Research Article |
Corresponding author: Alice Laciny ( alice.laciny@nhm-wien.ac.at ) Academic editor: Marek Borowiec
© 2018 Alice Laciny, Herbert Zettel, Alexey Kopchinskiy, Carina Pretzer, Anna Pal, Kamariah Abu Salim, Mohammad Javad Rahimi, Michaela Hoenigsberger, Linda Lim, Weeyawat Jaitrong, Irina S. Druzhinina.
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:
Laciny A, Zettel H, Kopchinskiy A, Pretzer C, Pal A, Salim KA, Rahimi MJ, Hoenigsberger M, Lim L, Jaitrong W, Druzhinina IS (2018) Colobopsis explodens sp. n., model species for studies on “exploding ants” (Hymenoptera, Formicidae), with biological notes and first illustrations of males of the Colobopsis cylindrica group. ZooKeys 751: 1-40. https://doi.org/10.3897/zookeys.751.22661
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A taxonomic description of all castes of Colobopsis explodens Laciny & Zettel, sp. n. from Borneo, Thailand, and Malaysia is provided, which serves as a model species for biological studies on “exploding ants” in Southeast Asia. The new species is a member of the Colobopsis cylindrica (COCY) group and falls into a species complex that has been repeatedly summarized under the name Colobopsis saundersi (Emery, 1889) (formerly Camponotus saundersi). The COCY species group is known under its vernacular name “exploding ants” for a unique behaviour: during territorial combat, workers of some species sacrifice themselves by rupturing their gaster and releasing sticky and irritant contents of their hypertrophied mandibular gland reservoirs to kill or repel rivals. This study includes first illustrations and morphometric characterizations of males of the COCY group: Colobopsis explodens Laciny & Zettel, sp. n. and Colobopsis badia (Smith, 1857). Characters of male genitalia and external morphology are compared with other selected taxa of Camponotini. Preliminary notes on the biology of C. explodens Laciny & Zettel, sp. n. are provided. To fix the species identity of the closely related C. badia, a lectotype from Singapore is designated. The following taxonomic changes within the C. saundersi complex are proposed: Colobopsis solenobia (Menozzi, 1926), syn. n. and Colobopsis trieterica (Menozzi, 1926), syn. n. are synonymized with Colobopsis corallina Roger, 1863, a common endemic species of the Philippines. Colobopsis saginata Stitz, 1925, stat. n., hitherto a subspecies of C. badia, is raised to species level.
autothysis, behavioural ecology, Camponotini , Colobopsis , Formicidae , integrative taxonomy, male morphology, molecular biology, morphometry, new species, new status, new synonymy, phylogeny, Southeast Asia, taxonomy
The Colobopsis cylindrica (COCY) group likely represents a monophyletic clade containing Southeast Asian ant species with distinctive hypertrophied mandibular gland reservoirs. In territorial combat, minor workers use the sticky and irritant contents of their enlarged mandibular gland reservoirs to kill or repel rival arthropods. In species where this defensive behaviour is more advanced, this happens via the characteristic suicidal “exploding” by voluntary rupture of the gastral integument (autothysis) (
The Bornean members of the COCY group have been the subject of various ecological (e.g.,
The primary field research took place in the lowland dipterocarp rainforest at the Kuala Belalong Field Studies Centre (KBFSC), Temburong District, Brunei Darussalam (4°32'48.2"N, 115°09'27.9"E), where Colobopsis explodens sp. n. was sampled during five collecting trips (each of 30 days duration) encompassing different seasons from 2014 to 2016.
The behaviour of C. explodens sp. n. was observed at multiple nesting sites on several height-levels, starting from the forest floor and understory up to the canopy and emergent layer. The activity of ants was recorded in situ and in vitro using a CANON 70D Digital SLR Camera with a CANON EF 100 mm macro lens and a Tamron AF 28–200 mm F/3.8–5.6 XR Di aspherical (IF) macro zoom lens (Suppl. material
Sampling of Colobopsis badia in southern Thailand was conducted by H. Zettel and W. Jaitrong in June 2016. The sampling site was located in the Khao Chong Botanical Garden, near the Ton Pliw Waterfall (07°32'34"N, 99°47'33"E); a single male specimen was caught at a light at the Botanical Garden headquarters.
Nesting habits of C. explodens sp. n. were observed based on the model colony occupying several trees and an artificial nest (nest #38, Fig.
The activity of C. explodens sp. n. occupying artificial nest #38 was observed from 14th to 30th November of 2015 at different times during the day, for 30 minutes each by counting the ants entering and leaving the nest. Temperature, barometric pressure, and weather conditions were recorded, as well as any observed noteworthy behaviour (see Fig.
DNA extraction, gene fragment amplification, and sequencing were performed for minor worker ants of five different taxa (C. explodens sp. n., C. badia, C. nr. saundersi, C. aruensis Karawajew, 1933, and C. cylindrica (Fabricius, 1798)), as well as for mandibular gland reservoir content of C. explodens sp. n., one symbiotic cricket (Camponophilus sp.) from artificial nest #38, and the host plant of C. explodens sp. n. (Shorea johorensis).
For DNA barcoding of ant specimens, DNA was extracted from ant legs using Qiagen’s tissue QIAamp DNA Micro kit following the manufacturer’s protocol (Qiagen, Venlo, Netherlands). To obtain sufficient DNA quantity for further processing, the amount of legs used per sample varied. For the DNA extraction of queens, a minimum of three legs of one individual were transferred into one 1.5 ml microcentrifuge tube and frozen with liquid nitrogen. Three legs of one individual was also the minimum amount for males and major workers. For minor workers, all legs from two to four individuals were pooled (see Suppl. material
For DNA barcoding of symbiotic crickets, DNA was extracted from one whole specimen applying the same procedure as used for ants legs, but with a pretreatment with an enzymatic lysis buffer (Tris·Cl 20 mM, pH 8.0, sodium EDTA 2 mM, Triton X-100 1.2%, add lysozyme to 20 mg/ml for 60 min).
For DNA barcoding of the host plant, 100 mg of a leaf were ground with mortar and pestle under the use of liquid nitrogen and DNA was extracted using Qiagen’s DNeasy Plant Mini Kit according to manufacturer’s instructions.
For the ants, the gene fragments cytochrome C oxidase subunit I and II (COI, COII), cytochrome B (cytB), and carbamoyl-phosphate synthase II (cad) were amplified, for the cricket only COI. Additionally, a fragment of 16S rRNA was amplified from the DNA extracted from the mandibular gland content of C. explodens sp. n. minor workers, to assess the presence of bacteria. For the plant, the gene fragment maturase K (matK) was amplified. Primer sequences and specific annealing temperatures are given in Tab.
Sequences are deposited in NCBI GenBank. Accession numbers for ant specimens are given in Table
Gene | Name | Sequence 5’–3’ | Length [bp] | GC content [%] | Fragment Length [bp] | Annealing Temp [°C] | Reference |
---|---|---|---|---|---|---|---|
COI | LCO1490-F | GGTCAACAAATCATAAAGATATTGG | 25 | 32 | 709 | 45 |
|
HCO2198-R | TAAACTTCAGGGTGACCAAAAAATCA | 26 | 35 | ||||
COII | J2791-F | ATACCHCGDCGATAYTCAGA | 20 | 40–55 | 858 | 51 |
|
H3665-R | CCACARATTTCWGAACATTG | 20 | 35–40 | ||||
cytB | CB11400-F | TATGTACTACCHTGAGGDCAAATATC | 26 | 35–42 | 485 | 45 |
|
CB11884-R | ATTACACCNCCTAATTTATTAGGRAT | 26 | 27–35 | ||||
cad | CD1423EF | AGGTRATACRATCGGARAGRCCDGA | 25 | 40–60 | 800 | 55 |
|
CD1910R | CCGAGRGGRTCRACRTTYTCCATRTTRCAYAC | 32 | 38–63 | ||||
matK | 472F | CCCRTYCATCTGGAAATCTTGGTTC | 25 | 44–52 | 750 | 47 |
|
1248R | GCTRTRATAATGAGAAAGATTTCTGC | 26 | 31–38 | ||||
16S rRNA | fD1 | AGAGTTTGATCCTGGCTCAG | 20 | 50 | 1500 | 56 |
|
rP1 | ACGGTTACCTTGTTACGACTT | 21 | 43 |
List of sequence accession numbers in NCBI GenBank. * Nucleotide sequences from NCBI GenBank.
TUCIM | Other IDs | Organism | ng/µl | COI | COII | cytB | cad |
---|---|---|---|---|---|---|---|
5053 | C. explodens sp. n. | 14.7 | MF993252 | MF993269 | MF993286 | MF993304 | |
5056 | C. explodens sp. n. | 19.7 | MF993253 | MF993270 | MF993287 | MF993305 | |
5080 | C. explodens sp. n. | 10.4 | MF993254 | MF993271 | MF993288 | MF993306 | |
5098 | C. explodens sp. n. | 8.6 | MF993256 | MF993273 | MF993290 | MF993308 | |
5104 | C. explodens sp. n. | 16.6 | MF993257 | MF993274 | MF993291 | MF993309 | |
5148 | C. explodens sp. n. | 6 | MF993258 | MF993275 | MF993292 | MF993310 | |
5185 | C. explodens sp. n. | 7.3 | MF993259 | MF993276 | MF993293 | – | |
5205 | C. explodens sp. n. | 29.8 | MF993260 | MF993277 | MF993294 | MF993311 | |
6600 | C. explodens sp. n. | 8 | – | MF993284 | – | – | |
5855 | C. explodens sp. n. | 16.9 | MF993262 | MF993278 | MF993297 | MF993314 | |
5856 | C. explodens sp. n. | 28.2 | MF993263 | MF993279 | MF993298 | MF993315 | |
5942 | C. explodens sp. n. | 34.3 | MF993264 | MF993280 | MF993299 | MF993316 | |
5943 | C. explodens sp. n. | 142.1 | MF993265 | MF993281 | MF993300 | – | |
YG* | C. explodens sp. n. | n.a. | EF634201 | – | – | ||
6461 | C. badia | 21.3 | MF993266 | MF993282 | MF993301 | MF993317 | |
6463 | C. badia | 5.4 | MF993267 | MF993283 | MF993302 | MF993318 | |
6601 | C. badia | 17.91 | MF993268 | MF993285 | MF993303 | MF993319 | |
5698 | C. nr. saundersi | 50.1 | KU975365.1 | KU975366.1 | MF993296 | MF993313 | |
CH* | C. cf. cylindrica | n.a. | EF634198 | – | – | ||
5086 | C. cylindrica | 26 | MF993255 | MF993272 | MF993289 | MF993307 | |
5300 | CAMP004 | C. aruensis | 169.1 | MF993261 | – | MF993295 | MF993312 |
Cflor36* | Camponotus floridanus | n.a. | AY334397 | – | – | – |
GapStreeze v. 2.1.0 (https://www.hiv.lanl.gov/content/sequence/GAPSTREEZE/gap.html) was used for COI gene alignment with 95 % gap tolerance in order to retain only the conserved region. The individual gene alignments were subjected to best substitution model selection using the BIC criterion in SMS (
All specimens used for morphometry were card-mounted, individually numbered, and measured at magnifications from 25.6× up to 256× with a Nikon SMZ1500 binocular microscope. Genital structures of two male specimens were dissected and mounted separately. Results represent minimum and maximum values for each morph; in cases where a character could not be measured in all individuals, the number of measured specimens is given in parentheses. The complete dataset of measurements is provided in Suppl. material
Measurements and indices (* = only gynes and males)
EL Eye length. Maximum diameter of compound eye, measured in lateral view.
FeL Femur length. Maximum length of metafemur, measured from base to apex.
FWL* Forewing length. Length of forewing, measured from tegula to distal tip.
HaL Hair length. Length of the longest standing hair on first gastral tergite, measured from base to apex.
HL Head length. Maximum length of head in full-face view, excluding mandibles, measured from anteriormost point of clypeus to posterior-most point of head vertex, parallel to midline.
HS Head size. (HW + HL) / 2.
HW Head width. Maximum width of head in full-face view (including eyes if protruding; only in gynes).
ML Mesosoma length. Measured laterally from anterior surface of pronotum proper (excluding collar) to posterior extension of propodeal lobes.
MSW* Mesoscutum width. Maximum diameter of mesoscutum, measured dorsally.
NH Node height. Height of petiolar node, measured laterally, from the intersection point of the axes of maximum height and length to dorsal apex
OcD* Ocellar distance. Minimum distance between lateral ocelli, measured between median borders.
OcW* Ocellus width. Maximum diameter of median ocellus.
OED* Ocellar eye distance. Minimum distance between lateral ocellus and outer border of compound eye.
PH Petiole height. Maximum height of petiole in lateral view, measured from ventral-most point of petiolar sternum to dorsal apex.
PL Petiole length. Maximum length of petiole in lateral view, measured from inflexion point of anterior constriction to posterior margin, perpendicular to axis of maximum height.
PS5 Length of maxillary palp segment V, measured from base to apex.
PS6 Length of maxillary palp segment VI, measured from base to apex.
SL Scape length. Maximum length of antennal scape in dorsal view excluding basal neck and condyle.
SW Scape width. Maximum width of antennal scape, measured dorsally.
TL Total length. The added lengths of head (excluding mandibles), mesosoma, petiole, and gaster.
2r* Maximum length of 2nd radial crossvein (see Figs
4Rs+M* Length of 4th radial sector fused with median (see Figs
CI Cephalic index. HW / HL × 100
EI Eye Index. EL / HW × 100
FeI Femur Index. FeL / HW × 100
OI* Ocellar Index: OED / OcD × 100
PI Petiole Index. PH / PL × 100
PSI Palp Segment Index. (PS5 + PS6) / HS × 100
SI Scape index. SL / HW × 100
SWI Scape width index. SW / SL × 100
WVI* Wing Vein Index. 4RsM / 2r × 100
Digital stacked images of most specimens (Figs
Type material of C. explodens: Holotype (minor worker): Brunei, Temburong, Kuala Belalong Field Studies Centre, 04°33'N, 115°09'E, 60 m a.s.l., 10.XI.–5.XII.2015, leg. A. Laciny & A. Kopchinskiy (“YG Vienna Colony”, specimen number COCY 01565).
Paratypes (59 minor workers, 8 major workers, 16 gynes, and 6 males dry mounted; > 500 imagines stored in 96 % ethanol): 19 minor workers, 2 major workers, 12 alate gynes, 4 dealate gynes, 6 males (including allotype) (all dry mounted), as well as 8 males, 2 alate gynes, ca. 500 minor workers (in alcohol) from the same nest sample as holotype; 1 major worker, same locality and collector as holotype, 17.IV.2015, “YG 373 main natural nest”; 1 major worker, same data as holotype (“YG doorkeeper #19”); 2 major workers, same data as holotype except 20.IV.2015, leg. A. Kopchinskiy (“cf. YG 39 (351) artificial nest”); 8 minor workers, 2 major workers, same locality as holotype, 2002, leg. D.W. Davidson (“YG KB02-108”); 4 minor workers, same locality and collector as previous, no collection date, “YG 2025”; 5 minor workers, same locality and collector as previous, I.2012, “YG T-trail (202)”; 5 minor workers, same locality and collector as previous, 15.V.2014, “YG-2 (73)”; 7 minor workers, same locality and collector as previous, 15.V.2014, “YG-2 (49)”; 2 minor workers, same data as previous except Batu Apoi Forest Reserve, N04°32', E115°10', 200 m a.s.l., 25.XI.2004, (“CAYG A-370”); 15 minor workers (on 5 pins), same data as previous, except N04°55', E115°19', 60 m, 3.VII.2002, (“YG KB02-108 voucher”); 4 minor workers, Thailand, Chumphon Province, Krom Luang Chumphon W.S, 3.II.2002, leg. W. Jaitrong (“WJT02-TH-0116”); 5 minor workers, West Malaysia, Kelantan, 60 km NE Tanah Rata, Tanah Kerajaan, 1000 m a.s.l., 12.–30.IV2007, leg. P. Cechovský.
Additional material: 3 pupae (Suppl. material
For unique identification numbers of all 90 dry mounted specimens (60 minor workers, 8 major workers, 16 gynes, and 6 males), as well as information on caste and colony affiliation, see Suppl. material
The holotype and a portion of the paratypes will be deposited at the Brunei Museum; additional paratypes will be housed in the Universiti Brunei Darussalam, the Natural History Museum Vienna, the University of California (Davis, USA), the Natural History Museum of Los Angeles County (Los Angeles, USA), the Thailand Natural History Museum (Technopolis, Thailand), and the collection of H. Zettel (Vienna, Austria).
The topology of the phylogram based on the concatenated alignment of 2757 bp was concordant with topologies of COI and COII and not contradicted by the topology of cytB. The phylogram based on cad was statistically unresolved (data not shown). The obtained Bayesian consensus tree (Fig.
Bayesian consensus phylogram of C. explodens sp. n. and related taxa based on the concatenated alignments (2757 bp) of the three mitochondrial (COI, COII, cytB) and one nuclear (cad) phylogenetic markers. Nodes with posterior probabilities above 0.94 are marked with black dots. Specimens from Borneo are shown in green, from Thailand in blue. TUCIM 6463 corresponds to a male specimen.
Camponotus (Colobopsis)
sp. Yellow Goo:
Camponotus (Colobopsis)
sp. YG:
Colobopsis
sp. YG:
Present participle of Latin explodere, referring to the “exploding”-like autothysis behaviour.
Minor worker (Figs
Measurements of holotype minor worker: TL 6.78; HW 1.48; HL 1.67; HS 1.58; PS5 0.23; PS6 0.25; EL 0.42; SL 1.33; SW 0.14; ML 2.05; HaL 0.15; PH 0.55; PL 0.47; NH 0.33; FeL 2.05. Indices: CI 88; SI 90; SWI 11; EI 29; PI 116; FeI 139; PSI 30.
Measurements of paratype minor workers: (n = 59): TL 4.74–7.21; HW 1.22–1.57; HL 1.30–1.78; HS 1.27–1.67; PS5 0.21–0.25 (20); PS6 0.20–0.26 (21); EL 0.33–0.43; SL 1.21–1.39; SW 0.11–0.16; ML 1.50–2.22; HaL 0.08–0.19; PH 0.41–0.56 (44); PL 0.33–0.49 (47); NH 0.24–0.38 (52); FeL 1.73–2.10. Indices: CI 85–94; SI 87–104; SWI 9–12; EI 27–29; PI 112–133 (41); FeI 123–151; PSI 28–35 (20).
Structures: Head (Fig.
Colour: Body mainly reddish brown. Vertex of head, margins of clypeus, masticatory and lateral margins of mandibles, dorsum and ventral margins of mesosoma, mid portion of gastral tergites I–III, and legs slightly darker brown in most specimens; some specimens with darker area extending medially from head vertex to frons. Gastral tergites and sternites with very narrow hyaline margins. All gastral sternites, lateral fourths and posterior margins of tergites I–III, as well as entire tergites IV and V black.
Pilosity: Dorsum of head with very short, inconspicuous, appressed and subdecumbent setae; a few very long, standing setae on frons near declivity to vertex, medial of frontal carinae, and on lateral portions of clypeus. Mesosoma and petiole with fine and short, whitish, velvety pilosity; long, standing, slightly undulated setae restricted to pronotum; declivity of propodeum and node of petiole with few very short standing setae. Gastral tergites with moderately dense, short whitish, decumbent setae and few slightly darker, longer standing setae, most of them in transverse rows near hind margins. Longest setae in transverse rows near hind margins of sternites and at base of gastral tergite V.
Notes: Minor workers of Colobopsis explodens sp. n. show a continuous size variation across a remarkably wide range, similar to that found in the undescribed Colobopsis sp. nrSA (Fig.
(Figs
Measurements of paratype major workers (n = 8): TL 7.30–8.71; HW 1.72–1.89; HL 2.25–2.58; HS 1.99–2.20; PS5 0.15–0.17 (6); PS6 0.15–0.17 (6); EL 0.50–0.56; SL 1.15–1.26; SW 0.17–0.20; ML 2.22–2.74; HaL 0.11–0.20; PH 0.59–0.69 (6); PL 0.45–0.51 (6); NH 0.40–0.45 (6); FeL 1.50–1.70. Indices: CI 71–77; SI 64–69; SWI 14–17; EI 28–31; PI 125–143 (6); FeI 87–95; PSI 14–17 (6).
Structures: Integument mostly dull, only head and legs shiny. Head (Fig.
Colour: Overall slightly darker than minor worker; head, legs and mesosoma reddish brown; gaster slightly darker chocolate-brown, becoming darker towards caudal apex, hyaline margins yellowish; elevated crest of frontal shield, anterior clypeal margin, frontal carinae, and masticatory and lateral margin of mandibles blackish brown.
Pilosity: As in minor worker, except long setae on clypeus sides restricted to the area behind clypeal shield; mesonotum with standing setae which are approx. half the length of those on pronotum.
Notes: The head shield with a sharp, elevated crest is typical for majors of the Colobopsis saundersi complex (Fig.
(Fig.
Measurements of paratype gynes (n = 16): TL 10.50–12.16; HW 1.74–1.83; HL 2.28–2.45; HS 2.02–2.14; PS5 0.19–0.21 (13); PS6 0.19–0.23 (13); EL 0.62–0.66; SL 1.33–1.45; SW 0.20–0.22; ML 4.11–4.63; HaL 0.14–0.29 (15); PH 0.77–0.92 (11); PL 0.54–0.67 (10); NH 0.40–0.54 (11); FeL 2.25–2.35; OcW 0.13–0.16; OED 0.34–0.38; OcD 0.54–0.64; FWL 9.72–10.50 (11); MSW 1.68–2.15; 2r 0.50–0.64 (12); 4Rs+M 0.14–0.32 (12). Indices: CI 73–77; SI 75–80; SWI 15–16; EI 35–37; PI 123–150 (8); FeI 125–132; PSI 18–20 (13); OI 54–69; WVI 26–58 (12).
Structures: Head (Fig.
Colour: Chiefly as in major worker. Head and pronotum reddish brown; ventral and posterior mesosoma, petiole, legs and gaster somewhat darker chocolate-brown; mandibles and ridges of clypeal shield blackish brown. Pronotum and mesonotum with very narrow yellow margins. Gastral tergites medially with very narrow hyaline margins; sternites with relatively broad posterior margins. Wings hyaline, but forewing cells along veins, as well as pterostigma darkened to brownish. On hind wing all veins pale yellow (Fig.
Pilosity: Short pilosity and distribution of long setae on head, petiole, and gaster similar as in major worker, but that of mesosoma different; pronotum with few long, undulated setae. Medial part of mesonotum (between parapsidal furrows) with numerous long erect setae, scutellum with few long erect setae; lateral part of mesonotum in front of tegulae without setae.
Notes: The head shield with a sharp, elevated crest is typical for gynes of the Colobopsis saundersi complex (Fig.
(Figs
Measurements of allotype male: TL 7.11; HW 1.26; HL 1.20; HS 1.23; PS5 0.20; PS6 0.15; EL 0.44; SL 0.84; SW 0.11; ML 2.54; HaL n.a.; PH 0.46; PL.40; NH 0.29; FeL 1.83; OcW 0.18; OED 0.27; OcD 0.43; FWL 6.33; MSW 1.37; 2r 0.38; 4Rs+M 0.27. Indices: CI 105; SI 66; SWI 13; EI 35; PI 116; FeI 145; PSI 28; OI 62; WVI 70.
Measurements of paratype males (n = 5): TL 6.46–6.85; HW 1.24–1.29 (4); HL 1.14–1.24; HS 1.20–1.27; PS5 0.17–0.21 (4); PS6 0.13–0.17 (4); EL 0.43–0.46; SL 0.80–0.85; SW 0.10–0.12; ML 2.38–2.87; HaL n.a.; PH 0.45–0.49 (4); PL 0.38–0.40 (4); NH 0.26–0.33 (4); FeL 1.71–1.86; OcW 0.18–0.19; OED 0.25–0.27; OcD 0.43–0.46; FWL 5.87–6.33; MSW 1.17–1.50; 2r 0.38–0.47; 4Rs+M 0.14–0.22. Indices: CI 104–110 (4); SI 64–67 (4); SWI 12–15; EI 35–36 (4); PI 113–123 (4); FeI 136–151 (4); PSI 27–30 (4); OI 53–62; WVI 31–53.
Structures: Head (Fig.
Genital structures (Fig.
Variation of relevant morphometric measurements for the distinction of castes and species; a Metafemur length (FeL) in relation to head width (HW) for minor workers of C. explodens sp. n., as well as minor workers of C. badia and C. saundersi (type specimens plotted separately) b Scape length (SL) in relation to head width (HW) for minor workers, major workers, gynes and males of C. explodens sp. n., as well as male of C. badia.
Colour: Mainly dark chocolate-brown. Head somewhat darker; eyes pale grey to blackish; ocelli translucent, ranging from almost clear to reddish amber. Antennae and legs lighter brown, fading into yellowish towards apices. Margins of mesoscutum, scutellum, and metanotum lighter yellowish brown. Gastral tergites medially with very narrow hyaline margins; sternites with relatively broad, indistinctly separated posterior margins. Wings almost hyaline, with a slight whitish tinge, but forewing cells along veins, as well as pterostigma darkened to brownish, all veins pale yellowish brown. On hind wing all veins pale yellow.
Pilosity: On head setae sparsely distributed, short, inconspicuous, appressed, subdecumbent; a few very long standing setae on frons near vertex, and on anterior and posterior clypeal margins. Mandibles with dense short pilosity on lateral face, and few moderately long setae on anterolateral margin. Short pilosity and distribution of long setae on mesosoma, petiole, and gaster similar as in gyne, but pronotum lacking long, undulated setae. Medial part of mesonotum (between parapsidal furrows) with numerous long erect setae, scutellum with few long erect setae; lateral part of mesonotum in front of tegulae without setae. Tegulae with dense brush of setae. Petiole with a few stout setae anteroventrally (see insert Fig.
Colonies of C. explodens sp. n. observed in the Ulu Temburong National Park are commonly polydomous and polygynous. This species was selected as a model for the study of the “exploding ants” because among the species with advanced autothysis behaviour it was the most abundant COCY taxon in the vicinity of KBFSC.
Colobopsis explodens sp. n. frequently nests on dipterocarp trees and its colonies can contain thousands of individuals. The largest part of the studied colony lived on a 60 m tall Shorea johorensis Sym. (Dipterocarpaceae) tree identified morphologically and by DNA barcoding (matK, identical to GenBank accession number KY973022, E-value is zero;
The colonies are distributed three-dimensionally, occupying any suitable nesting ground within the colony boundaries. On the main tree, we found four nesting sites of the examined colony in dead branches at heights ranging between 35 and 55 m above ground and two nesting sites in the living stem 50–60 m above ground. No nests in living branches were observed. At least five nest entrances were also seen in the stem of S. maxwelliana. No signs of necrosis of the plant tissue were observed around stem entrances.
The translocation of a nest fragment in a fallen branch to the laboratory’s terrace, 30 m away from its original location, resulted in the expansion of the colony’s foraging ground to a neighbouring Shorea sp. tree where these ants were not previously present, while the connection to the colony on the original host tree was maintained.
If provided with an appropriate artificial nest (Fig.
Activity of C. explodens sp. n. at the entrance to the artificial nest #38. a Scatter plot of the number of minor workers entering (black dots) and leaving (open dots) the nest depending on the time of day b Scatter plot of the number of minor worker ants entering (black dots) and leaving (open dots) the nest depending on the air temperature. Polynomial trend lines on a and b are shown for the numbers of entering (black) and exiting (grey) minor workers c and d show high and low worker activity near the entrance of nest #38, respectively; white bars denote 1 cm.
Remarkably, during all observations the numbers of the minor workers leaving and entering the nest were almost equal. The fact that this proportion did not change over the day (Fig.
After dusk, other species of Camponotini such as Polyrhachis spp., Camponotus spp., and Dinomyrmex gigas (Latreille, 1802) were observed on the trees in the vicinity of the artificial nest.
Within the colonies, minor workers were by far the most abundant caste of C. explodens sp. n., whereas major workers (soldiers) were rare and almost never seen outside the nest. Alate gynes and males were observed leaving the nest during nuptial flight after dusk on two occasions during our field observations (Suppl. material
Observations have shown that minor workers of C. explodens sp. n. display a characteristic, possibly defensive pose with raised gaster (Suppl. material
Minor workers of C. explodens sp. n. spend significant time on leaves, which has previously been hypothesized to contribute to their nutrition (compare with
Another very specific behaviour was exhibited on the tree bark: Minor workers “graze” on the layers of epiphytes (mosses, lichens, algae, filamentous fungi, and yeasts) with their mandibles, often for up to 60 minutes. This behaviour differs from the cleaning behaviour on leaves and presumably contributes to the ants’ nutrition.
Preliminary feeding experiments using cultures of filamentous fungi isolated from the phyllosphere of the host trees remained unsuccessful, no fungal feeding was observed. Only a suspension of yeast in water was accepted in vitro (M. Rahimi, pers. obs.). However, minor workers of C. explodens sp. n. have been observed to feed on small dead insects, fruit, and fish when offered on the foraging grounds (A. Kopchinskiy, A. Laciny & M. Hoenigsberger pers. obs.).
Commonly observed modes of behaviour of C. explodens sp. n. in situ and in vitro as well as a variety of nesting sites are documented in the Suppl. material
The molecular analysis of the mandibular gland (MG) content of C. explodens sp. n. resulted in PCR amplification and sequencing of the 16S rRNA fragment of the bacteria Blochmannia sp. (Gammaproteobacteria), a genus of obligate symbiotic bacteria found in carpenter ants (
More detailed data on autothysis, composition of mandibular gland secretion, biodiversity of the COCY-associated microorganisms, and experimental assessment of nutrition will be presented in future publications.
Formica
badia
:
Camponotus
badius
:
Colobopsis
badia
:
examined. 1 lectotype minor worker (Oxford University Museum of Natural History, present designation), Singapore, “Formica badia”, “Syntype”, CASENT 0901897, “Lectotypus Formica badia Smith, 1857 des. Laciny & Zettel, 2017”, 2 paralectotype minor workers (Oxford University Museum of Natural History) mounted on the same card, Sarawak (“Sar 32”), “Formica badia”, “Syntype”, “Paralectotypes Formica badia Smith, 1857”.
1 male (Natural History Museum Vienna), Thailand, Trang Province, Nayong District, Khao Chong Botanical Garden, at light of head quarter, 7°33'N, 99°46'E, 60 m a.s.l., 1–7.VI.2016, leg. H. Zettel (68); 10 minor workers (Natural History Museum Vienna), Thailand, Trang Province, Nayong District, Khao Chong Botanical Garden, trail to Ton Pliw Waterfall, N07°32'34", E99°47'33", 150 m a.s.l., 1–7.VI.2016, leg. H. Zettel (66-4).
Lectotype: Minor worker glued to a square cardboard, in relatively good condition; right hind leg missing; tarsi of middle legs and left hind leg broken; erect setae on dorsum probably lost. Structures agree well with other species of the C. saundersi complex, a few characteristic features are given: Setae on scape more decumbent than in C. explodens sp. n. Dorsal outline of mesosoma almost straight, only with weak indentation at meso-metanotal suture. Propodeum forming a distinct obtuse angle in lateral view. Petiolar node relatively short, apex acute in lateral view, its crest slightly indented in middle. Tergites I–III with very fine, strongly transverse microsculpture (lateral parts not visible). Colour relatively dark brown; appendages strongly infuscate; antennal segments III–XII, meso- and metafemora almost black.
Paralectotypes: Two minor workers glued to the same square cardboard, in relatively poor condition. Left specimen with damaged head and gaster, lacking right middle leg; major parts of body covered by dirt or glue; most erect setae probably lost. Right specimen with slightly damaged head, lacking gaster and right hind leg; some parts of body covered by dirt or glue; most erect setae probably lost. The two specimens are probably conspecific, but conspecificity with the lectotype is uncertain. The combination of morphological features is intermediate between C. badia and C. explodens sp. n.: setae on scape similar to C. explodens sp. n., more erect than in the lectotype; dorsal outline of mesosoma intermediate, more structured than in the lectotype, but propodeum with angle; shape of node intermediate, apex more acute than in C. explodens sp. n. Colour almost as dark as in the lectotype.
Measurements of lectotype minor worker: TL 6.13; HW 1.51; HL 1.63; HS 1.57; PS5 n.a.; PS6 n.a.; EL 0.40; SL 1.43; SW 0.15; ML 1.96; HaL 0.17; PH 0.54; PL 0.36; NH 0.32; FeL 2.22. Indices: CI 93; SI 95; SWI 10; EI 26; PI 150; FeI 147; PSI n.a.
Measurements of paralectotype minor workers* (n = 2): TL 6.13, n.a.; HW n.a., 1.36; HL n.a., 1.52; HS n.a., 1.44; PS5 n.a., 0.21; PS6 n.a., 0.25; EL 0.36, 0.37; SL 1.39, 1.40; SW 0.12, 0.13; ML 1.89; HaL 0.13, n.a.; PH n.a., 0.51; PL 0.37, 0.42; NH 0.27; FeL 2.15. Indices: CI n.a., 89; SI n.a., 103; SWI 8, 9; EI n.a., 27; PI n.a., 121; FeI n.a., 158; PSI n.a., 32. *One specimen with strongly damaged head, one with missing gaster.
Measurements of non-type minor workers (n = 10): TL 5.64–6.23; HW 1.46–1.59; HL 1.63–1.72; HS 1.54–1.65; PS5 0.24–0.25 (3); PS6 0.24 (3); EL 0.38–0.40; SL 1.37–1.43; SW 0.13–0.14; ML 1.96–2.22; HaL 0.13–0.19; PH 0.51–0.56 (7); PL 0.41–0.45; NH 0.31–0.37 (9); FeL 2.09–2.28. Indices: CI 88–92; SI 90–96; SWI 9–10; EI 25–27; PI 118–130 (7); FeI 141–148; PSI 30–31 (3).
Notes on collecting and identification: A single male collected at light was identified as a specimen of the C. cylindrica group. DNA barcoding revealed specific identity with a nest series of C. badia from the same botanical garden. The morphological identification of this nest series (Col. 66-4) was carried out by direct comparison to the lectotype of C. badia.
Description (Fig.
Measurements of male (n = 1): TL 8.28; HW 1.35; HL 1.26; HS 1.30; PS5 0.20; PS6 0.15; EL 0.48; SL 0.83; SW 0.10; ML 3.07; HaL n.a.; PH 0.47; PL 0.41; NH 0.31; FeL 1.96; OcW 0.19; OED 0.29; OcD 0.46; FWL 7.43; MSW 1.37; 2r 0.41; 4Rs+M 0.37. Indices: CI 107; SI 61; SWI 13; EI 36; PI 116; FeI 145; PSI 26; OI 61; WVI 91.
Structures: Size larger (TL ca. 8.3 mm). Integument rather shiny (Fig.
Genital structures (Fig.
Colour: Head chiefly dark brown, with lighter area comprising frons between antennal insertions and clypeus. Eyes grey. Ocelli clear, almost colourless. Posterior and anterior clypeal margins, as well as proximal fourth of clypeal carina black. Gaster dark brown. Mesosoma, petiole, mandibles, antennae, and legs lighter brown, appendages becoming yellowish towards apices. Antennal insertions, mandibular bases, margins of thoracic sclerites (especially below tegulae) creamy yellow. Gastral tergites medially with very narrow hyaline margins; sternites with relatively broad posterior margins. Wings hyaline, forewing with a slight brownish tinge and cells along veins, as well as pterostigma darker brownish, all veins pale brown. On hind wing all veins pale yellow.
Pilosity: Appressed and subdecumbent setae comparatively shorter and sparser, but difference less obvious on gaster. Standing setae on mesonotum and gaster shorter, on mesonotum less numerous.
Comparative notes: The male of C. badia can be distinguished from males of C. explodens sp. n. by larger body size, differing colour pattern, more shiny integument, well-developed clypeal carina, differing proportions of wing venation, and relatively shorter scapes (Fig.
In this study, three mitochondrial DNA loci and one nuclear DNA fragment were applied for the construction of a molecular phylogenetic tree (Fig.
The DNA extraction from the gastral parts of the mandibular gland reservoirs of C. explodens sp. n. minor workers resulted in drastically low yields indicating no abundant microbial symbionts present there. The successful 16S rRNA PCR amplification gave a sharp band that was sequenced with high reproducibility. The 16S rRNA fragment corresponding to the whole genome sequenced Blochmannia endosymbiont of North American Colobopsis obliquus strain 757 (NCBI GenBank accession number CP010049,
The treatment of Colobopsis as a genus separate from Camponotus is supported by molecular, morphological, and biological data (
A first attempt of a comprehensive classification of the species of Colobopsis (as a subgenus of Camponotus) was done by Carlo Emery. In his outstanding treatment of Formicinae (
More species of the COCY group were subsequently described by
According to our morphological studies the COCY group can be defined as such: polymorphic Colobopsis with at least three distinct female castes: (i) winged, phragmotic gynes, (ii) phragmotic soldiers (doorkeepers), and (iii) minor workers with a considerable size variation; intermorphic workers may occur in addition (
Following this definition, the COCY group presently comprises 17 names in the rank of species, subspecies or variations, which are partly in synonymy to each other. The strong intraspecific variation of minor workers, the frequently lacking knowledge on soldiers (or gynes), and the historical descriptions of taxa from different morphs (either workers or gynes) make the species taxonomy a true challenge. A preliminary analysis of morphological and molecular data (unpublished) supports the division of the group into four species complexes (molecular data of one complex not available). We restrict the following analysis to the species of the C. saundersi complex, which includes C. explodens sp. n. and can be defined by the following combination of characters observable in minor workers and soldiers: head always red or brown (not black); mesosoma moderately elongated and dorsally with some long undulated setae, at least on pronotum; node of petiole without long setae; gastral tergites with dense (in most species strongly transverse) micro-reticulation and with small hair pits (without large grooves). Soldiers and gynes (not known of all species) have a strongly truncated head, in most species with a well-defined (crested) head shield. The following names are available in this group (listed chronologically): Colobopsis badia (Smith, 1857), C. corallina Roger, 1863, C. saundersi Emery, 1889, C. badia var. krama Forel, 1912, C. badia saginata Stitz, 1925, C. solenobia (Menozzi, 1926), and Colobopsis trieterica (Menozzi, 1926), comb. n.
Colobopsis corallina (=C. solenobia syn. n., =C. trieterica syn. n.) is an endemic species from the Philippines. Soldiers and gynes differ strongly from C. explodens sp. n. and other taxa of the complex (as far as such morphs are known) by a very obtusely margined, not crested head shield. Minors have a bright orange colour on head, mesosoma, and petiole, often extending to gastral tergite I. Morphometrically, the examined minor workers of C. corallina (n = 31) mainly differ from those of C. explodens sp. n. by a greater average length of appendages (SI 92–109 vs. 87–104; FeI 136–159 vs. 123–151; PSI 30–39 vs. 28–35).
The greatest similarity is observed between C. explodens sp. n. and C. saginata (stat. n.), a taxon only known from a single alate gyne from Northern Borneo. The important structures of the head shield are almost identical. Although strongly different from C. explodens sp. n. by pale orange brown colour, this gyne differs only by subtle morphometric characters of which the long and distally wide scape seems to be the most reliable (SI 83 vs. 78–80). The length of vein 4Rs+M is considerably longer in C. saginata than in C. explodens sp. n. (WVI 65 vs. 26–58).
Colobopsis badia var. krama, described from a soldier from Java (
Colobopsis badia was described by
We examined two syntype minor workers of Colobopsis saundersi from Myanmar (“Tenasserim, Thagata”, one illustrated by AntWeb.org under CASENT0905463). Morphometric analysis revealed no differences between the types of C. saundersi and C. badia, suggesting that the two species should be synonymized. Colobopsis saundersiwas considered a junior synonym of C. badia by Carlo Emery himself (
The morphology of males of the tribe Camponotini is insufficiently studied, so that a complete comparison at generic level is not possible. The modified (enlarged) first funicular segment is presumably characteristic for males of Colobopsis. This characteristic has been described in the type species, Colobopsis truncata (Spinola, 1808), by
Males of the COCY group have previously been described for three species (see below). However, these descriptions largely lack the necessary details to meaningfully compare taxa. No previous accounts of genital morphology or illustrations of male specimens have been found in the literature.
Colobopsis badia:
Colobopsis severini (Forel, 1909): The extremely brief description of a male from the island Labuan (near Borneo) is not sufficient to draw any meaningful taxonomic conclusions.
Colobopsis leonardi (Emery, 1889):
According to our knowledge, males of the C. cylindrica group can be distinguished from other Southeast Asian Colobopsis species by the relatively rich subdecumbent pilosity and the dense microreticulation of gastral tergites.
The behavioural observations on C. explodens sp. n. conducted at KBFSC revealed multiple modes of behaviour that are either poorly studied or new to science.
The diurnal activity pattern, as well as the positive influence of high temperatures correspond to the results of previous studies in related taxa (see
A further noteworthy activity observed in foraging workers was so-called “grazing” behaviour, in which minor workers were frequently seen using their mandibles to pluck and chew at various mosses, lichens, and other epiphytes on the bark of trees or other surfaces. While this activity can last for up to one hour at a time, its exact purpose remains unclear. It is possible that minor workers cut and consume parts of the plants and microorganisms or merely ingest fluids. As previous analyses of nitrogen isotopes (
We are thankful to Diane W. Davidson for introducing us to the diversity of Colobopsis cylindrica ants at KBFSC and for passing down her knowledge on the ecology of these organisms. We thank the administration of KBFSC and Universiti Brunei Darussalam (UBD) for project approval and Brunei’s Forestry Department for permission to collect ants and use canopy walkways. The authors appreciate the help of UBD and KBFSC staff, especially Salleh Abdullah Bat, Teddy Chua, Masnah Mirasan, Rafhiah Kahar, Roshanizah Rosli, Rodzay Wahab, Chan Chin Mei, and Kushan Tennakoon for facilitating research and fieldwork in many ways. Sincere thanks are due to the authorities of the Khao Chong Botanical Garden, Trang, Thailand, for support and permission for fieldwork. We acknowledge the efforts of Komal Chenthamara (TU Wien, Austria) for her assistance in phylogenetic analysis and Marco Prusa (TU Wien, Austria) for the molecular DNA analysis of MG products. Thanks are owed to Heinz Wiesbauer (Vienna) for providing photographs of living specimens. We thank Sigfrid Ingrisch (Bad Karlshafen) for identifying the ant crickets. Finally, thanks are owed to James Hogan (Oxford University Museum of Natural History), Giar-Ann Kung (Natural History Museum of Los Angeles County), Phil Ward (University of California, Davis), and Roberto Poggi and Maria Tavano (Museo Civico di Storia Naturale “Giacomo Doria”, Genova) for contributing important specimens. The research was supported by the WWTF LS13-048 grant to ISD.
Figure S1. Living workers of C. explodens sp. n. on a detached branch containing a nest fragment
Figure S2. Natural nest of C. explodens sp. n. in a dead branch in the high canopy of S. johorensis (the main host tree for the model colony)
Figure S3. Interior of the natural nest of C. explodens sp. n. found in a dead tree branch of S. johorensis on the forest floor
Figure S4. Autothysis as defensive behaviour in an experimental setting
Figure S5. Pupa of C. explodens sp. n. found inside the opened natural nest
Table S6.
Video S7. Video depicting habitat, nesting sites and behaviour of C. explodens sp. n.