Acerentuloides bernardi Shrubovych, 2017

Shrubovych, Julia, Starý, Josef & D’Haese, Cyrille A., 2017, Molecular phylogeny of Acerentomidae (Protura), with description of Acerentuloides bernardi sp. nov. from North America, Florida Entomologist 100 (2), pp. 433-443 : 434-438

Acerentuloides bernardi Shrubovych sp. nov., 2016 ( Figs. 1–34 ; Table 2)

TYPE MATERIAL

The type materials were deposited in the collection of the Institute of Systematics and Evolution of Animals of the Polish Academy of Sciences ( ISEA), Cracow, Poland , and in the collection of the State Museum of Natural History ( SMNH) of the National Academy of Sciences of Ukraine, L’viv, Ukraine .

HOLOTYPE 1 female ( ISEA 6647) USA: Indiana, Oakland City, Hugh Boyd, Patoka River National Wildlife Refuge and Management Area, 9-V-2008, latitude 38.3539°N, longitude 87.3156°E, elevation 133 m, deciduous growth with dominant black locust tree ( Robinia pseudoacacia L.; Fabaceae ) and red oak, sample of litter and rhizosphere, collected by J. Tajovský & J. Frouz.

PARATYPES 2 females ( ISEA 6648 , SMNH 89.1 ), data same as holotype .

Subfamily Berberentulinae Berberentulinae Berberentulinae Berberentulinae Berberentulinae Berberentulinae Berberentulinae Berberentulinae Berberentulinae Berberentulinae Nipponentominae .) available not was sequence Family Acerentomidae Acerentomidae Acerentomidae Acerentomidae Acerentomidae Acerentomidae Acerentomidae Acerentomidae Acerentomidae Acerentomidae Acerentomidae the that indicates gene S 18 rRNA aExtension afer the corresponds name specimen voucher 17 008 - ABPE — 557247 EU — 010 ABPE 17 - — — 037169 AY EU 557246 557251 EU — species to the code. dash a (analysis COI 210842 210838 210839 882820 395311 395312 395313 — 882817 882818 882823 phylogenetic KJ KJ KJ HQ KJ KJ KJ HQ HQ HQ the in 2

D used sequences S 1 D – rDNA — — — 557260 EU ABPE 009 - ABPE 010 17 — 376049 EU EF 192433 EF 192435 EU 557263 28 gene of numbers D – 3 6

rDNA

D 007 17

ABPE

- 17 - 008

ABPE — 557260 —

-

ABPE

010 17

— 376049 192433 192435 557263

. code accession S 28 EU EU EF EF EU voucher and specimen proturans 17 17 17 17 to the investigated 17 - 007 008 10

- ABPE 11 ABPE 009 - 16 ABPE - 010 19 011 ABPE - 20 corresponds Continued List of () a name ABPE csp 14 noeli pyreneicus csp csp pyreneicus sinensis csp rapoporti rapoporti csp csp rapoporti densus tienmushanensis maijiawensis sinensis name species afer the. 1 Table Species Acerentulus Acerentulus Acerentulus Acerentulus Andinentulus Andinentulus Andinentulus Baculentulus Baculentulus Gracilentulus Nosekiella a Extension

DESCRIPTION

Habitus typical for members of Acerentomidae ( Fig. 24 ). Head setae short, cephalic seta l3 setiform ( Figs. 1 and 2 ), seta sd5 thick, sensilliform ( Figs. 1 and 3 ), setae sd4 and d6 absent, length ratio of posterior setae d7:sd7 as 1.0:1.5 ( Fig. 1 ). Pseudoculus circular, with short posterior extension, PR = 14 ( Fig. 2 ). Maxillary palpus apically with tuft of setae and 4 single setae, basal sensilla slender, subequal in length ( Fig. 4 ). Labial palpus well developed with 4-branched apical tuft and sausage-shaped basal sensillum ( Figs. 5 and 25 ). Maxillary gland with small elongated and weakly granulated calyx and small globular vesicles on calyx, long posterior filament with row of small globules posteriorly ending in bilobed dilation, CF = 3.7 ( Fig. 6 ).

Foretarsus with sensillum b’ present, t1 claviform, t3 long and finger-like, all other sensilla slender and parallel-sided ( Figs. 9, 10 , and 26). Sensillum a reaching base of t2. Sensillum b longer than c, extending past base of sensillum e. Sensillum c reaching base of sensillum e. Sensillum b inserted proximally to c, sensillum d inserted between sensilla c and t2. Sensillum a’ short, extending past base of t2 ( Fig. 26 ). Sensillum b’ reaching base of sensillum c’, apex of sensillum c’ not reaching base of claw. Length formula of sensilla: t1 <t3 <(a’ = b’) <(g = c’) <(c = t2) <e <(a = d) <b <f. Setae β1 and δ4 modified, sensilliform, shorter than other δ-setae (length of β1 = 4 μm, δ4 = 6 μm). Pores on foretarsus near base of sensillum t3 and between bases of sensillum c and seta α3. Claw short, without inner tooth, empodial appendage short. BS = 0.6, TR = 3.6, EU = 0.12.

Formula of chaetotaxy given in Table 2. Setae on nota strongly differing in length. Pronotal seta 1 twice the length of seta 2 ( Fig. 11 ). Meso- and metanotal setae P1a and P2a very short, oblong ( Fig. 27 ), P2a situated close to P3, seta P5 gemmate ( Figs. 11 and 12 ). Length ratio of mesonotal setae P1:P2 = 1.0:1.6. Seta P4 on metanotum a short sensillum, length 3 μm ( Fig. 12 ). Meso- and metanota with sl and al pores ( Figs. 11 and 12 ). Thoracic sterna without pores. Setae A2 and M2 on prosternum and seta A2 on thoracic sterna short, sensilliform; length of prosternal A2 = 3 μm, of prosternal M2 = 1 μm ( Figs. 28 and 29 ), of meso- and metasternal A2 = 1 μm ( Figs. 13–15 ).

Setae A5 and P2a on tergite I, setae P2a and P4a on tergites II–VI short and sensilliform, their length 3 μm ( Figs. 16, 17 , and 30). Accessory setae P1a, P2a, and P4a on tergite VII setiform, longer, length 4 μm ( Fig. 18 ). Seta P3 on tergites II–VI inserted anteriorly to other setae of P-row ( Fig. 17 ), P3 on tergites I and VII level with oth- er setae of P-row ( Figs. 16 and 18 ). Tergites VI–VII with connecting line in anterior part ( Figs. 17 and 18 ). Pores psm present on tergites I–VII between setae P1 and P2, and close to seta P1a on tergite VII, al pores on tergites II–VII, psl pores on tergite VI only ( Figs. 16–18 ).

Abdominal legs with 4, 3, 3 setae. Subapical seta on 2nd and 3rd pairs of abdominal legs nearly twice the length of apical lateral seta (21 and 12 μm, respectively). Apical median seta minute, length 1 μm ( Figs. 21 and 31 ). Accessory setae P1a on sternites I–VI sensilliform, on sternite VII setiform, short, P1a length on sternites I‒VII 3 μm ( Figs. 20, 22, 23 , and 31–33). Sternites VI–VII with a connecting line in anterior region ( Figs. 22 and 23 ). Sternites II–V with asymmetrical spsm pore ( Fig. 20 ). Sternite VI with spsm pores, 2 near each other and 1 close to base of seta P1 ( Figs. 22 and 32 ); sternite VII with spm pore ( Fig. 23 ) or in 1 paratype with asymmetrical spsm pore between setae P1 and P1a ( Fig. 33 ).

Abdominal segment VIII with well-developed striate band ( Figs. 19 and 34 ). Pore psm without accompanying teeth ( Fig. 19 ). Posterior margin of sternite VIII and laterotergites smooth ( Figs. 23 and 34 ). Comb on tergite VIII with 8–9 short teeth ( Fig. 7 ). Sternites IX–XII with smooth hind margin; setae 1 about one-third length of setae 2 ( Figs. 23 and 34 ). Segment XII with medial pore on dorsal lobe and pair of sternal anterolateral pores.

Female squama genitalis with short, forked acrostyli ( Fig. 8 ). Males unknown.

Body measurement (3 females): body length 1,100 μm; head 130– 134 μm; pseudoculus 9 μm, lever 1 μm; posterior part of maxillary gland 35 μm; head setae d7 = 11 μm, sd7 = 16 μm, l3 = 6 μm; pronotal setae 1 = 25 μm, 2 = 12 μm, mesonotal setae P1 = 19–20 μm, P1a = 1 μm, P2 = 30–31 μm; foretarsus 90 μm, claw 25 μm, empodial appendage 3 μm.

ETYMOLOGY

The new species is cordially dedicated to our colleague and eminent zoologist, Prof. Dr. Ernest C. Bernard.

REMARKS

The habitus of the new species is very similar to A. confinis ( Figs. 24 and 35 ). The species is characterized by absence of seta sd4 and additional seta d6 on head, absence of seta P1a on abdominal tergites I–VI, presence of seta Pc on sternite VII, short sensillum a reaching to the base of sensillum t2, long foretarsal sensilla b and f, sensillum c shorter than b, short and slender sensilla a’, b’ and c’, and presence of psl pores on tergite VI only. Acerentuloides bernardi sp. nov. is similar to A. americanus in the shape of the maxillary gland, presence of a minute apical medial seta on abdominal legs II and III, shape of the comb and the length of foretarsal setae a, b’ and c’ ( Ewing 1921, 1940; Bonet & Tuxen 1960). The new species differs in possessing seta Pc on sternite VII and in the shape of foretarsal sensillum t3 (lancet-like in A. americanus , finger-like in the new species). Foretarsal sensillum b is longer than c and clearly surpasses the base of seta γ 3 in the new species, whereas in A. americanus sensillum b is shorter and does not reach the base of γ3; in Canadian specimens of A. americanus sensilla b and c are of equal length ( Nosek & McEwan Kevan 1984) and sensillum a’ is longer and extends past the base of sensillum t2. In addition, the new species has a smooth, globular vesicle on the maxillary gland, whereas Canadian specimens of A. americanus have a granulated appendix on the calyx ( Nosek & McEwan Kevan 1984).

MOLECULAR ANALYSIS

Unfortunately, DNA was not retrieved from specimens of A. bernardi sp. nov. However, the sequencing of A. confinis , a species from a closely related genus, was successful. A 658 bp fragment of the COI gene (DNA barcode) as well as 1,541 bp of the 18S rRNA and 971 bp of the 28S rDNA D1–D2 were amplified and sequenced from a specimen collected in the USA. To complete the COI data sets previously published ( Shrubovych et al. 2012, 2014b, 2014c), other Acerentulus and Andinentulus species were amplified for 18S rRNA: Acerentulus charrieri Shrubovych, Schneider & D’Haese, 2012 , A. noeli Shrubovych, Schneider & D’Haese, 2014 ., A. pyreneicus Shrubovych, Schneider & D’Haese, 2014 , and Andinentulus rapoporti (Condé, 1963) ; 28S rDNA D1–D2: A. noeli and Andinentulus rapoporti (2 different specimens); 28S rDNA D3–D6: A. charrieri (2 different specimens), A. noeli (2 different specimens), A. pyreneicus , and Andinentulus rapoporti . The sequences were deposited in BOLD under accession numbers ABPE002- 17-ABPE011-17 (see Table 1).

The concatenated data (COI, 18S rRNA, 28S rDNA D1–D2, and 28S rDNA D3–D6) consist of a total alignment length of 4,438 bp for 55 terminal taxa (27 species).The ML inference and direct optimization parsimony phylogenies all showed consistent agreement among topologies. The only real difference is the position of Nosekiella + Huashanentulus sister group of Acerella in the ML analysis and sister group of the rest of Acerentominae in parsimony. Here we show the ML tree ( Fig. 41 ).

Figures

Figs. 1–10. Acerentuloides bernardi sp. nov. 1. Head, right side; 2. pseudoculus with seta l3; 3. cephalic seta sd5; 4. maxillary palpus; 5. labial palpus; 6. maxillary gland; 7. comb; 8. female squama genitalis; 9. foretarsus, exterior view; 10. foretarsus, interior view. Arrows indicate pores (cp = clypeal pore, fr = frontal pore). Scale bars: 20 µm.

Figs. 11–15. Acerentuloides bernardi sp. nov. 11. Pronotum and mesonotum, right side; 12. lateral part of metanotum; 13. anterior part of prosternum; 14. anterolateral part of mesosternum; 15. anterolateral part of metasternum. Arrows indicate pores (al = tergal anterolateral, sl = tergal sublateral). Scale bars: 20 µm.

Figs.16–23. Acerentuloides bernardi sp. nov. 16.Tergite I, right side;17.tergite VI, right part; 18. tergite VII, right part; 19.tergite VIII;20.sternite II; 21. abdominal leg of sternite II; 22. sternite VI; 23. sternites VII–IX. Arrows indicate pores (psm = tergal posterosubmedial, psl = tergal posterosublateral, spsm = sternal postero- submedial, spm = sternal posteromedial). Scale bars: 20 µm.

Figs. 24–34. Acerentuloides bernardi sp. nov. scanning electron microscopy photographs. 24. Habitus; 25. labial palp (apical tuf with 4 setae broken; s = labial sensillum); 26. foretarsus, interior view: sensilla t1, t3 and a’; 27. modified seta P2a on mesonotum; 28. modified seta A2 on prosternum; 29. modified seta M2 on prosternum; 30. modified seta A5 on tergite I and P1a on sternite I; 31. abdominal leg on segment III (sa = subapical, am = apical medial, al = apical lateral setae; apical lateral seta is broken); 32. hind margin of sternite VI; 33. hind margin of sternite VII; 34. sternites VIII–XII. Scale bars = 300 µm (Fig. 24), 4 µm (Figs. 25 and 27–29), 10 µm (Fig. 31), and 20 µm (Figs. 26, 30, and 32–34).

Figs.35–40. Acerentulus confinis (Berlese,1908),American specimen scanning electron microscopy photographs.35. Habitus; 36. pseudoculus and cephalic setae sd4 and l3; 37. foretarsus, exterior view: sensilla t1, t3 and a’; 38. labial palpi with apical tuf of setae and basal sensillum (s); 39. sternite VIII; 40. modified seta P4 on metanotum. Scale bars = 300 µm (Fig. 35), 20 µm (Figs. 36 and 38), and 40 µm (Figs. 37, 39, and 40).

Fig. 41. Molecular phylogeny of Acerentomidae inferred from concatenated COI, 18S rRNA, 28S rDNA D1–D2, and 28S rDNA D3–D6 sequences with maximum likelihood (ML). ML bootstrap values shown above the branches.