A new Typhlocharis (Coleoptera: Carabidae: Anillina) from Spain: combining adult and larval morphological data with DNA information Author Andújar, Carmelo Author Grebennikov, Vasily V. Author Lencina, José Luís Author Serrano, José text Zootaxa 2010 2485 47 57 journal article 10.5281/zenodo.195522 eef3b882-6784-4746-a381-69c9821989e5 1175-5326 195522 Typhlocharis toletana Lencina & Andújar new species ( Figs. 1–5 ) Type series. Holotype : 1ɗ, 19–VI–2007 , Arroyo de Vitoria, Villarrubia de Santiago (province of Toledo, Spain ), 590 m (N 40, 0 2 W –3,30) C. Andújar and J.L. Lencina leg., deposited in the Departamento de Zoología y Antropología Física, Universidad de Murcia (ZAF–UMU). Paratypes : 52ɗ and 21Ψ, same locality and date, C. Andújar and J.L. Lencina leg. (2ɗ and 2Ψ gold coated); 79ɗ and 69Ψ, 15–IX–2007 same locality, C. Andújar, J.L. Lencina and J. Sánchez leg. Two paratypes , one male and one female are deposited in the collection of Museo Nacional de Ciencias Naturales, Madrid. The other paratypes are deposited in the Departamento de Zoología y Antropología Física, Universidad de Murcia (ZAF–UMU), and in J. L. Lencina collection, Museo de Ciencias Naturales de Jumilla (Murcia). Diagnosis : Anophthalmous. Body long, depressed and parallel ( Fig. 1 ), light brownish. Integument microreticulate with scattered short setae. Antennomeres 3–11 moniliform ( Fig. 2 c). Clypeus without median tooth. Pronotum ( Fig. 2 d) relatively slender (L/W = 1.2) with minute dentiform projection at hind angle and base arcuate. Each elytron with seven setae in the lateral umbilicate series distributed in two groups of 4+3 ( Fig. 2 j); no dentiform projection is found at apex or laterally at the end of the slight carina corresponding to seventh stria ( Fig. 2 h). Female with lateral slight fovea on first ventrite ( Fig. 2 i). Male genitalia ( Fig. 3 b) with median lobe slightly arcuate and ventral side almost straight. Female genitalia ( Fig. 3 a) with gonocoxite 2 of IX gonocoxa unguiform, with one internal seta and two external nematiform setae. Description of the adult : Length of holotype : 1.2 mm . Length of paratypes : 1.1–1.3 mm . Body long, depressed and parallel, light brownish. Integument microreticulate, mesh pattern isodiametric, with scattered short setae. Head capsule ( Fig. 2 a) robust, slightly narrower than pronotum and almost as wide ( 0.27 mm .) as long ( 0.29 mm .), slightly depressed in the middle of the frons, with two frontal divergent depressions delimiting it. Dorsal surface reticulate, except stridulatory area without microlines ( Fig. 2 f). Clypeus without median tooth. Anophthalmous. Appendages: antennomeres 1–2 cylindrical, 3–11 moniliform ( Fig. 2 c); labial mentum with one tooth and lateral lobes rounded ( Fig. 2 b); penultimate labial palpomere thickened, ultimate palpomere minute. Setation: clypeus with two setae, the external pair longer than the internal; cephalic capsule with 3–4 frontal setae, two median, and three large periorbital setae (supraorbital plus temporal); cephalic capsule and lateral carina with some other minor setae; labrum with five setae, three short and medial, one long and lateral and one external to the latter; labial mentum with two pairs of long setae, one close to the base of tooth and another on the lateral lobes, prebasally there are two other lateral pairs of setae. FIGURE 1. Typhlocharis toletana n. sp. , adult, habitus, dorsal view, total body length 1.2 mm. FIGURE 2. Typhlocharis toletana n. sp. , adult, SEM photographs, ( a) details of head, dorsal view; ( b) mouth parts, ventral view; ( c ) antenna; ( d ) pronotum, dorsal view; ( e ) right protibia, posterior view; ( f ) stridulatory area, internal view; ( g ) basal portion of elytra, dorsal view; ( h ) apex of elytra, dorsal view; ( i ) metathorax, ventral view; ( j ) left elytron, lateral view. FIGURE 3. Typhlocharis toletana n. sp. , adult, ( a ) ovipositor sclerites, ventral view; ( b ) male genitalia, left lateral view. Prothorax. Pronotum ( Fig. 2 d) longer ( 0.35 mm ) than wide ( 0.29 mm ), surface microreticulate, slightly trapezoidal with maximum width close to the apical margin. Disc flattened. Anterior margin slightly arcuate. Sides slightly crenulate only in the posterior quarter. Hind angle obtuse and distinctly marked with a small acute tooth. Posterior margin arcuate. One seta in anterior quarter of lateral margin and another on hind angle. Three longitudinal series of minute setae. Six pairs of strong setae on the anterior margin and two pairs of setae on the posterior margin. Both anterior and posterior margins with a row of tomentose setae. Prosternum with sparse and short setae. Elytra ( Fig. 2 g; 2h; 2j) twice longer ( 0.60 mm ) than wide ( 0.28 mm ), totally reticulate, with longitudinal carina at seventh stria slight and short, not extended to the apical margin of elytron (it disappears in the posterior fifth), and thus without forming there a small dentiform projection. Basal and lateral margins perpendicular, humeral region clearly marked and humeral angle smoothly rounded. Sides parallel with lateral margin slightly serrate to posterior quarter. Sutural angle not dentate ( Fig. 2 h). There are four longitudinal series of setae. Lateral margin with four setae in the anterior umbilicate group and three setae in the posterior group ( Fig. 2 j). First ventrite of female with slight lateral fovea ( Fig. 2 i). Legs similar in both sexes without special features such as dentiform projections or distally dilated femora. Front tibia with antenna cleaner (toilette organ) long and regularly arcuate, with one securiform clip seta at its proximal border ( Fig. 2 e), also reported in other species (e.g., T. gonzaloi : Ortuño 2005 ; T. martini Andujar et al. 2008 ). Trochanter long with rounded apex ( Fig. 3 a), similar in male and female. Median lobe of male genitalia ( Fig. 3 b) slightly curved, ventral side almost straight and dorsal side with a membranous appearance to the apex. Apex short and acute. Internal sac with a small sclerotized piece. Parameres with two apical setae. Gonocoxite 2 of IX gonocoxa of female genitalia unguiform ( Fig. 3 a), with one internal seta and two external nematiform setae. Spermatheca rounded, sclerotized, spermathecal gland distally dilated. Description of the larva: Material studied . Twenty apparently older-instar larvae were collected in the type locality, ten in June 19, 2007 and another ten in November 12, 2007 . Larvae were labelled “ SPAIN Villarrubia de Santiago Toledo 19.VI-–2007 Spring on a hillside 590m N 40, 0 2 W -3,30 Andújar and Lencina leg., and “ SPAIN Villarrubia de Santiago Toledo 15.IX-2007 Spring on a hillside 590 m N 40, 0 2 W -3,30 Andújar, Lencina and Sánchez leg.. Five larvae of these two dates were mounted on Euparal microscope slides and stored at the Canadian National Collection of Insects, Arachnids and Nematodes, Ottawa, Canada . Larvae are not designated as type specimens. Diagnosis . Three-segmented antennae is a unique feature readily distinguishing Typhlocharis from those of the vast majority of Carabidae genera, including the genus Geocharidius Jeannel, 1963 , the only other member of Anillina with described larvae ( Grebennikov 2002 ). Larval characters of T. toletana are similar to those described for Tyhlocharis sp. ( Arndt et al. 1999 ). FIGURE 4. Typhlocharis toletana n. sp. , older-instar larva, habitus, dorsal ( a, c ) and lateral ( b ) views, head, dorsal ( d ) and left dorso-lateral ( e ) views. Chaetotaxy nomenclature follows Bousquet and Goulet (1984). Description of the larva . Body length about 2.2 mm ( Figs. 4–5 ); maximal head width 0.173 mm ( 0.172– 0.175 mm , n = 5). Cephalic capsule almost parallel-sided and slightly elongated ( Figs. 4 , 5 a), without epicranial suture ( Fig. 4 d) and stemmata ( Fig. 4 e), mandible with a wide base, terebrum apically of retinaculum with one apical and large tooth, and one medial and small. All other characters of these larvae, including details of the chaetotaxy, are identical with a Typhlocharis larva previously described by Arndt et al. (1999) . Molecular analysis: ModelTest showed that GTR+G was the best-fit substitution model, with a γ value of 0.1603. A pairwise divergence analysis conducted in PAUP with this parameter showed that divergences between the adults and the putative larva of T. toletana are low, between 0 and 0.013%, while divergences among different Typhlocharis species vary from 15% to 26%. Divergences between Typhlocharis and Hypotyphlus are around 30% ( Table 2 ). FIGURE 5 . Typhlocharis toletana n. sp. , older-instar larva, ( a ) head, thorax and abdominal segments I–II, ventral view; ( b ) pro-, meso- and metathorax, left dorso-lateral and ( c ) dorsal view; ( d ) abdominal segments VII–X and urogomphi, dorsal view. Chaetotaxy nomenclature follows Bousquet & Goulet (1984). Habitat : The type locality of Typhlocharis toletana n. sp. is a spring on a lateral hillside facing Arroyo de Vitoria, a stream with permanent water in white loamy soil area close to Villarrubia de Santiago (Toledo, Spain ). Type locality is 1.5 km apart from the main course of river Tajo. There was only herbaceous vegetation, mainly composed of grass, like Brachypodium sp. TABLE 2. Pairwise distances between cox1 sequences of 5 species of Typhlocharis and one Hypotyphlus . Analyses were conducted in PaupUp using the nucleotide substitution model GTR+G (γ= 0.1602), calculated with ModelTest 3.7. T. toletana T. toletana T. toletana T. diecki T. estrellae T. armata T. martini larva adult 1 adult 2 T. toletana larva - T. toletana adult 1 0.000 - T. toletana adult 2 0.003 0.001 - T. diecki 0.205 0.201 0.210 - T. estrellae 0.209 0.209 0.209 0.200 - T. armata 0.265 0.265 0.265 0.216 0.201 - T. martini 0.257 0.257 0.256 0.232 0.156 0.169 - Hypotyphlus navaricus 0.319 0.320 0.331 0.302 0.289 0.288 0.248 Etymology : The specific epithet is a noun in genitive case deriving from Toletum (=Toledo), the province in central Spain where the new species has been found. Affinities: Adults of T. toletana n. sp. share with those of T. monastic a Zaballos and Wrase 1998 and T. peregrina Zaballos and Wrase 1998 the unguiform type genitalia. However, T. toletana can be easily distinguished from them by the absence of tooth at the posterior margin of elytra and the umbilicate series of 4+3 type (as compared to 4+2 type found in the cited species). These and other characters suggest the inclusion of T. toletana in the T. diecki species group, whereas the lack of teeth in the posterior half of the lateral margin plus the sutural angle of elytra is characteristic to the T. silvanoides species group. Presently described T. toletana larva cannot be reliably distinguished from the congeneric larvae described by Arndt et al. (1999) and, therefore, are not informative in clarifying species relationships. Affinities of the new species are, therefore, ambiguous.