Revision of the genus Euagathis Szépligeti ( Hymenoptera , Braconidae , Agathidinae ) from Thailand , with description of three new species

The species of the genus Euagathis Szépligeti (Hymenoptera, Braconidae, Agathidinae) from Thailand are revised. Eight species are treated, three new species are described, i.e. Euagathis breviantennata sp. n., E. setosimaculata sp. n. and E. pallitarsis sp. n. Disophrys sogdiana Fahringer, 1937, D. chinensis Fahringer, 1937, and Euagathis sentosus Chen & Yang, 1995, are new junior synonyms of Euagathis chinensis (Holm‐gren, 1868). Euagathis guangxiensis (Chen & Yang, 2006) is a new combination. Lectotypes are designated for Disophrys sogdiana Fahringer, 1937, and D. chinensis Fahringer, 1937. A dichotomous illustrated key to species is presented; links to electronic interactive keys and to distribu‐ tion maps are also included.


introduction
Agathidinae is a moderately large subfamily of medium-sized to fairly large Braconidae with 1,154 described species worldwide and 316 in the Oriental Region (Yu et al. 2012), although there are an estimated 2,000-3,000 species awaiting description worldwide (Sharkey et al. 2006).The subfamily has a worldwide distribution, but its members are more common in subtropical and tropical regions than in temperate areas.The history of the classification of the Agathidinae was summarized by Sharkey (1992) and Sharkey et al. (2006) conducted phylogenetic analyses based on morphology and the D2-D3 regions of 28S rDNA.The Oriental fauna of Agathidinae was first revised by Bhat and Gupta (1977), who provided a detailed history of taxonomic research for the area.Keys to the Oriental genera of Agathidinae were published by Sharkey et al. (2009), van Achterberg and Long (2010) and Sharkey and Clutts (2011).The first key to the Oriental species of the genus Euagathis Szépligeti, 1900 was provided by Bhat and Gupta (1977).The Indo-Australian species of the genus Euagathis were revised by Simbolotti and van Achterberg (1990, Sulawesi;1995, Sunda islands), van Achterberg and Chen (2002, China and Vietnam), van Achterberg (2004, Wallacea, Australian region), van Achterberg and Raychaudhuri (2004, India) and van Achterberg and Long (2010, Vietnam).Chen and Yang (2006) provided a key to the Chinese species of Euagathis.The genus Euagathis was shown in the phylogenetic analysis by Sharkey et al. (2006) to be firmly nested within the tribe Disophrini and closely related to the genus Coccygidium de Saussure, 1892.

Methods
As part of the TIGER (Thailand Inventory Group for Entomological Research) NSFfunded entomological inventory of Thailand, three Malaise traps (per locality) were used at 30 different localities throughout Thailand from 2006-2010, comprising approximately 90 Malaise trap years.The specimens dealt with here are primarily from these traps.Species concepts are based on morphological and molecular data from COI and 28S.
Morphological terms follow van Achterberg (1988) and van Achterberg and Long (2010).Distributional data are listed for all new species and a Google map with associated distributional data is included for all species.
Phylogenetic methods: Regions D2-D3 of 28S rDNA (roughly 560 base pairs) were sequenced using the primers 28SD2hymF (5'-AGAGAGAGTTCAAGAGTACGTG-3') and 28SD3hymR (5'-TAGTTCACCATCTTTCGGGTC-3').Sequences were edited using Geneious Pro v4.7.5 (Drummond et al. 2009) and aligned using MAFFT (Katoh et al. 2006) through the GUIDANCE server (Penn et al. 2010) which was used to assess confidence scores for each column in the alignment.Columns with confidence scores < 93% (default) were removed prior to all phylogenetic analyses.COI sequences were generated with the primers LepF1 (5'-ATTCAACCAATCATAAAGATATTGG-3') and LepR1 (5'-TAAACTTCTGGATGTCCAAAAAATCA-3').MAFFT was used to align the COI sequences and no regions of ambiguous alignment were detected.Three permutations of the molecular data were phylogenetically analyzed herein: (1) a 49-OTU 28S-only data set, (2) a 31-OTU COI-only data set and (3) a 30-OTU data set in which all OTUs contain both COI and 28S sequences.The data sets were analysed under Bayesian Inference (BI) with MrBayes (v3.2;Huelsenbeck andRonquist 2001, Ronquist andHuelsenbeck 2003) under the GTR+I+G model of evolution (Rodriguez et al. 1990), partitioned by gene in the 2-gene data set, and conducted for 10 million generations.Additionally, the data sets were analyzed under maximum likelihood (ML) using Garli (v1.0;Zwickl 2006), using the default settings and the GTR+I+G model for best-tree searches and 100-replicate bootstrap analyses.Finally, 100 maximum parsimony (MP) bootstrap replicates were conducted on each data set using PAUP* (v4.0b10;Swofford 2001).Herein, we present the tree with the highest log-likelihood from each ML analysis, with nodal support values obtained from each method.Rooting the analyses with Disophrys spp.(GenBank accession numbers: COI: KC899814-KC899816; 28S: HQ667969-HQ667971, JF506257, KC867209) was based on the close relationship between these two genera recovered from analyses of large agathidine data sets (Sharkey, unpublished).
Distribution data, pdf 's of non-copyright references, images, notes, and host and type information can be found by searching TaxaBank (a combined specimen and taxonomic database; http://purl.org/taxabank.Codes beginning with an "H" and followed by numbers are unique identifiers used for specimens in the HIC (below), and in the specimen database TaxaBank (e.g., H 647).

HIC
Hymenoptera Institute Collection, University of Kentucky, Department of Entomology, Lexington, Kentucky, USA.

Species delimitation.
Both morphological and molecular data, specifically COI and 28S, were used to determine species limits.Our original morphological species concepts were tested against the molecular data.Most of these morphological concepts were corroborated, including the rather subtle morphological differences between E. abbotti and E. forticarinata, which are distinguished with both COI (Fig. 2) and 28S (Fig. 3).The COI ML tree (Fig. 2) shows two distinct lineages of E. forticarinata and one specimen (H004) that is an outlier from both of these clades.The sole specimen of the morphologically distinct species E. setosimaculata lies between the two clades.This strongly suggests that E. forticarinata may be comprised of more than one species.However we could discover no consistent morphological differences between the two lineages.The COI ML tree (Fig. 2) also shows considerable variation within E. abbotti, but again we could not discern morphological characters consistent with these lineages.The 28S ML tree (Fig. 3) is more conservative and provides different information for our purposes, it separates all of our morphologically based species concepts and all members within these species have identical sequences, with the following two exceptions.First, E. ophippium and E. pallitarsis are distinct morphologically but have identical 28S sequences; nonetheless based on the morphological data we chose to suggest species status for both.Second, the two E. forticarinata specimens, H004 and H743, are identical and distinct from the other E. forticarinata specimens.We do not have COI data for H743, however COI data for H004 is distinct and widely separated from all other E. forticarinata specimens (Fig. 2).Unfortunately, both H004 and H743 are male specimens and they both appear identical to other melanic males of E. forticarinata (see Fig. 9k).It is our opinion that these two specimens probably represent a new species, but due to the lack of female specimens, the lack of diagnostic characters to distinguish the putative species from E. forticarinata and our rather small sample size, we have decided against proposing a new species.Figure 4 is a ML tree of the combined COI and 28S data.Since the 28S data are largely monotonous within species the topology mostly reflects that of the COI tree (Fig. 2).Notes.The pterostigma, mesosoma, metasoma and hind leg of the male are usually largely dark brown or black.Diagnosis.This new species keys to E. fuscinotum Enderlein, 1920, from the Sunda area (Java, Sumatra, Borneo, West Malaysia) in the key by Simbolotti and van Achterberg (1995).Females of both species have the segments of the apical half of the antenna about as long as wide.E. breviantennata sp.n. differs by having the hind femur punctate-rugose ventrally (coarsely punctate in E. fuscinotum), third antennal segment of female about twice as long as wide (about 3 times), setae of middle tarsus about as long as width of tarsal segments (about half as long as width of segments), anterior crenulae of precoxal sulcus long (short to medium-sized), head normally triangular in anterior view and about 1.2 times wider than high (eyes strongly protruding, head about 1.4 times wider than high in female), apical half of fore wing with yellowish tinge (without yellowish tinge) and first tergite comparatively elongate and 1.7-2.2times as long as wide apically (comparatively short and 1.4-1.9times as long as wide apically).
Head.Antennal segments 45, length of third segment 1.3 times fourth segment, length of third, fourth and penultimate segments 2.2, 1.8 and 1.0 times their width, respectively; apical antennal segment as long as penultimate segment; maxillary palp 0.6 times height of head; malar space 2.8 times as long as basal width of mandible; length of eye 2.1 times temple; temple directly narrowed posteriorly and slightly concave laterally; POL:OD:OOL= 6:5:9; face shiny with shallow medial groove dorsally, spaced punctulate and setose; frons, vertex and temple shiny and smooth (Fig. 6d); temple concave near lower level of eye.
Mesosoma.Length of mesosoma 1.4 times its height; pronotum smooth, but setose and punctulate dorsally and finely crenulate posteriorly; area near lateral carina of mesoscutum smooth; mesoscutum shiny, with spaced and rather coarse punctures and lateral lobes distinctly convex posteriorly (Fig. 6g); notauli complete, smooth or nearly so; scutellum convex and densely coarsely punctate, antero-dorsal margin rounded and without transverse carina; precoxal sulcus complete and anterior crenulae long (Fig. 6f ); mesopleuron and metapleuron medially coarsely punctate with interspaces equal to diameter of punctures or wider; propodeum coarsely areolate, anterior face about as long as posterior face.
Legs.Length of hind femur, tibia and basitarsus 4.9, 8.3 and 7.6 times their width, respectively; hind femur punctate-rugose ventrally; setae of middle tarsus about as long as width of tarsal segments; fore and middle tarsal segments moderately slender; length of outer and inner spur of middle tibia 0.6 and 0.8 times middle basitarsus, respectively; outer side of middle tibia without pegs, except for 1 apical peg; length of outer and inner spur of hind tibia 0.4 and 0.6 times hind basitarsus.
Biology.Unknown.Distribution.Only known from Thailand.For a distribution map, see Appendix I. Molecular data.Genbank accession numbers: KC867218 (28S).Etymology.From "brevis" (Latin for "short") and "antenna" (Latin for "sailyard, feeler") because of the short antenna of the female.

Euagathis dravida
Notes.This is a variable species, females vary in sculpture of the mesosoma and the males both in colour and sculpture.Males may have the body yellow (as female) up to largely black (except head, anterior part of mesosoma, fore and middle legs; Fig. 9k); intermediates occur and melanic males are more common in Thailand than are yellow ones.The area below the precoxal sulcus varies from widely spaced punctate, densely punctate, punctate-rugulose to densely obliquely rugulose with dense puncta-   crenulae of the precoxal sulcus coarse (medium-sized), the third and fourth segments of the fore tarsus normal (shortened) and the head, hind femur and mesoscutum largely brown (black).
Head.Antennal segments 49, length of third segment 1.4 times fourth segment, length of third, fourth and penultimate segments 2.8, 2.0 and 1.2 times their width, respectively; apical antennal segment 1.3 times as long as penultimate segment; maxillary palp 0.7 times height of head; malar space 2.8 times as long as basal width of mandible; length of eye 1.8 times temple; temple directly narrowed posteriorly, with long setae and slightly concave laterally; POL:OD:OOL= 5:5:8; face shiny with shallow medial groove dorsally, punctulate, finely rugulose medio-ventrally and long setose; frons, vertex and temple shiny and smooth (Fig. 11d); temple nearly straight near lower level of eye.
Mesosoma.Length of mesosoma 1.3 times its height; pronotum largely smooth, but with some curved striae anteriorly, punctulate dorsally and finely crenulate posteriorly; area near lateral carina of mesoscutum smooth anteriorly and finely crenulate posteriorly; mesoscutum shiny, with spaced and rather coarse punctures and lateral lobes distinctly convex posteriorly (Fig. 11f ); notauli complete, anterior half finely crenulate and posterior half smooth or nearly so; scutellum tuberculate, with long setae and densely coarsely punctate, antero-dorsal margin rounded and with irregular transverse rugae; precoxal sulcus complete and all crenulae long and connected to rugae ventrally, area below it punctate-rugose (Fig. 11h); metapleuron coarsely punctaterugose; propodeum coarsely areolate-rugose, anterior face much shorter than posterior face and with many long setae.
Legs.Length of hind femur, tibia and basitarsus 5.3, 8.0 and 10.2 times their width, respectively; hind femur superficially pimply and largely smooth ventrally; setae of middle tarsus shorter than width of tarsal segments; third and fourth fore and middle tarsal segments shortened; length of outer and inner spur of middle tibia 0.4 and 0.5 times middle basitarsus, respectively; outer side of middle tibia without pegs, except for 2 apical pegs; length of outer and inner spur of hind tibia 0.3 and 0.6 times hind basitarsus.
Metasoma.First tergite 2.3 times as long as wide apically, gradually widened apically, with short dorsal carinae basally and smooth (Fig. 11g); second metasomal suture faintly impressed; ovipositor sheath 0.06 times as long as fore wing, truncate apically and widened.
Variation.Length of fore wing 8.6-9.8 mm; length of ovipositor sheath 0.06 times as long as fore wing; antennal segments of female 48 (1) or 49 (3); first metasomal tergite 1.9-2.3times as long as its apical width; dark brown patch of first tergite minute or large; third epipleuron large black or ivory anteriorly.
Distribution.Only known from Thailand.For a distribution map, see Appendix I. Molecular data.Genbank accession number KC867252 (28S).Etymology.From "pallidus" (Latin for "pale") and "tarsos" (Greek for "flat part of the foot between toes and heel") because of the pale hind tarsus.
Diagnosis.This new species keys to E. abbotti (Ashmead, 1900) from the Sunda area, Thailand, Laos and Vietnam in the key by Simbolotti and van Achterberg (1995).Females of both species have the third and fourth segments of the fore tarsus slender, vein 1-R1 of fore wing somewhat darker than the pterostigma, the precoxal sulcus comparatively narrow, the mesoscutum distinctly punctate and the scapus yellow.E. setosimaculata sp.n. differs by having the dorsal face of the propodeum much shorter than its posterior face (about as long as posterior face in E. abbotti); the hind femur about 4 times as long as wide (5-6 times); the area near vein cu-a of the hind wing glabrous (sparsely setose); vein cu-a of hind wing about as long as wide (distinctly longer than wide).
Head.Antennal segments 48, length of third segment 1.1 times fourth segment, length of third, fourth and penultimate segments 3.1, 2.8 and 1.2 times their width, respectively; apical antennal segment 1.8 times as long as penultimate segment; maxillary palp 0.6 times height of head; malar space 2.7 times as long as basal width of mandible; length of eye 1.8 times temple; temple directly narrowed posteriorly and slightly concave laterally (Fig. 12c); POL:OD:OOL= 12:10:21; face shiny with shallow medial groove dorsally, punctulate and short densely setose; frons, vertex and temple shiny and smooth (Fig. 12c); temple concave near lower level of eye.
Legs.Length of hind femur, tibia and basitarsus 3.9, 7.3 and 8.4 times their width, respectively; hind femur reticulate-rugose ventrally; setae of middle tarsus shorter than width of tarsal segments; fore and middle tarsal segments moderately slender; length of outer and inner spur of middle tibia 0.5 and 0.7 times middle basitarsus, respectively; outer side of middle tibia without pegs, except for 2 apical pegs; length of outer and inner spur of hind tibia 0.25 and 0.55 times hind basitarsus.
Metasoma.First tergite twice as long as wide apically, gradually widened apically, without dorsal carinae and smooth (Fig. 12h); second metasomal suture absent; ovipositor sheath 0.08 times as long as fore wing, truncate apically and widened.
Distribution.Known only from Thailand.For map showing the locality of the sole specimen, see Appendix I.

Figure 1 .
Figure 1.Map showing the collection sites in Thailand.

Figure 2 .
Figure 2. ML tree from the analysis of the COI-only data set with BI posterior probabilities (×100) and ML bootstrap values above the branches (left to right) and MP bootstrap values below the branches.Arrow points to a rogue exemplar of E. forticarinata.

Figure 3 .
Figure 3. ML tree from the analysis of the 28S-only data set with BI posterior probabilities (×100) and ML bootstrap values above the branches (left to right) and MP bootstrap values below the branches.Top to bottom, arrows point to a sequence of E. breviantennata identical to those of E. abbotti b rogue exemplars of E. forticarinata that may indicate a new species and c sequences of E. ophippium and E. pallitarsis which are identical to one another.

Figure 4 .
Figure 4. ML tree from the analysis of the COI+28S data set in which every taxon has both genes.BI posterior probabilities (×100) and ML bootstrap values are above the branches (left to right) and MP bootstrap values are below the branches.Arrow points to a rogue exemplar of E. forticarinata.

Figure 5 .
Figure 5. Euagathis abbotti (Ashmead), female.A lateral habitus B wings C dorsal habitus D dorsal mesothorax e propodeum F lateral mesosoma G base of hind wing H male, dorsal habitus i female, 10 th flagellomere from apex J male, lateral habitus.

Figure 6 .
Figure 6.Euagathis breviantennata sp.n., female, paratype.A lateral habitus B fore wing C hind wing D dorsal head e lateral head F lateral mesosoma G dorsal thorax H postero-lateral propodeum and anterior metasoma.
Bhat & Gupta, 1977 http://species-id.net/wiki/Euagathis_dravidaDistribution.For a map showing the locality of the sole Thai specimen, see Appendix I. India; Vietnam.New for Thailand.Molecular data.Genbank accession number 28S: DQ201905.

Figure 8 .
Figure 8. Euagathis dravida Bhat & Gupta, female.A Dorsal head B lateral head C wings D Lateral habitus e dorsal habitus F lateral mesosoma G dorsal scutellum and propodeum.

Figure 9 .
Figure 9. Euagathis forticarinata (Cameron).A female, lateral habitus B fore wing C hind wing D dorsal head e dorsal head F lateral mesosoma G dorsal thorax H propodeum i first metasomal tergite J female, lateral habitus showing variation, note color of hind tibia K male, lateral habitus l fore wing variation.

Figure 11 .
Figure 11.Euagathis pallitarsis sp.n., female, paratype.A lateral habitus B fore wing C hind wing D dorsal head e lateral head F dorsal thorax G dorsal first metasomal tergite H lateral mesosoma i dorsal propodeum.