, solid black dot meaning absence) 27 trochanteral organ 28 hind foot complex (lateral view) 29 SEM photomicrograph of fore foot complex (lateral view) 30 SEM photomicrograph of middle foot complex (lateral view) 31 SEM photomicrograph of hind foot complex (lateral view). Scale bars: 20 μm.
Variation of tergal chaetotaxy of the new species (?, chaetotaxy not seen clearly).
Species | Specimen number | Th. II | Th. III | Abd. I | Abd. III | Abd. IV | |
---|---|---|---|---|---|---|---|
posterior | posterior | lateral | medial | posterior | |||
Callyntrura xinjianensis sp. nov. | 1242-2A | 9+? | 12+13 | 7+7 | ?+? | 16+17 | 23+24 |
1242-2B | 8+8 | 14+14 | 7+7 | 12+12 | 16+17 | 23+24 | |
1242-2C | 8+8 | 12+13 | 7+7 | 13+13 | 16+17 | 17+17 | |
1242-3A | 9+9 | 12+13 | 7+7 | 12+14 | 17+? | 18+21 | |
1242-3B | 9+9 | 13+13 | 7+7 | 11+13 | 15+16 | 17+17 | |
1242-3C | 9+9 | 14+14 | 8+8 | 11+11 | 16+16 | 23+24 | |
1243-3A | 9+9 | 13+13 | 7+7 | 12+? | 15+? | 14+18 | |
1243-3B | 9+? | 14+14 | 7+8 | 10+12 | 16+16 | 16+19 | |
1243-3C | 9+9 | 12+13 | 8+8 | 14+? | ?+? | 18+23 | |
1243-3D | 8+9 | 13+14 | 7+7 | 11+? | 15+16 | 13+14 | |
C. tongguensis sp. nov. | 1235-3A | 10+10 | 15+15 | 11+11 | 16+16 | 15+15 | 19+22 |
1235-3B | 10+10 | 14+15 | 11+11 | 14+14 | 17+17 | 20+23 | |
1235-3C | 10+10 | 15+15 | 10+11 | 16+? | 17+18 | 18+19 | |
C. raoi sp. nov. | 1244-1A | 10+10 | 14+14 | 8+9 | 10+? | 17+17 | 10+11 |
1244-1B | 10+10 | 14+14 | ?+? | 15+15 | 15+15 | 14+18 | |
1244-1C | 10+10 | 14+14 | 9+? | 14+18 | 18+18 | 19+27 | |
1244-3A | 9+9 | 14+14 | 9+9 | 14+? | 15+15 | 22+24 | |
1244-3B | 10+10 | 14+14 | 8+8 | 12+13 | 15+15 | 17+19 | |
1244-3C | 10+10 | 14+14 | 8+8 | 12+? | 13+14 | 10+11 | |
1244-4A | 9+10 | 14+14 | 9+9 | 13+? | 12+16 | 15+16 | |
1244-4C | 9+9 | 14+14 | 9+9 | 17+? | 13+14 | 12+12 | |
1244-4D | 10+10 | 14+15 | 9+9 | 15+? | 15+15 | 14+16 |
Abdomen. Range of Abd. IV length as 6.71–13.75 times as dorsal axial length of Abd. III. Abd. I usually with seven (a3, a5, m2, m3, m4, m4i, p5, a1 rarely present) mac and one ms (Figs
Scales. Scales present on head, body, legs (Figs
Callyntrura xinjianensis sp. nov. 32 cheatotaxy of Abd. I–III (right side, solid black dot meaning absence) 33 SEM photomicrograph of lateral Abd. I (right side) 34 SEM photomicrograph of central Abd. II (right side) 35 SEM photomicrograph of lateral Abd. III (right side). Scale bar: 20 μm.
Callyntrura xinjianensis sp. nov. 39 anterior face of ventral tube 40 posterior face of ventral tube apically 41 lateral flap of ventral tube 42 manubrial plaque (dorsal view) 43 SEM photomicrographs of mucro (lateral view from internal side) 44 SEM photomicrographs of mucro (lateral view from external side) 45 mucro (upper view) 46, 47 scales on body 48 scale on antenna and furcula. Scale bars: 20 μm.
Named after its locality: Xinjian District.
Found in the leaf litter, mainly composed of bamboo.
Holotype. ♀ on slide, China, Jiangxi Province, Yichun City, Tonggu County, Tonggu Park, 9-XI-2020, 31°54'50"N, 114°22'36"E, 239 m asl, sample number 1235. Paratypes. 2 ♀♀ on slides, same data as holotype. All collected by Y-T Ma.
Size. Body length up to 2.23 mm.
Coloration. Ground colour pale yellow; antennae with scattered brown pigment; eye patches dark blue; brown stripe present from head to Abd. III laterally; middle part of Abd. II–III with brown pigment; medial and posterior margin of Abd. IV with pair of irregular brown patches, respectively; brown pigment scattered on antennae and legs (Figs
Head. Antenna not annulated and 1.45–1.49 times length of body. Ratio of Ant. I–IV as 1.00/0.90–0.98/0.58–0.63/1.50–1.54. Distal part of Ant. IV with many sensory chaetae and normal ciliate chaetae, without apical bulb (Fig.
Thorax. Tergal ms formula on Th. II–Abd. V as 1, 0/1, 0, 1, 0, 0, sens as 1, 1/0, 2, 2, 37–41, 3 (Figs
Abdomen. Range of Abd. IV length as 10.00–12.00 times as dorsal axial length of Abd. III. Abd. I usually with 11 (a1–3, a5i, a5, a5p, m2–4, m4i, p5 rarely absent) mac and one ms. Abd. II with central six (a2, a3, m3, m3e, m3ei, m3ep), lateral three (m5, a6, p6) mac. Abd. III with central two (a2, m3), lateral three (am6, pm6, p6) mac and 10–14 mes (Fig.
Scales. Scales present on head, body, legs (Figs
Callyntrura tongguensis sp. nov. 63 anterior face of ventral tube 64 posterior face of ventral tube apically 65 lateral flap of ventral tube 66 manubrial plaque (dorsal view) 67 mucro (lateral view from internal side) 68, 69 scales on body 70 scale on antenna and furcula. Scale bars: 20 μm.
Named after its locality: Tonggu County.
Found in the leaf litter.
Holotype. 1♀ on slide, China, Jiangxi Province, Shangrao City, Dexing City, Raoshoukun Memorial Park, 13-XI-2020, 28°57'20"N, 117°34'08"E, 88 m asl, sample number 1244. Paratypes. 8 ♀♀ on slides, same data as holotype. All collected by Y-T Ma.
Size. Body length up to 2.45 mm.
Coloration. Ground colour pale yellow; eye patches dark blue; head almost brown entirely; brown stripe present from Th. II to Abd. III laterally; ventral tube, middle part of Abd. II–III brown pigmented; medial and posterior margin of Abd. IV with pair of irregular brown patches, respectively; brown pigment present also on legs, distal Ant. IV and distal dentes (Figs
Head. Antenna not annulated and 1.32–1.53 times length of body. Ratio of Ant. I–IV as 1.00/0.92–1.00/0.60–0.75/1.60–2.04. Distal part of Ant. IV with many sensory chaetae and normal ciliate chaetae, without apical bulb (Fig.
Thorax. Tergal ms formula on Th. II–Abd. V as 1, 0/1, 0, 1, 0, 0, sens as 1, 1/0, 2, 2, 29–42, 3 (Figs
Abdomen. Range of Abd. IV length as 7.02–10.67 times as dorsal axial length of Abd. III. Abd. I with 8–9 (a3, a5, a5p, m2–4, m4 always present, a1, a2 or a5i sometimes absent) mac and one ms. Abd. II with central six (a2, a3, m3, m3e, m3ei, m3ep), lateral three (m5, a6, p6) mac. Abd. III with central two (a2, m3), lateral three (am6, pm6, p6) mac and 8–13 mes (Fig.
Scales. Scales present on head, body, legs (Figs
Callyntrura raoi sp. nov. 87 anterior face of ventral tube 88 posterior face of ventral tube apically (partially) 89 lateral flap of ventral tube 90 manubrial plaque (dorsal view) 91 mucro (upper view) 92 mucro (ventral view) 93, 94 scales on body 95 scale on antenna and furcula. Scale bars: 20 μm.
“raoi” (in apposition) refers to Lieutenant General Shoukun Rao, who made immortal achievements in the Chinese People’s War of Resistance against Japanese Aggression and the War of Liberation.
Found in the leaf litter.
The three new species are very similar in overall chaetotaxy, colour pattern and other characters. The chaetotaxy of each studied specimen is listed in Table
The subgenus Japonphysa contains four species at present: C. (J.) japonica (Kinoshita, 1917), C. (J.) oligosetosa Kim, Rojanavongse & Lee, 1999, C. (J.) semilineata Yosii, 1961 and C. (J.) unilineata Yosii, 1961. The differences between the three new species and the four known species are great, especially in chaetotaxy of body (Table
Comparison between the new species and all known species of Callynthrura (Japonphysa).
Characters | C. (J.) xinjianensis sp. nov. | C. (J.) tongguensis sp. nov. | C. (J.) raoi sp. nov. | C. (J.) japonica | C. (J.) oligosetosa | C. (J.) semilineata | C. (J.) unilineata |
---|---|---|---|---|---|---|---|
Brown pigment on head | laterally | laterally | almost entirely | entirely | laterally | laterally | laterally |
Brown pigment on ventral tube | anteriorly | absent | almost entirely | entirely | absent | absent | absent |
Chaetae on labial base | MREL1L2 | MREL1L2 | MREL1L2 | MRel1l2* | not known | not known | MRel1L2 |
Posterior mac on Th. II | 8–9 (p2p absent) | 10 (p2p present) | 9–10 (p2p present) | 7* | 0 | 4 | 4 |
Posterior mac on Th. III | 12–14 | 15 | 14 | 10* | 0 | 9 | 7 |
Mac on Abd. I | 7 (rarely 8) | 11 (rarely 10) | 8–9 | 6* | 7 | 7 | 7 |
Central mac on Abd. II | 6 | 6 | 6 | 5* | 4 | 5 | 5 |
Central mac on Abd. III | 2 | 2 | 2 | 2* | 1 | 2 | 2 |
Inner teeth on unguis | 4 | 4 | 4 | 3* | 4 | 3 | 3–4 |
Anterior chaetae on ventral tube | 18–22 | 25–29 | 17 | not known | 7 | not known | not known |
Sequenced individuals in the present study had a mean K2-P distance of COI sequences between 0.190–0.197 (about 19%). The shortest interspecific distance was 0.190 between C. tongguensis sp. nov. and C. raoi sp. nov. and the longest was 0.197 between C. tongguensis sp. nov. and C. xinjianensis sp. nov. (Table
Genetic distances (mean K2-P divergence) of the COI sequences between the new described species.
Colour pattern usually plays a very important role in the classification of Collembola and many species were described based on it previously. Although colour pattern is a good character and intraspecific variability is low in Callyntrura taxa, it is sometimes very difficult for taxonomists to distinguish those different species who share similar colouration. DNA barcoding is a good tool to separate species with a similar colour pattern and well used in classification in some genera of Collembola, such as Coecobrya (
1 | No labral chaetae modified (subgenus Japonphysa) | 2 |
– | Part or all chaetae on the first row of labrum modified | 5 |
2 | Pigment present on Th. II–Abd. III entirely | C. (Japonphysa) japonica (Kinoshita, 1917) |
– | Pigment mainly present on Th. II–Abd. III laterally | 3 |
3 | Pigment present on head entirely, Abd. I with 8–9 mac | C. (J.) raoi sp. nov. |
– | Pigment present on head only laterally, Abd. I usually with 7 or 11 mac | 4 |
4 | Abd. I usually with 7 mac, p2p mac absent | C. (J.) xinjianensis sp. nov. |
– | Abd. I usually with 11 mac, p2p mac present | C. (J.) tongguensis sp. nov. |
5 | Three median chaetae on the first row of labrum modified | 6 |
– | All chaetae on the first row of labrum modified (subgenus Gunungphysa) | 7 |
6 | Abd. III with 1 dorsal mac | C. (Javaphysa) guangdongensis Ma, 2012 |
– | Abd. III with 2 dorsal mac | C. (Istanaphysa) hainanensis Ma, 2013 |
7 | Dens with spines | C. (Gunungphysa) spinidentata Lee & Park, 1989 |
– | Dens without spines | 8 |
8 | Abd. I with 3 mac | C. (G.) taiwanica Yosii, 1965 |
– | Abd. I with more than 3 mac | 9 |
9 | Abd. I with 5 mac | 10 |
– | Abd. I with 9 mac | C. (G.) affinis Lee & Park, 1989 |
10 | A longitudinal stripe present from Th. II to Abd. III laterally | C. (G.) striata Yosii, 1965 |
– | Pigment diffused on body | C. (G.) microphysarum Yosii, 1965 |
The authors have declared that no competing interests exist.
No ethical statement was reported.
This research was funded by the Second Tibetan Plateau Scientific Expedition and Research Program (2019QZKK05010303).
Mei-Dong Jing: Sorting of specimens and writing of manuscript.Yin-Huan Ding: Molecular analysis.Yi-Tong Ma: Observing of specimens under microscope.
Mei-Dong Jing http://orcid.org/0000-0002-8498-9498
Yin-Huan Ding https://orcid.org/0000-0002-8002-7056
Yi-Tong Ма http://orcid.org/0000-0002-8660-0503
All of the data that support the findings of this study are available in the main text.