A revision of Malagasy species of Anochetus Mayr and Odontomachus Latreille (Hymenoptera: Formicidae). Author Fisher, B. L. Author Smith, M. A. text PLoS ONE 2008 3 1 23 http://dx.doi.org/10.1371/journal.pone.0001787 journal article 21401 Anochetus grandidieri Forel Figures: worker 3a-d, 5c; queen 3e-h; male 3i-j, 8b; map 6b Type material: Anochetus grandidieri Forel, 1891 : 108 [ 21 ] . Lectotype: worker, Madagascar , Forest of the east coast ( M. Humblot ) ( MHNG ), present designation [examined], AntWeb CASENT0101819 . Brown, 1978: 606 [ 2 ] (description of worker) . Anochetus madecassus Santschi, 1928 : 54 [ 22 ] . Lectotype: dealate queen, Madagascar , Nossi-Be( Descarpentries ) ( NHMB ) Lectotype by present designation [examined] AntWeb CASENT0101098 . Synonymized with grandidieri by Brown, 1978 : 557 [ 2 ]. Worker measurements: maximum and minimum based on all specimens, n = 20. HL 0.79-1.19, HW 0.71-1.06, CI 85-95, EL 0.08-0.13, ML 0.33-0.57, MI 41-54, SL 0.57-0.88, SI 78-86, WL 0.87-1.35, FL 0.57-0.90, PW 0.44-0.62. Figure 8. Anochetus males, terminalia, lateral view. A, boltoni CASENT0063847. B, grandidieri CASENT0080660. B, madagascarensis CASENT0063421. D, pattersoni CASENT0172617. doi:10.1371/journal.pone.0001787.g008 Figure 9. NJ tree of K2P for five species of Anochetus in Madagascar, Comoros and Aldabra (all specimens with.500 bp). Deep divergences evident between madagascarensis , grandidieri , and goodmani are evident. Deep divergences within A. goodmani are evident (In this tree, A. boltoni falls within goodmani ). The rightmost column of colors differentiate which biogeographical groupings of Wilme et al. [ 29 ] these populations fall. WCE-1 = Binara, Antsahabe. WCE-12 = Andavakoera, Ankarana. WCE-7= Kirindy Mite. WRDW-B = Vazimba, Androngonibe, Andranopasazy. doi:10.1371/journal.pone.0001787.g009 Table 2. Anochetus goodmani within-species pair-wise partitioning of genetic variance for the CO1 DNA barcode. K2P distances are beneath the diagonal and the number substitutions are above the diagonal. Table 3. Comparison of the utility of various complimentary nuclear markers for species diagnosis in the ponerine ants of the Malagasy.

Taxa	18S	28S	ITS1	Comments
Anochetus goodmani	Intra - no variation. nter - 2 bp from A. boltoni , and 3 bp from O. troglodytes , O. coquereli	Intra - no variation across north nter -.15 bp divergent from A. madagascarensis .	Intra - extreme variation (length and substitution) across range. Some corresponding to deep CO1 splits - provisionally orthologous. However, deep paralogous divergences have been sequenced within single individuals through different amplifications and extractions.	rRNA is, a priori, difficult to differentiate orthologous from paraologous. Not as immediately useful as an independent marker without cloning.
Anochetus boltoni	Intra - no variation. nter - 2 bp from A. goodmani , 2 bp from O. troglodytes and no difference from O. coquereli .	N/A	Intraspecific variation of 1% (indels and substitutions) between the two sampled populations.
Anochetus madagascarensis	N/A	Intra - no variation. nter -.15 bp divergent from A. goodmani .	Intra - variation that does NOT reflect CO1 variation.	rRNA is, a priori, difficult to differentiate orthologous from paraologous. Not as immediately useful as an independent marker without cloning. -Positive Wolbachia test.
Anochetus grandidieri	N/A	N/A	Low iIntraspecific variation that does reflect CO1 geographic variation.	- Positive Wolbachia test.
Anochetus pattersoni	N/A	N/A	N/A
Odontomachus coquereli	Intra - no variation. nter - 2 bp from O. troglodytes . 2bp from A. boltoni ,and 3 bp from A. goodmani .	Intra - variation. Large variation at geographically distal ends of distribution. nter - differentiates between three Malagasy species.	Intraspecific variation that only partially reflects geography and CO1 variation - while some clearly does not. Paralogous and orthologous
Odontomachus troglodytes	Intra - no variation. nter - 2 bp from O. coquereli . 3 bp from A. boltoni ,and3bp from A. goodmani .	Intra - some variation that does not correspond to geography or CO1. nter - does not differentiate between O. simillimus	Intraspecific variation that only partially reflects geography and CO1 variation	All specimens tested positive for Wolbachia.
Odontomachus simillimus	N/A	Intra - no variation. nter - does not differentiate O. troglodytes	N/A

doi:10.1371/journal.pone.0001787.t003 Queen measurements: maximum and minimum based on n = 5. HL 0.88-1.15, HW 0.81-1.07, CI 92-96, EL 0.17-0.23, ML 0.39-0.56, MI 44-49, SL 0.62-0.87, SI 77-81, WL 1.08- 1.46. FL 0.68-0.96, PW 0.60-0.78. Male measurements: maximum and minimum based on n = 5 from Madagascar: HL 0.58-0.73, HW 0.78-0.94, CI 129- 135, EL 0.37-0.46, SL 0.10-0.15, SI 13-16, WL 1.17-1.52, FL 0.78-1.08 Worker diagnosis: Inner blade of mandible without teeth and denticles; apical end of inner blade without a notched semicircular concavity (Fig. 2a). Eyes small (0.05-0.11 mm), projecting dorsolaterally. In full face view, antennal scape usually not reaching, and not surpassing posterior margin of occipital lobe. Dorsal surface of head with numerous short setae. Pilosity and sculpture as in Figures 3 a-d. Queens alate: Very similar to workers, only slightly larger than respective size class (Figs 3e-h). Ergatoid queens not recorded. Within a single locality, two size classes of workers, queens and males are present in this species, but the differences within a site do not hold up when variation across all sites is included. These differences suggest that two reproductive and developmental pathways can occur in this species. Further work is needed to explore the biotic or abiotic factors that trigger the development of small and large castes. The species is most similar to A. madagascarensis but can be easily distinguished by its small eyes and scape that does not surpass the occipital lobe. A. madagascarensis has large eyes (0.24-0.26 mm), and scapes that surpass occipital lobes. Distribution and biology. A. grandidieri is endemic to Madagascar and is widespread throughout Madagascar in forest and shrubland habitats below 1,550 m elevation (Fig 4b). It has been collected in gallery, dry, littoral, lowland, and montane forest, in desert spiny bush thicket in the southwest, and Uapaca woodland in the ceIntral plateau. As in many soil dwelling ants, A. grandidieri has reduced eyes (EL/HW 0.11-0.13) and short scapes . A. grandidieri is the only Anochetus in Madagascar with these soil nesting modifications. The subterranean habitat of this species may allow it to survive in a wide range of habitats in Madagascar from desert to woodland to montane forest. Out of 453 collecting events, A. grandidieri was most often recorded in sifted litter (97 collection records), rotten logs (96), and Malaise traps (155). CO1. Shallow iIntraspecific (average within species sequence divergence of 2.72, SE = 0.048) and deep interspecific divergences(9.4% SE = 0.05) between A. grandidieri and the other species. Small and large castes had identical DNA barcodes. (Figs 9, 16). Diagnostic barcoding loci. A grandidieri : T-273, T-282, T- 306, A-312, (shared with one population of A. goodmani ), A-312, T- 333, A-483, T-528 (all 3rd base pair positions). Specimens examined for Anochetus grandidieri : Specimens from 456 separate collection events from the following 140 localities were examined. MADAGASCAR : Province Antsiranana : Sakalava Beach ; Montagne des Francais , 7.2 km 142° SE Antsiranana ; Antsiranana II Pref: Antsahampano S.-Pref : Montagne d'Ambre. Site MD2 ; Parc National Montagne d'Ambre , 3.6 km 235° SW Joffreville ; Reserve Speciale de l'Ankarana , 13.6 km 192° SSW Anivorano Nord ; Foret d'Ampondrabe , 26.3 km 10° NNE Daraina ; Foret d' Antsahabe , 11.4 km 275° W Daraina ; Foret de Binara , 7.5 km 230° SW Daraina ; Foret de Binara , 9.1 km 233° SW Daraina ; Nosy Be , Lokobe Forest ; Foret Ambato , 26.6 km 33° Ambanja ; Ambondrobe , 41.1 km 175° Vohemar ; Ampasindava, Foret d'Ambilanivy , 3.9 km 181° S Ambaliha ; R.S. Manongarivo , 10.8 km 229° SW Antanambao ; R.S. Manongarivo , 12.8 km 228° SW Antanambao ; R.S. Manongarivo , 14.5 km 220° SW Antanambao ; Foret d'Anabohazo , 21.6 km 247° WSW Maromandia ; Parc National de Marojejy , Manantenina River , 27.6 km 35° NE Andapa , 9.6 km 327° NNW Manantenina ; Parc National de Marojejy, Manantenina River , 28.0 km 38° NE Andapa , 8.2 km 333° NNW Manantenina ; Parc National Marojejy ; Marojejy R.N.I. #12 ; Foret Ambanitaza , 26.1km 347° Antalaha ; 9.2 km WSW Befingotra , Res. Anjanaharibe-Sud ; 6.5 km SSW Befingotra , Res. Anjanaharibe-Sud ; 17 km W Andapa , Res. d' Anjanaharibe-Sud ; 5 km SW Antalaha ; 14 km W Cap Est , Ambato ; Fotodriana , Cap Masoala . Province Mahajanga : Mahavavy River , 6.2 km 145° SE Mitsinjo ; Reserve d'Ankoririka , 10.6 km 13° NE de Tsaramandroso ; Ampijoroa National Park , 160 km N Maevatanana , Mahajanga Prov ., deciduous forest ; Parc National de Namoroka , 17.8 km 329° WNW Vilanandro ; Parc National de Namoroka , 16.9 km 317° NW Vilanandro ; Parc National de Namoroka , 9.8 km 300° WNW Vilanandro ; Reserve Speciale de Bemarivo , 23.8 km 223° SW Besalampy ; Parc National Tsingy de Bemaraha , 10.6 km ESE 123° Antsalova ; Foret de Tsimembo , 8.7 km 336° NNW Soatana ; Parc National Tsingy de Bemaraha , 2.5 km 62° ENE Bekopaka , Ankidrodroa River ; Parc National Tsingy de Bemaraha , 3.4 km 93° E Bekopaka , Tombeau Vazimba ; Province Toamasina : Montagne d'Anjanaharibe , 19.5 km 27° NNE Ambinanitelo ; Montagne d'Anjanaharibe , 18.0 km 21 ° NNE Ambinanitelo ; Montagne d'Akirindro 7.6 km 341 ° NNW Ambinanitelo ; 19 km ESE Maroantsetra ; 6.9 km NE Ambanizana , Ambohitsitondroina ; Ambanizana , Parc National Masoala ; 5.3 km SSE Ambanizana , Andranobe ; 6.3 km S Ambanizana , Andranobe ; 1 km W Andampibe , Cap Masoala ; Parc National Mananara-Nord , 7.1 km 261° Antanambe ; Foret d'Analava Mandrisy , 5.9 km 195° Antanambe ; Res. Ambodiriana , 4.8 km 306° Manompana , along Manompana River ; Ile Sainte Marie , Foret Ambohidena , 22.8 km 44° Ambodifotatra ; Ile Sainte Marie , Foret Kalalao , 9.9 km 34° Ambodifotatra ; Parcelle E3 Tampolo ; S.F. Tampolo, 10 km NNE Fenoarivo Atn . ; Bridge at Onibi , NW of Mahavelona ; Mahavelona ( Foulpointe ) ; 2.1 km 315° Mahavelona ; Foulpointe ; Reserve Betampona , Camp Vohitsivalana , 37.1 km 338° Toamasina ; Reserve Betampona , Camp Rendrirendry 34.1 km 332° Toamasina ; F.C. Sandranantitra ; F.C. Didy ; F.C. Andriantantely ; P.N. Mantadia ; Analamay ; Foret Ambatovy , 14.3 km 57° Moramanga ; Torotorofotsy ; Andasibe National Park , botanic garden near eIntrance, West of ANGAP office ; Res. Analamazaotra , Parc National , Andasibe ; Fianarantsoa : Foret d'Atsirakambiaty, 7.6 km 285° WNW Itremo ; Ranomafana Nat. Park , Miaranony Forest ; Vohiparara broken bridge , Fianarantsoa Prov . ; Parc National de Ranomafana , Sahamalaotra River , 6.6 km 310° NW Ranomafana ; Parc Nationale Ranomafana : Talatakely ; 3 km W Ranomafana , nr. Ifandiana ; research cabin at Talatakely , Ranomafana National Park ; radio tower, Ranomafana National Park , Fianarantsoa Prov . ; Namorona River at footbridge, Ranomafana National Park ; Ranomafana National Park , Tavolo tree ; Belle Vue trail , Ranomafana National Park , Fianarantsoa Prov . ; 7 km W Ranomafana ; Vatoharanana ; Parc National de Ranomafana , Vatoharanana River , 4.1 km 231 ° SW Ranomafana ; P.N. Ranomafana , Vatoharanana-Ankovoka ; 8 km E Kianjavato , Vatovavy Forest ; 7.6 km 122° Kianjavato , Foret Classee Vatovavy ; 2 km W Andrambovato , along river Tatamaly ; Foret d'Ambalagoavy Nord , Ikongo , Ambatombe ; 45 km S. Ambalavao ; 45 km S Ambalavao ; 43 km S Ambalavao , Res. Andringitra ; Parc National d'Isalo , Ambovo Springs , 29.3 km 4° N Ranohira ; 8.0 km NE Ivohibe ; 9.0 km NE Ivohibe ; R.S. Ivohibe , 7.5 km ENE Ivohibe ; Parc National d'Isalo , 9.1 km 354° N Ranohira ; Foret d'Analalava , 29.6 km 280° W Ranohira ; Foret de Vevembe , 66.6 km 293° Farafangana ; Province Toliara : Reserve Speciale d'Ambohijanahary , Foret d'Ankazotsihitafototra , 34.6 km 314° NW Ambaravaranala ; Reserve Speciale d'Ambohijanahary , Foret d'Ankazotsihitafototra , 35.2 km 312° NW Ambaravaranala ; Vohibasia Forest , 59 km NE Sakaraha ; southern Isoky-Vohimena Forest , 59 km NE Sakaraha ; Foret Classee d'Analavelona , 33.2 km 344° NNW Mahaboboka ; Foret Classee d'Analavelona , 29.2 km 343° NNW Mahaboboka ; Foret Classee d'Analavelona , 29.4 km 343° NNW Mahaboboka ; Foret de Tsinjoriaky , 6.2 km 84° E Tsifota ; Parc National de Zombitse , 19.8 km 84° E Sakaraha ; Parc National de Zombitse , 17.7 km 98° E Sakaraha ; 15 km E Sakaraha ; Foret de Mite , 20.7 km 29° WNW Tongobory ; Sept Lacs; Beza-Mahafaly, 27 km E Betioky ; Ehazoara Canyon , 26 km E Betioky ; 70.7 km NNE Tolanaro , Mahermano Mt .; 11 km NW Enakara , Res. Andohahela ; 10 km NW Enakara , Res. Andohahela ; Res . Andohahela , 6 km SSW Eminiminy ; Parc National d'Andohahela , Col du Sedro , 3.8 km 113° ESE Mahamavo , 37.6 km 341° NNW Tolagnaro ; Parc National d'Andohahela , Manampanihy River , 5.4 km 113° ESE Mahamavo , 36.7 km 343° NNW Tolagnaro ; 2.7 km WNW 302° Ste. Luce ; 9.2 km N Tolanaro , Ilapany Mt . ; 29.5 km WNW Tolanaro , Vasiha Mt . ; Parc National d'Andohahela , Foret d'Ambohibory , 1.7 km 61° ENE Tsimelahy , 36.1 km 308° NW Tolagnaro ; Mandena , 8.4 km NNE 30° Tolagnaro ; Reserve Prive Berenty , Foret de Bealoka , Mandrare River , 14.6 km 329° NNW Amboasary ; Reserve Prive Berenty , Foret de Malaza , Mandrare River , 8.6 km 314° NW Amboasary ; Reserve Berenty; Foret de Petriky , 12.5 km W 272° Tolagnaro ; 4.4 km 148° SSE Lavanono ; Reserve Speciale de Cap Sainte Marie , 14.9 km 261° W Marovato ; near road, Zombitse National Park , Tulear Prov .; near ANGAP office, Zombitse National Park , Tulear Prov .; Parcel I, Beza Mahafaly Reserve , near research station, Tulear Province ; Tsimelahy - Parcel II , Andohahela National Park , transition forest, Tulear Province .