The Venom Apparatus and Other Morphological Characters of the Ant Martialis Heureka (Hymenoptera, Formicidae, Martialinae)
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Volume 50(26):413‑423, 2010 The venom apparaTus and oTher morphological characTers of The anT Martialis heureka (hymenopTera, formicidae, marTialinae) carlos roberTo ferrreira brandão1,3 Jorge luis machado diniz2 rodrigo dos sanTos machado feiTosa1 AbstrAct We describe and illustrate the venom apparatus and other morphological characters of the recently described Martialis heureka ant worker, a supposedly specialized subterranean predator which could be the sole surviving representative of a highly divergent lineage that arose near the dawn of ant diversification. M. heureka was described as the single species of a genus in the subfamily, Martialinae Rabeling and Verhaagh, known from a single worker. However because the authors had available a unique specimen, dissections and scanning electron microscopy from coated specimens were not possible. We base our study on two worker individuals collected in Manaus, AM, Brazil in 1998 and maintained in 70% alcohol since then; the ants were partially destroyed because of desiccation during transport to São Paulo and subsequent efforts to rescue them from the vial. We were able to recover two left mandibles, two pronota, one dismembered fore coxa, one meso-metapropodeal complex with the median and hind coxae and trochanters still attached, one postpetiole, two gastric tergites, the pygidium and the almost complete venom apparatus (lacking the gonostylus and anal plate). We illustrate and describe the pieces, and compare M. heureka worker morphology with other basal ant subfamilies, concluding it does merit subfamilial status. Keywords: Ant phylogeny; Formicidae; Martialis; Ultrastructure; Venom Apparatus. IntroductIon dusk in a primary lowland rainforest walking on the leaf litter. The ant Martialis heureka was recently described The phylogenetic position of this ant was inferred by Rabeling and Verhaagh (in Rabeling et al. 2008) from several nuclear genes, sequenced from one front based on a single worker collected by Christian Ra‑ leg (Rabeling et al., 2008). On the basis of morphol‑ beling in Manaus (Embrapa Headquarters 02°53’S, ogy and phylogenetic evidence the authors suggested 59°59’W), Amazonas state, Brazil, 28 km of Highway this specialized subterranean predator is the sole sur‑ AM 010 in May 9, 2003. The specimen was found at viving representative of a highly divergent lineage that 1. Museu de Zoologia, Universidade de São Paulo, Caixa Postal 42.494, 04218‑970, São Paulo, SP, Brasil. 2. Campus Jataí, Unidade Jatobá, Laboratório de Hymenoptera Frei Kempf, Universidade Federal de Goiás, BR 364, km 192, nº 3.800, 75801‑615, Jataí, GO, Brasil. 3. Corresponding author, E‑mail address: [email protected]. 414 Brandão, C.R.F. et al.: Venom apparatus and morphology of Martialis heureka ant workers arose near the dawn of ant diversification. Accordingly MAterial And Methods they described it as a species of a monotypic genus in the subfamily Martialinae Rabeling and Verhaagh. Photographs taken under the MZSP scan‑ This very important ant was regrettably de‑ ning electron microscope (SEM) (LEO 440®) were scribed from a single worker, which thus did not allow used to record morphological details of M. heu- for a more detailed morphological study, for instance, reka workers. The recovered parts were previously under electron scanning microscope, and of the cleaned in acetone, critical‑point dried in a Balzer mouth parts or the venom apparatus. The M. heureka (Bal‑Tec® CPD 030), and sputtered over with gold holotype is deposited in Museu de Zoologia da Uni‑ (Bal‑Tec® SCD 050). After that, the fragments were versidade de São Paulo. Rabeling et al. (2008) briefly fixed on the stubs using silver glue. The images were mention that five years before the discovery of the obtained under several magnifications, according to holotype of M. heureka, two workers of the same spe‑ the size of the observed structure. Finally, the images cies were discovered by M. Verhaagh in a soil sample were edited (Adobe Photoshop CS2®) to enhance taken by the SHIFT‑project team under the guidance brightness and contrast. of Marcos Garcia (Embrapa, Manaus) in a different We studied also the sting apparatus of one of but nearby patch of primary rainforest from where the the fragmented specimens. The apparatus was cleared holotype came from. in NaOH for 12 hours, and then in lactophenol at Indeed, M. Verhaagh brought several ant samples 45‑50°C for 12 more hours (or longer if necessary), from Manaus to the Museu de Zoologia at that time, rinsing it twice in distilled water, and then transferred including the one with two M. heureka workers and to Hoyer’s fluid. After the clearing process, the sting several other ants found in the same spot. The label apparatus was dismembered, and soaked in Hoyer’s information found along with this sample says: “Flo. fluid for observation and illustration under optical Solo Berl. AN. 06, 02/09/1998, SHIFT 52, Manaus, light microscopes. The terms for sting apparatus mor‑ leg. M. Garcia et al.”, meaning forest soil submitted phology follow Kugler (1978) and Diniz (1997). to Berlese, sample 06, collection date September 02, 1998. SHIFT project 52. Manaus. M. Garcia et al. leg. Unfortunately, the vial which included the results M. heureka workers dried up during the trip from Karl‑ sruhe to São Paulo, because the lid opened. Brandão The fragments we were able to recover did not tried to recover the dried specimens by filling the vial allow for a complete reconstruction of a M. heureka with 70% alcohol and adding a drop of detergent; worker. Notwithstanding, we recuperated important even allowing for several days, the ants did not free pieces from the two individuals: two left mandibles, themselves from the vial walls and from the other ants two pronota, a dismembered fore coxa, one meso‑ that dried up with them. In a desperate move, he gen‑ metapropodeal complex with the right median and tly let the vial with the ants to come in contact with an hind coxae with the trochanters attached, and the left ultrasound apparatus, just to watch most ants in the median coxa with the trochanter, one postpetiole, one vial shatter into small pieces. He collected the pieces gastric tergite, the pygidium and the almost complete and kept them in 70% alcohol until now. This ant venom apparatus. From the original Automontage proved so important and interesting that we decided picture and description, it is possible to see that the to be worthwhile recovering the pieces and studying metasoma (abdominal segments IV‑VIII visible) is the different morphological aspects in detail, in order laterally compressed and drop shaped in lateral view to compare this ant to other formicid subfamilies. (Rabeling et al., 2008). Unfortunately, the pieces we were able to recov‑ The pictures (Figs. 1‑3) we show do not display er did not allow a full reconstruction of the M. heu- similar pilosity in comparison to the original Auto‑ reka workers, but we present here details of some montage pictures; summing our information with parts of the exoskeleton and of the complete venom that presented by Rabeling et al. (2008), we can say apparatus, aiming to stimulate future research on this that in general, pronotum and legs are densely covered spectacular ant. Finally we compared our observations with erect to suberect hairs and sparsely with longer with those of Ettershank (1966) and Gotwald (1969) erect setae, although most of the hairs were lost during on the mandibles, with Kugler’s (1978) records of SEM preparation. The scars, however, are perfectly vis‑ the venom apparatus, with the character coding em‑ ible under the SEM. Most hairs are erect, thin, flexu‑ ployed by Bolton (1990), Baroni Urbani et al. (1992), ous, and follow no definite direction. Dense appressed Astruc et al. (2004), and Brady & Ward (2005). pubescence is absent from entire body. Special features Papéis Avulsos de Zoologia, 50(26), 2010 415 of surface sculpture and hairs, when conspicuous, will of short suberect hairs (better seen in Automontage be described below, in the appropriate sections. pictures; Rabeling et al., 2008). The mandible is not cylindrical in cross section (Fig. 1A); the internal face of the mandible is almost Mandibles (Fig. 1) flat throughout its length, vertical, and bears oblique rugulae that arise from the ventral margin and gently As described in Rabeling et al. (2008), Martialis curve upwards, although never attaining the dorsal heureka worker mandibles are forceps‑like, slender, ex‑ margin (Fig. 1D). The mandibular base is triangu‑ tremely elongate, and even longer than the head cap‑ lar when seen from behind (Fig. 1B); the mandalus sule. The mandibles are inserted at the far lateral sides (MA) is also triangular, and, according to Ettershank of head. Mandibular pubescence is dense, consisting (1966) may contain the mandibular gland orifice; the FIgure 1: Electron micrographs of the left mandible of a Martialis heureka worker. A. Internal view of the whole mandible. b. Detail of mandibular base: CA, canthellus, GI, ginglymus, MA, mandalus, TR, trulleum. c. Detail of mandibular apical portion. d. Preapical teeth, showing, at left, the beginning of the two dorsal ridges and their teeth. 416 Brandão, C.R.F. et al.: Venom apparatus and morphology of Martialis heureka ant workers FIgure 2: Electron micrographs of the mesosoma and legs of a Martialis heureka worker. A. Pronotum in lateral view. b. Meso‑ metapropodeal complex with middle coxa and trochanter in profile. c. Detail of metapleural gland (MG) and propodeal spiracle openings (PS), and petiolar foramen (FR). d. Meso‑metapropodeal complex in dorsal view. e. Meso‑metapropodeal complex in ventral view. F. Detail of metacoxal cavity (MC), propodeal spiracle opening (PS), and petiolar foramen (FR). g. Fore coxa in profile. Papéis Avulsos de Zoologia, 50(26), 2010 417 trulleum (TR) is open and rather large; the canthellus the mandible is falcate (Fig.