Review Article |
Corresponding author: Clément Schneider ( clement.schneider@mnhn.fr ) Academic editor: Wanda M. Weiner
© 2016 Clément Schneider, David Porco, Louis Deharveng.
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:
Schneider C, Porco D, Deharveng L (2016) Two new Megalothorax species of the minimus group (Collembola, Neelidae). ZooKeys 554: 37-68. https://doi.org/10.3897/zookeys.554.6069
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Two new Megalothorax species, M. potapovi sp. n. from the Russian Far East and M. sanguineus sp. n. from the French Pyrénées are described. The two new species have a set of morphological characters (including a smooth mucro) that places them among the minimus group sensu
Springtails, Neelipleona , description, chaetotaxy, taxonomy, DNA barcodes, labrum
During an expedition organized by Mikhail Potapov in Primorye (Russian Far East) in 2004, a large number of samples yielded a huge diversity of Collembola. Several of the most remarkable taxa collected have been recently described, like the new genus Sensillonychiurus with three new species (
Megalothorax potapovi sp. n. The specimens were obtained from a 2000 cm3 sample of forest litter from Primorye, that were processed for fauna extraction in a field laboratory of Anisimovka. The litter sample was dried for 8 days on a Berlese funnel without heating.
Megalothorax sanguineus sp. n. The two sampled sites are located at medium elevation in Ariège Pyrenees (France): in very humid mossy habitats near the peat-bog of Bernadouze, and from humid litter at Osque du Couret. Samples were processed in the lab on Berlese funnels in the same conditions as above.
Specimens were preserved in 95% ethanol then cleared in lactic acid and finally mounted on microscope slides in Marc André II medium. They were examined using a Leica DMLB compound microscope with differential phase contrast optics at magnifications ranging from 250 to 1000. Drawings were made with a drawing tube and vectorized with Inkscape. For Scanning Electronic Microscope (SEM) observations, specimens were dehydrated in 100% ethanol, before critical point drying (Emitech K850) and gold coating (Jeol JFC-1200) and observations were performed with a SEM Jeol 840A. Megalothorax sanguineus sp. n. was not observed with SEM, however we were able to locate all the positions of the τ-chaetae (trichobothria) with the optical microscope.
The standard DNA barcode (658bp of the COI–mitochondrial cytochrome oxidase subunit I gene,
DNA was extracted from entire specimens in 30μl of lysis buffer (http://www.ccdb.ca/docs/CCDB_DNA_Extraction.pdf) and proteinase K incubated at 56 °C overnight. DNA extraction followed a standard automated protocol using 96-well glass fibre plates (
Forty-nine specimens of Megalothorax dataset representing 14 morphologically recognized species were selected, 36 from
Distance analyses were performed with MEGA6 (
A nomenclature for the integumentary crests on the labrum is introduced (Fig.
Head chaetotaxy (Fig.
Crests on the anterior process of the labrum: al2, 3 = anterior longitudinal, ml1–3 = posterior longitudinal, amt0–2 = antero-median transversal, mt2 = posterior transversal. Antenna: Ant. I–IV = antennomere I to IV, S1–S4 = S-chaetae of Ant. III; or = Ant. IV organite; S, Sx, Sy = S-chaetae of Ant. IV. Trunk: Th. I–III = thoracic segment I to III; Abd. I–VI = abdominal segment I to VI; av = chaetae of anal valve; s1, s2 = s-chaetae; sm = special chaetae of male Abd. VI sternum; τ = τ-chaetae; wrc1–wrc8 = free wax rod secretory element 1 to 8. Claw: la, lp, Ba, Bp = auxiliary lamellae and crest of unguis; Ca, Cp = anterior and posterior crests of unguiculal lamella. Misc: dp = dens proximal, dd = dens distal, sf1–6 = sensory field 1 to 6.
Holotype: male on slide (
Whitish in alcohol. Presence of median integumentary protuberance in front of chaeta a0 on forehead. Presence of chaeta X on Ant. IV. Labium: basomedian fields with 3 + 3 chaetae, basolateral fields with 1 + 1 chaetae. Integumentary channels as a paired tree on posterior part of the head, absent on anterior part, connection of channels with linea ventralis circular. Some chaetae enlarged and lanceolate, of which 5 + 5 dorsal, posterior on head and 2 + 2 on Th. II tergum. Inner chaeta of sensory field 2 slender with blunt apex, all inner chaetae of sensory fields 3–6 short flam-shaped. Dorsal abdominal s-chaetae s2 globular, absence of dorsal abdominal s-chaetae s3. Abd. I to V terga with 18 + 18 ordinary chaetae. Each claw of ordinary morphology, subequal. Tenaculum with 3 + 3 teeth. Abd. IV sternum with 2 + 2 chaetae. Mucro lamellae smooth, thin.
General aspect. Habitus and segmentation typical of the genus (Fig.
Integument. Secondary granulation made of the usual dorsal rough granules (e.g. Fig.
Sensory fields and wax rods. A total of 14 + 14 wax rod secretory crypts (2 + 2 on head, 12 + 12 on body), including the ones inserted in each 6 + 6 sensory fields (Figs
Megalothorax potapovi sp. n. Chaetotaxy of head A dorsal side B ventral side; chaetotaxy of antenna C anterior side D posterior side; labrum and maxillary outer lobe E anterior side F posterior side; palp of labium G focused on ventral chaetae H focused on distal chaetae I hypostomal papillate chaeta. * indicates a supplementary chaeta, absent in other specimens.
Labrum. Chaetae (Figs
Other mouth parts. Oral fold with 2 + 2 mesochaetae. Maxillary outer lobe: palp with subapical mesochaeta and apical papillate macrochaeta, edge of apical papilla with three strong integumentary lobes (Figs
Head chaetotaxy. Trend for posterior chaetae to be longer and stronger than anterior chaetae, with 5 + 5 remarkable posterior lanceolate macrochaetae (up to 18 μm, Figs
Antennal chaetotaxy. Illustrated in Figs
Ant. | I | II | III | IV | ||||
---|---|---|---|---|---|---|---|---|
chaetae | chaetae | chaetae | S-chaetae | chaetae | S-chaetae | Organit | Sensory rods | |
M. potapovi sp. n. | 1 | 4 | 8 | S1–S4 | 6 (X incl.) | 12 (10 S, Sx, Sy) | Or | 2 (a, sa) |
M. sanguineus sp. n. | 9 | 7 (X incl.) |
Thoracic terga chaetotaxy. Th. II with 12 + 12 chaetae of variable length, 1 + 1 s-chaetae s1 tubular and curved and 3 + 3 τ-chaetae (Figs
Legs chaetotaxy. Legs with ordinary chaetae of variable size as in Fig.
Leg | I | II | III | |||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Segment | Sc1 | Sc2 | Cx | Tr | Fe | Ti | Sc1 | Sc2 | Cx | Tr | Fe | Ti | Sc1 | Sc2 | Cx | Tr | Fe | Ti |
M. potapovi sp. n. | 1 | 0 | 1 | 3 | 8 | 12 | 1 | 1 | 1 | 3 | 8 | 12 | 2 | 1 | 1 | 4 | 8 | 11 |
M. sanguineus sp. n. |
Claws. Ratio unguis length : pretarsus width on leg I–III respectively as 2.2, 2, 1.73, each claw with ordinary morphology, claw III bulkier than claw I and II (Fig.
Abd. I–V terga chaetotaxy. With a total of 18 + 18 chaetae, 1 + 1 τ-chaetae, 2 + 2 free wax-rod generating crypts (wrc7, 8), 1 + 1 globular s-chaetae s2 (Figs
Abd. VI and genital chaetotaxy. Abd. VI: with nine dorsal mesochaetae (Fig.
Abd. IV sternum and furca. Abd. IV sternum with 2 + 2 neosminthuroid chaetae (Figs
Tenaculum and ventral tube. Tenaculum with 3 + 3 hook-like teeth (Fig.
Megalothorax potapovi sp. n. has the characteristics of the minimus group species (
The integumentary structure on forehead and the lanceolate macrochaetae clearly separate M. potapovi sp. n. from M. minimus, M. aquaticus Stach, 1951, M. svalbardensis, M. willemi Schneider and D’Haese, 2013, M. tuberculatus Deharveng and Beruete, 1993, M. carpaticus Papáč & Kováč, 2013 and M. tatrensis. Other peculiar characteristics of the species are shape of hypostomal papillate chaeta, presence of a strong lobe protecting S2, S3 on Ant. III, and S4 in apical position on Ant. III. On the basis of labral features M. minimus (Fig.
The species was collected in lowland forest litter, and only found so far in the southern part of Primorye.
Megalothorax potapovi sp. n. is dedicated to Mikhail Potapov, who led the 2004 field trip in Primorye which allowed to discover the new species.
A 658bp fragment of the COI gene was amplified and sequenced from paratype (
5’– TAAGTTTTTGACTTCTTCCACCTTCTCTCACCCTTCTACTTTCAAGAGGTCTAGCAGAATCAGGTGCTGGAACAGGTTGAACTGTTTATCCTCCTTTATCTTCAAATATTTCCCATGCAGGAGCCTCTGTCGACTTAACTATTTTCAGTTTACATTTAGCTGGTATGTCATCAATTTTAGGAGCTATTAATTTTATTACAACTATCTTTAACATACGATCCCCAGGAATAACATGAGATCAAACTTCACTATTTATTTGATCTGTTTTAATTACATCAATTTTACTTCTCTTGTCTCTTCCAGTTCTAGCAGGAGCTATCACCATACTTTTAACCGACCGAAATTTAAATACTTCATTTTTTGACCCCGCTGGGGGTGGTGACCCAATTTTATACCAACACCTATTC–3’
Type material. Holotype: female on slide (
4 females on slides (
Reddish in alcohol. Absence of median integumentary protuberance in front of chaeta a0 on forehead. Presence of chaeta X on Ant. IV. Labium: basomedian fields with 3 + 3 chaetae, basolateral fields with 1 + 1 chaetae. Integumentary channels as a paired network on posterior part of the head and a simple channel on anterior part, connection of channels with linea ventralis circular. Chaetae on head and trunk with ordinary shape. All inner chaetae of sensory fields 2–6 short flam-shaped. Dorsal abdominal s-chaetae s2 bean-shaped, absence of dorsal abdominal s-chaetae s3. Abd. I to V terga with 20 + 20 ordinary chaetae. Slightly elongated claws. Tenaculum with 3 + 3 teeth. Abd. IV sternum with 2 + 2 chaetae. Mucro lamellae smooth, moderately enlarged.
General aspect. Habitus and segmentation typical of the genus. Length from labrum to anus: ~500 μm. Specimens with pale to deep red pigmentation in alcohol. Body chaetotaxy sparse including chaetae, s-chaetae, τ-chaetae as trichobothria, neosminthuroid chaetae, wax rod secretory elements and special swollen chaetae within sf2–6. Length of chaetae ranging from microchaetae [<6 μm] to mesochaetae [6–10 μm] and macrochaetae [11–15 μm]. Chaetae simple, without any remarkable development.
Integument. Secondary granulation made of the usual dorsal rough granules (Fig.
Megalothorax sanguineus sp. n. Chaetotaxy of head A dorso-posterior side B dorso-anterior side C ventral side; anterior process of labrum D posterior side E anterior side; maxillary outer lobe F dorsal side G ventral side; labium H focused on ventral chaetae and basolateral field I focused on distal chaetae J hypostomal papilla. Legend: blf = basolateral field of labium, of = oral fold.
Sensory fields and wax rods. A total of 14 + 14 wax rod secretory crypts (2 + 2 on head, 12 + 12 on body), including the ones inserted in each 6 + 6 sensory fields (Figs
Labrum. Chaetae (Fig.
Other mouth parts. Oral fold with 2 + 2 mesochaetae (Fig.
Head chaetotaxy. Dorsally and laterally with mesochaetae, posterior and anterior mesochaetae subequal with a slight trend for posterior chaetae to be stronger than anterior chaetae (Fig.
Antennal chaetotaxy. Illustrated in Fig.
Thoracic terga chaetotaxy. Th. II with 12 + 12 chaetae of variable length, 1 + 1 s-chaetae s1 tubular and curved and 3 + 3 τ-chaetae (Figs
Legs chaetotaxy. Legs with ordinary chaetae of variable size as in Fig.
Claws. Ratio unguis length : pretarsus width on leg I–III respectively as 3.2, 2, 1.87, claw I with rather slender morphology, claw III bulkier than claw I and II. Claw I with longer unguis and each claw with subequal length of unguiculus, ratio unguiculus : unguis for claw I, II, III as ~ 0.43, 0.5, 0.5 (Fig.
Abd. I–V terga chaetotaxy. With a total of 20 + 20 chaetae, 1 + 1 τ-chaetae, 2 + 2 free wax-rod generating crypts (wrc7, 8), 1 + 1 s-chaetae s2 shaped as a bean (Figs
Abd. VI and genital chaetotaxy. Abd. VI: with nine dorsal chaetae (6–7 μm) (Fig.
Abd. IV sternum and furca. Abd. IV sternum with 2 + 2 neosminthuroid chaetae and 2 + 2 posterior mesochaetae (Fig.
Megalothorax sanguineus sp. n. A Chaetotaxy of Abd. IV and Abd. VI sterna with furca, female B ventral tube and tenaculum, lateral side C focus on tenaculum D focus on tenaculum axial lobes E chaetotaxy of Abd. IV–VI sterna, male F focus on genital plate and Abd. IV posterior chaetae, male.
Tenaculum and ventral tube. Tenaculum with 3 + 3 hook-like teeth (Fig.
Megalothorax sanguineus sp. n. has the characteristics of the minimus group species (
The species is known from humid micro-habitats in Pyrenees, though it was absent from the Bernadouze peat-bog itself. Other Megalothorax found in moist mosses in mountains are M. aquaticus (1750m in High Tatras Mountains) (Stach 1957) and M. minimus (up to 1500m in Pyrenees Mountains) (pers. obs.). The combination of morphological features shared with Megalothorax aquaticus seems to be related to hygrophilous ecology. In that regard, M. sanguineus sp. n. would remain less morphologically specialized than M. aquaticus but more than M. minimus. “Red” Megalothorax are present across the whole Pyrenean range (pers. obs.), and might be M. sanguineus sp. n., but identification has only been confirmed so far for Ariège and Pyrénées-Atlantique specimens.
Megalothorax sanguineus sp. n. is named after the deep red pigmentation of the species.
A 658bp fragment of the COI gene was amplified and sequenced from five specimens from the Saint-Lary locality. Specimens were unfortunately lost, sequences identification is based on consistency between: the peculiar pigmentation of the species observed on specimens before destruction, the genetic similarity of the five specimens and the morphological identification of four other specimens with the same pigmentation from the same sample. The sequences are deposited into the GenBank database under accession numbers JN298074–JN298078.
Four sequences are identical (JN298074–JN298077, provided below), base composition is A = 29.6%, C = 17.5%, G = 15.8%, T = 37.1% (A + T = 66.7%). The fifth sequence (JN298078) differs in 11 sites (= 98.3% pairwise identity), base composition is A = 29.5%, C = 17.6%, G = 15.7%, T = 37.2% (A + T = 66.7%).
5’–AACCTTATATTTAATTTTTGGAGTATGATCTGCTATAGTTGGAACAGCATTTAGAGTTTTAATTCGGTTAGAATTAGGACACCCAGGAAGCTTAATTGGAAACGATCAAATCTATAATGTAATAGTTACGGCCCATGCATTTGTAATAATTTTTTTTATAGTAATACCAATAATAATTGGAGGCTTTGGTAATTGATTAGTACCTTTAATAATTGGAGCACCTGATATAGCATTTCCTCGAATAAACAATTTAAGATTCTGACTTTTACCACCATCTTTAATCTTATTACTATCCAGAGGGTTAGTTGAAACTGGTGCTGGCACAGGATGAACAGTATATCCCCCTCTATCGTCTAATATTTCTCATAGAGGAGCTTCTGTAGATTTAACTATTCTTAGACTTCATTTAGCTGGGATATCTTCTATTCTTGGGGCAATTAATTTTATTACAACTATTCTTAATATACGAATACCAGGAATAACATGAGACCAAACTTCTTTATTTGTATGATCAGTTTTTATTACCTCAATTTTATTACTCCTCTCGCTTCCAGTGCTTGCTGGAGCAATTACTATACTTTTAACTGACCGTAACCTGAATACCTCATTTTTTGATCCTGCGGGAGGAGGAGACCCTATTCTATATCAACATTTATTT–3’.
Fig.
Intra and inter MOTUs genetic distances estimated by Neighbor-Joining with Kimura-2 parameter model.
Intra-MOTU distances | ||||||||||||||||||||
Megalothorax cf. interruptus L1 | 10,70 | |||||||||||||||||||
Megalothorax cf. interruptus L2 | - | 22,98 | ||||||||||||||||||
Megalothorax granulosus | - | 30,02 | 27,10 | |||||||||||||||||
Megalothorax minimus | 3,22 | 38,58 | 36,01 | 34,99 | ||||||||||||||||
Megalothorax nigropunctatus | 2,31 | 28,97 | 26,91 | 31,34 | 29,01 | |||||||||||||||
Megalothorax perspicillum | 0,00 | 31,11 | 30,23 | 24,91 | 31,72 | 26,11 | ||||||||||||||
Megalothorax potapovi sp. n. | 0,00 | 29,66 | 22,89 | 29,02 | 27,19 | 27,91 | 29,10 | |||||||||||||
Megalothorax sanguineus sp. n. | 0,95 | 29,87 | 23,49 | 28,96 | 33,90 | 28,19 | 27,25 | 25,79 | ||||||||||||
Megalothorax sp1 | 3,75 | 35,18 | 27,46 | 30,39 | 25,25 | 27,74 | 30,40 | 23,39 | 23,36 | |||||||||||
Megalothorax sp2 | - | 31,42 | 26,42 | 26,02 | 32,87 | 29,78 | 31,61 | 31,92 | 32,21 | 30,97 | ||||||||||
Megalothorax sp3 | - | 29,17 | 27,52 | 31,81 | 32,61 | 31,16 | 27,84 | 27,62 | 27,21 | 29,64 | 30,30 | |||||||||
Megalothorax sp4 | - | 28,74 | 27,23 | 25,65 | 27,20 | 21,90 | 23,31 | 24,78 | 28,56 | 26,25 | 23,93 | 29,61 | ||||||||
Megalothorax svalbardensis | - | 28,61 | 26,08 | 26,72 | 29,39 | 26,47 | 25,67 | 19,36 | 22,00 | 23,83 | 28,43 | 29,10 | 28,38 | |||||||
Megalothorax willemi L1 | 0,00 | 27,39 | 28,66 | 26,36 | 27,88 | 26,69 | 28,63 | 22,84 | 26,89 | 22,87 | 32,22 | 32,80 | 26,58 | 23,49 | ||||||
Megalothorax willemi L2 | 0,00 | 33,34 | 32,28 | 32,22 | 29,68 | 32,97 | 27,64 | 29,27 | 29,17 | 23,80 | 34,81 | 33,19 | 28,69 | 26,29 | 21,89 | |||||
Megalothorax willemi L3 | 0,52 | 34,70 | 35,89 | 34,40 | 26,48 | 31,97 | 29,36 | 27,97 | 34,84 | 30,04 | 32,00 | 37,62 | 26,26 | 29,31 | 24,15 | 25,08 | ||||
Megalothorax willemi L4 | - | 35,46 | 28,38 | 37,99 | 28,84 | 30,68 | 32,34 | 26,23 | 30,68 | 24,34 | 35,00 | 34,42 | 27,54 | 30,71 | 25,13 | 24,26 | 25,45 | |||
Megalothorax willemi L5 | - | 30,24 | 31,81 | 32,52 | 27,27 | 33,44 | 29,24 | 26,12 | 31,69 | 26,56 | 35,48 | 29,72 | 30,02 | 26,05 | 23,31 | 26,32 | 27,08 | 28,17 | ||
Megalothorax willemi L6 | 0,56 | 30,64 | 29,85 | 28,76 | 25,65 | 26,88 | 26,02 | 22,39 | 28,10 | 21,84 | 30,24 | 31,64 | 27,26 | 25,06 | 20,80 | 25,99 | 24,20 | 22,98 | 20,99 | |
Megalothorax willemi L7 | - | 32,26 | 33,03 | 34,25 | 26,14 | 30,37 | 31,42 | 23,60 | 31,72 | 21,83 | 31,09 | 29,61 | 27,70 | 26,98 | 23,08 | 27,64 | 26,01 | 26,68 | 23,68 | 20,67 |
Specimen | MOTU name | Barcode GenBank accession number |
---|---|---|
Megalothorax potapovi 10770C01 RU120 | Megalothorax potapovi sp. n. | KR736064 |
Megalothorax potapovi 10770C02 RU120 | Megalothorax potapovi sp. n. | KR736063 |
Megalothorax potapovi 00620C03 RU120 | Megalothorax potapovi sp. n. | KR736067 |
Megalothorax potapovi 00620C04 RU120 | Megalothorax potapovi sp. n. | KR736070 |
Megalothorax potapovi 00620C05 RU120 | Megalothorax potapovi sp. n. | KR736069 |
Megalothorax potapovi 00620C06 RU120 | Megalothorax potapovi sp. n. | KR736068 |
Megalothorax potapovi 00620C07 RU120 | Megalothorax potapovi sp. n. | KR736065 |
Megalothorax potapovi 00620C08 RU120 | Megalothorax potapovi sp. n. | KR736066 |
Megalothorax sanguineus 6139D02 09761 | Megalothorax sanguineus sp. n. | JN298074 |
Megalothorax sanguineus 6139D03 09761 | Megalothorax sanguineus sp. n. | JN298075 |
Megalothorax sanguineus 6139D04 09761 | Megalothorax sanguineus sp. n. | JN298076 |
Megalothorax sanguineus 6139D05 09761 | Megalothorax sanguineus sp. n. | JN298077 |
Megalothorax sanguineus 6139D06 09761 | Megalothorax sanguineus sp. n. | JN298078 |
Megalothorax cf. interruptus GUF 1 | Megalothorax cf. interruptus L1 | JN970929 |
Megalothorax cf. interruptus GUF 2 | Megalothorax cf. interruptus L1 | JN970928 |
Megalothorax cf. interruptus GUF 3 | Megalothorax cf. interruptus L2 | JN970910 |
Megalothorax granulosus cs110_CHL021 | Megalothorax granulosus | KC900204 |
Megalothorax minimus BEL 1 | Megalothorax minimus | JN970925 |
Megalothorax minimus cs70_Be001 | Megalothorax minimus | KC900191 |
Megalothorax minimus cs71_Be001 | Megalothorax minimus | KC900192 |
Megalothorax minimus cs93_Fr38 | Megalothorax minimus | KC900195 |
Megalothorax nigropunctatus cd345c | Megalothorax nigropunctatus | KC900196 |
Megalothorax nigropunctatus cs104_CHL102 | Megalothorax nigropunctatus | KC900197 |
Megalothorax nigropunctatus cs118_CHL033 | Megalothorax nigropunctatus | KC900198 |
Megalothorax nigropunctatus cs119_CHL205 | Megalothorax nigropunctatus | KC900199 |
Megalothorax perspicillum cs121_Fr114 | Megalothorax perspicillum | KC900200 |
Megalothorax perspicillum cs122_Fr114 | Megalothorax perspicillum | KC900201 |
Megalothorax perspicillum cs123_Fr114 | Megalothorax perspicillum | KC900202 |
Megalothorax perspicillum cs124_Fr114 | Megalothorax perspicillum | KC900203 |
Megalothorax sp. ARG 1 | Megalothorax sp2 | JN970916 |
Megalothorax sp. ARG 2 | Megalothorax sp1 | JN970926 |
Megalothorax sp. CHL 1 | Megalothorax sp1 | JN970927 |
Megalothorax sp. FRA 8 | Megalothorax sp4 | JN970913 |
Megalothorax sp. USA 1 | Megalothorax sp3 | JN970909 |
Megalothorax svalbardensis cs40_sva19 | Megalothorax svalbardensis | KC900205 |
Megalothorax willemi ARG 3 | Megalothorax willemi L6 | JN970918 |
Megalothorax willemi ARG 4 | Megalothorax willemi L6 | JN970919 |
Megalothorax willemi cs91_Be005 | Megalothorax willemi L6 | KC900193 |
Megalothorax willemi cs92_Be005 | Megalothorax willemi L4 | KC900194 |
Megalothorax willemi FRA 1 | Megalothorax willemi L5 | JN970912 |
Megalothorax willemi FRA 2 | Megalothorax willemi L6 | JN970917 |
Megalothorax willemi FRA 3 | Megalothorax willemi L1 | JN970911 |
Megalothorax willemi FRA 4 | Megalothorax willemi L3 | JN970920 |
Megalothorax willemi FRA 5 | Megalothorax willemi L3 | JN970921 |
Megalothorax willemi FRA 6 | Megalothorax willemi L3 | JN970922 |
Megalothorax willemi FRA 7 | Megalothorax willemi L4 | JN970915 |
Megalothorax willemi FRA 9 | Megalothorax willemi L2 | JN970924 |
Megalothorax willemi FRA 10 | Megalothorax willemi L2 | JN970923 |
Megalothorax willemi FRA 11 | Megalothorax willemi L7 | JN970914 |
Twenty MOTUs were delineated using a conservative 19.5% threshold based on the higher bound of the barcode gap (not shown). Over the seven species for which several specimens were sequenced, five were represented by a single MOTU (M. nigropunctatus, M. perspicillum Schneider & D’Haese, 2013, M. minimus, M. potapovi sp. n., M. sanguineus sp. n.) and two exhibited several discrete MOTUs each (M. willemi—7 MOTUs, Megalothorax cf. interruptus—2 MOTUs). Deep genetic divergences showed among the MOTUs (Fig.
The two new species M. potapovi sp. n., M. sanguineus sp. n. are both supported by differences in morphological and molecular characters. The sequencing of COI for a paratype for M. potapovi sp. n. is critical as it will prevent ambiguities if a case of cryptic diversity is discovered in this species (
Labrum.
Integument. The pairs of pseudopore-like elements at the base of antenna were never reported in the Megalothorax genus but seem to be common features of the genus. We observed them clearly on M. sanguineus sp. n. and M. carpaticus, they are visible but faint in the following Megalothorax species: potapovi sp. n., minimus, granulosus, nigropunctatus, willemi, svalbardensis and also in French specimens of M. sanctistephani. They were observed in SEM in M. perspicillum and M. aquaticus, where the dorsal one consists of a ring of primary grain and a small integumentary duct and the lateral one of a simple ring of primary grain. At the species level, those characters do not yield evident systematics value. They could not be observed in Neelus murinus and Neelides folsomi but the presence of dermastrons could blur the observation of the integument. In the state of knowledge, it is a putative apomorphy of the genus Megalothorax.
τ-chaetae.
Position of the most lateral τ-chaeta: (i) between p2 and p3 (M. perspicillum), (ii) in lateral position and close to p4 with 2 or less granules between the chaetae (M. minimus, M. svalbardensis, M. carpaticus, M. granulosus), (iii) in lateral position and more or less far from p4 with 5 or more granules between the chaetae (the two new species and M. nigropunctatus). M. draco, M. tatrensis and M. hipmani were also observed but the τ-chaetae could not be spotted at the exception of the most dorsal τ-chaeta in M. tatrensis.
Antenna. The homology of the chaeta directly below S4 (m5) on Ant. III of M. potapovi is ambiguous; an alternative hypothesis is provided in Fig.
Claws. The apparent elongation of the claws in M. sanguineus sp. n. called for a comparison with the other species of Megalothorax. The ratio of unguis I length : tibiotarsus I width was used by
Special thanks to Mikhail Potapov for having organized the field trip to Primorye (Russia), and to Jacek Pomorski and Anne Bedos for their contribution to specimen collection and extraction. The study on the Bernadouze peat-bog (France) was supported by grants of the LABEX DRIIHM - OHM Vicdessos for field trips and the laboratory work of Lorène Marchal. We are grateful to Vladimir Papáč and to anonymous reviewer for the thorough review of this manuscript and their valuable corrections and recommendations.