or JHR 50: 25-79 (20 I 6) JOURNAL CF Apeenreviewed open-access journal AA Po eter (f-) Hymenoptera http://jhr.pensoft.net The insertional Society of Hymenoptersts. RESEARCH Revision of the genus Promicrogaster (Hymenoptera, Braconidae, Microgastrinae) from Area de Conservacion Guanacaste, Costa Rica, with a key to all species previously described from Mesoamerica Jose Fernandez-Triana', Caroline Boudreault', Tanya Dapkey’, M. Alex Smith’, J. Rodriguez*, Winnie Hallwachs’, Daniel H. Janzen? | Canadian National Collection of Insects, 960 Carling Ave., Ottawa, ON KIA 0C6 Canada 2. Department of Biology, University of Pennsylvania, Philadelphia, PA 19104-6018 USA 3 Department of Integrative Biology, University of Guelph, Guelph, ON NIG 2W1 Canada 4 Dept. of Natural Sciences, University of Virginias College at Wise, Wise, VA 24293 USA Corresponding author: Jose Ferndndez-Triana (jose.fernandez@agr.gc.ca) Academic editor: G. Broad | Received 22 February 2016 | Accepted 19 May 2016 | Published 27 June 2016 http.//zoobank. org/889928 9 F-7707-4666-9D57-068469D75918 Citation: Fernandez-Triana J, Boudreault C, Dapkey T, Smith MA, Rodriguez J, Hallwachs W, Janzen DH (2016) Revision of the genus Promicrogaster (Hymenoptera, Braconidae, Microgastrinae) from Area de Conservacién Guanacaste, Costa Rica, with a key to all species previously described from Mesoamerica. Journal of Hymenoptera Research 50: 25-79. doi: 10.3897/JHR.50.8220 Abstract The genus Promicrogaster (Hymenoptera, Braconidae, Microgastrinae) from Area de Conservacién Gua- nacaste (ACG), Costa Rica, is revised. A key is provided to all new species as well as those previously described from Mesoamerica. A total of 21 species, all authored by Fernandez-Triana & Boudreault, are described as species nova: alexmartinezi, andreyvallejosi, brandondinartei, daniellopezi, daretrizoi, eddycas- troi, eimyobandoae, fabiancastroi, fabriciocambroneroi, hillaryvillafuerteae, kevinmartinezi, kiralycastilloae, leilycastilloae, liagrantae, luismendezi, monteverdensis, naomiduarteae, pablouzagai, ronycastilloi, sebastian- cambroneroi, tracyvindasae. A species previously described from India is considered as incertae sedis. Promicrogaster is considered to be restricted to the New World, with the vast majority of the species found in the Neotropics and a few extending north to the Nearctic. Almost 60% of the known species in ACG are found in cloud forests at over 1,000 m altitude. All of the verified and authenticated host records for Promicrogaster are from caterpillars living more deeply inside plant tissue than simply in rolled leaf struc- tures — although no host data from ACG is available. Copyright Jose Ferndndez-Triana et al. 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. 26 Jose Fernindez-Triana et al. / Journal of Hymenoptera Research 50: 25—79 (2016) Keywords Promicrogaster, Microgastrinae, Neotropics, Area de Conservacién Guanacaste, taxonomic revision, para- sitoid wasps, DNA barcoding Introduction The genus Promicrogaster was described by Brues and Richardson (1913) to accom- modate a single female specimen from Guyana, which they considered to be unique based on several morphological characters. Half a century later Muesebeck (1958) and Nixon (1965) added 10 more new species of Promicrogaster, all from the Neo- tropics. An additional species was described from India by Sathe and Bhoje (1998), but we consider that a incertae sedis (see below for details). The true diversity of the genus remains unknown, although Mason (1981) considered that, in spite of being rare in collections, it might have more than 100 species overall. The highest diversity is in the Neotropics but a few, undescribed species reach the Nearctic. Biological information for the genus is very scarce: two host records from concealed caterpillars in the Lepidoptera families Sesiidae and Tineidae (Davis 1996, Garcia and Mon- tilla 2010), an unidentified caterpillars infesting bracket fungus (Muesebeck 1958, Mason 1981). This paper revises the genus Promicrogaster in Area de Conservacién Guanacaste (ACG), northwestern Costa Rica (Janzen et al. 2009, Janzen and Hallwachs 2011), as part of comprehensive studies on the fauna of Microgastrinae from that region (e.g. Fernandez-Triana et al. 2013, 2014a—d). A key to all known species from Mes- oamerica is also included, but the North and South American species are not dealt with in this paper. Methods Promicrogaster is a rarely collected genus (Mason 1981), and it is poorly represented in collections. This study is based on 120+ specimens, most of them collected in ACG, with a few additional specimens from other Mesoamerican countries available for study in the Canadian National Collection of Insects (CNC) in Ottawa. Some para- types of the new species being described will be deposited in other collections with the following acronyms used: BMNH (The Natural History Museum, London, United Kingdom), INBio (Instituto Nacional de Biodiversidad, Santo Domingo de Heredia, Costa Rica) and NMNH (National Museum of Natural History, the Smithsonian Institution, Washington DC, United States). Four species of Promicrogaster had been described previously from Mesoamerica. ‘The original descriptions and illustrations for three of them were sufficiently detailed to allow us to describe the new species with confidence. The only exception was Promicrogaster apharea Nixon, 1965 (Brazil and highlands of southwestern Mexico), which could not be reliably defined based on the original description alone, but in that case we examined the holotype. Revision of the genus Promicrogaster (Hymenoptera, Braconidae, Microgastrinae)... 27 Morphological terms and measurements of structures are mostly as used by Mason (1981), Huber and Sharkey (1993), Whitfield (1997), Karlsson and Ronquist (2012), and Fernandez-Triana et al. (2014a). Mediotergites 1, 2, etc., are abbreviated as T1, T2, etc. Flagellomere 1 is the closest to pedicel while flagellomere 16 is the most apical one. Because the ovipositor in Promicrogaster is curved and crooked at the very apex, its length is difficult to measure accurately; the ovipositor length measurements provided for each new species are only intended as an approximation. In any case, the ovipositor and its sheaths are some of the longest observed in any Microgastrinae genera; they are usually two times longer than the metatibia length. The dichotomous key and descriptions of the new species are based on the study of all available female specimens, so as to reflect intraspecific variation, but always include data from the holotype for the new species. Males of most species cannot be readily identified unless associated with females via rearing or molecular data. The dichotomous key is mostly based on morphological characters, but in one cou- plet we used molecular characters to differentiate species that are morphologically similar to each other. In that case we used characteristic loci in the DNA barcoding region. The bases are numbered from the start of the COI gene according to the reference sequence U37541 (Drosophila melanogaster), and are only diagnostic within that couplet. ‘The let- ters A, C, G, and T correspond to adenine, cytosine, guanine, and thymine respectively. The descriptions include 21 characters that are commonly used in describing Mi- crogastrinae (e.g., body measurements such as length of body and fore wing, ovipositor sheath; and also color of particular body areas). These characters follow a recent revi- sion of the related genus Sendaphne (Fernandez-Triana et al. 2014d), so as to facilitate future comparisons between species in those two genera. Photos were taken with a Keyence VHX-1000 Digital Microscope, using a lens with a range of 13—130x. Multiple images through the focal plane were taken of a structure and these were combined to produce a single in-focus image, using the soft- ware associated with the Keyence System. Together with morphological studies, we also analyzed DNA barcodes (the 5’ re- gion of the cytochrome c oxidase I (CO1) gene, Hebert et al. 2003) whenever availa- ble. DNA barcodes were obtained using DNA extracts prepared from single legs using a glass fibre protocol (Ivanova et al. 2006). Briefly, total genomic DNA was re-sus- pended in 30 yl of dH20, and a 658-bp region near the 5’ terminus of the CO1 gene was amplified using standard primers (LepF1—LepR1) following established protocols (Smith et al. 2006, 2007, 2008). If the initial 658 bp amplification was unsuccessful, smaller sequences were generated using internal primers. If each amplification worked, a composite sequence was generated, however in cases where only one read amplified, this shorter sequence was used. All information for the sequences associated with each individual specimen can be retrieved from the Barcode of Life Data System (BOLD) (Ratnasingham and Hebert 2007). A Neighbor-Joining tree based on Kimura 2-pa- rameter was also generated (Fig. 160). The new species described below received patronyms based on the winners of a school child nature awareness competition conducted by the Programa de Educacién Biolégica de ACG in the last half of 2015 (Kazmier 2015). 28 Jose Fernindez-Triana et al. / Journal of Hymenoptera Research 50: 25-79 (2016) Results Characterization of the genus Promicrogaster Promicrogaster Brues & Richardson, 1913 Promicrogaster: Brues and Richardson 1913: 499. Diagnosis. Glossa elongate and bilobate (Figs 4, 10, 16, 22, 28, 34, 40, 46, 70, 76, 82, 101, 107, 118, 130, 149). Lateral face of scutellum with polished area (=lunules) occupying most of the lateral face (Figs 12, 18, 24, 32, 36, 42, 62, 66, 72, 78, 84, 110, 114, 120, 126, 138, 147, 151). Propodeum clearly sculptured and usually with some carination (Figs 12, 18, 36, 42, 66, 90, 99, 114, 120, 132, 133, 157). Metacoxa very long, 0.8—1.0 x metafemur length and 0.6—0.8 x metatibia length (Figs 7, 15, 38, 45, 51, 57, 63, 74, 87, 106, 117, 129, 135, 146, 148, 154). Mediotergite 1 parallel-sided (Figs 8, 13, 19, 32, 60, 140) to slightly narrowing towards posterior margin (Figs 38, 44,55, 67, 73, 85, 91, 105, 127, 132, 147). Mediotergite 2 transverse, its width at posterior margin 3.0—4.5 x (rarely 2.0 x) its length medially. Ovipositor very long for a microgastrine wasp (approximately two times longer than metatibia length), strongly curved and with its apex sinuate (Figs 1, 5, 14, 15, 20, 26, 30, 37, 43, 50, 51, 56, 61, 63, 68, 69, 74, 86, 87, 92, 98, 104, 108, 116, 121, 123, 128, 139, 140, 144, 148, 154, 159). Fore wing usually with a small areolet, which is sometimes poorly defined (Figs DP 2S. 29. 35 P47 SIS O93- (Ly 7-7 OO OI 1, LOB; 1s LD) Sl SF 375156) but some small species have no trace of areolet whatsoever (Figs 41, 102, 125, 150). Body and fore wing lengths ranging from 1.8—2.0 mm in the smallest species up to 4.9—5.3 mm in the largest species. Body length varying from slightly longer to slightly shorter than fore wing length. Promicrogaster is a very distinctive genus as defined by the combination of elongate and bilobate glossa, large polished areas on lateral face of scutellum, and ovipositor shape and length. Within Microgastrinae, it can only be confused with Sendaphne, but the later has a much less transverse mediotergite 2, the apex of ovipositor is not sinu- ate, the propodeum is entirely or mostly smooth, and the first discal cell is much wider (e.g., Fernandez-Triana et al. 2014d). We consider the described ‘Promicrogaster’ from India as incertae sedis (see be- low for a detailed discussion on that species). Thus, Promicrogaster as defined here is restricted to the New World, with the vast majority of the species found in the Neotropics and a few extending north to the Nearctic (Mason 1981, and unpublished data from the CNC collection). This pattern of distribution is remarkably similar to that found in other genera recently revised in the New World, such as Pseudapanteles and Venanus (Fernandez-Triana et al. 2013, 2014b, 2014c). A total of 21 new species are described below, increasing the total known Mesoamerican species from 4 to 25, and the total number of described species for the genus from 11 to 32 (Table 1). We are aware of many additional undescribed species in collections, from North America (Canada/US) and South America, which will be dealt with in future papers. Revision of the genus Promicrogaster (Hymenoptera, Braconidae, Microgastrinae)... 29 As for habitat preference, 82% of the ACG species were collected in rain forests and cloud forests (with almost 60% restricted to cloud forests), while only 18% were found in dry forests. Around 40% of the species were found at low-mid elevation (0-500 m) whereas almost 60% of the species were collected at altitudes over 1,000 m. This strongly contrasts with the closely related genus Sendaphne, which has been mostly found at altitudes between 100-900 m with just a few species found in cloud forests (Fernandez- Triana et al. 2014d). All of the verified and authenticated host records for Promicrogaster are from cat- erpillars living more deeply inside more or less woody plant material than simply in rolled leaf structures. It is possible that the sinuate ovipositor tip in all known species of Promicrogaster is an adaptation allowing the ovipositor to be steered through fissures in harder/more woody structures than mere leaf rolls (e.g., Quicke 2015). Muesebeck (1958) described Promicrogaster polyporicola as reared from unidenti- fied Lepidoptera larvae infesting a bracket fungus (Fomes sp.) in Panama. Davis (1996) reared an unidentified species of Promicrogaster from caterpillars of Prosetomorpha fal- Table |. Mesoamerican species of Promicrogaster. The presence of species in Area de Conservacién Gua- nacaste is indicated below by adding ‘ACG’ after “Costa Rica’. New country records for species previously described are marked with an asterisk (*). Species Distribution Promicrogaster alexmartinezi Fernandez- Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster andreyvallejosi Fernandez-Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster apharea Nixon, 1965 Brazil, Mexico Promicrogaster brandondinartei Fernandez- Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster daniellopezi Fernandez-Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster daretrizoi Fernandez-Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster eddycastroi Fernandez-Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster eimyobandoae Fernandez- Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster fabiancastroi Fernandez- Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster fabriciocambroneroi Fernandez- Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster hillaryvillafuerteae Fernandez-Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster kevinmartinegzi Fernandez- Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster kiralycastilloae Fernandez-Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster leilycastilloae Fernandez-Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster liagrantae Fernandez-Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster luismendezi Fernandez- Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster miranda Muesebeck, 1958 Panama, Trinidad (*) Promicrogaster monteverdensis Fernandez- Triana & Boudreault, sp. n. Costa Rica, ACG and other localities Promicrogaster munda Muesebeck, 1958 Costa Rica, Honduras, Mexico, Panama (*) Promicrogaster naomiduarteae Fernandez- Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster pablouzagai Fernandez- Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster polyporicola Muesebeck, 1958 Panama Promicrogaster ronycastilloi Fernandez- Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster sebastiancambroneroi Fernandez-Triana & Boudreault, sp. n. Costa Rica, ACG Promicrogaster tracyvindasae Fernandez-Triana & Boudreault, sp. n. Costa Rica, ACG and other localities 30 Jose Fernindez-Triana et al. / Journal of Hymenoptera Research 50: 25—79 (2016) cata (Tineidae) in Colombia; the larvae of this small moth are scavengers feeding on frass within the burrows made by larval Curculionidae in the stems of Solanum qui- toensis (Solanaceae: “Naranjilla’). Garcia and Montilla (2010) reared Promicrogaster as a parasitoid of cocoa fruit borers, Carmenta spp. (Sessidae) in Venezuela; Carmenta lar- vae mine through the pericarp of cocoa (Malvaceae: Theobroma cacao) fruit, and their damage leads to secondary phytopathogenic fungal infections (such as Phytophthora sp.) that cause the fruit to rot (e.g., Morrilo et al. 2009). The 21 species of Promicro- gaster found in ACG have only been collected in Malaise traps. It is likely that those wasp species have not been reared yet because they are parasitizing small moth larvae concealed inside more or less woody plant or fungal tissues - the ACG inventory, while being the most comprehensive effort ever done to rear tropical caterpillars, has only focused on exposed feeders and rollers of green leaves. Key to Promicrogaster species in Mesoamerica (female specimens) i Head entirely yellow-orange to orange-brown; anteromesoscutum and scutel- lar disc mostly orange-brown (Figs 129, 130, 133); antenna mostly orange- yellow (only apical 4—5 flagellomeres dark brown)... eee eeeseeeeeeeeeeeeees ms Promicrogaster pablouzagai Fernandez-Triana & Boudreault, sp. n. - Head entirely black to dark brown (except for clypeus and labrum orange- yellow in some species); anteromesoscutum and scutellar disc black; antenna dark brown to black (rarely some species with apical 5—7 flagellomeres yel- lewew ites ss, La eee de eee ee da Bs eng ene eA ere Se 2 2(1) Lighter coloured species; clypeus entirely or mostly yellow to orange-yellow (as in Figs 4, 107, 118); all sternites and hypopygium yellow to orange-yellow (as in Figs 3, 5, 7,57, 61, 106, 108, 117, 121); metasoma dorsally with T1—-T4 entirely (rarely mostly) yellow, orange or red (as in Figs 8, 60, 109, 110, 119, 120)...... 3 — Darker coloured species; most sternites (usually) and hypopygium (partially to entirely) dark brown (as in Figs 14, 20, 26, 30, 43, 56); metasoma dorsally mostly dark brown to black (rarely some tergites with small areas orange to light brown) (as in Figs 13, 19, 25, 31, 38, 44, 48, 55); clypeus coloration Natiable; usually dark: Drow1y touDlAGk tacaJ caneustsepe Son neneth abyttavahednea insbioonpestiee 6 3(2) Propodeum with complete, raised, and strongly defined median carina (par- tially visible in Fig. 110); and T1 entirely smooth (Fig. 109); and malar distance more than half eye length (Fig. 107); and clypeus large and entirely Oratices yellows Fig. 4h 7): (Panaria,aPrinidad |. cst.dktteimamiectmagwwecestevas ee. ee ee ee ee Promicrogaster miranda Muesebeck, 1958 — Propodeum without a median carina but with central depression (Figs 62, 120, 122); either with T1 mostly rugose (Figs 119, 120) and with malar distance less than half eye length (Figs 4, 118); or with clypeus smaller and not entirely yellow- red (as in Fig. 58) [other Mesoamerican countries, mostly Costa Rica].............. 4 4(3) 5(4) 8(6) Revision of the genus Promicrogaster (Hymenoptera, Braconidae, Microgastrinae)... 31 Metasoma dorsally entirely yellow-orange (Figs 119, 120); T1 with coarse sculpture on 0.7 or more its length; T2 relatively narrow, its width at pos- terior margin more than 3.0 x its length centrally (Figs 119, 120); scape entirely to mostly black to dark brown (partially visible in Fig. 118) [Costa Rica, Honduras, Mexico, Panama, mostly found at elevations over 1000m]. ree 6 eee ae ey Od ROE Promicrogaster munda Muesebeck, 1958 Metasoma dorsally with some red, brown or black coloration (Figs 8, 60); T1 smoother (Fig. 60), at most with sculpture on posterior 0.5 of its length (Figs 8); T2 broader, its width at posterior margin usually less than 3.0 its length centrally (Figs 8, 60, 62); scape mostly yellow (Fig 4, 58) [Costa Rica, ACG, Gis rore stead er ss OO ma irs cules Sethe ohagea lose onal etree aaah: Meats deta Aes oe 5 Posterior margin of clypeus strongly concave (Fig. 58); lower face clearly elongate, malar distance at least 1.5 x mandible width (Fig. 58); fore wing vein 1M transparent (Fig. 59); T1 shape more or less rectangular (width at middle length, anterior or posterior margin about the same); T1—T4 entirely orange-yellow, 15+ mostly dark brown to black (Fig. 60)... Promi- crogaster fabriciocambroneroi Fernandez-Triana & Boudreault, sp. n. Posterior margin of clypeus very slightly concave, almost straight (Fig. 4); lower face of normal length, malar distance 1.0 x mandible width (Fig. 4); fore wing with vein 1M brown (Fig. 3); Tl narrowing towards posterior margin; T1 mostly red with black margins (posterior 0.3 of T1 black some- times), [2—T4 red-orange or yellow-orange, [5+ yellow with small central Dane Drow iri GZGyus Acti, Mee ae lies acele bb eealnatag ane tonnettas slik stalin SR .... Promicrogaster alexmartinezi Fernandez-Triana & Boudreault, sp. n. Antenna with most flagellomeres 10—16 yellow-white (Figs 33, 87)............ 7, Antenna entirely dark brown to black (rarely with flagellomeres 1-4 lighter itpbrerel Fob vag dneevawmt ceciciey Cooip ive) ay 80:9 ae ePREe Ee mn aT a EN 8 Antenna with flagellomeres 11-16 yellow-white (Fig. 87); propleuron, an- terior 0.5 of metacoxa, sternites and hypopygium dark brown (Figs 87, 89); ovipositor strongly down curved on posterior 0.2 (Fig. 92) [Costa Rica, ACG, cloud forest over LOOOm]........... cee eeeeeeeeccceccccsssscccccccccssssssccceceeeeesseeess lowe Promicrogaster leilycastilloae Fernandez-Triana & Boudreault, sp. n. Antenna with flagellomeres 1-8 and 15-16 dark brown, and flagellomeres 9-14 (sometimes only 10-14) yellow white (Fig. 33); propleuron, metacoxa, sternites and hypopygium mostly yellow to orange-yellow (Figs 33, 37); ovi- positor relatively straight (Fig. 37) [Costa Rica, ACG, mid-elevation rain FOrestse DUO SOOO Ts Lae UU cue esi S ARNE A, cho Pah RRS Lome ta Bn le Senge Promicrogaster daretrizoi Fernandez-Triana & Boudreault, sp. n. Fore wing without areolet (as in Figs 41, 53, 102, 125, 150), and hypopyg- ium and sternites almost always entirely dark brown (at most hypopygium with small pale spot basally, and/or anterior 2-3 sternites yellow) (Figs 30, 43, 52 11(10) 12(11) 13(9) 14(13) Jose Fernindez-Triana et al. / Journal of Hymenoptera Research 50: 25—79 (2016) 56, 100, 104, 128, 144, 153), and smaller size (body length 1.8—2.4 mm, foreay ine: lene thee sG: yah hahaa dtearecceve re atapladlett woke Ne telat ciate are 9 Fore wing with small areolet (as in Figs 137), and/or hypopygium and ster- nites usually mostly to entirely yellow, and/or larger size (body and fore wing Tenors ‘usd ally<52O ANOS ht tes ecsvtaecteRynceeasteticlRal ala Wena ect nibccne 15 Propodeum mostly sculptured (except for polished areas postero-laterally) (Pigs 49. 6945 OD SPOS) lS... oncaeid neue selius sd uosihsh Sees woh cewut ae 10 Propodeum mostly smooth (except for small striae around nucha) (Figs 32, PA ge Ae AUD i ee re acre sedctace oe netos seve teas ae teeth ces eon Vecceancreuasteh was 13 Smaller size, body length 2.0—2.1 mm, fore wing length 2.2 mm, metacoxa 0.45—0.46 mm; ocular—ocellar line 0.08—0.09 mm; T1 width/length 0.4—0.5 x; ten diagnostic characters in the DNA barcoding region: 79C, 235C, 346C, 364C> 386A 415A, 42 eS O2A60/G O22 Cathe ates Seen ee cates Promicrogaster fabiancastroi Fernandez-Triana & Boudreault, sp. n. Larger size, body length 2.3-2.4 mm, fore wing length 2.4—2.6 mm, meta- coxa 0.47-0.56 mm; ocular—ocellar line 0.11-0.15 mm (rarely 0.08); T1 width/length 0.5—0.7 x; different base pairs in the barcoding region : 79A or 79T, 235T, 346A or 346T, 364A or 364T, 386T, 415G, 421A or 421T, 5626-0162 1, 60 7A-orG07 li, O22 OF G22. levis A cccuceantlawiueiiece Moun 11 Ocular—ocellar line shorter than interocellar distance (0.9 x); T1 posterior width 1.3 x T2 central length; T2 width 3.2 x its length centrally... fees Promicrogaster luismendezi Fernandez-Triana & Boudreault, sp. n. Ocular—ocellar line longer than interocellar distance (1.4—1.6 x); T1 poste- rior width 1.8—2.0 x T2 central length; T2 width 3.5—4.9 x (usually more than: 4:0:x) "its lenge, cemtrall ys. id wccnn cei kinwre trates secant once depsocotetaneettnae 12 T2 width 4.9 x its length centrally; T1 length 1.9 x its posterior width; larg- er species, fore wing length 2.6 mm, metacoxa length 0.6 mm, metafemur lengths): 7a1nina,, mietapibiayO: 9; maint... uct eeetprevesdycsmsnitiames heen ake sovixaaueetasvele suhtas Promicrogaster eddycastroi Fernandez-Triana & Boudreault, sp. n. T2 width 3.5-4.0 x its length centrally; T1 length 1.5—1.7 x its posterior width; smaller species, fore wing length 2.3—-2.4 mm, metacoxa length 0.5 mm, metafemur length 0.5—0.6 mm, metatibia 0.7—0.8 mm... eee ..Promicrogaster naomiduarteae Fernandez-Triana & Boudreault, sp. n. Pterostigma with anterior 0.3 or more white, most veins of fore wing trans- parent or white (Fig. 29) [Costa Rica, ACG, dry forest under 300m] ........... ON Promicrogaster daniellopezi Fernandez-Triana & Boudreault, sp. n. Pterostigma entirely brown or at most with anterior 0.1 pale, fore wing with veins mostly brown (Figs 143, 150) [Costa Rica, ACG, cloud forests over LG) OC Tia teeta etree tec aan eect cee a an eres ec re Pee Od ee eR Pee 14 Posterior 0.6 of T1 sculptured (Fig. 147); clypeus, procoxa, most of sternites and laterotergites brown (Figs 141, 142, 144)... eee eeeeeeeeseeeeeeneeeees satahe Promicrogaster ronycastilloi Fernandez-Triana & Boudreault, sp. n. Revision of the genus Promicrogaster (Hymenoptera, Braconidae, Microgastrinae)... Ei. - Posterior 0.6 of T1 mostly smooth (at most with sculpture restricted to margins) (Fig. 152); labrum yellow-orange, procoxa yellow, sternites and laterotergites mostly to partially yellow (Figs 148, 149, 153)... Promi- crogaster sebastiancambroneroi Fernandez-Triana & Boudreault, sp. n. 15(8) Posterolateral corners of anteromesoscutum orange (Fig. 138); hypopygium and sternites dark brown (Figs 137, 139); Tl hardly narrowing towards pos- terior margins; T1 relatively wide, its medial length barely longer than its widthatanteriormarcin tics 58) 140): (Patiatina) ...cveconcasntanesthesaaiuets caiias sean Peet tape Me oadie gudieset Masa Promicrogaster polyporicola Muesebeck, 1958 — Anteromesoscutum entirely black; hypopygium and sternites usually mostly to entirely yellow; T1 usually narrowing towards posterior margin; T1 rela- tively narrower, its medial length much longer than its width at anterior mar- git [Brazil *@osta, Racay Mexico |p. nia cetitee it ters cee cebtaest Sus daa tae aae 16 16(15) Propleuron almost entirely yellow (Figs 45, 69, 70, 76), except for anterior 0.2 near head yellow-white (rarely propleuron partially yellow, partially light-brown); hypopygium, sternites and most laterotergites entirely yellow (hypopygium may have a dark spot on posterior 0.1 or less) (Figs 50, 69, 74, 81, 86)... eee 17 — Propleuron almost entirely dark brown to black, except for anterior 0.2 near head yellow (Figs 9, 63, 111); hypopygium, sternites and laterotergites at leastapattially-Gabkyerow fees, n.o eee eat. ee roles eee ee 19 17(16) Metacoxa dark brown on anterior 0.6 (Fig. 86); tegula brown; propleuron partially yellow, partially light-brown (partially visible in Figs 81, 82) .......... ..Promicrogaster kiralycastilloae Fernandez-Triana & Boudreault, sp. n. = Metacoxa, tegula and propleuron entirely yellow (propleuron with anterior 0.2 near head yellow-white) (Figs 45, 46, 50, 69, 74, 76) ...eeeeseeseeseeeees 18 18(17) T3 mostly yellow-white, with anterior 0.4 light brown, T4—7 brown anteri- orly, white on posterior 0.3—0.5 (Figs 71, 73) [Costa Rica, ACG, cloud forest OVEt a OOO WTA ae te le ut Peed ee Onn ate er Re as) eee Ret Menke it ees late ai ... Promicrogaster kevinmartinezi Fernandez-Triana & Boudreault, sp. n. = All tergites dark brown to black (Fig. 48) [Costa Rica, ACG, mid-elevation EAMMPIGFOTESTs 29 OO vctasn oan cuegevaden, release ne venice miesla Esteli vets ... Promicrogaster eimyobandoae Fernandez-Triana & Boudreault, sp. n. LOGG)s_ Hore wring without areolen (RIG. 1) ciascus.cct ahecncssusncened apeteinta ates cadena teats .. Promicrogaster andreyvallejosi Fernandez-Triana & Boudreault, sp. n. - Forewing with emiall areGlet icccccun cost ceeveweecevewsvancsuvsunenevecessvceeuvensentuccs 20 20(19) Flagellomeres 1—4 yellow-brown, clearly paler than rest of entirely brown flagel- lomeres (partially visible in Figs 63, 64); orange-yellow areas on metapleuron posterior 0.4, T1 anterior 0.6, T3 anterior 0.5 and small spot on mesopleuron POSteHOrly EFI gs:G3 GO—O8) Medtech weston tn oro ete acre naa seta incr ps eedlstuldaeh Sate ..Promicrogaster hillaryvillafuerteae Fernandez-Triana & Boudreault, sp. n. = All flagellomeres same color (brown to dark brown); meso and metapleuron entirely dark brown to black, coloration of T1 and T3 variable but not as AD OMEN = 40k Retreat Bret sr IR tu eee eo DN ih ee, ee Oe re Re Zl 34 Jose Fernindez-Triana et al. / Journal of Hymenoptera Research 50: 25—79 (2016) 21(20) Tegula and wing base dark brown to black (Fig. 114); clypeus black (same color 22(21) 23(22) 24(23) as face) (Fig. 112); clypeus slightly protruding and labrum slightly depressed, giving the appearance of a circular opening between the margin of clypeus and mandibles (barely distinguishable im Fig. VID) si..cacsssmesstas obervenannsronctaeds ..Promicrogaster monteverdensis Fernandez-Triana & Boudreault, sp. n. Tegula and wing base yellow; clypeus entirely to partially orange-yellow or orange-brown (clearer than face) (Figs 16, 155); clypeus not protruding and labrum not depressed, not giving the appearance of a circular opening .....22 Flagellomere 15 length 1.6—1.7 x its width; and ovipositor tip strongly bent downwards; and clypeus entirely orange-yellow (different from dark brown to black face); and body length and fore wing length 4.0 mm; and T1 al- most parallel-sided, very slightly narrowing towards posterior margin; and T2 mostly sculptured; and T3 with small yellow spot laterally [Brazil, Mexico]... Be A ae bale Ea atten ik bas ad ln ca Poh Promicrogaster apharea Nixon, 1965 Flagellomere 15 length 1.0-1.3 x its width; and/or ovipositor tip less strong- ly bent downwards; and/or clypeus entirely to partially dark brown to black (same color than face); and/or body length and fore wing length less than 4.0 mm; and/or T1 clearly narrowing towards posterior margin; and/or 12 mostly smooth; and/or T3 entirely dark brown to black [Costa Rica] ...... 23 Metacoxa with anterior 0.3—0.6 black (Figs 15, 19) [Costa Rica, ACG, cloud TOrestsovetaleOOt ia pees teeter ee aoe NUL 1 oe tre TL Oe ..Promicrogaster brandondinartei Fernandez-Triana & Boudreault, sp. n. Metacoxa entirely yellow (at most with small dark spot on anterior 0.1 or less, barely visible) (Figs 95, 154, 156) [Costa Rica, ACG, dry forest or mid- elevation rainforests, 300=SO0na] bocca. ones Skcue cade dof beceds cashoantess esenedeeucsaness 24 T1 clearly narrowing towards posterior margin, its length 2.0 x its width at posterior margin; [2 mostly sculptured; T3 with yellow spots laterally (Figs 94, 2) anaee Promicrogaster liagrantae Fernandez-Triana & Boudreault, sp. n. T1 almost parallel-sided, very slightly narrowing towards posterior margin, its length 1.7 x its width at posterior margin; T2 mostly smooth; T3 entirely dark brown to black (partially visible in Figs 156-158)... cee eeeeeeeeeeeeees .... Promicrogaster tracyvindasae Fernandez-Triana & Boudreault, sp. n. Taxonomic treatment of species Promicrogaster saraswatti Sathe & Bhoje, 1998, incertae sedis Promicrogaster saraswatti Sathe & Bhoje, 1998: 105. Original description. Holotype. Female, depository unknown. INDIA, Maharashtra, Kolhapur. Holotype not examined. Revision of the genus Promicrogaster (Hymenoptera, Braconidae, Microgastrinae)... 35 Comments. There are several issues that will require further study to clarify the status and identity of this species. First, all other Promicrogaster specimens that we have seen in collections (from either described or undescribed species) are restricted to the New World, and all published evidence (e.g., Mason 1981, Whitfield 1997) does not support this genus to be present in India. Second, Promicrogaster saraswatti was described by Sathe and Bhoje (1998) as a parasitoid of Phytomyza atricornis Meigen (Diptera: Agromyzidae), which is very suspicious, as all authenticated records of Microgastrinae are from Lepidoptera (e.g., Shaw and Huddleston 1991, Whitfield 1997, Quicke, 2015). Third, Sathe and Bhoje (1998) recorded in their paper the body size of the parasitoid wasp to be 4.64 mm and the wasp cocoon to be 4.5 mm; this is in strong contrast to a much smaller size for its supposed host: larvae of Phytomyza atricornis are only known to reach a maximum length of 3.5 mm and their puparia are even smaller at 2.1—2.5 mm (Cohen 1936). All of that points towards this species not being Promicrogaster but some other Microgastrinae (impossible to tell based on the unclear original description, which lacked any illustration); and the parasitoid’s biology might well have also been misunderstood and should be considered as highly questionable. To complicate things further, no details about the institution storing the holotype and 33 paratypes (reportedly from the same locality and data as holotype) were provided by the authors. Because the original description of P. saraswatti is the only source of information currently available, this species remains unrecognizable. Promicrogaster alexmartinezi Fernandez-Triana & Boudreault, sp. n. http://zoobank.org/2392E300-06B5-44CB-B4DE-D54F90EF21E1 Figs 1-8 Holotype. Female, CNC. COSTA RICA, Guanacaste, Area de Conservacién Guan- acaste, Sector Santa Rosa, Area Administrativa, 295m, 10.83764, -85.61871. DNA Voucher code: DHJPAR0031827. Paratype. 12 (CNC), same locality as holotype. DNA voucher code: DHJPARO031814. Description. Head: mostly black, clypeus, labrum and mandibles yellow. Fla- gellomeres: dark brown to black. Mesosoma: black. Tegula: white. Metasoma (dorsal- ly): T1 mostly red with black margins (posterior 0.3 of T1 black sometimes), T2—T4 red-orange or yellow-orange, 15+ yellow with small central band brown. Metacoxa: Or- ange. Malar distance: 0.3 x eye length. Fore wing areolet: present. T1 sculpture: mostly smooth, with posterior 0.3 sculptured. T2 sculpture: mostly smooth. Ocular—ocellar line: 0.14 mm. Interocellar distance: 0.12 mm. Posterior ocellus diameter: 0.11 mm. Body length: 4.20-4.97 mm. Fore wing length: 4.17—4.83 mm. Ovipositor length: 2.40—3.50 mm. Metacoxa length: 1.10—1.41 mm. Metafemur length: 1.11—1.32 mm. Metatibia length: 1.44—-1.89 mm. T1 length/width at posterior margin: 0.56—0.73mm/ 0.32-0.40 mm. T2 length/width at posterior margin: 0.18—0.20 mm/ 0.42—0.63 mm. 36 Jose Fernindez-Triana et al. / Journal of Hymenoptera Research 50: 25-79 (2016) Figures |-8. Promicrogaster alexmartinezi sp. n. holotype. Revision of the genus Promicrogaster (Hymenoptera, Braconidae, Microgastrinae)... ae Distribution. Known only from the holotype locality in ACG, dry forest, Costa Rica. Comments. The female holotype is larger and slightly darker than the paratype. Etymology. Promicrogaster alexmartinezi is named in honor of 13-year-old Alex Geovanny Martinez Lopez from the Colonia Bolafos school for his growing enthusi- asm for understanding and protecting the wild nature that occurs in his homeland. Promicrogaster andreyvallejosi Fernandez-Triana & Boudreault, sp. n. http://zoobank.org/42C8D3E2-34E6-424B-B594-F3CEB5AC3FBO Figs 9-14 Holotype. Female, CNC. COSTA RICA, Guanacaste, Area de Conservacién Gua- nacaste, Sector Cacao, Sendero Cima, 1460m, 10.93328, -85.45729. DNA Voucher code: DHJPAR0034152. Description. Head: mostly black, labrum and mandibles yellow-orange. Fla- gellomeres: dark brown to black. Mesosoma color: black. Tegula: dark brown. Meta- soma (dorsally): black to dark brown. Metacoxa: dark brown on anterior 0.5, yellow on posterior 0.5. Malar distance: less than 0.2 x eye length. Fore wing areolet: absent. T1 sculpture: mostly sculptured. T2 sculpture: mostly sculptured. Ocular—ocellar line: 0.15 mm. Interocellar distance: 0.06 mm. Posterior ocellus diameter: 0.08 mm. Body length: 3.46 mm. Fore wing length: 3.34 mm. Ovipositor length: 2.18 mm. Metacoxa length: 0.76 mm. Metafemur length: 0.89 mm. Metatibia length: 1.06 mm. T1 length/width at posterior margin: 0.45mm/ 0.27 mm. T2 length/width at posterior margin: 0.14 mm/ 0.44 mm. Distribution. Known only from the holotype locality in ACG, cloud forest, Costa Rica. Etymology. Promicrogaster andreyvallejosi is named in honor of 11-year-old Andrey Vallejos Lépez from the La Garita Vieja school for his growing enthusiasm for understanding and protecting the wild nature that occurs in his homeland. Comments. [he ovipositor was too curved and the metacoxa was partially hidden in the available specimen, so those measurements should be considered as approximate. Promicrogaster apharea Nixon, 1965 Promicrogaster apharea Nixon, 1965: 234. Original description. Holotype. Female, BMNH (examined). MEXICO, Guerrero, Omilteme, 2400 m. Distribution. Mexico, Brazil. The two Mexican specimens were collected at alti- tudes of 1,400 and 2,400 m respectively, whereas the Brazilian specimen was collected at around 500 m (Nixon 1965). Comments. The relatively long flagellomere 15, as well as body length and fore wing length over 4.0 mm are the most useful characters to separate the species from a 38 Jose Fernindez-Triana et al. / Journal of Hymenoptera Research 50: 25-79 (2016) Figures 9-14. Promicrogaster andreyvallejosi sp. n. holotype. Revision of the genus Promicrogaster (Hymenoptera, Braconidae, Microgastrinae)... 39 few Costa Rican species that are otherwise similar morphologically (e.g., P. brandond- inartei, P. liagrantae and P. tracyvindasae). P. apharea is only known from three female specimens, as mentioned in the original description Nixon (1965). The strange geo- graphic and ecological distribution reported suggests it might actually comprise two different species, one from Mexico and one from Brazil. Because we only examined the holotype we cannot conclude about that. Promicrogaster brandondinartei Fernandez-Triana & Boudreault, sp. n. http://zoobank.org/BC14B161-CE15-4AA2-B24C-E9019D20759E Figs 15-26 Holotype. Female, CNC. COSTA RICA, Guanacaste, Area de Conservacién Guan- acaste, Sector Cacao, Sendero Arenales, 1080m, 10.92471, -85.46738. DNA Voucher code: DHJPAR0031290. Paratypes. 12 (NMNH), same locality as holotype; 22. 1d (CNC), Costa Rica, Guanacaste, ACG, Sector Cacao, Sendero Circular, 1185m, 10.92714, -85.46683; 13 (CNC). Panama, Cerro Campana, 850m, 8° 40’ N 79° 55’ W. DNA Voucher codes: CNCHYM 01985, DHJPAR0031203, DHJPAR0031206, DHJPAR003 1258, DHJPARO003 1326. Description. Head: mostly black, labrum and mandibles light brown to yellow. Flagellomeres: dark brown to black. Mesosoma: black. Tegula: brown. Metasoma (dorsally): dark brown to black. Metacoxa: yellow. Malar distance: less than 0.2 x eye length. Fore wing areolet: present. T1 sculpture: mostly sculptured. T2 sculpture: most- ly sculptured, except for smooth central area. Ocular—ocellar line: 0.15 mm. Interocel- lar distance: 0.10 mm. Posterior ocellus diameter: 0.08 mm. Body length: 3.54 mm. Fore wing length: 3.70 mm. Ovipositor length: 2.89 mm. Metacoxa length: 0.85 mm. Metafemur length: 0.97 mm. Metatibia length: 1.27 mm. T1 length/width at posterior margin: 0.50 mm/ 0.31 mm. T2 length/width at posterior margin: 0.11 mm/ 0.47 mm. Distribution. Costa Rica (ACG, cloud forest), Panama. Etymology. Promicrogaster brandondinartei is named in honor of 12-year-old Brandon Josué Dinarte Barrientos from the Huacas school for his growing enthusiasm for understanding and protecting the wild nature that occurs in his homeland. Comments. There are two barcode divergent groups within what we call Prom- icrogaster brandondinartei (Fig. 160). The holotype (DHJPAR0031290, from the bar- code BIN BOLD:ABZ2999) and three of the paratypes (DHJPAR0031203, DHJ- PAR0031258, DHJPAR0031206, from the barcode BIN BOLD:AAM9535) form two sequence clusters that are not monophyletic within the genus. A fourth paratype specimen (DHJPAR0031326) is represented by a short sequence that clusters with the holotype. These two BINS are separated by 2.3 % sequence divergence, four amino acid differences and are derived from specimens collected from Malaise traps placed at different elevations along Volcan Cacao (Arenales and Circular at approximately 1,000m and 1,200m respectively). This may suggest that there are multiple species Jose Fernindez-Triana et al. / Journal of Hymenoptera Research 50: 25—79 (2016) Figures 15-20. Promicrogaster brandondinartei sp. n. holotype. Revision of the genus Promicrogaster (Hymenoptera, Braconidae, Microgastrinae)... 4] Figures 21-26. Promicrogaster brandondinartei sp. n. paratype. 42 Jose Fernindez-Triana et al. / Journal of Hymenoptera Research 50: 25—79 (2016) within this name. However, the trace files for the BIN associated with the holotype (BOLD:ABZ2999) contain ambiguities at the bases associated with the amino acid changes between the two BINS, a potential signal of co-amplification of a pseudogene or numt (Zhang and Hewitt 1996). Since the specimens are morphologically indistin- guishable, we consider it likely that the barcode records for P. brandondinartei contain variation derived from a pseudogene rather than representing a true mitochondrial variant. However, clearly delineating between the hypotheses of morphologically cryptic species-level variation (two BINS) or pseudogene-derived barcode variation within brandondinartei will require further collections and specimens to be barcoded. Promicrogaster daniellopezi Fernandez-Triana & Boudreault, sp. n. http://zoobank.org/79 CB6DA4-9678-408D-8 10F-DB276467FF2E Figs 27-32 Holotype. Female, CNC. COSTA RICA, Guanacaste, Area de Conservacién Guan- acaste, Sector Santa Rosa, Area Administrativa, 295m, 10.83764, -85.61871. DNA Voucher code: DHJPAR0031641. Paratype. 12 (CNC), same locality as holotype. DNA voucher code: DHJPARO031819. Description. Head: mostly black, labrum and mandibles light brown to yellow. Flagellomeres: dark brown to black. Mesosoma: black. Tegula: dark brown. Meta- soma (dorsally): black to dark brown. Metacoxa: mostly black to dark brown (posterior 0.1—0.2 yellow). Malar distance: less than 0.2 x eye length. Fore wing areolet: present. T1 sculpture: mostly sculptured. T2 sculpture: mostly sculptured, except for smooth central area. Ocular—ocellar line: 0.10 mm. Interocellar distance: 0.05 mm. Poster- ior ocellus diameter: 0.03 mm. Body length: 1.81 mm. Fore wing length: 2.02 mm. Ovipositor length: 1.47 mm. Metacoxa length: 0.31 mm. Metafemur length: 0.47 mm. Metatibia length: 0.58 mm. T1 length/width at posterior margin: 0.27 mm/ 0.13 mm. T2 length/width at posterior margin: 0.04 mm/ 0.24 mm. Distribution. Known only from the holotype locality in ACG, dry forest, Costa Rica. Etymology. Promicrogaster daniellopezi is named in honor of 13-year-old Migdonio Daniel Lépez Martinez from the Colonia Bolafios school for his growing enthusiasm for understanding and protecting the wild nature that occurs in his homeland. Promicrogaster daretrizoi Fernandez-Triana & Boudreault, sp. n. http://zoobank.org/F9 1 D4DBE-0D0C-457E-9CF2-FD82F92E6523 Figs 33-38 Holotype. Female, CNC. COSTA RICA, Alajuela, Area de Conservacién Guan- acaste, Sector San Cristobal, Bosque Trampa Malaise, 815m, 10.86280, -85.38460. DNA Voucher code: DHJPAR0025899. Revision of the genus Promicrogaster (Hymenoptera, Braconidae, Microgastrinae)... 43 f / helt. tal