Taxonomy, Host-Plant Associations and Phylogeny of African Crotalaria

Zootaxa 3895 (2): 238–256 ISSN 1175-5326 (print edition) www.mapress.com/zootaxa/ Article ZOOTAXA Copyright © 2014 Magnolia Press ISSN 1175-5334 (online edition) http://dx.doi.org/10.11646/zootaxa.3895.2.6 http://zoobank.org/urn:lsid:zoobank.org:pub:ACF6C45F-E37F-4483-9001-17634FCE5990 Taxonomy, host-plant associations and phylogeny of African Crotalaria-feeding seed beetles (Coleoptera, Chrysomelidae, Bruchinae): the Conicobruchus strangulatus (Fåhraeus) species group

BRUNO P. LE RU1,2, ALEX DELOBEL3, ZOLTÁN GYÖRGY4, GWENAËLLE GENSON5 & GAEL J. KERGOAT5,6 1Unité de Recherche IRD 072, African Insect Science for Food and Health (icipe), PO Box 30772, Nairobi, Kenya. E-mail: [email protected] 2IRD/CNRS, Laboratoire Evolution Génomes Spéciation, Avenue de la terrasse, BP1, 91198, Gif-sur-Yvette, France and Université Paris-Sud 11, 91405 Orsay, France 3Muséum National d’Histoire Naturelle, 45 rue Buffon, 75005 Paris, France. E-mail: [email protected] 4Department of Zoology, Hungarian Natural History Museum, H-1088 Budapest, Baross u. 13, Hungary. E-mail: [email protected] 5INRA - UMR 1062 CBGP (INRA/IRD/Cirad, Montpellier SupAgro), 755 Avenue du Campus Agropolis, 34988 Montferrier-sur-Lez, France. E-mail: [email protected] 6Corresponding author

Abstract

A small group of six morphologically related seed beetles (Coleoptera: Chrysomelidae: Bruchinae) belonging to the Con- icobruchus genus is reviewed. Species in this group for which host-plants are known feed on various species of Crotalaria (Fabaceae, Crotalarieae). Here we provide diagnoses and a dichotomous key for all six species. The following synonymies are proposed: Conicobruchus cicatricosus (Fåhraeus, 1839) (= Bruchus cicatricosus pallidioripennis Pic, 1941) syn. nov.; Conicobruchus strangulatus (Fåhraeus, 1839) (= Bruchus hargreavesi Pic, 1933) syn. nov. The corresponding Conico- bruchus strangulatus species group is hereby designated. New host-plant data are also included, which correspond to the results of recent collections of legume pods in East Africa. In addition we carried out molecular phylogenetic analyses on a representative sampling of Conicobruchus species (including the six species of interest). The latter allow us to assess the monophyly of the group of interest and to unravel their evolutionary relationships. Molecular phylogenetic analyses also indicate that at least two lineages of Conicobruchus successfully shifted toward Crotalarieae during the course of their diversification.

Key words: Crotalarieae, host-plants, molecular phylogenetics, morphology, systematics

Introduction

Seed beetles (Coleoptera, Chrysomelidae, Bruchinae) constitute a subfamily of moderate size that encompass about 1,700 species (Johnson et al. 2004). Bruchines are found in all continents (except Antarctica) and are more diverse in tropical regions (Southgate 1979). Their larvae are endophagous and usually develop inside seeds. Though collectively seed beetles are known to be able to attack more than 32 plant families, the majority of species are specialized on legume (Fabaceae) seeds (Johnson 1970, 1981, 1989). Almost all seed beetle species have a narrow host-plant range, and feed on a restricted set of plant species that usually belong to the same genus or botanical tribe (Johnson 1989). Phylogenetically related bruchine species usually feed on plants from the same tribe, a pattern referred as taxonomic conservatism in host-use (Kergoat et al. 2004, 2007). The history of bruchine taxonomy and systematics is complex (Borowiec 1987), especially because seed beetle groups have been usually defined on the basis of external morphological characters that are often homoplasious (Kergoat & Silvain 2004). It is only in last three decades that internal morphology (especially male genitalia) has been systematically taken into

238 Accepted by M. Schoeller: 17 Nov. 2014; published: 15 Dec. 2014 account in species description and classification. With the advent of molecular methods, significant advances have also been made as sequences are available for more and more species (Kergoat et al. 2008), allowing a better assessment of species boundaries and evolutionary relationships (e.g. Delobel et al. 2013). Decelle (1951) erected the Conicobruchus genus for a small number of African species previously assigned to the Bruchus genus: B. atrosuturalis Pic, B. bedfordi Pic, B. flabellicornis Boheman, and B. strangulatus Fåhraeus, the type species. This genus is characterized by a specific shape of the pronotum, which is conical with sides slightly concave in C. strangulatus and related species (Decelle 1951; Borowiec 1987). Since then, a number of additional species have been described (Decelle 1958) or included (Arora 1977; Singal & Pajni 1986; Borowiec 1987) in the genus (see Kergoat et al. 2011 for details). Kergoat et al. (2011), based on molecular, morphological and host association data, specified the limits of the genus and transferred 27 species from Bruchus and Bruchidius to Conicobruchus. Conicobruchus species with known host-plants are generally associated with Indigofereae (Fabaceae) but four species develop on Crotalaria (Fabaceae: Crotalarieae); three in Africa and one in the Oriental region (C. indicus (Pic)). Molecular phylogenetic analyses (Kergoat et al. 2011) underscored the monophyly of a group of three African Crotalaria-feeding species: C. atrosuturalis (Pic), C. cicatricosus (Fåhraeus), and C. strangulatus (Fåhraeus). However, in the corresponding study Kergoat et al. (2011) confounded C. flabellicornis (Boheman) with C. atrosuturalis because they overlooked the taxonomic significance of white pronotal spots in C. flabellicornis. Specimens of C. flabellicornis were thus labeled as C. atrosuturalis while the only specimen of C. atrosuturalis was considered as an unknown species related to C. strangulatus. At the time, the host of C. flabellicornis (“Indigofera inyazura” according to Decelle 1975) was considered doubtful because I. inyazura is not a valid botanical name (ILDIS 2014; TROPICOS 2014). Recently, an original set of ecological data was collected in East Africa by one of us (BLR). It enables us to review, precise and emend previously published results on species that are associated with Crotalarieae. Collections and sequencing of additional specimens (including species that have not been sequenced previously) also allow us to provide a more comprehensive phylogenetic framework that encompasses all known Crotalaria-feeding Conicobruchus seed beetles. A key for the identification of adults is also presented based on external morphology, in particular on color patterns. Abbreviations: CBGP, Centre de Biologie pour la Gestion des Populations, Montferrier/Lez, France; HNHM, Hungarian Natural History Museum, Budapest, Hungary; MNHN, Muséum National d’Histoire Naturelle, Paris, France; OÖLM, Oberösterreichisches Landesmuseum, Linz, Austria

Material and methods

Sampling. African Conicobruchus specimens were obtained through large-scale collection of Crotalaria pod samples in Kenya (C. incana L. subsp. purpurascens (Lam.) Milne-Redh., C. laburnifolia subsp. tenuicarpa Polhill, C. polysperma Kotschy, C. uguenensis Taub. and a number of unidentified Crotalaria species), in Mali (C. glaucoides Baker f., C. cf goreensis Guill. & Perr.), in Senegal (C. comosa Baker; C. glaucoides; C. perrottetii DC.; C. podocarpa DC.), in Togo (C. goreensis Guill. & Perr.), and in Zambia (C. aculeata De Wild. subsp. claessensii (De Wild.) Polhill). Adult seed beetles were reared from the collected pods and later preserved in 95- 100% ethanol. Our sampling also includes an old dried specimen of C. indicus that was kindly loaned to us by the HNHM. The inclusion of this species was important because C. indicus specimens have been reared from Crotalaria juncea L. seeds (Arora 1978). Morphological studies. All specimens were compared with type specimens preserved in MNHN collections. Examination of external structures was carried out under a stereoscopic microscope (Wild MZ8). After dissection, genitalia were heated in hypertonic NaOH solution, and examined under a light microscope (Leitz Laborlux K). For figures, digital photographs of microscope preparations were taken using a hand-held Canon Powershot G3 camera, and transferred to a vector graphics-editing program. After extraction, genitalia were cleared in heated hypertonic NaOH solution and temporarily mounted in water-soluble DMHF (dimethyl hydantoin formaldehyde). DNA extraction and sequencing. For this study, 24 specimens from 13 Conicobruchus species were included in the molecular dataset (see Table 1), of which three that were not represented in Kergoat et al. (2011): C. decoratus (Fåhraeus), C. medaniensis (Decelle) and C. indicus. As underlined beforehand all species that were known to feed on Crotalaria are included (C. atrosuturalis, C. cicatricosus, C. indicus and C. strangulatus). We also used the study of Kergoat et al. (2011) to select a balanced set of outgroup taxa. This set included two taxa that

THE CONICOBRUCHUS STRANGULATUS GROUP Zootaxa 3895 (2) © 2014 Magnolia Press · 239 are closely related to the Conicobruchus genus (Bruchidius biguttatus (Olivier) and Bruchidius cisti (Fabricius)), four taxa that are more distantly related (Bruchidius atrolineatus (Pic), Bruchidius villosus (Fabricius), Bruchus pisorum (L.) and Megabruchidius tonkineus (Pic)) and a representative of a distinct tribe (Pachymerus cardo (Fåhraeus), from tribe Pachymerini), which was used to root the tree. Non-destructive DNA extractions of new specimens were conducted using Qiagen DNAeasy tissue kits (Qiagen, Hilden, Germany). Polymerase chain reaction (PCR) amplifications were conducted for four mitochondrial gene fragments: 1,008 bp of the cytochrome oxidase I (COI), 778 bp of the cytochrome b (Cytb), 406 bp of the ribosomal 12S RNA (12S), and 542 bp of the ribosomal 16S RNA (16S). Four nuclear gene regions were also sequenced: 778 bp of the 28S ribosomal DNA domain D2-D3 (28S-D2D3), 698 bp of the 28S ribosomal DNA domain D4-D5 (28S-D4D5), 687 bp of the 28S ribosomal DNA domain D6-D7 (28S-D6D7), and 1,836 bp of the 18S ribosomal DNA (18S). For the 28S-D6D7 domain we used the set of primers and the settings detailed in Ceotto et al. (2008). For the 18S gene, three overlapping regions were sequenced following Kergoat et al. (2014). For all remaining genes, we used the primers and settings detailed in Kergoat et al. (2011). Newly generated PCR products were processed by Eurofins MWG Synthesis GmbH (Ebersberg, Germany). Both strands were sequenced for all specimens to minimize PCR artefacts and ambiguities. Sequences of complementary strands were edited and reconciled using Geneious v5.1 software (available at: www.geneious.com/). All the sequences generated in this study were deposited in GenBank (see Appendix S1 for the accession numbers). Unlike the sequences of coding genes (COI and Cytb), the sequences of ribosomal genes were variable in length. Their alignment was accomplished using MAFFT 7 (Katoh & Standley 2013) with default option settings. For all protein-coding genes, we used Mesquite 2.75 (available at: www.mesquiteproject.org) to check the coding frame for possible errors or stop codons. The combination of the eight gene fragments resulted in a combined matrix of 31 specimens and 6,733 aligned characters. Phylogenetic analyses. Phylogenetic analyses were conducted using Maximum likelihood (ML), as implemented in RAxML v.7.4.2 (Stamatakis 2006). We used partitioned analyses (Nylander et al. 2004) with one partition for the mitochondrial genes and one partition for the nuclear genes. For each partition, we combined a general time reversible (GTR) substitution model with a CAT (category) model, which optimizes the evolutionary rate of individual sites using a fixed number of rate categories. The best tree was obtained using a heuristic search implementing 100 random-addition replicates. Clade support was then assessed using non-parametric bootstrap values (BV) (1,000 replicates were used). Nodes supported by BV > 70% were considered as strongly supported following Hillis and Bull (1993).

Results

Ecological data and morphological studies. A total of six distinct species was reared from the sampled Crotalaria pods. Three of them (Conicobruchus atrosuturalis, C. cicatricosus, and C. strangulatus) were known to feed on Crotalaria but for the remaining three (C. decoratus, C. flabellicornis and C. rubricollis) our records constitute their first verified host-plant records. All these species are morphologically related and are assigned to the Conicobruchus strangulatus species group. The group is defined by the following set of characters: (i) integument from largely red with various black markings to entirely black (dark red brown in some specimens); (ii) antenna black (rarely reddish basally), sexually dimorphic, with segments 4 or 5 to 10 serrate in female, pectinate in male; (iii) eyes bulging, frons with strong median carina; (iv) pronotum subtriangular, with largest width at base, anteriorly produced into a neck or not (C. decoratus); (v) elytra without teeth at base of interstriae; (vi) legs not sexually dimorphic; (vii) hind femur with small preapical denticle, its ventral side more or less flattened and limited by external and internal carinae; (viii) hind tibial apex with mucro shorter than lateral coronal tooth; (ix) abdomen simple, ventrite 1 without basocentral brush of erect setae; (x) last visible tergite shield-shaped, subvertical in male, slanted in female; (xi) median lobe moderately stout, widened apically or not (C. decoratus); (xii) ventral valve subtriangular; (xiii) internal sac with spines, often with dorsal and/or ventral dented sclerites (absent in C. decoratus and C. rubricollis); (xiv) no hinge sclerites; (xv) lateral lobes very briefly fused basally, their apex modified, with more or less defined dorsal and ventral lips; (xvi) tegminal strut without median longitudinal carina. All known species from the group are Afrotropical and are strictly associated with the genus Crotalaria (Crotalariae).

240 · Zootaxa 3895 (2) © 2014 Magnolia Press LE RU ET AL. Morphological traits (both external and genital) that enable discrimination between the species in the group are outlined in the following paragraphs. For each species of the group we also provide bibliographic references corresponding to original descriptions, synonymies and new combinations.

TABLE 1. Taxon sampling. If known, rearing information is provided (otherwise the abbreviation N/A is used). Species voucher code Country Reared from

Conicobruchus albopubens (Pic 1931) BRU.Si11 Kenya Indigofera aspera Conicobruchus atrosuturalis (Pic 1939) GK35 Kenya Crotalaria uguenensis Conicobruchus cicatricosus (Fahraeus 1839) GK171 Zimbabwe N/A Conicobruchus cicatricosus (Fahraeus 1839) GK195 Kenya Crotalaria sp. Conicobruchus cicatricosus (Fahraeus 1839) GK448 Kenya Crotalaria laburnifolia Conicobruchus decoratus (Fahraeus 1871) GK446 Zambia Crotalaria aculeata Conicobruchus flabellicornis (Boheman, 1829) GK200 Kenya Crotalaria incana Conicobruchus flabellicornis (Boheman, 1829) GK243 Kenya Crotalaria incana Conicobruchus flabellicornis (Boheman, 1829) GK442 Kenya Crotalaria brevidens Conicobruchus flabellicornis (Boheman, 1829) GK452 Kenya Crotalaria incana Conicobruchus indicus (Pic 1909) GK695 India N/A Conicobruchus kidevuensis (Delobel & Le Ru 2010) GK156 Tanzania Indigofera basiflora Conicobruchus lineatopygus (Pic 1924) BRU.L21 Senegal Indigofera tinctoria Conicobruchus medaniensis (Decelle 1982) GK447 Tanzania Indigofera tanganyikensis Conicobruchus medaniensis (Decelle 1982) GK575 Tanzania N/A Conicobruchus nigricornis (Fabricius 1801) GK107 Kenya Indigofera sp. Conicobruchus nigricornis (Fabricius 1801) GK198 Cameroon Indigofera arrecta Conicobruchus rubricollis (Pic 1903) GK196 Kenya N/A Conicobruchus rubricollis (Pic 1903) GK406 Kenya Crotalaria cf. polysperma Conicobruchus rubricollis (Pic 1903) GK440 Kenya Crotalaria cf. polysperma Conicobruchus skaifei (Pic 1928) GK92 Kenya N/A Conicobruchus strangulatus (Fahraeus 1839) BRU.Xh11 Senegal Crotalaria podocarpa Conicobruchus strangulatus (Fahraeus 1839) GK116 Mali Crotalaria cf. goreensis Conicobruchus subdolus (Delobel & Le Ru 2010) GK245 Kenya Indigofera arrecta

Bruchidius atrolineatus (Pic 1921) BRU.Aa11 Senegal Vigna unguiculata Bruchidius biguttatus (Olivier 1795) BRU.B21 Greece N/A Bruchidius cisti (Fabricius 1775) BRU.Ch21 Italy N/A Bruchidius villosus (Fabricius 1792) BRU.V21 France Laburnum anagyroides

Bruchus pisorum (Linnaeus 1758) BRU.Zp11 Italy Pisum sativum

Megabruchidius tonkineus (Pic 1904) BRU.Xi11 Vietnam Gleditsia australis

Pachymerus cardo (Fahraeus 1839) BRU.Xj14 Fr. Guiana Elaeis guineensis

Bruchus atrosuturalis Pic, 1939:31. Conicobruchus atrosuturalis: Decelle 1951:181.

Material examined. Paratype (male) of Bruchus atrosuturalis var. Pic: Ethiopia: ix.1936 (genitalia on card in drop of DMHF) [MNHN] Other material: Kenya: 1♂, Chyulu Hills, vii.1938 [1♂ 00514] Coryndon Museum /Expdt Chyulu Hills /July 38 alt. 5600, Imp. Inst./Entom [MNHN]; 1♂, Kakamega, 00°10.930N 34°56.137E, 1693m, ex Crotalaria incana purpurescens, 0011.xi.2008 [1♂ 01109] (B. Le Ru) [MNHN]; 1♂, 2♀, Magadi, 01°46.142S 36°20.657E, 856m, i.2007, ex Crotalaria uguenensis [1♂ 02307, specimen GK35 used for DNA extraction] (B. Le Ru) [CBGP].

FIGURE 1. Dorsal view of specimens belonging the to six species of the Conicobruchus strangulatus species group.

242 · Zootaxa 3895 (2) © 2014 Magnolia Press LE RU ET AL. FIGURE 2. Conicobruchus atrosuturalis: 2—median lobe (Paratype, Ethiopia); arrows were used to show the following structures: 2A—minute spinules; 2B—ctenoid scales; 2C—small sclerotized teethes; 2D—strong ventro-lateral dented rods; 2E—teethes; 2F—dented sticks or masses; 2G—apical ampoule.

Body black, except elytra bright yellowish-red with black markings along suture (Fig. 1), also along apex of elytra in some specimens; ventrites dark reddish brown to black in typical form, red in paratype and Kenyan specimens reared from Crotalaria uguenensis seeds; last visible tergite red or black.

THE CONICOBRUCHUS STRANGULATUS GROUP Zootaxa 3895 (2) © 2014 Magnolia Press · 243 Major morphological traits are as follows: pronotum 1.3 times wider at base than long, its sides straight basally, strongly convergent anteriorly, with a distinct neck; lack of white hair spots on pronotum (setation uniformly pale greyish or yellowish); elytral striae with small, closely spaced punctuation, interstriae with punctures varying from strong to shallow. Male genitalia. Median lobe (Fig. 2) similar with that of Conicobruchus strangulatus (Fig. 10), stout (maximum width excluding basal hood / total length = 0.19), slightly widened apically, basal hood moderately widened, concave posteriorly; ventral valve large, subtriangular, with apex acute, bearing numerous sensillae and two lateral groups of 5 to 8 setae; no hinge sclerite. Internal sac densely lined anteriorly with minute spinules (Fig. 2A) and ctenoid scales (Fig. 2B), followed by an area almost devoid of ornamentation, then dense scales and spines blending dorsally into a short series of well sclerotized small teeth (Fig. 2C); a pair of strong ventro-lateral dented rods (Fig. 2D) similar to those observed in Conicobruchus nodieri and C. astragalinae, and between them zero to three isolated teeth (Fig. 2E); posteriorly a zone with a few strong, isolated teeth (Fig. 2E) and a number of setae; then again two or more groups of dented sticks or masses (Fig. 2F); apical ampoule (Fig. 2G) devoid of any ornamentation, gonopore not sclerotized; basal strut narrow, without keel. Lateral lobes cleft to nearly 90% of their length; apex modified, with two lips, the dorsal one bearing about 12-15 long setae, the ventral one with a dense group of short sensillae. Biology. Examined material was reared from pods of Crotalaria uguenensis and C. incana subsp. purpurescens in Kenya. Distribution. Ethiopia and Kenya. Discussion. Conicobruchus atrosuturalis has a very specific color pattern with a black body and bright orange elytra (Fig. 1). This pattern can only be found in red specimens of C. flabellicornis; the two species can be easily distinguished based on the presence of brushes and spots of dense white setae on the pronotum (absent in C. atrosuturalis, present in C. flabellicornis).

Conicobruchus cicatricosus (Fåhraeus, 1839)

Bruchus cicatricosus Fåhraeus, 1839:39. Bruchus cicatricosus pallidioripennis Pic, 1941:12, syn. nov. Conicobruchus cicatricosus: Kergoat et al., 2011:756

Material examined. Kenya: 2♂, Josa Wundanyi, 03°25.863S 31°21.314E, 1378m, 13.iv.2012, ex Crotalaria laburnifolia tenuicarpa [1♂ 02212, specimen GK448 used for DNA extraction] (B. Le Ru) [CBGP]. Tanzania: 1♂, Kidevu, 03°09.402S 36°41.058E, 1818m, 25.ii.2008, ex Crotalaria sp. [specimen GK195 used for DNA extraction] (B. Le Ru) [CBGP]. Republic of South Africa: 1♂, 1♀, Port Elisabeth [1♂ 04908]; 1♀, East London, ix.1915 (R. Ellenberger) [MNHN]. Tanzania: 1♂, xii 7n fv, ex Crotalaria sp. [1♂ 00508] (C. Conrads) [MNHN]. Zimbabwe: 1♂, 1♀, Masvingo, 22.i.1998 [1♂ 19007, specimen GK171 used for DNA extraction] (M. Halada) [OÖLM]. Body entirely black, or black with elytra and legs more or less light reddish brown (Fig. 1). The black form corresponds to variety A of Fåhraeus, the paler form to his variety B, as well as to Pic’s variety pallidioripennis. These are mere colour forms of the same species. Other important morphological traits are as follows: pronotum 1.2 times wider at base than long, its sides straight basally, strongly convergent anteriorly, then constricted into a distinct neck; lack of white hair spots on pronotum (setation uniformly yellowish); elytral striae thin and narrow, interstriae alternating greyish setose spots with bare areas surrounding large ocellate punctures, elytral disc thus showing a checkered pattern, less visible laterally and posteriorly. Male antennae strongly pectinate. Male genitalia. Median lobe (Fig. 3) of moderate length, stout (maximum width excluding basal hood / total length = 0.20), slightly widened apically, basal hood moderately widened, concave posteriorly; ventral valve large, subtriangular, bearing apically numerous sensillae and basally two lateral groups of 6 to 9 setae; no hinge sclerite; internal sac densely lined anteriorly with minute spinules and ctenoid scales, followed by strands of hyaline spicules and tubercles, progressively transformed laterally into a pair of groups of very dense setae; posteriorly numerous strong teeth, associated or not in two to four dented sticks or masses; apical ampoule devoid of any ornamentation, gonopore not sclerotized; basal strut (Fig. 4) narrow, without keel; lateral lobes cleft to about 90%

244 · Zootaxa 3895 (2) © 2014 Magnolia Press LE RU ET AL. their length; apex modified, with two lips, the dorsal one bearing 15-20 long setae, the ventral one densely lined with long setation. Biology. The type series was reared from Crotalaria volubilis seeds in ‘Caffraria’ (now Republic of South Africa). The name C. volubilis Thunberg is not valid (ILDIS 2014; TROPICOS 2014), so that the actual host remains unknown (but C. capensis according to De Luca, 1965). Examined material was reared from pods of Crotalaria laburnifolia subsp. tenuicarpa in Kenya and of Crotalaria sp. in Tanzania. Distribution. Kenya, Republic of South Africa, Tanzania and Zimbabwe. Discussion. Conicobruchus cicatricosus can be distinguished from other members of the group by its elytral setation (Fig. 1), in which bare spots alternate with densely setose patches (in other species elytra are uniformly covered with setae). Contrary to C. decoratus and C. rubricollis, the male genitalia of C. cicatricosus includes large sclerites. However, in contrast with C. atrosuturalis, C. flabellicornis and C. strangulatus, proximal sclerites are absent.

FIGURES 3–4. Conicobruchus cicatricosus: 3—median lobe; 4—basal strut and lateral lobes (specimen 02212, Kenya).

Bruchus decoratus Fåhraeus, 1871:448 Bruchus decoratus atrodorsalis Decelle, 1960:137, syn. nov. Bruchus nigromaculatus Pic, 1929 (synonymy in Decelle 1961: 8) Bruchidius decoratus Decelle, 1975:21 Conicobruchus decoratus: Kergoat et al., 2011:756

Material examined. Type (male) of Bruchus nigromaculatus: Republic of South Africa: Port St John, v.1923, R.E. Turner [dissected, genitalia on card in DMHF]. Other material: Botswana: 1♀, Leribé, 1923 (R. Ellenberger) [MNHN]. Republic of Mozambique: 1♂, Nova Chupanga, v.1928 (P. Lesne) [MNHN]. Tanzania: 2♂ (C. Conrads) [MNHN]. Zambia: 1♂, 2♀, Samununga, 13°37.156’S 24°07.349’E, 1091m, 19.iii.2012, ex Crotalaria aculeata claessensii, [1♂ 02012, specimen GK446 used for DNA extraction] (B. Le Ru) [CBGP].

FIGURES 5–7. Conicobruchus decoratus: 5—median lobe; 6—basal strut and lateral lobes (specimen 02012, Zambia); Conicobruchus flabellicornis: 7—median lobe (specimen 01209, Kenya).

246 · Zootaxa 3895 (2) © 2014 Magnolia Press LE RU ET AL. Body black, with large red parts; head (including antennae), thoracic sternites and legs always black, pronotum and abdomen always red. Elytra entirely black in some specimens, but usually red with more or less extensive black markings (Fig. 1): usually base, humerus, a round spot at basal fourth in interstriae 2 to 4, apical fourth to third of elytra, often also along suture. Other important morphological traits are as follows: pronotum 1.5 times wider at base than long, its sides almost straight, slightly sinuated, not constricted anteriorly into a neck; moderately lobed posteriorly and laterally, with dense white setation on lobes; strial punctures small, circular, closely spaced, interstriae finely imbricate, shining, with minute punctures; male antennae serrate. Male genitalia. Median lobe (Fig. 5) of moderate length, stout (maximum width excluding basal hood / total length = 0.15), not widened apically, basal hood moderately widened, not concave posteriorly; ventral valve large, subtriangular, bearing apically numerous sensillae and basally two lateral groups 9–12 setae; no hinge sclerite; internal sac anteriorly with minute spinules and scales, then densely lined with strands of hyaline and sclerotized spines; apical ampoule devoid of any ornamentation, gonopore not sclerotized; basal strut (Fig. 6) narrow, without keel; lateral lobes cleft to about 90% their length; apex modified, with two lips, the dorsal one bearing about 15 setae, the ventral one densely lined with long setation. Biology. Zambian material was reared from pods of Crotalaria aculeata subsp. claessensii. This constitutes the first report on its biology. Distribution. Botswana, Democratic Republic of Congo (Decelle, 1960), Republic of Mozambique, Republic of South Africa, Tanzania and Zambia. Discussion. Conicobruchus decoratus can be distinguished from other members of the C. strangulatus species group by the shape of the pronotum and structure of male genitalia. Contrary to the other species of the group, in C. decoratus the pronotum is short, its side non-concave and it is not compressed anteriorly into a neck (Fig. 1). Male genitalia are also very distinctive, being less widened anteriorly and the basal strut not concave posteriorly (Fig. 5). Specimens with black elytra were considered as a subspecies (atrodorsalis) by Decelle (1960). However the pattern of elytral coloration is extremely variable, ranging from red with few black markings to entirely black. Because of this high level of variability we chose to establish the synonymy Bruchus decoratus atrodorsalis Decelle = Conicobruchus decoratus (Fåhraeus).

Conicobruchus flabellicornis (Boheman, 1829)

Bruchus flabellicornis Boheman, 1829:105 Bruchus antonioi Pic, 1943:6 (synonymy in Decelle, 1975:18) Conicobruchus flabellicornis: Decelle, 1951:181

Material examined. Holotype (female) of Bruchus antonioi: Angola: San Antonio (now Soyo), 1929 (H. Brauns) [MNHN]. Other material: Kenya: 3♂, 4♀, Suam, 01°11.982’N 34°49.538E, 1951m, 12.xi.2008, ex Crotalaria incana [2♂ 01209, 02312, specimen GK200 used for DNA extraction] (B. Le Ru) [CBGP]; 1♀, Kakamega, 00°10.990’N 34°58.137E, 1693m, 11.xi.2008, ex Crotalaria incana purpurascens [specimen GK243 used for DNA extraction] (B. Le Ru) [CBGP]; 1♂, 3♀, Burguret, 00°04.926’N 37°02.203E, 1915m, 21.xi.2011, ex Crotalaria incana purpurascens [1♂ 02112, specimen GK452 used for DNA extraction] (B. Le Ru) [CBGP]; 2♂, 1♀, same data, but ex Crotalaria brevidens intermedia [1♂ 02412, specimen GK442 used for DNA extraction] (B. Le Ru) [CBGP]. Togo: 1♀, Lomé, 28.viii.1995, ex Crotalaria goreensis (I.A. Glitho) [MNHN]. Black, but Kenyan specimens often with elytral disc dark red. A pod sample collected in Suam yielded two black and five “red” specimens, whereas one sample from Burguret yielded only “red” specimens. Color is apparently unrelated with sex. Other major morphological traits are as follows: pronotum 1.37 to 1.43 times wider at base than long, with oblique lateral impression, constricted anteriorly into a wide neck, strongly rugose-punctate, with several white hair spots. These were described in detail by Fåhraeus (1839:4): a small spot in the ante-scutellar fossa, a short line just before apex, at posterior angles, and a lateral spot on each side; an additional pair of white spots is often visible on sides of basal lobes, scutellum with similar white setation. Elytral striae strong, with small punctures, interstriae flat, with series of shallow circular punctures.

Male genitalia. Median lobe (Fig. 7) of moderate length, stout (maximum width excluding basal hood / total length = 0.17), widened apically, basal hood widened, concave posteriorly; ventral valve large, subtriangular, with apex acute, bearing a few sensillae and two lateral groups of 3–5 setae; no hinge sclerite; internal sac without central column of tubercules, but with rather dense hyaline scales and tubercles, ending posteriorly in a dorsal mass of small, poorly sclerotized teeth, blending into a short series of well sclerotized teeth of variable length; a pair of strong ventro-lateral dented rods; between these a few isolated teeth; posteriorly two poorly defined groups of dented masses; apical ampoule devoid of any ornamentation, gonopore unsclerotized; basal strut narrow, without keel; lateral lobes cleft to 85% their length; apex modified, with two lips, the dorsal one bearing about twelve long setae, the ventral one with a dense group of short sensillae.

248 · Zootaxa 3895 (2) © 2014 Magnolia Press LE RU ET AL. Biology. Examined material was reared from pods of Crotalaria goreensis and Crotalaria incana subsp. purpurascens. This constitutes the first verified record on its biology. Distribution. Angola, Burundi (Decelle, 1956), Democratic Republic of Congo (Decelle, 1951), Kenya, Sierra Leone (Fåhraeus, 1839) and Togo. Discussion. Conicobruchus flabellicornis can be distinguished form all other members of the group based on the presence of brushes and spots of dense white setae on the pronotum (present in C. flabellicornis, absent in other species)(Fig. 1).

Conicobruchus rubricollis (Pic, 1903)

Bruchus rubrithorax Pic, 1903:169 Bruchus rubricollis Pic, 1913:45 (name preoccupied) Conicobruchus rubricollis: Kergoat et al., 2011:756

Material examined. Male paratype, Zimbabwe: Harare (with label “Bruchidius rubrithorax Pic” handwritten by J. Decelle) [MNHN]. Other material: Kenya: 2♂, Thange Town, 02°34.738’N 37°57.129’E, 904m, 24.iv.2011, ex Crotalaria cf. polysperma [1♂ 03211, specimens GK406 and GK440 used for DNA extraction] (B. Le Ru) [MNHN]; 1♂, SE Kenya, Voi (Tsavo), 23.3.-4.4.1997; 2♂, Kenya mer., Tsavo East, Buchuma env., 28.xi.1997 [specimen GK196 used for DNA extraction] (M. Snizek) [OÖLM]; 1♀, Kenya, S. Taita, Mwatate, 30.xi.1997 (M. Snizek) [OÖLM]. Zimbabwe: 1♂, Masvingo, 22.i.1998 [1♂ 18907] (M. Halada) [OÖLM]. Large species well defined by its mainly red colour, with major part of antennae, elytra, and often disc of mesothoracic sternite, black (Fig. 1); pubescence white, denser on median longitudinal line of pronotum, scutellum, base of elytral suture; pygidium with two lateral spots near base and a longitudinal line. Other major morphological traits are as follows: pronotum 1.5 times wider at base than long, without oblique lateral impression, its sides concave, produced anteriorly into a neck; elytral striae narrow and deep, with small punctures, interstriae wide and flat, with small punctures on shagreened background; antennae serrate. Male genitalia. Median lobe (Fig. 8) of moderate length (maximum width excluding basal hood / total length = 0.23), strongly widened and sclerotized apically; basal hood oval, apically indented; ventral valve large, subtriangular, with apex acute, with numerous sensillae, bearing two lateral groups of 10–12 setae; dorsal valve unsclerotized; no hinge sclerites; anterior part of the internal sac lined with hyaline spinules becoming denser and larger distally, then large ctenoid scales, posterior saccus with numerous sclerotized teeth; apical ampoule unarmed. Tegminal strut (Fig. 9) without keel, triangularly pointed apically, lateral lobes cleft to base; apex of parameres with numerous setae, slightly modified, with dorsal velum. Biology. Specimens from Thange were reared from a sample of Crotalaria sp. near polysperma pods. This constitutes the first report on its biology. Distribution. Kenya and Zimbabwe. Discussion. The red color of the last visible tergite distinguishes Conicobruchus rubricollis from C. cicatricosus, C. flabellicornis and C. strangulatus (Fig. 1). It cannot be confounded with C. decoratus based on differences on the shape of the pronotum. Finally the presence of a longitudinal line of whitish setae on the pronotum separates C. rubricollis from dark specimens of C. atrosuturalis. Male genitalia (Fig. 8) are similar in shape with other members of the group (except C. decoratus) but completely lack large sclerites.

Conicobruchus strangulatus (Fåhraeus, 1839)

Bruchus strangulatus Fåhraeus, 1839:4 Bruchus hargreavesi Pic, 1933:133, syn. nov. Bruchidius hargreavesi: De Luca, 1965:58 Conicobruchus strangulatus: Decelle, 1951:181 Bruchidius strangulatus Prevett, 1967:176

Material examined. Paratype (female) of Bruchus hargreavesi: Uganda: Kampala, ii.1932, ex pods of Crotalaria [MNHN] Other material: Burkina-Faso: 1♂, Bobo-Dioulasso, 1.xi.2003, light trap [1♂ 00714] (H. Perrin) [MNHN]. Mali: 2♂, 7♀, Bafing, vi.2008, ex C. cf goreensis [1♂ 00614, specimen GK116 used for DNA extraction] (G.J. Kergoat) [MNHN]. Senegal: 18♂, 13♀, 5km N. Missira, 25.xi.1995, ex C. glaucoides [1♂ 14495, 1♀ 02210] (H. & A. Delobel) [CBGP]; 4♂, 11♀, Nianing, xi.1998, ex C. perrottetii [1♂ 19207, 1♀ 11307] (H. & A. Delobel) [CBGP]; 2♂, 2♀, Dakar, 15.ix.1999, ex C. podocarpa [1♂ 02699, specimen Xh1 used for DNA extraction] (H. &

250 · Zootaxa 3895 (2) © 2014 Magnolia Press LE RU ET AL. A. Delobel) [CBGP]; 1♂, 1♀, 5 km N. Missira, 11.xi.1995, ex C. comosa (H. & A. Delobel) [CBGP]; 1♂, Joal - Samba Dia, 19.xi.1995, ex C. cf goreensis [1♂ 14395] (H. & A. Delobel) [CBGP]. Color variable, with two distinct forms: darker specimens can be entirely black whereas lighter specimens are dark reddish brown, with antennae black. Specimens of the lighter form were formerly considered as a distinct species (hargreavesi). Other important morphological traits are as follows: pronotum 1.6–1.7 times wider at base than long; lack of white hair spots on pronotum (setation uniformly pale greyish or yellowish); strong ocellate punctation on interstriae, more or less aligned, particularly deep on elytral base in some specimens; hind femur with small acute tooth. Male genitalia. Median lobe (Fig. 10) of moderate length, stout (maximum width excluding basal hood/ total length = 0.16), strongly widened apically, basal hood moderately widened, concave posteriorly; ventral valve large, subtriangular, with apex acute, bearing numerous sensillae and two lateral groups of 5 to 10 setae; no hinge sclerite; internal sac without central column of tubercles, but with rather dense hyaline scales and tubercles, ending posteriorly in a dorsal mass of small, poorly sclerotized teeth, blending into a short series of well sclerotized teeth of variable length; a pair of strong ventro-lateral dented rods similar to those observed in Conicobruchus nodieri and C. astragalinae, varying in size and shape, from short and straight (Fig 12) to long and crescent (Fig. 13) or rod-shaped (Fig. 14), sometimes very long and articulated (Fig. 15); between these from zero to three isolated teeth; posteriorly a zone with a variable number of strong, isolated sclerotized teeth: absent in specimens from Dakar (Fig. 13) and Bafing (Fig. 15), numerous in male from Missira (Fig. 16) ; then again two groups of dented sticks or masses, clearly rod-shaped as in Fig. 16 to ill-defined as in male from Nianing (Fig. 17); apical ampoule devoid of any ornamentation, gonopore without sclerotization; basal strut narrow, without keel (Fig. 11); lateral lobes cleft to 85% their length; apex modified, with two lips, the dorsal one bearing about twelve long setae, the ventral one with a dense group of short sensillae.

FIGURES 12–17. Conicobruchus strangulatus: Ornamentation of saccus, ventral view: 12—specimen 00714, Burkina Faso; 13—specimen 02699, Senegal, Dakar; 14—specimen 14395, Dakar (Joal); 15—specimen 00614, Mali; 16—specimen 14495, Senegal (Missira); 17—specimen 19207, Senegal (Nianing).

Biology. Examined material was reared from pods of Crotalaria comosa, C. glaucoides Baker f., C. cf goreensis, C. perrottetii, C. podocarpa. Recorded from other Crotalaria species: C. calycina Schrank, C. leprieurii Guill. & Perr., C. ochroleuca G. D on, and C. subcapitata De Wild. (Gillon et al., 1992). Crotalaria pods usually contain a large number of small seeds; Conicobruchus strangulatus larvae are external feeders, at least during the last phases of their development, because their body is much larger than the size of individual seeds; when mature, larvae weave a white cocoon inside the inflated pod, attached to its wall, and emergence occurs through a circular hole. Development of young larvae seems impossible in mature, dry seeds.

THE CONICOBRUCHUS STRANGULATUS GROUP Zootaxa 3895 (2) © 2014 Magnolia Press · 251 Distribution. Fåhraeus' types are from Senegal and Republic of South Africa; Pic’s typical series of Bruchus hargreavesi is from Uganda; material seen is from Burkina Faso, Mali, Senegal; also reported from Angola (Decelle, 1975), Congo (Decelle, 1951, 1960, Rasplus, 1989), Eritrea (Zampetti, 1988), Ethiopia (Decelle 1971), Ivory Coast (Rasplus, 1989; Gillon et al., 1992), Nigeria (Prevett, 1971), Senegal (Decelle, 1969) and Togo (Woegan et al., 1997). Discussion. Conicobruchus strangulatus cannot be confounded with the three species that have a red last visible tergite (C. atrosuturalis, C. decoratus, C. rubricollis). Differences in elytral setation easily separate C. strangulatus from C. cicatricosus (uniform in C. strangulatus, mix of bare spots and densely setose patches in C. cicatricosus). Finally it can be distinguished from C. flabellicornis based on the presence of brushes and spots of dense white setae on the pronotum (absent in C. strangulatus, present in C. flabellicornis). Conicobruchus strangulatus is the type species of genus Conicobruchus Decelle. Senegalese specimens are either of the lighter (hargreavesi) or of the darker (strangulatus) form. The fact that specimens of both forms emerged from pods of the same host-plant in the same location provides support to establish the synonymy Bruchus hargreavesi Pic = Conicobruchus strangulatus (Fåhraeus).

FIGURE 18. Best tree (L= -25368.23) from the partitioned maximum likelihood analyses of the dataset. Bootstrap values > 50% are figured on nodes.

252 · Zootaxa 3895 (2) © 2014 Magnolia Press LE RU ET AL. Phylogenetic analyses. Maximum likelihood analyses yield a best ML tree with a likelihood score of - 25368.23 (Fig. 18). The genus Conicobruchus is recovered monophyletic with a high support (BV of 97%). As in the study of Kergoat et al. (2011), Bruchidius biguttatus and B. cisti appear closely related to the genus Conicobruchus (BV of 93%). Within the genus Conicobruchus the relationships are very similar to those inferred in Kergoat et al. (2011). The six species corresponding to the Conicobruchus strangulatus species group cluster together, with a high bootstrap support (BV of 94%). The newly sequenced species Conicobruchus indicus is found sister to C. kidevuensis, and appear unrelated to the species of the C. strangulatus species group. Another newly sequenced species, C. medaniensis is found sister to C. albopubens with a high support (BV of 100%). Within the C. strangulatus species group a clade composed of C. decoratus and C. rubricollis is sister (BV of 94%) to a well- supported (BV of 89%) clade of four species. The latter is encompasses C. cicatricosus on the one hand and a subclade that groups C. flabellicornis sister to C. atrosuturalis and C. strangulatus on the other.

Key to the Conicobruchus strangulatus species group based on external morphology of the adults (see Fig. 1)

1. Pronotum short, its sides not concave, not compressed anteriorly into a neck. Mostly red, with dark markings on elytra ...... C. decoratus - Pronotum long with sides concave, produced anteriorly into a neck ...... 2 2(1). Elytra with bare spots alternating with densely setose patches...... C. cicatricosus - Elytra uniformly covered with setae ...... 3 3(2). Pronotum bearing brushes and spots of dense white setae, quite noticeable on black background ...... C. flabellicornis - Lack of white setae spots on pronotum...... 4 4(3). Last visible tergite and elytra black or dark reddish-brown ...... C. strangulatus - Last visible tergite red ...... 5 5(4). Body mostly deep reddish brown, elytra mainly black, pronotum with a longitudinal line of whitish setae . . . . . C. rubricollis - Body black, elytra bright orange with suture black ...... C. atrosuturalis

Discussion

For the species in the C. strangulatus species group there is an excellent congruence between the male genital morphology and the results of the molecular phylogenetics analyses. More specifically, the saccus ornamentation is identical in the three species that are grouped together in the molecular phylogenetics analyses: C. atrosuturalis, C. flabellicornis and C. strangulatus. Both show two pairs of strong rod-shaped or crescentic dented sclerites in the proximal part of the saccus (Figs. 2, 7, 10). The phylogenetically related C. cicatricosus differs from these in the lack of proximal sclerites (Fig. 3), but it is otherwise quite similar. By contrast, the two remaining species C. decoratus and C. rubricollis exhibit more marked differences in their male genital morphology. Conicobruchus rubricollis completely lacks large sclerites (Fig. 8) while C. decoratus differs from all others species in the general shape of the median lobe (less widened anteriorly, basal strut not concave posteriorly; Fig. 5), as well as of parameres (Fig. 6). In addition, the pronotum is wider, not narrowed anteriorly into a “neck” (Fig. 1). Interestingly the molecular phylogenetics analyses do not support the hypothesis of a close relationship between all Crotalaria-feeders, as C. indicus is not monophyletic with the C. strangulatus species group. This result is also supported by morphological evidence. In fact, the median lobe in C. indicus is characterized by a very large and acutely triangular ventral valve, and the absence of a dented sclerite in the saccus [Plate 8, p.35 in Arora 1977](Anton pers. comm.). The apex of parameres is hardly modified, being rounded and bearing elongated setae, and the basal strut shows a small but distinct keel. All these differences in genital morphology point to a different species group than C. strangulatus. A number of additional Oriental species, including C. decellei (according to Anton (pers. comm.), not a synonym of indicus, contrary to Arora‘s opinion) and other undescribed species, belong to the same group of species as C. indicus. Nothing is presently known of their biology, but they could possibly also be Crotalaria-feeders. The latter suggests that there may have been multiple radiations of Conicobruchus species on Crotalarieae during the course of their evolution, one in Africa and at least one in the Oriental region.

We would like to thank the editor Matthias Schöller and two anonymous referees for their constructive comments on a previous version of the manuscript. Thanks are due to the late N. Berti, to H. Perrin (MNHN) and F. Gusenleitner (OÖLM) for the loan of important material to AD. We also thank K.-W. Anton for all the information on Conicobruchus indicus and other related Oriental species. The authors thank Alexandre Dehne Garcia for his help on the CBGP HPC computational platform. GJK also thanks Jean-Marc Duplantier and Laurent Granjon for their assistance in the field in Mali. Sequencing of new specimens was supported by the Plant Health and Environment Division of INRA (project INRA-SPE EVOSEED).

References

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18S

ng- -missing- D6-D7 ssing- -missing- ssing- -missing- ssing- -missing- -missing- -missing- -missing- -missing- -missing- -missing- 28S -missing- -missing- -missing- -missing- -missing- -missing- -missing- -missing- -missing- -missing- -missing- -missing- -missing- -missing- -missing- -missing- -missing- -missing-

ng- -missing- KM378135 D4-D5 ssing- -missing- -missing- ssing- -missing- KM378134 issing- -missing- -missing- HQ178547 KM378157 -missing- HQ178547 KM378157 HQ178520 -missing- -missing-

28S D2-D3 issing- KM378148 -missing- KM378131 178383 HQ178474 -missing- KM378133 -missing- -missing- 28S AY625370 -missing- AY625365 HQ178451 -mi AY625365 HQ178451 -missi HQ178407 HQ178500 -missing- HQ178521 -missing- KM378126

487 -missing- HQ178522 177475 -missing- -missi 177488 -missing- HQ178523 HQ177440 -missing- -missing- HQ177471 -missing- -missing- HQ177416 HQ178367 HQ178456 -mi HQ178456 HQ177416 HQ178367 HQ177450 -missing- -missing-

- HQ177439 -missing- HQ178478 -missing- HQ178478 - HQ177439 -missing- KM378118 -missing- -m HQ178239 AY625470 AY625421 AY625371 -mi AY625421 HQ178239 AY625470 AY625371

AY390636 HQ178242 AY390700 AY390668 AY625378 HQ178551 HQ178013 HQ178241 HQ177912 HQ177508 HQ178447 HQ178550 HQ177947 -missing- KM378111 -missing- HQ177948 KM378136 KM378124 KM378143 KM378150 -missing- -missing KM378102 KM378095 -missing- KM378125 KM378112 KM378103 KM378119 -missing- KM378137 KM378144 KM378151 KM378127 -missing- KM378113 HQ177994 HQ178212 HQ177891 HQ177486 KM378138 KM378145 KM378152 KM378105 KM378096 KM378128 KM378097 KM378106 KM378120 KM378114 KM378139 KM378146 KM378153 -missing- KM378155 KM378130 -missing- KM378129 KM378098 KM378107 KM378121 KM378115 -missing- KM378141 HQ177860 KM378140 KM378147 KM378154 KM378099 KM378108 KM378122 HQ177958 -missing- HQ177882 HQ HQ177970 HQ178190 HQ177872 HQ177462 HQ177983 KM378100 KM378109 KM378132 KM378101 KM378110 KM378123 KM378117 -missing- KM378142 KM378149 KM378156 KM378116 -missing- HQ177847 HQ177435 -m HQ HQ177995 HQ178215 HQ177942 HQ177892 HQ177 HQ177979 -missing- -missing- AY625318 HQ178146 AY625464 AY625415 AY625323 HQ178214 AY625469 AY625420 AY625323 HQ178214 HQ178001 HQ178222 HQ177897 HQ177493 -missing- HQ177893 HQ HQ177996 -missing- HQ177485 HQ178427 HQ177993 KM378104 -missing- HQ177827 -missing- HQ177922 -missing- DQ307631 HQ178237 DQ307672 HQ177506 DQ307644 -mi DQ307631 HQ178237 DQ307672 HQ177506 DQ307644 AY625324 AY390655 HQ178236 AY390719 AY390687 HQ178445 AY625293 HQ178151 AY625438 AY625389 AY625341 HQ178457 AY390639 HQ178217 AY390703 AY390671 HQ178429 HQ178525

BRU.Si11 GK35 GK171 GK195 GK448 GK446 GK200 GK243 GK442 GK452 GK695 GK156 BRU.L21 GK447 GK575 GK107 GK198 GK196 GK406 GK440 GK92 BRU.Xh11 GK116 GK245 BRU.Aa11 BRU.B21 BRU.Ch21 BRU.V21 BRU.Zp11 BRU.Xi11 BRU.Xj14

Ru 2010)

(Boheman, 1829) (Boheman, 1829) (Boheman, 1829) (Boheman, 1829) (Boheman, 1829) (Boheman, 1829) (Boheman, 1829) (Fahraeus 1839) (Fahraeus 1839) (Fahraeus 1839) (Fahraeus 1839) (Pic 1939) (Pic 1924) (Decelle 1982) (Decelle 1982) (Decelle 1982) (Decelle 1982) (Fahraeus 1839) (Fahraeus 1839) (Fahraeus 1839) (Fahraeus 1839) (Pic 1931) (Delobel (Fabricius 1801) (Fabricius 1801) (Fabricius 1801) (Fabricius 1801) (Pic 1904) (Pic 1903) (Pic 1903) (Pic 1903) (Fahraeus 1871) (Delobel (Pic 1921) (Pic 1909) (Pic 1928) (Olivier 1795) (Fabricius 1792) (Fabricius 1792) (Fahraeus 1839) (Fahraeus 1839) (Linnaeus 1758) (Linnaeus 1758) (Fabricius 1775) (Fabricius 1775) S1. Accession numbers. PPENDIX Conicobruchus rubricollis Pachymerus cardo Conicobruchus subdolus Bruchidius atrolineatus Conicobruchus flabellicornis Conicobruchus flabellicornis Conicobruchus indicus Conicobruchus medaniensis Conicobruchus nigricornis Conicobruchus skaifei Conicobruchus cicatricosus Conicobruchus cicatricosus Conicobruchus decoratus Conicobruchus flabellicornis Conicobruchus strangulatus A Species Conicobruchus albopubens Voucher 12S 16S Cytb COI GenBank accession No. Conicobruchus strangulatus Conicobruchus cicatricosus Conicobruchus kidevuensis Conicobruchus lineatopygus Megabruchidius tonkineus Bruchidius biguttatus Conicobruchus atrosuturalis Conicobruchus rubricollis Conicobruchus medaniensis Bruchidius cisti Conicobruchus nigricornis Conicobruchus rubricollis Bruchidius villosus

Bruchus pisorum Conicobruchus flabellicornis

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