Phylogenomic Species Delimitation, Taxonomy, and ‘ Bird Guide’ Identification for the Neotropical Ant Genus Rasopone (Hymenoptera: Formicidae) Longino, John T. Branstetter, Michael G. Insect Systematics and Diversity 2020 2020-03-31 4 2 1 1 33 Longino & Branstetter, 2020 [818,1084,1465,1488] Insecta Formicidae Rasopone GBIF Animalia Hymenoptera 10 11 Arthropoda species costaricensis sp. nov.  ( Figs. 8–11; Supp Figs. S3–S 7[online only]) (Zoobank LSID: urn:lsid:zoobank.org:act: 3B2E87D5-CD8A-4CBE- B6B9-BDC604DC9500)    HOLOTYPE: 1 worker, Costa Rica, Puntarenas: Monteverde, 10.30892 −84.78498±  50 m, 1,617 m,  13-v-2014, cloud forest, ex sifted leaf litter, J. Longino, JTL8628-s [UCR, unique specimen identifier CASENT0635809].  PARATYPES: same data as holotype [ 1 worker, CAS, CASENT0644841]; same data except 10.29905 −84.78292±  200 m, 1,570 m,  30-iv-1989, JTL2486-s [ 1 worker, MCZC, INBIOCRI001281356; 1 worker, UCD, INBIOCRI001281355]; 10.3 −84.8 ± 2 km,  1,500 m,  26-vi-1984, JTL26Jun84/grou [ 1 worker, DZUP, INBIOCRI002278996];  10.30649 −84.81756±  100 m, 1,290 m,  21-xii-2013, evergreen forest, in clay bank soil, JTL8467 [ 1 worker, CAS, CASENT0635758]; 10.30719 −84.78627± 50 m, 1,670 m, 13-v-2014, JTL8627-s [1   Fig. 6.Distribution map, face view, and lateral view of petiole of  Rasopone pluviselva(holotype worker),  R. minuta(worker, Mexico, Chiapas,CASENT0609834), R. JTL043 (worker, Ecuador, MCZ-ENT00716620), and R. JTL048 (worker, French Guiana, CASENT0645961). On distribution maps, red dots are sites with UCE sequence data. Red boxes are type locality.   Fig. 7.Distribution map, face view, and lateral view of petiole of  RasoponeMAS010 (worker, Costa Rica, 08COSTA-1723),  R. cryptergates(worker, Costa Rica, INB0003660648), R. JTL042 (worker, Colombia, MCZ-ENT00716611),and R. JTL034 (worker, Mexico, CASENT0640282). On distribution maps, red dots are sites with UCE sequence data. Red boxes are type locality.   Fig. 8.Distribution map, face view, and lateral view of petiole of  Rasopone lunaris(worker, Brazil, Minas Gerais CASENT0644556),  R. guatemalensis(holotype worker),  R. costaricensisform c(worker,Costa Rica,INB0003659307),and R.JTL027 (worker,Panama,CASENT0633216).On distribution maps,red dots are sites with UCE sequence data. Red boxes are type locality.   Fig. 9.Distribution map, face view, and lateral view of petiole of  Rasopone costaricensis(holotype worker),  R. politognatha(holotype worker), R.JTL035 (worker, Mexico, CASENT0640453), and  RasoponeJTL049 (worker, Colombia, CASENT0644557). On distribution maps, red dots are sites with UCE sequence data. Red boxes are type locality.   Fig. 10.Distribution map,face view, and lateral view of petiole of  Rasopone costaricensisform b(worker,Costa Rica,INB0003223929), R.JTL047 (worker,French Guiana, CASENT0645960),  R. panamensis(worker,Costa Rica, CASENT0644252), and  R. subcubitalis(holotype worker). On distribution maps,red dots are sites with UCE sequence data. Red boxes are type locality.   Fig. 11.Distribution map, face view, and lateral view of petiole of  Rasopone ferruginea(worker, Nicaragua, CASENT0644264), R.JTL040 (worker, Colombia, MCZ-ENT00716609), R.JTL041 (worker,Colombia, MCZ-ENT00716614),and  R. costaricensisform a(worker, Costa Rica, INB0003659312).On distribution maps, red dots are sites with UCE sequence data. Red boxes are type locality (type locality for  R. ferrugineais ‘Mexico’).   Fig. 12.Distribution map, face view, and lateral view of petiole of  RasoponeJTL030 (worker, Panama, CASENT0633075), R.JTL037 (worker, Guatemala, CASENT0625283),  R. mesoamericana(holotype worker), and R.JTL029 (worker, Panama, CASENT0633053). On distribution maps, red dots are sites with UCE sequence data. Red boxes are type locality.   Fig. 13.Distribution map,face view,and lateral view of petiole of  Rasopone cubitalis(holotype worker),  R.titanis(worker,Mexico,Chiapas,MCZ-ENT00716640),  R.breviscapa(holotype queen),and  R.rupinicola(worker,Colombia,face view MCZ-ENT00716589,petiole CASENT0217567).On distribution maps,red dots are sites with UCE sequence data. Red boxes are type locality.   Table 2.  Rasoponespecies in order of mean HW of worker (queen values substituted for  R. breviscapa)    Species HW CI SI PTI Clypeus Petiole Mandible Range Figures   Pluviselva 0.76–0.93 79–97 66–81 57–70 Truncate Intermediate Striate Honduras- Panama Fig. 6; Supp Figs. S26 and S27 (online only)    minuta 0.79–0.96 78–96 70–85 51–63 Truncate Tapered Smooth Chiapas- Honduras Fig. 6; Supp Figs. S22 and S23 (online only)  JTL043 0.85–1.04 81–98 72–87 54–65 Truncate Cuboidal Smooth Ecuador Fig. 6; Supp Fig. S43 (online only)  JTL048 0.88–1.08 80–98 73–89 49–59 Truncate Intermediate Smooth French Guiana Fig. 6; Supp Fig. S45 (online only)  MAS010 0.90–1.10 79–97 72–87 53–64 Truncate Tapered Striate Costa Rica Fig. 7; Supp Fig. S47 (online only)    cryptergates 0.96–1.18 77–95 71–87 51–63 Truncate Cuboidal Striate Costa Rica Fig. 7; Supp Figs. S8 and S9 (online only)  JTL042 0.99–1.21 81–99 71–87 49–60 Truncate Intermediate Striate Colombia- Brazil Fig. 7; Supp Fig. S42 (online only)  JTL034 0.99–1.22 83–102 69–85 52–64 Truncate Intermediate Smooth Puebla- Honduras Fig. 7; Supp Fig. S37 (online only)    lunaris 1.00–1.22 82–100 70–85 54–66 Truncate Intermediate Smooth South America Fig. 8; Supp Figs. S16–S18 (online only)    guatemalensis 1.00–1.22 77–94 77–95 51–62 Truncate Tapered Smooth Chiapas- Guatemala Fig. 8; Supp Figs. S14 and S15 (online only)    costaricensis, Form c 1.00–1.23 79–96 80–97 48–58 Truncate Tapered Striate Costa Rica Fig. 8; Supp Fig. S7 (online only)  JTL027 1.00–1.23 75–91 86–106 58–70 Truncate Cuboidal Striate Panama Fig. 8; Supp Fig. S34 (online only)    costaricensis 1.03–1.26 78–95 76–93 48–59 Truncate Tapered Striate Costa Rica Fig. 9; Supp Figs. S3 and S4 (online only)    politognatha 1.05–1.28 83–101 74–90 50–62 Truncate Intermediate Smooth Chiapas- Nicaragua Fig. 9; Supp Figs. S28 and S29 (online only)  JTL035 1.05–1.28 87–107 69–85 48–58 Truncate Intermediate Smooth Veracruz Fig. 9; Supp Fig. S38 (online only)  JTL049 1.07–1.31 82–100 69–85 55–67 Truncate Intermediate Smooth Colombia Fig. 9; Supp Fig. S46 (online only)    costaricensis, Form b 1.07–1.31 77–94 82–100 49–60 Truncate Tapered Striate Costa Rica Fig. 10; Supp Fig. S6 (online only)  JTL047 1.09–1.33 86–105 66–81 53–65 Truncate Intermediate Smooth French Guiana Fig. 10; Supp Fig. S44 (online only)    panamensis 1.11–1.35 79–97 75–92 52–63 Truncate Cuboidal Striate Costa Rica- Panama Fig. 10, Supp Figs. S24 and S25 (on- line only)    subcubitalis 1.13–1.38 80–98 76–93 50–61 Truncate Intermediate Striate Chiapas- Honduras Fig. 10, Supp Fig. S32 (online only)    ferruginea 1.14–1.39 82–101 74–90 48–59 Sinuous Tapered Striate Mexico- Nicaragua Figs. 4 and 11; Supp Figs. S12 and S13 (online only)  JTL040 1.16–1.42 82–100 66–80 53–65 Truncate Cuboidal Smooth Colombia Fig. 11; Supp Fig. S40 (online only)  JTL041 1.19–1.45 83–101 74–91 50–61 Truncate Tapered Smooth Colombia Fig. 11; Supp Fig. S41 (online only)    costaricensis, Form a 1.19–1.46 76–93 79–97 50–61 Truncate Intermediate Smooth Costa Rica Fig. 11; Supp Fig. S5 (online only)  JTL030 1.26–1.54 79–97 74–90 51–63 Truncate Tapered Striate Panama Fig. 12; Supp Fig. S36 (online only)  JTL037 1.27–1.55 80–98 83–101 46–56 Truncate Intermediate Striate Guatemala Fig. 12; Supp Fig. S39 (online only)    mesoamericana 1.33–1.62 80–98 76–93 50–61 Truncate Tapered Striate Chiapas-Costa Rica Figs. 5 and 12; Supp Figs. S19–S21 (on- line only)   Table 2.Continued    Species HW CI SI PTI Clypeus Petiole Mandible Range Figures  JTL029 1.37–1.67 84–102 67–81 50–61 Truncate Intermediate Striate Panama Fig. 12; Supp Fig.  S35 (online only)    cubitalis 1.40–1.71 82–100 82–100 50–61 Sinuous Cuboidal Smooth Nicaragua- Fig. 13; Supp Figs.  Costa Rica S10 and S11 (on-  line only)    titanis 1.51–1.84 84–102 78–96 52–64 Sinuous Tapered Smooth Chiapas Fig. 13; Supp Fig.  S33 (online only)    breviscapa 1.53–1.87 90–110 71–87 Truncate Intermediate Striate Bolivia Fig. 13, Supp Fig. S2  (online only)    rupinicola 1.53–1.87 78–96 95–117 Sinuous Intermediate Smooth Panama- Fig. 13; Supp Figs.  Colombia S30 and S31 (on-  line only)  Metric characters are ±10% of mean.  worker, USNM, CASENT0635807];  10.31156 −84.80459±  50 m, 1,480 m,  31-xii-2015, cloud forest, under stone, JTL9474 [ 1 worker, USNM, CASENT0632484].   Geographic range. Costa Rica, Panama.   Diagnosis Montane; mandible striate or smooth; anterior clypeal margin truncate; side of head bare or with a few inconspicuous short setae; petiole scale-like. The species forms a clade of cryptic species that vary in size, mandibular sculpture, and petiole shape. They are all montane. Three unrelated species are within geographic and size range of the clade:   Rasopone cryptergates( Fig. 7; Supp Figs. S8and S 9[online only]): lowland; petiolar node more cuboidal; posterolateral margins of head more angular.   Rasopone panamensis( Fig. 10; Supp Figs. S24 and S25 [online only]): lowland; petiolar node cuboidal.   RasoponeMAS010 ( Fig. 7; Supp Fig. S47 [online only]): smaller, HW 1.00 for both of the measured specimens of R. MAS010 versus 1.09 for the smallest measured specimen of  R. costaricensis; petiole somewhat thicker (average PTI 59 vs 53).   Measurements, Barva Form a, worker:HW 1.32 (1.25–1.41, 6); HL 1.56 (1.51–1.65, 6); SL 1.20 (1.19–1.21, 3); PTH 0.90 (0.83–0.94, 5); PTL 0.50 (0.48–0.53, 5); CI 85 (83–86, 6); SI 88 (85–93, 3); PTI 56 (54–58, 5).  Measurements, Barva Form b, worker:HW 1.19 (1.14–1.25, 10); HL 1.39 (1.33–1.47, 10); SL 1.08 (1.04–1.11, 3); PTH 0.85 (0.78– 0.93, 7); PTL 0.46 (0.42–0.51, 7); CI 86 (84–89, 10); SI 91 (87–93, 3); PTI 54 (52–61, 7).  Measurements, Barva Form c, worker:HW 1.11 (1.09–1.14, 2); HL 1.28 (1.26–1.29, 2); SL 0.99 (0.98–1.00, 2); PTH 0.78 (0.77–0.78, 2); PTL 0.41 (0.41–0.41, 2); CI 88 (86–89, 2); SI 89 (87–90, 2); PTI 53 (53–53, 2).  Measurements, other populations, worker:HW 1.15 (1.09–1.21, 12); HL 1.33 (1.26–1.43, 12); SL 0.98 (0.91–1.09, 7); PTH 0.85 (0.72–0.95, 9); PTL 0.45 (0.39–0.53, 9); CI 86 (83–92, 12); SI 85 (76–91, 7); PTI 54 (50–58, 9).  Measurements, other populations, queen:HW 1.18 (1.08–1.27, 2); HL 1.32 (1.26–1.38, 2); SL 0.91; PTH 0.75; PTL 0.42; CI 89 (86– 92, 2); SI 84; PTI 55.   Biology This species complex occurs in cloud forest habitats, from 1,000 to 2,000 melevation (although one enigmatic collection is from 250 melevation on the Osa Peninsula). Workers, and in one case a dealate queen, are collected in Winkler samples of forest floor leaf litter and rotten wood. Workers have been collected beneath epiphytes in treefalls and beneath rotten wood on the ground. Workers have been found both in closed-canopy cloud forest and in open pastures near cloud forest edges (beneath wood on the ground). In Monteverde, in addition to specimens from multiple Winkler samples, a worker was found in soil of a clay bank in a steep-sided ravine, and a worker was found beneath a stone.At Estación Pittier in the Talamanca range, a dealate queen was found beneath a stone. It is likely that the most common forms (of the various cryptic species) nest in the soil, with workers foraging in the litter. One nests beneath epiphytes. Males can be common in Malaise traps (associated with COI results). An alate queen was collected by an INBio Parataxonomist, at Tapantí National Park, on 30 October 1991. Cryptic species in the complex may specialize on particular microhabitats (e.g., beneath epiphytes; see Comments).   Comments DNA sequence data, both COI and UCE, support a clade endemic to Costa Rican and adjacent Panamanian cloud forest. Within the clade, genetic evidence also supports the occurrence of multiple sympatric species at two intensively sampled sites. But the genetic data are fragmentary and the relationships among species across sites is unclear. We take the approach of referring to the entire clade as  R. costaricensis, with the acknowledgment that further study will almost certainly result in the further splitting of the clade into component species. We describe here what we currently know of the sympatric forms at particular sites. There is evidence that three sympatric species occur on the Barva transect, on Costa Rica’s Caribbean slope. The three species differ in morphology, microhabitat, and COI sequence.  Form a( Fig. 11; Supp Fig. S5[online only]): This is the largest of the three. The mandibles are smooth and shiny; the other two forms have striate mandibles. It is known from five separate collections: 1) 2 workersunder epiphytes in an old treefall; 2) a worker and some brood in a rotten knot in a treefall; 3) a worker under epiphytes on dead wood at the edge between pasture and forest; 4) a Berlese sample of epiphytic material; and 5) a worker in a collection of mixed ants collected by hand. The first four collections were from a 1,500 msite and the last collection from an 1,100 msite. It is notable that no specimens were collected in the 350 miniWinkler samples of forest floor litter that were taken at the two sites.  Form b( Fig. 10; Supp Fig. S6[online only]): This and Form chave striate mandibles. Form bis very similar to Form c, but the petiole is somewhat more tapering and scale-like. It occurred at the 1,500 mand 2,000 msites on the Barva transect, where it was moderately abundant in miniWinkler samples. Parataxonomist Ronald Vargascollected a worker by hand at the 2,000 msite.  Form c( Fig. 8; Supp Fig. S7[online only]): This form has a relatively less scale-like node than Form b. It occurred at the 1,500 msite, from two collections. One worker was collected by hand from under rotten wood. Two workers were collected by student Andy Boring in an open pasture area, beneath rotten wood. These two collections were united by DNA sequence data (see below), which also supported their distinctness from Form b. However, a few specimens from miniWinklers from the 1,500 mand 2,000 msites lacked sequence data and were intermediate in petiole shape, and thus could not be assigned to one form or the other. There is evidence for at least three sympatric species, based on COI clusters in the BOLD database, on the peak of Volcán Cacao, a cloud forest site in Guanacaste, Costa Rica. The evidence comes mostly or entirely from males in Malaise traps, sampled by Alex Smith and others. Barva Form aand Form bcluster with one of the Cacao clusters. Form aand Form bdiffer from each other by about 5%, while each differs from the Cacao specimens by about 3%. Barva Form cclusters with the largest Cacao cluster, which contains over 120 specimens, with much less than 0.5% sequence divergence among them. A third Cacao cluster is small, and currently unassociated with any other specimens ( Fig. 3; Supp Fig. S1[online only]). Other populations are known from Monteverde (the typelocality) in the Cordillera de Tilarán, multiple cloud forest sites in the Cordillera de Talamanca and mountains of western Panama, and one lowland site near the Osa Peninsula ( Fig. 9; Supp Figs. S3and S 4[online only]). 2618791326 2014-05-13 CASENT J. Longino Costa Rica 0 10.30892 Monteverde 70 -84.78498 10 11 CASENT0635809 1 1 Puntarenas holotype 2618791302 2014-05-13 CAS J. Longino Costa Rica 0 10.30892 Monteverde 70 -84.78498 10 11 CASENT0644841 1 1 Puntarenas paratype 2618791323 1989-04-30 MCZC, UCD J. Longino Costa Rica 0 10.29905 Monteverde 281 -84.78292 10 11 INBIOCRI001281356, INBIOCRI001281355 2 2 Puntarenas paratype 2618791324 1984-06-26 DZUP Costa Rica 1500 10 11 INBIOCRI002278996 1 1 Puntarenas paratype 2618791322 2013-12-21 CAS Costa Rica 0 10.30649 140 -84.81756 10 11 CASENT0635758 1 1 Puntarenas paratype 2618791333 [146,477,644,663] USNM 20 21 CASENT0635807 1 2618791327 2015-12-31 USNM 0 10.31156 70 -84.80459 20 21 CASENT0632484 1 1