Six new species of Allorhogas (Hymenoptera, Braconidae, Doryctinae) from south and southeast Brazil with host-plant record

Six new Brazilian species of the gall-associated Doryctinae genus Allorhogas are described and illustrated: A. copaiba sp. nov., A. ilexaffinis sp. nov., A. inquilinus sp. nov., A. quarentenus sp. nov., A. vassununga sp. nov. and A. viridis sp. nov. We provide host plant records for five of these species, three and one of which are new host plant genera (Ilex L., Copaifera L. and Eugenia P. Micheli ex L.) and new host plant family (Aquifoliaceae) records, respectively. Allorhogas inquilinus sp. nov., whose biology was previously reported, represents the first confirmed case of phytophagous inquilinism in the genus. An updated key to Brazilian species of Allorhogas is provided.


Introduction
Allorhogas Gahan (1912) is a braconid genus belonging to the subfamily Doryctinae, whose species have been reported to be mainly distributed in the Nearctic and Neotropical regions from south and northeast U.S. to central Argentina (Yu et al. 2016;Zaldívar-Riverón et al. 2014, 2018Samacá-Sáenz et al. 2020). This genus currently contains 58 recognised species (Yu et al. 2016;Zaldívar-Riverón et al. 2018;Joele et al. 2019;Samacá-Sáenz et al. 2020), of which 50 are Neotropical and six Nearctic. The remaining two species were described from Iraq in the Middle East (A. semitemporalis Fischer) (Fischer 1960) and India (Shaikh and Chatterjee 2020), though their generic placement is considerably doubtful based on the provided information.
The Doryctinae is mostly represented by parasitoid species. However, Allorhogas belongs to a clade composed of nine genera whose species are associated with members of various vascular plant families (Zaldívar-Riverón et al. 2007, 2014. Species of Allorhogas whose host records have been confirmed or reported from reliable observations have been reared from leaf and stem galls and fruits of species belonging to eleven plant families (Moreira et al. 2017;Zaldívar-Riverón et al. 2018). These records have also revealed the existence of different strategies of phytophagy within the genus, including gall induction on seeds, stems, fruits and floral buds (Macêdo and Monteiro 1989;Marsh 2002;Centrella and Shaw 2010;Zaldívar-Riverón et al. 2018;Joele et al. 2019), seed predation (Zaldívar-Riverón et al. 2018) and phytophagous inquilinism on galls made by other insects (Moreira et al. 2017). Moreover, some species of Allorhogas have been suggested to be parasitoids of other insects associated to galls (Marsh 2002;Centrella and Shaw 2013;Martínez and Zaldívar-Riverón 2013), though this has not been based on reliable rearing records and alternatively they could also be phytophagous inquilines.
Recent molecular phylogenetic studies have shown that Allorhogas is polyphyletic (Zaldívar-Riverón et al. 2014;Samacá-Sáenz et al. 2019), and a taxonomic revision of the genus is in progress. Allorhogas, as currently known, appears to be particularly species-rich in Brazil based on material deposited in national collections, though only 11 species have been described to date for this country (Zaldívar-Riverón et al. 2018;Joele et al. 2019). Here we describe six new species of Allorhogas from south and southeast Brazil with host plant records for five of them, including three new host plant genera and a new host plant family (Aquifoliaceae). One of the new species was previously reported to be associated with stem galls made by an undetermined cecidosid lepidopteran on a species of the plant family Anacardiaceae, representing the only confirmed case of phytophagous inquilinism in Allorhogas (Moreira et al. 2017).

Methods
The type material of the six species described here was obtained after examination of specimens deposited in the Coleção Taxonômica do Departamento de Ecologia e Biologia Evolutiva (DCBU) at Universidade Federal de São Carlos (UFSCar) in São Carlos, SP, Brazil. All holotypes and half of paratypes are deposited in the DCBU, whereas the remaining paratypes are deposited in the Colección Nacional de Insectos (CNIN), Instituto de Biología, Universidad Autónoma de México (IBUNAM). The terminology employed for the body and wing venation features follows Sharkey and Wharton (1997), and for the surface sculpture features follows Marsh (2002). Colour digital pictures were taken with an MC170 HD video camera attached to a Leica M205C stereomicroscope and the Leica Application Suite version 4.12. SEM digital pictures of uncoated specimens were taken with a FEI Quanta 250 SEM in a low vacuum mode.

Taxonomic part
Allorhogas copaiba sp. nov. http://zoobank.org/A050367F-42BF-4D25-B73A-226E0BA48524 Diagnosis. This new species and A. inquilinus sp. nov. (described below) are the only two known species of Allorhogas from Brazil with vertex and frons rugose. However, A. copaiba sp. nov. can be distinguished from the latter and the remaining species of Allorhogas from this country by having the following combination of morphological features: 1) frons and vertex striate-rugose and coriaceous (strongly rugose-coriaceous in A. inquilinus sp. nov., usually coriaceous in the remaining species); 2) first metasomal tergite longitudinally costate-rugose and coriaceous (generally longitudinally costatecoriaceous in the remaining species); and 3) vein 2RS distal with m-cu (interstitial with m-cu in A. inquilinus sp. nov., variable in the remaining species).
Wings: forewing 3.0 times longer than wide (Fig. 1E). Pterostigma 2.2 times as long as wide and 0.7 times as long as R. Vein r 0.7 times as long as 3RSa, 0.2 times as long as 3RSb, and as long as r-m. Vein 2RS distal with m-cu, vein RS+Mb absent. Hindwing vein M + CU 0.6 times as long as 1 M, m-cu slightly curved towards wing apex.
Legs: hind coxa with a distinct basoventral tooth. Hind femur 3.0 times longer than wide.
Metasoma: first tergite 1.3 times wider than long, longitudinally costate-rugose and coriaceous; anteriorly delimited by an indistinct transverse carina, with two parallel longitudinal carinae running to apex (Fig. 1F). Second tergite longitudinally costatecoriaceous, line between second and third tergites distinct and straight; third tergite longitudinally costate-coriaceous on basal half, smooth and polished on apical half; remaining tergites smooth and polished. Ovipositor sheaths 0.9 times as long as metasoma.
Biology. The type specimens of A. copaiba were reared from spherical galls on stems and buds of Copaifera langsdorffii Desf. (Fabaceae) ( Fig. 2A). The inducer of this gall is unknown. These spherical galls are brown, glabrous and relatively big (about 2 cm of diameter). Copaifera langsdorffii has been reported to contain 23 different gall morphotypes (Costa et al. 2010).
Etymology. The name of this species refers to its association with the vascular plant genus Copaifera, whose common name in Brazil is Copaíba.
Allorhogas ilexaffinis sp. nov. http://zoobank.org/D50AF723-D084-49FC-A58F-72A9FCEDAD25 Diagnosis. This new species could be distinguished from the remaining species of Allorhogas from Brazil by having the following combination of features: 1) mesosoma brown to dark brown; 2) first and second metasomal tergites costate, smooth between carinae, remaining tergites smooth and polished (first and second tergites costate but with coriaceous sculpture between carinae and third tergite also sculptured in other Brazilian species with dark brown colour).
Description. Female. Body size 2.7 mm (Fig. 3A), forewing 2.4 mm. Colour: head brown, temple, gena and clypeus light brown; palpi pale yellow; scape and pedicel yellow; basal flagellomeres brown, turning black to apex; eyes dark brown to black;  mesosoma dark brown, lower part of mesopleuron brown; metasoma brown to dark brown; legs pale yellow; tarsal claws black; wings hyaline, forewing veins brown to light brown, stigma brown; hindwing veins pale yellow; ovipositor sheaths dark brown to black, ovipositor brown, apex strongly sclerotised.
Head: transverse in dorsal view, 1.5 times wider than its median length (dorsal view) (Fig. 3C), 0.6 times as long as high (lateral view); occipital carina complete and reaching hypostomal carina before mandible; post ocellar line (POL) as long than ocellar diameter (OD), 0.5 times ocular ocellar line (OOL); frons, vertex, temple, gena and clypeus coriaceous; face sparsely pilose, coriaceous, transversally rugose laterobasally ( Fig. 2B), with a smooth median area; area surrounding clypeus and gena with large pilosity; clypeus coriaceous; frons excavation distinct but not defined by sharp lateral margins; eye 1.3 times longer than wide; eye width 2.0 times longer than temple in dorsal view; malar space 0.3 times eye height and 1.7 longer than width of hypoclypeal depression; mandibles tridentate, teeth short and relatively equal in size; antenna with 26 flagellomeres, first flagellomere about 3.6 times longer than wide, 1.3 times longer than second one.
Wings: forewing 2.7 times longer than wide (Fig. 3F). Pterostigma 2.8 times as long as wide and 0.7 times as long as R. Vein r 0.9 times as long as 3RSa, 0.2 times as long as 3RSb, and as long as r-m. Vein 2RS interstitial with m-cu, vein RS+Mb absent. Hindwing vein M + CU 0.7 times as long as 1 M, m-cu slightly curved towards wing apex.
Legs: hind coxa with a distinct basoventral tooth. Hind femur 3.2 times longer than wide.
Metasoma: first tergite 1.3 times wider than long, longitudinally costate, smooth between carinae; with two subparallel longitudinal carinae running to apex, anteriorly delimited by an indistinct transverse carina (Fig. 3E). Second tergite longitudinally costate, smooth between carinae; line between second and third tergites distinct and straight; remaining tergites smooth and polished. Ovipositor sheaths 1.1 times longer than metasoma.
Biology. The type specimens of this species were reared from fruits of Ilex affinis Gardner (Aquifoliaceae).
Etymology. The name of this species refers to the host plant species where the type specimens were reared.

Allorhogas inquilinus sp. nov.
http://zoobank.org/75E631BD-773B-41E1-AE36-DC8621F2F4BF Diagnosis. This is a morphologically distinctive species, which could be distinguished from the remaining described members of Allorhogas from Brazil by having: 1) eyes small, malar space 0.8 times eye height (eyes larger, malar space less than 0.8 times eye height in the remaining species); 2) frons and vertex strongly rugose-coriaceous (generally coriaceous in the remaining species; striate-rugose and coriaceous in A. copaiba); 3) face coriaceous with longitudinal v-shape striation (variable but never with this sculpture in the remaining species); 4) propodeum entirely strongly rugose-areolate, with two indistinct diverging carinae (at least partially coriaceous and with distinct diverging carinae in the remaining species); 5) hindwing with vein m-cu almost straight, slightly curved towards wing base (slightly curved towards apex in the remaining species); 6) apical half of third and remaining metasomal tergites punctate-slightly costate (usually smooth in the remaining species).
Head: transverse in dorsal view, 2.4 times wider than its median length (dorsal view) (Fig. 4B, D), and 0.6 times as long as high (lateral view); occipital carina complete and reaching hypostomal carina before mandible; post ocellar line (POL) as long than ocellar diameter (OD), 0.3 times ocular ocellar line (OOL); frons and vertex strongly rugose-coriaceous, temple and gena coriaceous-slightly rugulose; face pilose, costate-rugose in longitudinal v-shape and coriaceous (Fig. 2D); area surrounding clypeus and gena with large, dense pilosity; clypeus transversally costate-rugose; frons excavation distinct but not defined by sharp lateral margins; eye 1.1 times longer than wide; eye width 1.9 times longer than temple in dorsal view; malar space 0.8 times eye height and 2.0 times longer than width of hypoclypeal depression; mandibles bidentate; antenna with 29 flagellomeres, first flagellomere about 3.0 times longer than wide, 1.3 times longer than second flagellomere.
Wings: forewing 3.2 times longer than wide (Fig. 4F). Pterostigma 4.0 times as long as wide and 0.7 times as long as R. Vein r as long as 3RSa, 0.3 times as long as 3RSb, and as long as r-m. Vein 2RS interstitial with m-cu, vein RS+Mb absent. Hindwing vein M + CU as long as 1 M, vein m-cu almost straight, slightly curved towards wing base.
Biology. A detailed description of the feeding biology of A. inquilinus was previously reported by Moreira et al. (2017). This species was confirmed by the authors as a gregarious inquiline of stem galls made by the cecidosid moth Cecidonius pampeanus Moreira & Gonçalves on Schinus weinmannifolius Mart. ex Engl. (Anacardiaceae).
Etymology. The epithet of this species is a reference to its inquiline biology.  (Fig. 5A), forewing 3.0 mm. Colour: body entirely honey yellow, scape honey yellow, pedicel brown, flagellomeres brown to dark brown; eyes silverish black; palpi yellow; legs yellow to honey yellow, tarsi dark brown; tarsal claws dark brown to black; wings hyaline, fore and hindwing veins dark brown basally, light brown apically, stigma brown; ovipositor sheaths dark brown to black, ovipositor honey yellow, apex strongly sclerotised.
Head: transverse in dorsal view, 1.8 times wider than its median length (dorsal view) (Fig. 5B, D), 0.7 times as long as high (lateral view); occipital carina complete and reaching hypostomal carina before mandible; post ocellar line (POL) as long than ocellar diameter (OD), 0.6 times ocular ocellar line (OOL); frons, vertex, temple and gena coriaceous; face sparsely pilose, rugose-coriaceous, raised and smooth-coriaceous medially (Fig. 2C); clypeus transversally rugose-coriaceous; area surrounding clypeus and gena pilose; frons excavation distinct but not defined by sharp lateral margins; eye 1.5 times longer than wide; eye width 2.0 times longer than temple in dorsal view; malar space 0.5 times eye height and 2.1 longer than width of hypoclypeal depression; mandibles tridentate, inner tooth considerably small and blunt; antenna with 27 flagellomeres, first flagellomere about 2.8 times longer than wide, 1.2 times longer than second one.
Wings: forewing 2.8 times longer than wide (Fig. 5F). Pterostigma 3.6 times as long as wide and 0.8 times as long as R vein. Vein r as long as 3RSa, 0.3 times as long as 3RSb, and as long as r-m. Vein 2RS interstitial with m-cu, vein RS+Mb absent. Hindwing vein M + CU 0.9 times as long as 1 M, m-cu slightly curved towards wing apex.
Holotype Biology. The type specimens of A. quarentenus were reared from legumes of an undetermined species of Inga.
Etymology. The name of this species refers to the COVID-19 pandemics with its subsequent undefined quarantine, which occurred while the authors were describing it.
Remarks. Specimens of A. quarentenus and A. vulgaris could be either uniformly honey yellow or with dark brown areas. However, in the former species they are only present along the lateral mesoscutal lobes (also present in the basal areas of propodeum and central areas of second to fourth tergites in A. vulgaris).

Allorhogas vassununga sp. nov. http://zoobank.org/91524D1C-2D7D-46AF-9F13-44A0DCFAF0DF
Diagnosis. This new species can be distinguished from the remaining described species of Allorhogas from Brazil by having the following combination of morphological features: 1) first metasomal tergite longitudinally costate-rugose, with two longitudinal carinae only distinct at base (sculpture variable, often with longitudinal carinae running along the entire tergite in the remaining species); 2) frons excavation defined by sharp lateral margins and a median longitudinal carina (frons excavation with or without sharp lateral margins and median longitudinal carina in the remaining species); 3) mesosoma and metasoma mostly dark brown to black, mesopleuron medially and posteriorly and sixth to remaining tergites honey yellow (always with a different colour pattern in the remaining species).
Description. Female. Body size 3.4 mm (Fig. 6A), forewing 2.7 mm. Colour: head honey yellow, frons and vertex brown; scape, pedicel and first and second flagellomeres honey yellow, remaining flagellomeres dark brown; palpi honey yellow; eyes black; mesoscutum, propodeum and metapleuron dark brown to black; pronotum honey yellow; mesopleuron dark brown to black anteriorly, turning honey yellow medially and posteriorly; venter of mesosoma dark brown to black; first metasomal tergite dark brown to black, second to fifth tergites dark brown to black, honey yellow along their edges; remaining tergites mostly honey yellow; legs honey yellow; tarsal claws dark brown to black; wings hyaline; forewing veins brown, stigma brown; hindwing veins yellow to light brown; ovipositor sheaths brown on two thirds, turning black to apex; ovipositor honey yellow, apex strongly sclerotised.
Head: slightly transverse in dorsal view, 2.0 times wider than its median length (dorsal view) (Figs 2E, 6B), and 1.6 times as long as high (lateral view); occipital carina complete and reaching hypostomal carina before mandible; post ocellar line (POL) as long than ocellar diameter (OD), 0.6 times ocular ocellar line (OOL); frons, vertex, temple and gena coriaceous; face coriaceous, laterally rugulose-coriaceous; area surrounding clypeus and gena with large, dense pilosity; frons excavation distinct, defined by sharp lateral margins and a median longitudinal carina; eye 1.4 times longer than wide; eye width 2.5 times longer than temple in dorsal view; malar space 0.4 times eye height and 1.8 times longer than width of hypoclypeal depression; mandibles bidentate; antenna with 28 flagellomeres, first flagellomere about 2.4 times longer than wide, 1.3 times longer than second flagellomere.
Wings: forewing 2.7 times longer than wide (Fig. 6E). Pterostigma 3.0 times as long as wide and 0.8 times as long as R. Vein r 0.5 times as long as 3RSa, 0.2 times as long as 3RSb, and 0.8 times as long as r-m. Vein 2RS interstitial with m-cu, vein RS+Mb absent. Hindwing vein M + CU 0.9 times as long as 1 M, m-cu distinctly curved towards wing apex. Legs: hind coxa with distinct, pointed basoventral tooth. Hind femur 4.1 times longer than wide.
Metasoma: first tergite 1.3 times wider than long, longitudinally costate-rugose, with two longitudinal carinae only distinct at base, anteriorly delimited by a transverse carina (Fig. 6C). Second and basal half of third tergite longitudinally costate, line between second and third tergites indistinct, slightly sinuate; apical half of third and remaining tergites slightly punctate. Ovipositor sheaths 0.6 times as long as metasoma.
Male. Similar to female. Body size 3.6 mm. Antenna with 29 flagellomeres. Hind femur swollen, about 3.6 times longer than wide.
Holotype Diagnosis. This new species could be distinguished from the remaining described species of Allorhogas by having the following combination of morphological features: 1) body greenish pale yellow (never greenish pale yellow in the remaining species); 2) claws white (dark in the remaining species); 3) anterior half of notauli strongly scrobiculate and distinct, posterior half less sculptured and distinct, not meeting, reaching the end of scutellum in a smooth-rugose area with two subparallel longitudinal carinae (notauli usually entirely distinct, without such longitudinal carinae in the remaining species); 4) vein 2RS distal with m-cu, vein RS+Mb absent (variable in the remaining species).
Head: transverse in dorsal view, 1.7 times wider than its median length (dorsal view) (Fig. 7D), 1.1 times as long as high (lateral view); occipital carina complete and reaching hypostomal carina before mandible; post ocellar line (POL) as long than ocellar diameter (OD), 0.5 times ocular ocellar line (OOL); frons, vertex, temple, gena and clypeus coriaceous; face sparsely pilose, coriaceous, slightly rugulose medially (Fig. 2F); frons excavation slightly distinct, not defined by sharp lateral margins; eye 1.2 times longer than wide; eye width 3.0 times longer than temple in dorsal view; malar space 0.4 times eye height and 1.4 longer than width of hypoclypeal depression; mandibles bidentate; antenna with 22 flagellomeres, first flagellomere about 4.3 times longer than wide, 1.2 times longer than second one.
Wings: forewing 3.0 times longer than wide (Fig. 7C). Pterostigma 2.4 times as long as wide and 0.6 times as long as R. Vein r 0.4 times as long as 3RSa, 0.1 times as long as 3RSb, and 0.8 times as long as r-m. Vein 2RS distal with m-cu, vein RS+Mb absent. Hindwing vein M + CU 0.7 times as long as 1 M, m-cu slightly curved towards wing apex.
Metasoma: first tergite 0.9 times as wide as long, longitudinally costate, with two indistinct longitudinal subparallel carinae, with a basal transverse carina (Fig. 7E). Second and basal two thirds of third tergite longitudinally costate, line between second and third tergites distinct and slightly sinuate, apical third of third and remaining tergites smooth and polished. Ovipositor sheaths as long as metasoma.
Biology. The type specimens of A. viridis were reared from conical galls apparently made by an undetermined cecidomyiid dipteran on leaves of Eugenia rotundifolia Casar. (Myrtaceae). These galls are cylindrical, unilocular, externally brown and internally white (Maia 1993).
Etymology. The epithet of this species derives from the Latin word viridi (green), in reference to its distinctive greenish colour.
Updated key to species of Allorhogas from Brazil

Discussion
The conserved external morphology in Allorhogas makes rearing records an important diagnostic feature to distinguish its species, since they appear to be highly specific to their host plants. This study increases to 17 the number of described species of Allorhogas from Brazil, and to 12 the number of host plant families that are associated with this genus. The plant association reported for A. ilexaffinis to fruits of Ilex affinis represents the first record for a member of the plant family Aquifoliaceae. Moreover, Copaifera L. (Fabaceae) and Eugenia P. Micheli ex L. (Myrtaceae), which are associated with A. copaiba and A.viridis, respectively, are new host genus records for Allorhogas. Of the six species described here, only the feeding biology of A. inquilinus could be confirmed. This represents the first confirmed record of a gregarious inquiline species in Allorhogas, and the only described species that is associated with the plant family Anacardiaceae. The galls where the type specimens of A. copaiba were reared also contained an undetermined lepidopteran moth species, and thus we presume that this could be the actual gall former in the system. Interestingly, A. inquilinus and A. copaiba are morphologically similar, sharing some unique features within the genus (e.g. vertex and frons with distinct rugose sculpture). Allorhogas viridis was, on the other hand, reared from leaf galls made by an undetermined cecidomyiid dipteran, though its feeding biology remains unknown.
Of all the previously proposed morphological synapomorphies of Allorhogas (Marsh 1993(Marsh , 2002 only two, a distinctly excavated frons and first metasomal tergite with a basal transversal carina, appear to occur in all of its species. The hindwing vein m-cu slightly curved towards its apex, on the other hand, is present in most Allorhogas species, though in some of them (e.g. A. marshi, A. inquilinus) could be straight or slightly curved towards the wing base. Further taxonomic studies will help to determine the actual species composition of this genus.