Description of three new species of Callyntrura (Japonphysa) (Collembola, Entomobryidae) from China with the aid of DNA barcoding Author Jing, Mei-Dong https://orcid.org/0000-0002-8498-9498 School of Life Sciences, Nantong University, Nantong, Jiangsu 226000, China Author Ding, Yin-Huan https://orcid.org/0000-0002-8002-7056 Department of Agronomy and Horticulture, Jiangsu Vocational College of Agriculture and Forestry, Jurong, Jiangsu 212400, China Author Ma, Yi-Tong https://orcid.org/0000-0002-8660-0503 School of Life Sciences, Nantong University, Nantong, Jiangsu 226000, China mayitong@ntu.edu.cn text ZooKeys 2023 2023-12-21 1187 237 260 http://dx.doi.org/10.3897/zookeys.1187.109608 journal article http://dx.doi.org/10.3897/zookeys.1187.109608 1313-2970-1187-237 4C4C934F4A2F4493ABB5C55604CE4A21 F218242BEDF554C7BA784C2BB909A11D Callyntrura (Japonphysa) raoi sp. nov. Figs 71-73 , 74-80 , 81-83 , 84 , 85, 86 , 87-95 Type material. 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. Description. 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 71-73 ). Figures 71-73. Habitus of Callyntrura raoi sp. nov. 71, 72 lateral view 73 dorsal view. Scale bars: 500 μm . 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. 74 ). Ant III organ with two rod-like chaetae (Fig. 75 ). Dorsal chaetotaxy of head as in Fig. 76 , An series with six mac, A with four mac or mes, S with eight mac, P with two mac. Eyes 8+8, G & H smaller than others; interocular chaetae as p, r, t; frontal spines 4+4, all serrate. Prelabral chaetae four, ciliate; labral chaetae 5, 5, 4, smooth and pointed, a0, a1 longer than a2; labral papillae absent (Fig. 77 ). Basal chaeta on maxillary outer lobe thick and blunt; sublobal plate with three smooth chaetae-like processes (Fig. 78 ). Lateral process (l. p.) of labial palp E differentiated, as thick as normal chaeta, with tip not reaching apex of papilla E (Fig. 79 ). Labial base with MREL1L2, all ciliate and R 0.51-0.77 length of M (Fig. 80 ). Figures 74-80. Callyntrura raoi sp. nov. 74 apex of Ant. IV (dorsal view) 75 distal Ant. III (ventral view) 76 dorsal head 77 prelabrum and labrum 78 maxillary palp and outer lobe (right side) 79 labial palp E (right side) 80 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, 29-42, 3 (Figs 81 , 84 - 86 ). Th. II with medial five mac, usually posterior 10 (p1i, p1, p2, p2a, p2a2, p2p, p2e, p2ep, p3, p4, p1i or p2ep rarely absent) mac, one ms and one sens. Th. III with anterior-lateral five (a4, a5, m5, a6i, a6), usually posterior 14 mac (p2a rarely present), one sens (Fig. 81 ). Trochanteral organ with 63-64 chaetae (Fig. 82 ). Tenent hair clavate, 1.06-1.20 as long as inner edge of unguis; unguis with four inner teeth, most distal tooth very faint, basal pair located at 0.31-0.42 distance from base of inner edge of unguis, unpaired teeth at 0.66-0.68 and 0.82-0.89 distance from base, respectively; unguiculus lanceolate, with one inner tooth and outer edge slightly serrate (Fig. 83 ). Figures 81-83. Callyntrura raoi sp. nov. 81 chaetotaxy of Th. II-III (right side, solid black dot meaning absence) 82 trochanteral organ 83 hind foot complex (lateral view). Scale bars: 20 μm . 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. 84 ). Abd. IV with 27-40 elongate and two (as, ps) normal sens, medial 14-18 and posterior 10-27 mac or mes, lateral 8-9 mac (Fig. 85 ). Abd. V with three sens (Fig. 86 ). Ventral tube with 17 ciliate chaetae on each side anteriorly (Fig. 87 ); numerous ciliate chaetae and two apical smooth chaetae posteriorly (Fig. 88 ); 14-21 smooth and 6-19 ciliate chaetae on lateral flap (Fig. 89 ). Manubrial plaque with four ciliate mac and one pseudopore (Fig. 90 ). Dens without spines. Mucro with six (v1, v2, v3, d1, d2, i.l.) teeth (Figs 91 , 92 ). Scales . Scales present on head, body, legs (Figs 93 , 94 ); Ant. I-III and ventral side of manubrium and dens with narrower scales (Fig. 95 ). Ant. IV, ventral tube and tenaculum without scales. Figure 84. Chaetotaxy of Abd. I-III of Callyntrura raoi sp. nov. (right side, solid black dot meaning absence). Scale bar: 20 μm . Figures 85, 86. Callyntrura raoi sp. nov. 85 chaetotaxy of Abd. IV (right side, solid black dot meaning absence) 86 chaetotaxy of Abd. V (right side). Scale bars: 20 μm . Figures 87-95. 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 . Etymology. " 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. Ecology. Found in the leaf litter. Remarks. 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 1 , and the differences between these three species are slight. One main difference in chaetotaxy is that Abd. I has 7(8), 11, 8-9 mac in C. (J.) xinjianensis sp. nov., C. (J.) tongguensis sp. nov. and C. (J.) raoi sp. nov., respectively. Another difference is p2p mac on Th. II is absent in C. (J.) xinjianensis sp. nov., but present in the latter two new species. One main difference in colour pattern between them is that only the lateral side of the head is brown pigmented in C. (J.) xinjianensis sp. nov., and C. (J.) tongguensis sp. nov., but almost the entire head is brown in C. (J.) raoi sp. nov. The brown pigment on the ventral tube is present anteriorly in C. (J.) xinjianensis sp. nov., absent in C. (J.) tongguensis sp. nov. and present almost entirely in C. (J.) raoi sp. nov. The subgenus Callyntrura 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 2 ). Table 2. 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

* based on Yoshii's description ( Yoshii 1982 ). Molecular results 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 3 ). Therefore, the interspecific distances of COI between the three new species were more than the accepted barcoding gap recently reported for the species of Entomobryidae ( Zhang et al. 2018b ) and Tomoceridae ( Yu et al. 2018 ). The molecular distances coincided with the morphological divergences, thus further supporting the distinction of the three species. Table 3. Genetic distances (mean K2-P divergence) of the COI sequences between the new described species.

Species	C. (J.) tongguensis sp. nov.	C. (J.) raoi sp. nov.	C. (J.) xinjianensis sp. nov.	GenBank Accession Numbers
C. (J.) tongguensis sp. nov.	-	-	-	OQ940723
C. (J.) raoi sp. nov.	0.190	-	-	OQ940724
C. (J.) xinjianensis sp. nov.	0.197	0.196	-	OQ940725