JHR 87: 523-573 (2021) er, JOURNAL OF eet ennacnn nn doi: 10.3897/jhr.87.7293 | RESEARCH ARTICLE () I Tymenopter a 4 https://jhr.pensoft.net Thelmernaonl Scie of Hymenoptxriss, RESEARCH A semantically enriched taxonomic revision of Gryonoides Dodd, 1920 (Hymenoptera, Scelionidae), with a review of the hosts of Teleasinae Istvan Mik6', Lubomir Masner’, Jonah M. Ulmer?, Monique Raymond, Julia Hobbie', Sergei Tarasov*, Cecilia Beatriz Margaria’, Katja C. Seltmann’®, Elijah J. Talamas’ | UNH Collection of Insects and other Arthropods, Department of Biology and Life Sciences, University of New Hampshire, Durham, NH, USA 2. Canadian National Collection of Insects and Arachnids, Ottawa, Canada 3 Staatliches Museum fur Naturkunde Stuttgart, Stuttgart, Germany 4 Natural History Museum, University of Helsinki, Helsinki, Finland § Zoologta Agricola, Centro de Investigacién en Sanidad Vegetal, Universidad Nacional de La Plata, La Plata, Argentina 6 Cheadle Center for Biodiversity and Ecological Restoration, Santa Barbara, CA, USA 7 Florida State Collection of Arthropods, Gainesville, FL, USA Corresponding author: Istvan Mik6 (istvan.miko@gmail.com) Academic editor: Zachary Lahey | Received 14 August 2021 | Accepted 10 November 2021 | Published 23 December 2021 http://zoobank.org/E48E5D39-12C3-40AF-A135-35F54C8BIE63 Citation: Miké I, Masner L, Ulmer JM, Raymond M, Hobbie J, Tarasov S, Margaria CB, Seltmann KC, Talamas EJ (2021) A semantically enriched taxonomic revision of Gryonoides Dodd, 1920 (Hymenoptera, Scelionidae), with a review of the hosts of Teleasinae. In: Lahey Z, Talamas E (Eds) Advances in the Systematics of Platygastroidea LI. Journal of Hymenoptera Research 87: 523-573. https://doi.org/10.3897/jhr.87.7293 1 Abstract Teleasinae are commonly collected scelionids that are the only known egg parasitoids of carabid beetles and therefore play a crucial role in shaping carabid populations in natural and agricultural ecosystems. We review the available host information of Teleasinae, report a new host record, and revise Gryonoides Dodd, 1920, a morphologically distinct teleasine genus. We review the generic concept of Gryonoides and provide diagnoses and descriptions of thirteen Gryonoides species and two varieties: G. glabriceps Dodd, 1920, G. pulchellus Dodd, 1920 (= G. doddi Ogloblin, 1967, syn. nov. and G. pulchricornis Ogloblin, 1967, syn. nov.), G. brasiliensis Masner & Miko, sp. nov., G. flaviclavus Masner & Mik6, sp. nov., G. fuscoclavatus Masner & Miko, sp. nov., G. garciai Masner & Miké, sp. nov., G. mexicali Masner & Miko, sp. nov., G. mirabilicornis Masner & Miké, sp. nov., G. obtusus Masner & Miké, sp. nov., G. paraguayensis Masner & Mik6, sp. nov., G. rugosus Masner & Miké, sp. nov., G. uruguayensis Masner & Miké, sp. nov. We treat Gryonoides scutellaris Dodd, 1920, as status uncertain. Gryonoides mirabilicornis Masner & Miké, sp. nov. is the only known teleasine with tyloids on two consecutive flagellomeres, a well-known trait of Copyright Istvan Miko 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. 524 Istvan Miké et al. / Journal of Hymenoptera Research 87: 523-573 (2021) Sparasionidae. An illustrated identification key to species of Gryonoides, a queryable semantic representa- tion of species descriptions using PhenoScript, and a simple approach for making Darwin Core Archive files in taxonomic revisions accessible are provided. Keywords Egg parasitoids, Carabidae, South America, color pattern evolution, biomedical anatomy ontology, ovi- positor, Darwin Core Archive Introduction Egg parasitoids are the most important egg mortality factors in insects (Fatouros et al. 2020). Members of the subfamily Teleasinae are the only known egg parasitoids of ground beetles (Coleoptera, Carabidae), and ground beetles and rove beetles are the only known hosts for Teleasinae (Table 1). Besides Teleasinae, there was only one prior report of a carabid egg parasitoid, Ooencyrtus alboantennatus (Subba Rao, 1971) (Chal- cidoidea, Encyrtidae) (Subba Rao 1971). A label of the holotype specimen states that it was reared from the eggs of Nothopeus hemipterus Olivier, a well-known cerambycid pest of the clove tree, Syzygium aromaticum (L.) Merr. and L. M. Perry (Myrtaceae) (Franssen 1937). However, in his original publication, Subba Rao (1971) reported the host of his new species as Nothopus Leconte (Coleoptera: Carabidae) instead of Nothopeus Pascoe, an error in the transcription of the label data. Carabid species play an important role in natural and agricultural ecosystems. Un- derstanding their population dynamics and mortality factors are thus essential for de- veloping sustainable ecosystem management programs (Lévei and Sunderland 1996; Kromp 1999; El-Danasoury and Iglesias-Pifeiro 2018). Teleasinae are common in yel- low pan traps (Mik6é 2016), screen sweeping, sifted litter, and pitfall trap samples, even in disturbed habitats. However, their abundance in the environment does not correlate to knowledge of their biology. Collecting small carabid eggs can be challenging as many of the species lay their eggs en- closed in mud cells, below the soil surface, in clusters of potential prey eggs, or inside prey nests (Claassen 1919; Brandmayr and Brandmayr 1979; Bin 1983; Moore and Di Giulio 2019). Teleasinae are the only scelionid subfamily that is not represented in the fossil record. Nel and Azar (2005) classified a fossil specimen from the early Cretacous in Teleasinae and erected a new genus, Cretaxenomerus Nel & Azar, 2005, as the oldest platygastroid fossil from Lebanese amber. The authors exclusively used the identifica- tion key and the brief description of Teleasinae from Goulet and Huber, 1993, to classify the new fossil in Platygastroidea, Scelionidae and Teleasinae. This approach lacks taxonomic rigor and unfortunately has been widely applied in paleoentomology (Miko et al. 2018). The line drawing provided by Nel and Azar (2005) does not depict a teleasine, and we consider very unlikely that it is a platygastroid. Proper placement of this taxon will require reexamination of the holotype specimen. Phylogenetic analyses (Austin and Field 1997; Murphy et al. 2007; Popovici et al. 2017; Chen et al. 2021) consistently retrieve Teleasinae as monophyletic and the sub- Review of Gryonoides 525 Table |. Species of Teleasinae with known hosts. Parasitoid species Host species Reference Teleas rugosus Kieffer, 1908 Zabrus tenebrioides, Harpalus sp. (Carabidae) Telenga 1959, Egorova 1967, Plastin and Kononova 1991 Teleas lamellatus Szab6, 1956 Zabrus tenebrivides Goeze, 1777 (Carabidae) Miké et al. 2005 Teleas rugosus Kieffer, 1908 Harpalus sp. (Carabidae) Telenga 1959 Teleas sp. Harpalus sp. (Carabidae) Telenga 1959 Teleas rugosus Kieffer, 1908 Amara sp. (Carabidae) Telenga 1959 Xenomerus canariensis Huggert, 1979 Dromius sp. (Carabidae) Bin 1983 Trimorus caraborum (Riley, 1893) Chlaenius impunctifrons Say, 1823 (Carabidae) Ashmead 1893 Trimorus mandibularis (Ashmead, 1887) Harpatlus rufipes (Degeer, 1774) (Carabidae) present paper Xenomerus orientalis Miké & Masner, 2010 arena nigrolineata (Chaudoir, 1852) (Carabidae) Miké et al. 2010 Trimorus fulvimanus Kieffer Acylophorus wagenshieberi Kiesenwetter, 1850 Staniec 2005 (Staphylinidae) family has clear morphological limits (Masner 1976, 1980, 1993). However, most of its constituent genera are not well-defined and require revision at the genus and species levels. The subfamily can be recognized by the long marginal vein of the fore wing (3-4 times the length of the stigmal vein), metasomal tergite 3 as the longest and widest, and the presence of acrosternal calyces. Although each of these characters can be found in other scelionids (Miko et al. 2010), this combination is unique to Teleasinae. Gryonoides is one of the few teleasine genera with well-defined morphological limits. The genus was erected by Dodd (1920) based on the presence of lateral spines on the mesos- cutellum. Although a few other teleasines also possess lateral mesoscutellar spines (e.g. Dvi- varnus), Gryonoides can be readily diagnosed from them via multiple characters (Talamas et al. 2016) including the extremely elongate male flagellomeres. Gryonoides is strictly Neo- tropical in distribution and although rarely represented in historic collections (e.g. only 15 specimens can be found in the Hymenoptera holdings of the United States National Museum), a significant number of specimens have been are present in the Canadian Na- tional Collection of Insects, Arachnids and Nematodes. Lubomir Masner has participated in more than 50 collecting trips to the Neotropics and gleaned specimens of Gryonoides from the collecting efforts of many colleagues Johnson 2009). The 1,126 specimens of Gryonoides amassed at CNC represent the bulk of the specimens used in this treatment. Recently it has come to our attention that substantial holdings of Gryonoides ex- ist in smaller collections. For example, over 400 specimens are deposited at the C. A. Triplehorn Insect Collection at The Ohio State University, USA (https://mbd-db.osu. edu/) and over 1,400 specimens are housed in the Museo del Instituto de Zoologia Agricola “Francisco Fernandez Yepez’ (Garcia and Montilla 2004). Materials and methods Specimens This revision is based on specimens housed in the following collections: BMNH (The Natural History Museum, London, United Kingdom), CNC (Canadian Nation- 526 Istvan Miké et al. / Journal of Hymenoptera Research 87: 523-573 (2021) al Collection of Insects, Ottawa, ON, Canada), FSCA (Florida State Collection of Arthropods, Gainesville, FL, USA), USNM (National Museum of Natural History, Washington, DC, USA), UFES (Universidade Federal do Espirito Santo, Departa- mento de Biologia, Colecao Entomologica, Vitéria, Brazil), MLP (Museo de La Plata, Universidad Nacional de La Plata, La Plata, Argentina). Specimen occurrence data are made available through a Darwin Core Archive (Wieczorek et al. 2012) published on GitHub (https://github.com/seltmann/taxonomy-darwin-core) using. Occurrence Core following the recent GitHub Approach to Publishing Darwin Core Formatted Occurrence Data for Taxonomic Studies (Appendix 1. Seltmann et al. 2021). Microscopy Morphological phenotypes were examined with an Olympus SZX16 stereo-microscope, with an Olympus SDF PLAPO 2x PFC objective (230x). Brightfield images of pinned specimens were taken with an Olympus BX43 compound microscope equipped with an Olympus DP72 digital camera. Extended-focus images were rendered with Zerene Stacker (Version 1.04 Build T201404082055; Zerene Systems LLC, Richland, WA), annotated, and modified with Adobe Photoshop 6 (Adobe Systems, San Jose, CA) using the Adjust/Filter/Unsharp mask and Image/Adjustments/Exposure (Gamma cor- rection) tools. Metasomata were removed from the specimens and placed in 20% KOH for 24 hours, rinsed in 20% acetic acid for 30 minutes then transferred to a glycerin droplet on a concavity slide (Sail Brand Ltd., West Yorkshire, United Kingdom) and dissected. Sample preparation and imaging with confocal laser scanning microscopy followed Miko et al. (2016). Ovipositors were mounted between two coverslips (1.5 um, 22 x 60) ina glycerin droplet, using Blu-tack (Bostik, Wauwatosa, WI) as a spacer. Specimens were imaged with an Olympus FV 10i desktop CLSM using a 60x objective. Terminology, natural language (NL) descriptions Morphological terms largely follow Mik et al. (2007) and were matched to the Hy- menoptera Anatomy Ontology (HAO) (Yoder et al. 2010) using the HAO portal (http://portal.hymao.org/projects/32/public/ontology/). The medial area of the lateral propodeal area (malp) is delimited laterally by a longitudinal carina (car1) that is me- dial to and parallel with the plica (plica) and delimited ventromedially by the L-shaped lateral propodeal carina (Ipc: Fig. 22). The malp can be either glabrous or setose and is usually divided by a longitudinal carina (car2) that is incomplete in most Gryonoides species (Fig. 22). Taxonomic treatments including NL phenotype representations were compiled in mx (http://purl.org/NET/mx-database). Terminology of the phe- notype statements used in descriptions are mapped to the HAO (available at http:// purl.obolibrary.org/obo/hao.owl), Phenotypic Quality Ontology (PATO, available at http://purl.obolibrary.org/obo/pato.owl), Biospatial Ontology (BSPO, available at http://purl.obolibrary.org/obo/bspo.owl), and Common Anatomy Reference Ontol- ogy (CARO, available at http://obofoundry.org/). NL phenotype representations are Review of Gryonoides 527 in ‘entity attribute: value’ format. Taxonomic nomenclature, specimen data, support- ing images, OTU concepts, and NL phenotypes were compiled in mx (http://purl.org/ NET/mx-database). Taxonomic histories, descriptions, and material examined sec- tions were also produced with this software. The semantic phenotype statements were created using PhenoScript (https://github.com/sergeitarasov/PhenoScript/wiki) that is a language for semantic description of phenotypes using instance-based approach; its grammar, inspired by the graph description language DOT (https://en.wikipedia.org/ wiki/DOT_(graph_description_language)), enables fast creation of semantically rich statements that can be directly imported into Web Ontology Language (OWL) using the PhenoScript compiler (https://github.com/sergeitarasov/PhenoScript/blob/mas- ter/Examples/Gryonoides/Workflow.R). The PhenoScript descriptions (Suppl. saterial 2) were generated using Atom (https://atom.io/) that help using controlled vocabular- ies, built from ontology terms, through respective snippets. PhenoScript descriptions, output ontologies and supporting scripts are available from https://doi.org/10.5281/ zenodo.5768770. The instance-based approach of PhenoScript is similar to the class- based one using Manchester Syntax (http://www.w3.org/TR/owl2-man-chester-syn- tax/). The difference between the two is that the first creates ABox expressions, while the second one TBox expressions. The ABox expressions (i.e., instance-based ones) are easier to write and interpret. The protocols for semantic phenotype annotation follow Balhoff et al. (2013), Miko et al. (2014). For more formal rdf representations of hyme- nopteran phenotypes see https://github.com/hymao/hymao-data. Distribution maps were generated using QGIS software (QGIS Development Team 2021). Rearing experiment Eges of Harpalus rufipes (Degeer, 1774) were collected during September 2016, in Boalsburg, PA, USA, by uprooting young shoots of Digitaria Haller (crabgrass) from gravel in the backyard of a suburban home. Females of H. rufipes and their freshly laid eggs were exposed when the plants were pulled out. The eggs were placed on wet filter paper in sterilized petri dishes and kept at 20 °C for two weeks. Following this pe- riod, the two initially brown eggs were dissected in 0.1 M phosphate buffer. Eggs and emerging larvae were fixed with 75% ethanol and transferred into a glycerol droplet on concave microscope slides. The reared specimens of Trimorus mandibularis (Ashmead) were identified by comparison with images of the holotype (USNMENT01059227) provided by Talamas et al. (2016). Results and discussion New host record Trimorus mandibularis were reared from the eggs of Harpalus rufipes. Of the 37 eggs, two were light brown and the rest were white at the time of collection. During egg 528 Istvan Miké et al. / Journal of Hymenoptera Research 87: 523-573 (2021) collection, we observed 10-30 H. rufipes females with enlarged abdomens digging in the substrate (gravel densely overgrown by crabgrass). Eye spots and mandibles ap- peared through the chorion in white eggs 2—3 days after collection, indicating that the eggs were freshly laid. These structures did not appear on the two light brown eggs (Suppl. material 1: Fig. 1) indicating that the development of the embryos inside the eggs was terminated. One week after the egg collection, carabid larvae started to emerge from the white eggs, while the chorion of the two light brown eggs gradually darkened and became less transparent. After 2 weeks, carabid larvae from all white eggs hatched and the chorions of the two initially light brown eggs became harder and opaque. At the end of the second week we dissected two pharate adults of 7 mandibularis from the parasitized, originally light brown eggs. Trimorus mandibularis is most similar to 7’ arenicola (Thomson 1859) (holotype examined by Istvan Miks), a Palearctic species. Both possess rugulose sculpture on the lateral portions of the mesonotum and the medial region of T3, have spines along the lateral surface of the mesotibia, and have the clypeus more than 2 times as wide as long. Many, but not all, specimens of 7’ mandibularis have rugae radiating posteriorly from the medial portion of the anterior margin of the mesoscutellum. Whether 7) arenicola and T. mandibularis represent two distinct species or are synonyms require further analysis. Some of the characters of 7’ mandibularis are shared with Teleas (enlarged mandibles, spines on the mesotibia) and were hypothesized to help females dig while searching for host eggs (Sharkey 1981). Our observation supports this hypothesis as H. rufipes females lay their eggs 2-3 cm under the soil surface, requiring females of 77 man- dibularis to dig to reach the eggs. The majority of teleasines, including Gryonoides, do not possess enlarged mandibles, spines on the mesotibia or rugulose sculpture on the mesosoma, suggesting that these insects do not dig in the soil. Although it is generally accepted that carabid beetles usually burrow their eggs in the soil, our knowledge on carabid egg laying behavior is limited, and based on some scattered information, the number of carabid species that lay their eggs above the soil surface may be substantial (Claassen 1919; Sasakawa 2017; Saska and Honek 2004). Distribution Gryonoides might be the most commonly collected teleasine genus in the Neotropical realm (Garcia and Moontilla 2005). Despite extensive Neotropical sampling efforts in the last 50 years Johnson 2009), Gryonoides specimens have not been recovered from the West Indies (Fig. 24). We do not know the host relationships for Gryo- noides, but, based on their body size (1900-3125 um), the eggs of its host species are predicted to be 3000-4000 um. Eggs of this size range are laid by larger carabid species (Casale et al. 1996; Gilgado and Ortufio 2011; Moore and Di Giulio 2019). The carabid fauna of the West Indies is almost exclusively composed of winged and small to medium size species less than 24 mm long that colon ized the islands from mainland Mesoamerica (Darlington 1970). We suspect that the absence of larger carabids, which are the putative hosts of Gryonoides, is the reason for the absence of Gryonoides in the West Indies. 929 Review of Gryonoides PhenoScript: from class-based to specimen-based semantic representations In the following addenda we propose the transversion of natural language (NL) state- ments into PhenoScript, an instance-based OWL/XMLI syntax (Table 2). We also provide a brief comparison between some PhenoScript-based and Manchester Syntax- based semantic statements (Balhoff et al. 2012; Mik6 et al. 2016). PhenoScript is generally simpler than Manchester Syntax, and since it is instance based it allows direct comparison between instances of anatomical structures within the same organism or between different organisms. It provides simpler, less abstract, expressions that are better suited for data min- ing applications than multi-level nested Manchester Syntax expressions which are complicated and sometimes impossible to reformat into a queryable triplestore (for further advantages of an instance based approach over a class based approach see Vogt 2021). Systematics of Gryonoides Gryonoides Dodd, 1920 Gryonoides Dodd, 1920: 360 (original description. Type: Gryonoides pulchellus Dodd, by original designation), Dodd, 1930: 42 (keyed); Fouts, 1948: 92 (keyed); Muese- beck & Walkley, 1956: 356 (citation of type species); Masner, 1976: 70, 74 (de- scription, keyed); Johnson, 1992: 513 (cataloged, catalog of world species); Austin Table 2. Instance versus class based semantic representations of Hymenoptera phenotypes. Statement Type NL Statement PhenoScript Manchester Syntax Compares relative positions of Head triangular from anterior view (longest head > anterior_side > maximum___ | ‘Has part’ some (head and ‘has part’ some (‘anterior width dorsal_to. horizontal_plane side’ and ‘has part’ some (‘maximum width and anatomical lines head width dorsal to < head; (‘dorsal to’ some (horizontal plane and (‘bearer of horizontal midline of some centered) and (‘inheres in’ some head)))))) head) Describes Torular triangle open | torular_triangle > (carinal, carina2); | ‘Has part’ some (‘torular triangle’ and ‘has part’ connectedness of | dorsally (left and right carinal >> left; carina2>> right; some (carina and (‘bearer of’ some left) and cuticular elements | carinae not continuous | carinal continuous_with. carina2 (continuous_with some (carina and ‘bearer of” dorsomedially) some right)))) Describes color shared by multiple anatomical structures Compares lengths of anatomical lines (morphometric indices) Expresses surface properties of cuticular regions (e.g. coloration, sculpture, pilosity) Interantennal prominence, mouthparts, antenna yellow Scape 4.5 times as long as wide Metanotal spine longer than striated proximal region on mesoscutellum (interantennal_prominence, ‘has part’ some ‘interantennal prominence’ mouthparts, antenna) >> yellow; and ‘bearer of’ some yellow; ‘has part’ some mouthparts’ and ‘bearer of’ some yellow; ‘has part’ some mouthparts and ‘bearer of’ some yellow; scape >> length is_quality_ ‘Has part’ some (scape bearer_of some (length and measured_as. measurement_datum | is_quality_measured_as some ((has_measurement_ has_measurement_unit_label. length unit_label. length inheres_in. radicle) and (has_ << radicle; measurement_datum measurement_value.(=4.0,=4. 5float)))); has_measurement_value. 4.2; metanotal_spine >> length |>| length ‘Has part’ some (‘metanotal spine’ and ‘bearer << proximal_region < mesoscutellum; | of’ some (length and increased_in_magnitude_ proximal_region >> striated relative_to some (length and inheres_in some (proximal region and (‘part of some mesoscutellum) and (‘bearer of’ some striated))))) 530 Istvan Miké et al. / Journal of Hymenoptera Research 87: 523-573 (2021) Ls \ ns “Se y) % ve SS ~~ 500 um Figure |. Gryonoides Dodd 1920, habitus A Gryonoides pulchellus var. pulchellus Dodd, 1920, female, lateral view (CNCHymen_132834) B Gryonoides glabriceps Dodd, 1920, male, dorsal view (CNCHy- men_132133). & Field, 1997: 46, 68 (structure of ovipositor system, discussion of phylogenetic relationships); Talamas, Miké and Copeland 2016: 7 (keyed). Diagnosis. Gryonoides was diagnosed from two other taxa with lateral mesoscutel- lar spines, Dvivarnus Rajmohana and Veenakumari and a species in the Trimorus carus group, by Talamas et al. (2016). The elongate clypeus and torular triangle of Gryonoides are shared with the undescribed species in the 7’ carus group, whereas the presence of the proximal projections of cercal plates (pac: Fig. 2A) is shared with Dvivarnus (Fig. 3A). Additional diagnostic characters can be found in the ovipositor assembly: Gryonoides differs from Dvivarnus and the T. carus species group in the presence of a resilin rich sclerotized bridge (based on its blue autofluorescence in Review of Gryonoides Bee Figure 2. Ovipositor assembly of Gryonoides Dodd, 1920 A Gryonoides glabriceps, T7+8, (CN- CHymen_132935) B Gryonoides pulchellus var. doddi Dodd, 1920, ovipositor assembly, (CNCHy- men_132961). Sclerotised and resilin rich bridge is present in the ovipositor assembly of Gryonoides (crc = cercus, pac = proximal projections on cercal plate, la = lateral apodemes). response to 405 nm excitation wavelength and strong red fluorescence in response to 488 nm laser) connecting the lateral arms of T7+8 (brg, la: Figs 2A, 2B). Gryonoides males and females exhibit a unique sexual dimorphism in the ratio of the length of 532 Istvan Miko et al. / Journal of Hymenoptera Research 87: 523-573 (2021) Figure 3.17+8 of Teleasinae with lateral mesoscutellar spines A, E, C, F brightfield B and D CLSM im- ages using 488 and 405 lasers with red, green and blue filters A, B, F Duivarnus agamades (USNM_96246). C, D, F Trimorus carus-group (CNCHymen_133695). Proximal projections of cercal plates (pac) are present in Dvivarnus and Gryonoides and absent from Trimorus sp. Sclerotised and resilin rich bridges are absent from both taxa illustrated on this figure (la = lateral apodemes, crc = sercus). the radicle and the scape. Whereas in other scelionid species, the ratio is the same in males and females, in Gryonoides, the female radicle is longer than that of the male relative to the length of the scape (Figs 1A, B). It should also be noted that Gryo- Review of Gryonoides DDD noides has a strictly Neotropical distribution whereas other teleasines with lateral mesoscutellar spines are not known from this region. Description. Shape of male flagellomeres 3-11: cylindrical. Visibility of frontal patch: obscured by facial striae. Erect whorl of setae on male flagellomeres: absent. Male antenna length: more than 4 times as long as the body length. Number of papillary sensilla on female A12: 1. Number of papillary sensilla on female A7: 0. Female radicle length: elongate (4—5.7 times as long as wide). Mandibular teeth: 3. Mandibular teeth length: dorsal tooth > ventral tooth > medial tooth. Genal patch: absent. Facial striae: present. Clypeus length versus mandible width: clypeus at least 3 times as long as the width of the mandible. Hyperoccipital carina: absent. Vertex patch: absent. Anterior process of pronotum structure: reduced. Epomial carina: present. Pronotal cervical sul- cus: present. Pronotal suprahumeral sulcus: present. Pronotal cervical sulcus sculpture: smooth. Pronotal suprahumeral sulcus sculpture: foveolate. Pronotal suprahumeral sul- cus versus pronotal cervical sulcus: Pronotal suprahumeral sulcus ends medially before reaching pronotal cervical sulcus. Netrion sulcus versus pronotal: netrion sulcus does not reach pronotal rim. Netrion sulcus: present. Netrion sculpture: foveolate. Netrion length: netrion exceeding 2/3" of pronoto-mesopectal “suture”. Posterior pronotal sulcus: present. Ventral propleural area: smooth. Propleural epicoxal sulcus sculpture: crenulate (scalloped). Subalar pit: present. Epicoxal sulcus sculpture: crenulate (scal- loped). Fovea of the foveolate scutoscutellar sulcus diameter: diameter of fovea decreas- ing towards midline. Medial area of the anteromesoscutum sculpture: areolate. Notau- lus anterior end: anterior to the transscutal line. Mesonotal humeral sulcus sculpture: crenulate. Mesonotal suprahumeral sulcus anteromedial end: extending to anterior ends of notauli. Mesonotal suprahumeral sulcus sculpture: crenulate (scalloped). Sc- utoscutellar sulcus sculpture: smooth medially, foveolate laterally. Scutoscutellar sulcus lateral end: reaching the axillula laterally. Mesoscutellum posterior margin in dorsal view: concave. Mesoscutellum medial spine: absent. Transaxillar carina: present. Poste- rior scutellar sulcus lateral end: reaching the axillula laterally. Posterior scutellar sulcus: present. Posterior scutellar sulcus sculpture: foveolate. Mesepisternum (area anteroven- tral to mesopleural depression) sculpture: areolate (irregular foveae around setal bases present). Mesopleural pit: present. Mesopleural carina: present. Postacetabular patch: absent. Acropleural sulcus length: elongate. Acropleural sulcus: present. Apical semi transparent lamella on the metanotal spine: absent. Metascutellum sculpture: striated proximally. Metanotal spine length: longer than proximal striated region of metascutel- lum. Metanotal trough sculpture: foveolate. Metanotal spine: present. Metanotal spine shape dorsal view: pointed. Metapleural pit: present. Metapleural sulcus sculpture: smooth. Metapleural sulcus: present. Ventral metapleural area sculpture: transverse carinae present. Central propodeal area pilosity: absent. Lateral propodeal carina: pre- sent. Lateral propodeal carina versus posterior propodeal projection: adjacent. Lateral propodeal carinae shape: inverted Y-shaped. Posterior propodeal projection: present. Hind wing largest width versus marginal ciliae length: hind wing is more than two times as wide as marginal cilia length. Dorsal margin of female T1 in lateral view shape: convex. Lateral setae on T1: 5 or more. Basal depressions of T1: present. Felt field: pre- sent. Lateral patch on T2: present. Basal depressions on T2: present. Basal depressions 534 Istvan Miké et al. / Journal of Hymenoptera Research 87: 523-573 (2021) on T3: present. Apical setae on T3 length: apical setae on T3 are not longer than non apical setae on T3. Posterodorsal patch on T3: present. Basal depression on S1: present. Posterior felt field in female: absent. Basal depression on S2: present. Basal grooves on S3: present. Acrosternal calyx: present. Acrosternal calyx shape: circular. Acrosternal calyces medially: separated. Medial extension of lateral apodemes (female): separated. Medial apodeme on SG (female): present. Identification key to Gryonoides species Gryonoides females can be divided into two distinct species groups: the G. glabriceps species group and the G. pulchellus species group. Females of the G. glabriceps species group have a dorsally closed torular triangle that does not extend above the horizontal midline of the upper face (Figs 7A, 5A), and a largely black or dark brown mesoscu- tum, pronotum and mesopectus (Figs 10—14). This contrasts with the females of the G. pulchellus species group, which possess either a dorsally opened torular triangle or a closed triangle that extends above the horizontal midline of the upper face (Figs 7B, 8C) and are characterized by the yellowish-orange mesoscutum, pronotum and meso- pectus (Figs 1A, 4-9, 27). Although males of the two species groups cannot be separated by any combina- tions of traits, male specimens of each species belonging to the G. glabriceps species group can be confidently identified as they possess multiple diagnostic characters. Although we found diagnostic characters on male specimens for some Gryonoides species, matching males and female specimens should be further tested using mo- lecular markers. We were not able to identify male specimens for G. brasiliensis, G. flaviclavus, G. fuscoclavatus, and G. pulchellus as their most important diagnostic characters (structure of the torular trinagle) cannot be scored in male specimens (in males of the G. pulchellus group the torular triangle is always open dorsally). Male specimens keyed out as “other G. pulchellus group males” most likely belong to these species. 1 Females: antenna clavate, A7—A12 wider than A3—AG (Fig. 1A); A7—A12 with papillary sensilla on ventral surface; metasoma with 6 visible tergites ............. 2 = Males: antenna filiform, with A3—A12 equal in diameter (F5—F10 as wide as F1—F4; Fig. 1B); A7—A12 without papillary sensilla on ventral surface; meta- SONAeWiITNe/ WISI De CSLOUESe eco sencencarauee cn cweccevewtv-ccven ver ur a uvateguseecedtberseeneurs 12 2 Torular triangle open dorsally, if closed then extending above horizontal mid- line of upper face (Figs 7B, 8C); Mesoscutum, pronotum and mesopectus largely yellow to orange (Figs LA, 4-9, 27) wee 3 (G. pulchellus group) - Torular triangle closed dorsally and not extending to horizontal midline of upper face (Figs 5A, 7A); Mesoscutum, pronotum and mesopectus black, sometimes with brown margins (Figs 10—14)........ 10 (G. glabriceps group) 3 Medial area of lateral propodeal area setose (Fig. 9B) oo... cece eeesseeseeseeeseeees Lor nr BP Anas arate a Gryonoides garciai Masner & Mik6, sp. nov. — Medial area of lateral propodeal area glabrous (Fig. 5A) wo... cee eeeeeeseeseeeeeeee 4 10 11 Review of Gryonoides 535 Dorsal metapleural area setose (Fig. 21C); Lateral propodeal area areolate- rugose (Fig. 21C); Scape in distal 2/3", pedicel, A3—A6 dark brown; clava yellow (Fig. 21A) ........ Gryonoides uruguayensis Masner & Miké, sp. nov. Dorsal metapleural area glabrous; Lateral propodeal area not areolate rugose, traversed by one or two longitudinal carinae; color of antenna variable, not as Lb ay cinta Ave AREPe Seer RAO A. Pn Mince Sect Lees (oe ERE Sake Rae ane oS SEP SR 5 Posterior propodeal projection short, as long as wide (Figs 4A, 16, 18A).....6 Posterior propodeal projection more than 1.5 times as long as wide SE EAS96 J lis Sa 9 0 Stereo ne ce ee 7 T3 with rugulose sculpture (Figs 4B, 6A); Setal bases on dorsal upper face pustulate (Fig. 7A); Vertex striate laterally, punctate medially (Fig. 4B); Toru- lartriangléclosed:dorsallys Central keel present: saaneenoncenarnesonrondionnestscnnevede DAT Pee yee. SRN ne ey fe Gryonoides brasiliensis Masner & Mik6, sp. nov. T3 smooth posterior to basal costae (Figs 5B, 7C); Setal bases on dorsal up- per face smooth (Fig. 7B); Vertex smooth (Fig. 16A); Torular triangle open dorsally: central ed abso et EIe 0G \iccesicuacctcucniicetcuaed ub edavee wenn su boehur acest Ph eRe tassel AN wih cera eb SR me) cases Gryonoides obtusus Masner & Mik6, sp. nov. Torular triangle open dorsally; Central keel absent (Figs 7A, B, 8C) ........... 8 Torular triangle closed dorsally; Central keel present (Fig. 19B)............0. 9 Longitudinal midline of T3 with striae extending posteriorly from basal cos- tae for half the length of the tergite (Fig. 8A); Clava gradually darkening api- callye(Fig.e8B)s-Posterior-verex ptinciate!( PIG tS A) a... on tec ucanvee onan teasinrsecovent eatin wine ecusn ts Beem Gryonoides fuscoclavatus Masner & Miko, sp. nov. Foveae along anterior T3 not extending into striation (Fig. 7C); Clava en- tirely yellow (Fig. 7D); Posterior vertex smooth (Fig. 7C) ......ceeeeeeeeseeseesees sed ca hetdetia Rca cir aks Gryonoides flaviclavus Masner & Mik6, sp. nov. EB TUCULOSE -as4s sete! Gryonoides paraguayensis Masner & Miko, sp. nov. i SicimOo thy tees OL ly te me Gryonoides pulchellus Dodd, 1920 Dorsal metapleural area setose (Fig. 11A); Upper face with two transverse patches of dense, white setae (Figs 11A, 12B); Anterior propodeal pits absent (e.g. Fig. 18A); Lateral mesoscutellar spines distally curving ventrolaterally (Cerrexsa BW, WA OED) ee ean eerie ayer Gryonoides glabriceps Dodd, 1920 Dorsal metapleural area glabrous (Fig. 13C); Upper face without transverse patches of dense setation (Figs 13E, 14D); Anterior propodeal pits present (Figs 13F, 20B); Lateral mesoscutellar spines straight apically (Figs 13B, 14A)..... eee 11 Punctures of upper face well separated and not contiguous (Fig. 13E); Com- pound eye in lateral view 2 times as high as wide (Fig. 13C); Head rounded in anterior view (longest head width in horizontal midline of head; 13E);’T3 without rugulose sculpture (Fig. 13D); Notaulus absent (Figs 13B, 14A); Anterior propodeal pits adjacent to anterior end of lateral propodeal carinae si tefeal hi 1) Soap ae Gryonoides mexicali Masner & Miko, sp. nov. Punctures of upper face contiguous (Fig. 20A); Eye 1.5 times as high as wide in lateral view (Fig. 20D); Head triangular in anterior view (longest head width dorsal to horizontal midline of head; Fig. 20A); T3 with rugulose 536 Istvan Miké et al. / Journal of Hymenoptera Research 87: 523-573 (2021) sculpture (Fig. 20C); Notaulus present (Fig. 20E, F); Anterior propodeal pits lateral to anterior end of lateral propodeal carinae (Fig. 20B) 0... eee resect gels eteivanaeieree stories arevines Gryonoides rugosus Masner & Mik6, sp. nov. 12 Spread and release structures (RSS) present on A3 and A4 (Figs 22A—C)...... cae tee hapete aitesntaete, Seer Gryonoides mirabilicornis Masner & Mik6, sp. nov. = Spread and release structure absent from A3 present on A4 (Figs 12A, 18B). Ret alee Nas see ee Aes ink ABA OMe Rl oO. ARES ie Rr ca oe ete 13 13 Notauli absent (Fig. 13B) .. Gryonoides mexicali Masner & Mik6, sp. nov. — Norattlinpresenit (Rios: LOA IB) nce. cox convonces tin cos se ices Pecoaten desea Bet sour te testomaaeat 14 14 Lateral mesoscutellar spines curved ventrolaterally (Figs 11A, 12C, D)......... bal Asbing seuhis! oe bta.cbteedSD eta, seb Suara Gryonoides glabriceps Dodd, 1920 — Lateral mesoscutellar spines straight or curved medially (Figs 13B, 14A) ...... SO eee ee oe ore eter Gee oe SY epee OL, Vaal, Ueee epee Dens Seen 15 ibs) Mesoscutellar spines curved medially (Fig. GA) ..... ce eeceeeseeseeseseeeseseeeeeeenees cduseatsstiewe tunevsgdvearenson Gryonoides paraguayensis Masner & Miko, sp. nov. — Mesoseutellar.spines straight (Pigs | SA y V9) .ccccevurcevsnesunssrveraesrevecesvenees 16 16 dS wathrruculosesculpeire (PIG DOG) sas. s.assane noleauhsuseguat Seas caetekneneaarraestiend DAB hechds Mts aas he heave pense Gryonoides rugosus Masner & Mik6, sp. nov. = T3 without rugulose sculpture (Figs 10A, B, 13D) oe eeeeeeeeeeeees 17 ie Dorsaltinetaplcurallares setose-( hig. 2 IC). peatnaraecm en eeaetetotac a antares oe Aa erat OA Loe I Gryonoides uruguayensis Masner & Mik6, sp. nov. ~ Dorsaltnietaplewralarea