Calacalles

Astrin, Jonas J. & Stüben, Peter E., 2009, Molecular phylogeny in ' nano-weevils': description of a new subgenus Nanoacalles and two new species of Calacalles from the Macaronesian Islands (Curculionidae: Cryptorhynchinae), Zootaxa 2300, pp. 51-67 : 53-56

Calacalles relationships and subgenera

Among the usually tiny species of the genus Calacalles , exoskeletal differences are usually marginal. Equally—like in many other western Palaearctic Cryptorhynchinae—the female genital characters cannot be reliably used in species diagnosis (e.g. high intraspecific variability, different degree of sclerotisation of the spiculum ventrale and the spermatheca). For morphological analysis, only the male genitalia (the structure of the internal sac of the aedeagus, cf. Figs. 9–26 ) offer the required stability (cf. Bahr 2000). In addition to the aedeagi, we here consider molecular genetic (mitochondrial) characters, which have not been used in Calacalles taxonomy before. Although we could not get hold of 3 of the so-far described Calacalles species for DNA sequencing, we see this paper (based on 15 out of 18 known Calacalles species) as a well-founded basis for a necessary revision including both morphological and molecular data.

Bodegas, N28 °33'38 " W16°0 9 '20",

500m, Sonchus spp., 2003, Stüben

Calacalles azoricus P.: Azores, Faial, Caldeira, N38°35'10" JJ0102, P-0125-azo EU286303 EU286468

W28°42'04", 886m, Tolpis azorica, 2003 ,

Stüben

Calacalles droueti P.: A z o r e s, P i c o, P ic o d a U r z e, JJ0100, P-0123-dro EU286302 EU286467

N 3 8 ° 2 7 ' 2 1 " W 2 8 ° 2 1 ' 0 4 ", 8 11 m,

Euphorbia stygiana, 2003 , Stüben

Calacalles exiguus E.: Tenerife, Anaga Mts., NE La Laguna, JJ0719, E-817-exi GQ332322 GQ332337

Bosque de Las Mercedes, N28°31'49"

W 1 6 ° 1 7 ' 1 2 ", 9 0 5 m, L a u r u s

no vo c ana rie nsis, 200 8, Flor en &

Machado

continued next page Taxon Collecting data Vouchers Acc.# 16S CO1

Calacalles palmensis E.: La Palma, 11 km NE El Paso: Mt. JJ0098, C-0121-set EU286301 EU286466

Tagoja, N28°43'18" W17°47'07", 1047m,

laurisilva, 2006, Stüben

Calacalles pumilio E.: La Gomera, S Hermigua , El Cedro: JJ0718, E-811-pum GQ332321 GQ332336

Las Mimbreras, N28°07'27" W17°13'26",

901m, Myrica faya, 2008 , Astrin, Stüben,

Behne & Floren

Calacalles pumilio E.: Tenerife, Teno Mts., Monte del Agua, JJ0721, E-812-pum GQ332324 GQ332339

N28°19'23" W16°49'15", 942m, Picconia

excelsa, Laurus novocanariensis, 2008 ,

Astrin, Stüben, Behne & Floren

Calacalles pusillus E.: Tenerife, Teno Mts., E Los Silos, El JJ0629, E-702-pus FJ716521 FJ716570

Tanque Bajo, N28°21'39" W16°46'19",

534m, plant detritus, 2008, Astrin &

Stüben

Calacalles seticollis E.: El Hierro, 4 km SW La Frontera: El JJ0099, C-0122-set GQ332311 GQ332328

Parque, N27°44'20" W18°01'30", 739m,

Sonchus hierrensis, 2006 , Stüben

Calacalles seticollis E.: El Hierro, 1 km SW Guarazoca: JJ0307, E-0265-set GQ332312 – C a m i n o d e l a Pe n a, N 2 7 ° 4 8 ' 0 6 "

W17°58'47", 746m, Carlina salicifolia,

2007, Stüben

Calacalles subcarinatus P.: Azores, São Miguel, Lagoa do Fogo, JJ0716, P-485-sub GQ332319 – a b o v e p o w e r p la n t, N 3 7 ° 4 6 ' 1 6 "

W25°29'07", 720m, Ilex perado, 2003 ,

Stüben

Calacalles theryi P.: N Caldas da Rainha, São Martinho do JJ0720, E-865-the GQ332323 GQ332338

Porto, N39°30 '24" W0 9 °0 8'48 ", 3m,

Crithmum maritimum, 2009 , Astrin

Calacalles theryi M O R O C C O: 2 k m S E s s a o u i r a, JJ0724, E-914-the GQ332326 GQ332341

N31°28'08" W09°45'27", 22m, Senecio

anteuphorbium at riverside, 2009, Stüben

Calacalles wollastoni P.:, Madeira, Santana, N 32° 48' 12" JJ0444, P-507-wol GQ332313 GQ332329

W16°52'53", 457m, Castanea sativa,

2008, Stüben

OUTGROUP:

Canariacalles alluaudi E.: Tenerife, Teno Mts., Casa Blanca near JJ0622, E-695-all GQ332314 GQ332330

Buenavista del Norte, N 28 °21 '36 "

W 1 6 ° 5 2 ' 1 0 ", 2 3 1 m, C e ro p e g i a

dichotoma, 2008, Astrin & Stüben

D ic h ro m a c a ll e s E.: Prov. Cádiz, 6,5 km S Algeciras, JJ0178, E-0204-dio EU286336 EU286500

diocletianus P u n ta d e l C a r n e r o, N 3 6 ° 0 4 ' 3 5 "

W05°25'46", 34m, thistles, 2007, Astrin

& Stüben

At this point it should be mentioned that even though the exoskeletal characters of the Calacalles species look very similar at first view, it is the clues gained from molecular analysis that enable a more differentiated morphological investigation.

This does not mean that morphological analysis was 'dirtier' before applying DNA sequence analysis, but it is much easier, for example, to interpret variation as interspecific variation when we are able to base ourselves on molecular data. Such was the case in the 'discovery' of the new species Calacalles hermigua described below.

Synonymy of the subgenus Saetiacalles Bahr, 2000 : Both mitochondrial genes unambiguously suggest joining Calacalles moraguesi (funicle with 7 antennomeres) with Calacalles theryi (6 antennomeres), as proposed by Stüben (2005). During the last years, the number of antennomeres played a decreasing role in the taxonomy of Curculionoidea (e.g. Tychius , Ceutorhynchus ; Clark 1976; Caldara 1990; Colonnelli 1984). It can be argued that structures repeated serially in an organism, i.e. homonomous segments like antennomeres, are often controlled by a single gene. While phenotypically 'feigning' a macromutation, the variation in antennomeres is in fact probably nothing more than the product of repeated gene activity. Such segmental duplications or reductions are – phylogenetically – of lesser importance (cf. Wägele 2005 on homonomies or serial homologies).

Based on these theoretical considerations as well as on the above-mentioned molecular findings ( Fig. 1 ), we reject a subgenus Saetiacalles Bahr, 2000 and abandon the use of the number of antennomeres as a taxonomic criterion. Instead, we propose a new subgeneric classification for the genus Calacalles as follows:

Calacalles s. str.: Figure 1 shows the consensus tree of the phylogenetic analyses. The continental species Calacalles theryi and C. moraguesi are recovered as sister to all other Calacalles species, from which they are separated by very high genetic distances (p - or uncorrected distance for e.g. C. moraguesi – C. atomarius : 10.9%). On the background of these molecular findings, we now also include Calacalles moraguesi with 7 antennomeres in the formerly monotypic subgenus Calacalles s. str. – misleadingly defined by Bahr (2000) as always possessing an antennal funiculus with 6 antennomeres (this change was already proposed by Stüben 2005: 123).

During typesetting of this article, we received another Calacalles sample from Libya that is morphologically very close to C. theryi , but molecularly distinct. It clearly belongs into the subgenus Calacalles s. str., but represents a distinct species from C. theryi and C. moraguesi . We suspect the sample to belong to the species C. kabylianus , of which only the damaged holotype from Algeria has been known so far. Therefore, in the Catalogue, we treat C. kabylianus as a Calacalles s. str. species. We will present the sequence of this sample in an upcoming publication.

Crateracalles Stüben, 2005: Sister to Calacalles s. str. stands a polytomy including, among others, an Azorean monophyletic lineage: the two large species of the subgenus Crateracalles plus the smaller C. subcarinatus . The two large Crateracalles species are nested within Calacalles , although morphologically, they are highly divergent. The two species are molecularly conspicuously close to each other (divergence of 1.1%) and it will have to be clarified whether they really represent two distinct species. The molecular and also new morphological evidence suggests the inclusion of C. subcarinatus into Crateracalles (see Taxonomy section).

Nanoacalles Astrin & Stüben subg. n. (see also Taxonomy section): Next to Crateracalles in the reconstruction stand the Canarian taxa: C. atomarius and C. pusillus by themselves, and the remaining species in a well-supported clade. This clade constitutes the new subgenus Nanoacalles, to which we also assign C. atomarius and C. pusillus due to their high morphological affinity to the other Madeiran-Canarian species. Hence Nanoacalles comprises the Madeiran-Canarian clade, but does not include the Azorean clade with C. droueti , C. azoricus , C. subcarinatus (all now Crateracalles) or the continental species of Calacalles s. str.: C. theryi , C. moraguesi , and C. kabylianus .

It can be noted that, according to our reconstruction, the Azores and the Canaries were colonised relatively early by Calacalles (Nanoacalles) species, while Calacalles (Nanoacalles) wollastoni from the

Madeiran archipelago evolved at a later time (this stands in contrast to the findings of Stüben & Astrin, in press, but the study included only six species of Calacalles ).

Figures

FIGURE 1. Bayesian 50 % majority rule consensus using the 16 S and CO 1 genes. Numbers indicate nodal posterior probabilities. The scale shows the expected nucleotide substitutions per site and the bars next to the tree denote subfamily assignment. Taxonomic changes are indicated (new subgeneric placement only through subgenus bars).

FIGURES 9 – 26. Comparison of the endophalli of all known Calacalles species.