JHR 97: 277-296 (2024) ee, JOURNAL OF Areminrcmasser nur doi: 10.3897/jhr.97.1 19470 RESEARCH ARTICLE () I Tymenopter a > https://jhr.pensoft.net Thelmernaonl Scie of Hymenopeariss, RESEARCH Discovery of a new Pseudalomya Telenga, 1930 (Hymenoptera, Ichneumonidae, Ichneumoninae) species from Taiwan and its implications for the systematic position of this genus Hsuan-Pu Chen', Namiki Kikuchi??, Shiuh-Feng Shiao! | Department of Entomology, National Taiwan University, No. 1, Sec. 4, Roosevelt Rd., Daan Dist., Taipei 106, Taiwan 2 Toyohashi Museum of Natural History, 1-238 Oana, Oiwa, Toyohashi, Aichi 441-3147, Japan 3 Systematic Zoology Laboratory, Department of Biological Sciences, Graduate School of Science, Tokyo Metropolitan University, 1-1 Minamiosawa, Hachioji-shi, Tokyo, 192-0397, Japan Corresponding author: Shiuh-Feng Shiao (sfshiao@ntu.edu.tw) Academic editor: Tamara Spasojevic | Received 25 January 2024 | Accepted 21 March 2024 | Published 9 April 2024 https:/zoobank. org/4C'5 LAI B9-275F-448E-B940-3D544E25B1FE Citation: Chen H-P, Kikuchi N, Shiao S-F (2024) Discovery of a new Pseudalomya Telenga, 1930 (Hymenoptera, Ichneumonidae, Ichneumoninae) species from Taiwan and its implications for the systematic position of this genus. Journal of Hymenoptera Research 97: 277-296. https://doi.org/10.3897/jhr.97.119470 Abstract The rare genus Pseudalomya Telenga comprises two species, which are found only in the Eastern Palaearc- tic region and high mountains of the Oriental region. The phylogenetic position of Pseudalomya remains unclear because of its intermediate morphology between two ichneumonine tribes, Alomyini and Phaeo- genini. This article reports the discovery of a new species of Pseudalomya: Pseudalomya truncaticornis sp. nov. Specimens were collected during a survey of insect fauna in the Dasyueshan area of Shei-Pa National Park, one of the high-altitude regions in Taiwan. The new species can be diagnosed by its body coloration, frontal horn shape, facial punctures, metasomal tergite sculpture, and wing venation. To the best of our knowledge, this is the first record of Pseudalomya in Taiwan. This article also presents a diagnostic key to the global spe- cies of Pseudalomya. In this study, molecular phylogenetic analyses were performed using one mitochondrial and two nuclear gene sequences from P truncaticornis sp. nov. and other members of the Ichneumoni- formes group. The results indicate that Pseudalomya should be classified within Phaeogenini, distinct from Alomyini, but more comprehensive phylogenomic studies are needed to confirm this placement. Copyright Hsuan-Pu Chen 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. 278 Hsuan-Pu Chen et al. / Journal of Hymenoptera Research 97: 277-296 (2024) Keywords COI, high-altitude, molecular phylogeny, 28S, taxonomy Introduction Pseudalomya Telenga, 1930 is a rare genus comprising two valid species: P. praevara Telenga, 1930 and P. nepalensis Riedel, 2019. Their distribution is restricted to the Eastern Palaearctic region and high mountains of the Oriental region (Yu et al. 2016; Riedel 2019). Another previously described Pseudalomya species from Japan — P. takeii Kusigemati, 1984 — was later confirmed to be a misidentification of an oxytorine spe- cies, Oxytorus corniger (Momoi, 1965) (Watanabe 2016). Because Pseudalomya exhibits an intermediate morphology between two ichneu- monine tribes Alomyini and Phaeogenini, it has generally been placed in both tribes (Laurenne et al. 2006; Quicke et al. 2009; Tereshkin 2009; Quicke 2015; Riedel 2019). Consequently, the tribal status of Pseudalomya remains a topic of debate. On one hand, this genus exhibits similarity with Alomyini because of the following characters: horned frons, enlarged vertex and genae, foramen with genae meeting (or almost meeting) ventrally, oval propodeal spiracle, metasomal tergite II without thyridium, and forewing with the second abscissa of M shorter than the first. On the other hand, Pseudalomya also exhibits similarity with Phaeogenini because of the fol- lowing characters: two-segmented trochanter, two-spurred mid tibia, and metasomal tergite I with spiracles located at apical 0.3 (Laurenne et al. 2006; Tereshkin 2009; Riedel 2019). Phylogenetic hypotheses reconstructed based on 28S D2—D3 rDNA sequences or combined (morphology and 28S) datasets suggested the tribal placement of Pseudalo- mya within the tribe Phaeogenini (Laurenne et al. 2006; Quicke et al. 2009). Conse- quently, the morphological similarities between Pseudalomya and Alomyini were inter- preted as either symplesiomorphic traits or the result of convergent evolution (Quicke 2015). However, because of the lack of sampling of this genus in subsequent compre- hensive phylogenetic studies (Bennett et al. 2019; Santos et al. 2021), the phylogenetic position of Pseudalomya remains incertae sedis (Santos et al. 2021). In this study, we analyzed three Pseudalomya specimens that were newly obtained from the high mountains of central Taiwan. The specimens were collected during fau- na inspection for the project SP110113: A survey for the selection of insect indicator species and their microhabitat usage in the Daxueshan area of Shei-Pa National Park. After morphological examinations, these specimens were discovered to be distinct from the known species of Pseudalomya. On the basis of morphological evidence, the specimens were subsequently validated as a new species, Pseudalomya truncaticornis sp. nov. This article describes the new species and presents a key to the global species of female Pseudalomya. In this study, the phylogenetic position of Pseudalomya was reas- sessed through multigene phylogenetic analyses. New species of Pseudalomya 279 Materials and methods Morphological examination The morphological terms used in this study were identified from Broad et al. (2018). Measurements were performed with reference to Kikuchi and Konishi (2021). The following abbreviations were used in this study: OOL, ocello-ocular line; POL, pos- tero-ocellar line; OD, ocellar diameter; PSI, propodeal spiracle index: major axis of propodeal spiracle/minor axis of propodeal spiracle; 1/M, the first abscissa of forewing M; 2/M, the second abscissa of forewing M; NI, nervellar index of hindwing: length of hindwing CU between M and cu-a/length of cu-a; T, metasomal tergite; $, metasomal sternite; and pS, posterior section of metasomal sternite. The measurements in parentheses represent the measurements of the holotype. ‘The cuticular microsculpture is described as per a study conducted by Eady (1968). The whole metasomal sternum was observed and dissected using a method developed by Ki- kuchi and Konishi (2021). The specimens were examined and measured using a micro- scope (Leica S8 APO; Leica Microsystems, Wetzlar, Germany) with a micrometer. Pho- tographs were taken using a Leica DMC 5400 camera integrated into a Leica Z16 APO microscope equipped with the auto-stacking system Leica LAS V4.13 (all from Leica Microsystems). Line drawings were generated using Procreate (Savage Interactive, Ho- bart, Australia). All figures were edited and arranged into figure plates by using Adobe Illustrator CC and Photoshop CC (Adobe Systems, San Jose, CA, USA). The specimens and their photos have been deposited at the following institutes: NMNS, National Mu- seum of Natural Science, Taichung, Taiwan; NARO, Institute for Agro-Environmental Sciences, National Agriculture and Food Research Organization, Tsukuba, Japan; and SDETI, Senckenberg Deutsches Entomologisches Institut, Miincheberg, Germany. The Latin term ibidem, meaning “same as previous except as follows,” was abbreviated as “ibid” to abridge information on the location of the materials examined. Taxon sampling To reassess the phylogenetic position of Pseudalomya, 52 operational taxonomic units (OTUs) from 38 ichneumonine genera were analyzed (Suppl. material 1). In addi- tion, other ichneumonids belonging to the Ichneumoniformes group were included as outgroups (Suppl. material 1). These outgroups were selected from the dataset of Santos (2017) and included 24 species from 23 genera of Cryptinae, 11 species from 11 genera of Phygadeuontinae, one species from one genus of Agriotypinae (Agriotypus armatus), and one species from one genus of Microleptinae (Microleptes splendidulus). For OTUs whose sequences were obtained from an online database, species identi- fication was double-checked using the Basic Local Alignment Search Tool (BLAST) (Altschul et al. 1990) and the position of preliminary phylogenetic reconstruction. Chimera OTUs were constructed for genera lacking available sequences of multiple markers within individual species. 280 Hsuan-Pu Chen et al. / Journal of Hymenoptera Research 97: 277-296 (2024) Molecular data collection and analysis Total genomic DNA was extracted from the right midleg of each specimen by using a DNeasy Blood and Tissue Kit (Qiagen, Diisseldorf, Germany). Partial sequences of the mitochondrial cytochrome c oxidase I gene (CO/) and two nuclear genes — D2—D3 re- gions of 28S ribosomal RNA gene (28S) and 18S ribosomal RNA gene (8S) — served as molecular markers for phylogenetic reconstruction. The sequences were retrieved from GenBank (Nation Center for Biotechnology Information). Target sequences were ampli- fied through polymerase chain reaction (PCR). The primer pairs and conditions for PCR are listed in Suppl. material 2. The reaction volume was 15 wL: 5.1 uL of sterile distilled water, 0.6 wL of each (forward/reverse) primer (10 uM), 7.5 wL of GoTaq Green Master Mix (Promega, Madison, WI, USA), and 1.2 pL of DNA template. PCR products were purified and sequenced at Tri-I Biotech (Taipei, Taiwan). Sequences were edited using CodonCode Aligner v.10.0.2 (CodonCode Corporation, Dedham, MA, USA). MAFFT v.7 (Katoh et al. 2019) was used for automated multiple sequence align- ments. The default setting was used for the alignment of CO/. By contrast, the E-INS- I algorithm was used for the alignment of the two nuclear genes (Santos 2017); the alignments of nuclear markers were manually optimized by removing regions with variable lengths and gaps. The translated alignment of CO/ was checked for stop co- dons by using MEGA v.11 (Tamura et al. 2021). All newly obtained sequences were uploaded on GenBank. The numbering of positions started from the gene’s first nu- cleotide (full-length CO/ sequences served as references for numbering: GenBank ac- cession JX131613 [Diadromus collaris] and MG923483 [Amblyjoppa sp.]). Molecular phylogeny To infer the phylogenetic position of Pseudalomya, phylogeny was reconstructed us- ing the following four datasets: COJ, 28S, 18S, and concatenated 18$+285+COI. The concatenated dataset was first partitioned by gene and then, for the protein-coding COP, also by codon position (first plus second versus third). ModelFinder (Kalyaana- moorthy et al. 2017) with “partition merging” was used for searching the optimal partitioning scheme and substitution models under the Bayesian Information Criteria. Maximum likelihood (ML) phylogenetic trees were reconstructed using IQ- TREE v.1.6.12 (Nguyen et al. 2015). This program was accessed through the web server W- IQ-TREE (Trifinopoulos et al. 2016), which is available at http://iqtree.cibiv.univie. ac.at/. Ten independent ML searches were performed for the concatenated dataset. After, the tree with the highest likelihood score was selected as the best-topology tree. Nodal support was assessed using the ultrafast bootstrap approximation (UFBoot2) method (Hoang et al. 2017) and SH-like approximate likelihood ratio test (SH-aLRT) (Guindon et al. 2010) under the default setting for the ML method. Nodes with an SH-aLRT value of 280% and a UFBoot value of 295% were considered to have strong support. For further interpretation of the tribal placement of Pseudalomya, the tribal classification system described by Santos et al. (2021) was applied herein. New species of Pseudalomya 281 Results Taxonomy Subfamily Ichneumoninae Latreille, 1802 Genus Pseudalomya Telenga, 1930 Pseudalomya Telenga, 1930: 107. Type: Pseudalomya praevara Telenga, 1930, by origi- nal monotypy. Pseudalomya truncaticornis Chen & Kikuchi, sp. nov. https://zoobank.org/5C674A42-79 DA-410C-A52C-25191684E0F6 Figs 1-4 Diagnosis. This species can be distinguished from other congeners in having the following combination of characters: frontal horn short and apically truncated; face sparsely punctate; metasomal tergites smooth with sparse and minute punctures; fore- wing with lcu-a distad to M&RS; head reddish brown; mesosoma and legs black; and metasomal tergites metallic-blue. Material examined. Holotype. Tarwan * 12; Miaoli County, Tai’an Township, Shei-Pa National Park, Mt. Huoshi; 24°22'47.78"N, 121°10'53.67"E (DMS); alt. 3160 m; 25 Aug. 2021-12 May. 2022; Jung-Chang Chen, Kuang-Yao Chen, Li-Jen Chang, Ta-Hsiang Li and Hung-Yang Shen leg.; Malaise Trap; GenBank: PP175350 (COI), PP188485 (28S), PP 188484 (18S); Sample ID: SP0060; Voucher: NMNS ENT 8836-1. Paratypes. Tarwan * 29; ibid; GenBank: PP175351 (COL, SP0061), PP175352 (COI, SP0062); Sample ID: SP0061-SP0062; Voucher: NARO (SP0061); NMNS ENT 8836-2 (SP0062). Description. Female. Head: 1.43-1.75 (1.75)x as wide as deep; antenna with 26-27 (27) flagellomeres; first flagellomere 1.13—1.22 (1.13)x as long as wide and 1.04-1.12 (1.04)x as long as second one; second flagellomere 1.08—1.19 (1.08)x as long as wide, remaining flagellomeres about 1.0x as long as wide; frons strongly and transversely striated; horn laterally depressed, apically truncated or slightly rounded (Figs 2A, 4A); face polished, 2.16—2.43 (2.16)x as wide as long, sparsely punctate (dis- tance between punctures 2.0—3.0x average puncture diameter) in middle and relatively dense (distance 1.0—2.0x average puncture diameter) laterally (Figs 2B, 4C), with se- tae; eyes bare, frontal orbit elevated; clypeus polished, 2.25—2.88 (2.88)x as wide as long, sparsely punctate with long setae, rounded in ventral margin and flat in lateral view (Figs 2A, B, 4C, F); malar space 1.29-1.73 (1.73)x as basal width of mandible, smooth in dorsal half and densely coriaceous in ventral half (Figs 2A, B, 4C); mandi- ble unidentate, rounded apically, broad, flat, polished and punctate with long setae at 282 Hsuan-Pu Chen et al. / Journal of Hymenoptera Research 97: 277-296 (2024) base (Fig. 4C); gena polished and sparsely punctate (Fig. 2C), pointed and strongly narrowed ventrally, meeting under the foramen; ocellar area and vertex rugose, rugose- punctate laterally; POL/OD=1.20-1.50 (1.50), OOL/OD=2.20-2.54 (2.36) (Figs 2A, 4E); occiput smooth; occipital carina strong and complete, genal carina meeting hypostomal carina far from mandible base at ventral tip of genae in 1.40—1.73 (1.73)x of basal width of mandible. A 3.0 mm Figure |. Pseudalomya truncaticornis sp. nov. holotype (NMNS ENT 8836-1) A lateral view of the habitus B dorsal view of the habitus. Photographed by Hsuan-Pu Chen. New species of Pseudalomya 283 hh Ag Vea o5mm | \ SS af f Figure 2. Pseudalomya truncaticornis sp. nov. holotype (NMNS ENT 8836-1) A dorsal view of the head B anterior view of the head C lateral view of the head D foreleg E dorsal view of the mesoscutum F dorsal view of the propodeum G dorsal view of the metasomal tergites. Photographed by Hsuan-Pu Chen. Mesosoma: polished; pronotum with setae, transversely and strongly strigose dor- sally, evenly and coarsely punctate dorsolaterally, and rugose-punctate ventrolaterally (Figs 2E, 3A); epomia present; propleuron coarsely punctate with setae; mesoscutum flat, 1.15-1.21 (1.21)x as long as wide, with median lobe densely and coarsely punc- tate, lateral lobes evenly punctate but polished and sparsely punctate posteriorly, with setae (Fig. 2E); notauli present anteriorly; scutellum flat, 1.09-1.18 (1.18)x as long as wide, sparsely punctate with setae, lateral carina absent (Fig. 2E); mesopleuron punc- tate, but rugose in dorso-anterior corner and middle posterior 0.5—0.8 (0.5), with mesopleural sulcus and furrow crenulate (Fig. 3A); epicnemial carina complete, extend to whole height of mesopleuron; sternaulus present in anterior 0.4 of mesopleuron; mesepisternum smooth; metapleuron evenly and coarsely punctate with setae in upper division and dorsal half, strongly and transversely strigose in ventral half; propodeal spiracle oval, 1.43-1.57 (1.43)x as long as wide; posterior transverse carina of mes- osternum interrupted anterior to mid-coxa; propodeum evenly and coarsely punctate with setae, with area basalis smooth, area superomedia and area petiolaris rugose-punc- tate, and area postero-externa rugose-foveolate (Fig. 2F); juxtacoxal carina present in 284 Hsuan-Pu Chen et al. / Journal of Hymenoptera Research 97: 277-296 (2024) 3.0 mm C S3 $4 S5 sclerite of $2 _ weakly sclerotized spots 1.0 mm = laterosternite Figure 3. Pseudalomya truncaticornis sp. nov.. NUNS ENT 8836-1 (A), NMNS ENT 8836-2 (B), and NARO (C) A lateral view of the mesosoma B wings C metasomal sternites. Photographed by Hsuan-Pu Chen (A, B) and Namiki Kikuchi (C). New species of Pseudalomya 285 E F 0.5mm 0.5mm Figure 4. Characters of two Pseudalomya species. Frontal horns: A Pseudalomya truncaticornis sp. nov. (holotype, NUNS ENT 8836-1) B Pseudalomya nepalensis (holotype, SDEI). Faces: C P truncaticornis sp. nov. (holotype, NUNS ENT 8836-1) D P nepalensis (holotype, SDEI) E darker specimen of P trun- caticornis sp. nov. (paratype, NMNS ENT 8836-2), dorsal view of the head F darker specimen of P trun- caticornis sp. nov. (paratype, NMNS ENT 8836-2), lateral view of the head. Illustrated and photographed by Hsuan-Pu Chen (A-C, E, F) and Matthias Riedel (D). 286 Hsuan-Pu Chen et al. / Journal of Hymenoptera Research 97: 277-296 (2024) basal 0.6; pleural and submetapleural carina present basally; lateromedian longitudinal carina weak; lateral longitudinal carina, anterior transverse carina, and propodeal apo- physis absent; posterior transverse carina present, but weak medially. Legs: evenly punctate with setae; coxa smooth dorso-apically; fore femur 2.63—2.84 (2.74)x as long as wide; hind femur 3.23—3.34 (3.34)x as long as wide, 0.21—0.24 (0.21)x as long as hind tibia; hind first tarsomere 1.75—1.92 (1.92)x as long as second tarsomere, and 0.45—0.49 (0.45)x as long as hind tibia; tibial spurs 2; tarsal claws normal. Wings: forewings narrowed, 3.80—4.22 (4.12)x as long as wide, and length 8.01—8.45 (8.45) mm (Fig. 3B); forewing lcu-a postfurcal, distad M & RS 0.23-0.33 (0.33)x by its length; areolet pentagonal, truncate anteriorly, with 2rs-m 1.10—1.26 (1.16)x as long as 3rs-m, and 1/M 1.05—1.14x as long as 2/M; 2 m-cu with two bulla; hindwing hamuli 7-9 (9), NI = 3.52-4.29 (4.29), CU and AA after cu-a weak. Metasoma: polished; T1 2.00—2.68 (2.01)x as long as its apical width, smooth, postpetiole sparsely and minutely punctate (Fig. 2G); spiracles of T1 located at about 0.7 of length of tergite (Fig. 3A); T2 0.91-0.95 (0.91)x as long as its apical width, with gastrocoeli and thyridium absent (Fig. 2G); T3 0.63—0.72 (0.63)x as long as its apical width; tergites after Tl smooth with sparse and minute puncture (Fig. 2G); pS1 and S2 separated by weak crease (Fig. 3C); laterosternites strongly sclerotized in S1—5, separately from median sternites by crease; median sternites strongly sclerotized, with weakly sclerotized median area in S2—4 and separated by creases; sclerites of S2 0.5x as long as S2, not touched its posterior margin, with two weakly sclerotized spots; median sternites of S5 and S6 complete; metasomal apex amblypygous (Fig. 1A), with apical margin of hypopygium pointed (Fig. 3C); ovipositor sheath 0.16—0.20 (0.17)x as long as hind tibia, smooth with setae; ovipositor with upper valve slightly longer than lower valve, and lower valve with fine teeth. Coloration: head mainly reddish brown, except central area of frons, horn, orbits, ventral margin of gena and clypeus, face below antennal sockets and around tentorial pits, ocellar area, apical half of mandible, occiput except its dorsal margin, and occipi- tal carina black (Fig. 2A—C); antenna mainly black except flagellomeres 8 (or 9)—13 ivory and ventral surface of scape reddish brown (Fig. 1A, B); mesosoma mainly black, except two stripes at dorsal surface of pronotum, posterior corner of pronotum, and postspiracular sclerite before tegula reddish brown (Figs 1A, B, 2E, 3A); metasomal tergites metallic dark blue (Fig. 2G), with sternites black tinged with reddish brown; ovipositor sheaths black and ovipositor reddish brown; legs mainly black except apical fore femur, ventral side of fore tibia, basal and apical basitarsus, apical tarsomeres red- dish brown (Fig. 1A); wings tinged with infuscate, with veins and pterostigma black (Fig. 3B). One specimen (NMNS ENT 8836-2; SP0062) with temple above eyes, whole occiput, and all legs black (Fig. 4E, F). Male. Unknown. Etymology. The specific name “truncaticornis’ is derived from the Latin “truncati-” (meaning “maimed” or “having appendages cut off”) plus “cornis” (meaning “horned”). It refers to the truncated apex of the horn on the frons in this species. Name is an adjective. New species of Pseudalomya 287 Distribution. Taiwan. Bionomics. Host and phenology unknown. The specimens were collected from Taiwan Hemlock (Zsuga chinensis) (Pinaceae) forest in the high-altitude area (alt. 3160 m) of central Taiwan, with Yushania niitakayamensis (Bambusoideae, Poaceae), Rhododendron species (Ericaceae), and moss as ground-cover plants (Fig. 5). Remarks. Comparisons of the photos of holotypes and descriptions of conge- ner species revealed the highest level of similarity between P truncaticornis sp. nov. and P nepalensis Riedel, 2019. However, unlike P nepalensis, the new species had the black coloration in mesosoma and legs (reddish brown in P nepalensis), the middle of face with sparse punctures with distance between punctures 2.0—3.0x their diameter (dense in P nepalensis, distance less than 1.0x their diameter), and a short and apically truncated frontal horn (long and apically rounded in P nepalen- sis). While variation in the color of the head was observed within P truncaticornis sp. nov., given the disjunct geographical distributions between P truncaticornis sp. nov. and P nepalensis (Taiwan and the Himalaya, respectively) and the presence of morphological differences beyond mere coloration, P truncaticornis sp. nov. is considered a distinct species. To the best of our knowledge, this is the first record of Pseudalomya in Taiwan. aa Re aT Figure 5. Habitat of Pseudalomya truncaticornis sp. nov. in Mount Huoshi (24°22'47.78"N, 121°10'53.67"E DMS), Shei-Pa National Park. Photographed by Ta-Hsiang Lee. 288 Hsuan-Pu Chen et al. / Journal of Hymenoptera Research 97: 277-296 (2024) Key to world species of female Pseudalomya Telenga, | 930 1 Metasomal tergites not smooth, with postpetiole of T1 rugose, and T2 co- riaceous; Forewing Icu-a interstitial, opposite to M&RS; metasomal tergites MO tot tallie= DIME wl hl A eR Rk UD P. praevara Telenga, 1930 = Metasomal tergites smooth (Figs 1B, 2G); Forewing 1cu-a postfurcal, distad to M&RS (Fig. 3B); tergites metallic-blue (Figs 1B, 2G) wees eeeeeeeeeee 2 2, Frontal horn long, rounded apically (Fig. 4B); punctures separated by less than 1.0x their diameter in middle of face (Fig. 4D); mesosoma and legs AO St Ted Chishs DROMuIN os ats, oa Re mas en ayes ede t P. nepalensis Riedel, 2019 - Frontal horn short, truncated apically (Fig. 4A); punctures separated by 2.0— 3.0x their diameter in middle of face (Figs 2B, 4C); mesosoma and legs black CQ errigs FAG 6) eceere hone Se oer Aen area ORES Ae P. truncaticornis sp. nov. Molecular dataset The dataset for molecular phylogeny comprised six newly obtained sequences: three COI sequences from the ichneumonine species Quandrus pepsoides (Smith, 1852), Cal lajoppa exaltatoria (Panzer, 1804), and Holcojoppa bicolor (Radoszkowski, 1887) and one sequence each of CO/, 28S, and 18S from P truncaticornis sp. nov.. In addition, the dataset included 250 sequences — 83 COJ, 88 28S, and 79 18S sequences — from GenBank (Suppl. material 1). No pseudogene, identifiable by the occurrence of stop codons in translated (ami- no acid) sequences, was detected in the protein-coding gene dataset. However, one 18S sequence from Pseudoplatylabus apicalis (GenBank accession KU753140) was eliminated because of its abnormal genetic distance in the 18S gene tree pretest. Table 1 presents the basic details of each aligned dataset. The information presented in this table includes the average length of unaligned sequences, length of aligned sequences, number of variable and parsimony-informative sites, and percentage of GC content. The datasets for the concatenated and individual markers are presented in Suppl. material 3. Molecular phylogeny Figs 6, 7 depict the ML phylogenetic trees reconstructed using the concatenated 188+28S+ COI dataset and the CO/ and 28S datasets, respectively. The 18S gene tree Table |. Summary of each aligned and trimmed molecular dataset. The table presents information on the average length of unaligned sequences, length of aligned sequences, number of variable and parsimony- informative (Pars-Inf) sites, and percentage of GC content. Number of sequences _ Average length Aligned length — Variable sites Pars-Inf sites GC (%) Cor 87 638.8 648 414 351 27.7 28S 89 613.3 625 283 169 59.7 18S 72. 890.9 1302 70 31 49.5 188+28S+COI 89 2028.6 2575 767 551 45.9 New species of Pseudalomya 289 could not satisfactorily resolve the phylogenetic relationships in the Ichneumoniformes group (see Suppl. material 4: fig. $3). All trees were rerooted using the outgroup Ag- riotypus armatus (Agriotypinae). The complete phylogenetic trees resulting from all datasets are presented in Suppl. material 4. a Aglaojoppa centummaculata '——— Coelichneumon bivittatus Le Protichneumon nakanensis @ — Protichneumon pisorius ~= = Amblyjoppa cognatoria — Amblyjoppa proteus i ‘“— Coelichneumon nigroindicum Tricholabus strigatorius coreanus Diphyus albicoxalis lr [© Ichneumon sexmaculatus —_——— Fupalamus japonicus Ichneumon sugiharai —— /chneumon sarcitorius chosensis Setanta apicalis i Amblyjoppa basalis Holcojoppa bicolor = ————_____——. Pseudoplatylabus apicalis Monontos niphonicus = Cobunus birmanicus a Cobunus sp. _—_________|schnojoppa ee Callajoppa exaltatoria —_—————._ Quandrus pepsoides Cratichneumon aspratilis Cratichneumon bifasciatus U— Fupalamus giganteus sinensis é Hoplismenus coreanus Hoplismenus infulatus r« Gavrana sp. Melanichneumon spectabilis Hymenura nigra ch —— eypinichiier Limonethe maurator Vulgichneumon suigensis .—_——._ Wulgichneumon lissolaba Listrodromus Neotypus nobilitator orientalis . Stenichneumon culpator Stenichneumon posticalis Virgichneumon albilineatus Togea albofasciatus Linycus exhortator Platylabus takeuchii Diadromus troglodytes rg r-———. Phaeogenes eguchii r = Pseudalomya truncaticornis sp. nov. Pseudalomya sp. NLS564 Lusius Phaeogenes sensu lato Dicaelotus sp. ————————— Eurylabus quadratus | Eurylabini Notosemus bohemani_ | Notosemini Alomya debellator | Alomyiini eg Cryptinae (22 species, 21 genera) — Litochila jezonica bis 82/89 99.6/97 fs e Platylabini 81.6/82 Phaeogenini 36.4/92 ~ Mansa sp. - Polytribax penetrator Phygadeuontinae Polyaulon sp. Bathythrix sp. Hemiteles sp. Microleptes splendidulus | Microleptinae Endasys sp. a Isdromas sp. ——____ Lienella sp. Paraphylax sp. i Phygadeuon sp. ily é Acrolyta sp. Gelis sp. //—— Agriotypus armatus | Agriotypinae r——® Phygadeuontinae 0.04 Ichneumonini Ichneumoninae Outgroups @ SH-aLRT = 80% & UFBoot = 95% SH-aLRT 2 80% XOR UFBoot 2 95% Figure 6. Maximum likelihood phylogenetic tree of Ichneumoninae reconstructed using the concatenated 18S+285+COI dataset (2575 bp; 18S: 1302 bp; 28S: 625 bp; COL 648 bp; SYM+I+G4 [1-1302, 1303- 1927, 1928-2575\3, and 1929-2575\3 bp]; HKY+F+I+G4 [1930-2575\3 bp]). The red and blue colors indicate Phaeogenini and Alomyini, respectively. Branch lengths of the phylogenetic tree are proportional to the infer number of nucleotide substitutions per site, except for the branch of the outgroup Agriotypus ar- matus. Circles on the nodes indicate different SH-aLRT/UFBoot values. Nodal support with an SH-aLRT value of <80% and a UFBoot value of <95% is not shown. Abbreviations: SH-aLRT, SH-like approximate likelihood ratio test; UFBoot, ultrafast bootstrap approximation; XOR, one or the other but not both. 290 Hsuan-Pu Chen et al. / Journal of Hymenoptera Research 97: 277-296 (2024) Hoplismenus infulotus | Aglaojoppa centummaculata - Protichneumon nakanensis Halcojoppa bicolor Hoplismenus coreanus Tricholabus strigatorius coreanus Coelichneumon nigroindicum | Protichneumon pisorius Polyaulon sp - Ambiyioppa pro roteus Amblyjoppa cognatoria Fe mceueen eae IF ambiyioppa proteus (= Protichneumon nakanensis ‘\ Coelichneumon nigroindicum —y emer blir Coelichneumon bivittatus Val jehnewmen lissalaba Amblyjoppa basalis = Phaeagenes eguchii Setanta apicalis Tae Sais Sse | Phacogenin eae sere a Pseudalomya trun runcaticornk is sp.nov. | (in part) I. f i ablepatie Seem Maris icaela neumon sugiharai | nist ah wogiedyies e ‘= Callajoppa exaltatoria or Platylabus fakeuehii Quandrus pepsoides viciehpemon Bb Heh coreanus Hy @ Cobunus birmanicus Manont 108 niahanicus Ichneumoninae = COR