Chemistry of the Postpharyngeal Gland Secretion and Its Implication
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Chemoecology 7:163-171 (1996) 0937 7409/96/040163 09 $1.50+0.20 © 1996 Birkh/iuser Verlag, Basel Chemistry of the postpharyngeal gland secretion and its implication for the phylogeny of Iberian Cataglyphis species (Hymenoptera: Formicidae) Abdallah Dahbi l, Alain Lenoir 2, Alberto Tinaut 3, Timo Taghizadeh 4, Wittko Franeke 4, and Abraham Hefetz s,* ~Laboratoire d'Ethologie Exp~rimentale et Comparee, URA 2214, Universit~ Paris Nord, F-93430 Villetaneuse, France 2Laboratoire d'Ethologie et Pharmacologie du Comportement, Facult6 des Sciences et Techniques, F-37200, Tours, France 3Departamento de Biologia Animal y Ecologia, Facultad de Ciencias, Universidad de Granada, E-18071 Granada, Spain 4Institute of Organic Chemistry, University of Hamburg, Martin-Luther-King-Platz6, D-20146 Hamburg, Germany SDepartment of Zoology, Tel Aviv University, Ramat Aviv 69978, Tel Aviv, Israel Summary. A comparative morphological and chemical classification, and often for reconstructing phylogenies. study of six endemic species of Cataglyphis from the Defensive secretions, for example were used in phylo- Iberian Peninsula: C. ibericus, C. rosenhaueri, C. hispani- genetic studies of various insect groups including cus, C. humeya, C. velox and C. Jtoricola and the staphylinid beetles (Steidle & Dettner 1993), chrysome- Moroccan species C. bombycinus is described. The mor- lid leaf beetles (Pasteels 1993), and fire ants of the genus phological study relied primarily on genitalia character- Solenopsis (Brand et al. 1973; Vander Meer 1986). Sex istics, whereas the chemical study concentrated on the pheromones were used in the classification and evolu- postpharyngeal gland constituents. Cladograms based tion of tortricide moths (Roelofs & Brown 1982). Du- on the morphological and chemical data were performed four's gland constituents served as character states for using Ward's method. The dendrogram based on mor- the reclassification of lactone producing bees (Cane phological features revealed that the Iberian Cataglyphis 1983a) and of andrenid bees (Cane 1983b). The cephalic can be classified into three species groups albicans, secretion of male bumble bees was used for numerical altisquamis and emmae. The same pattern occurred when taxonomy studies (Belles et al. 1987), and even for the the dimethylalkanes constituents of the postpharyngeal distinction between colour morphs within a species gland were utilised as character states, with a slight (Bergstr6m et aI. 1973). Cuticular hydrocarbons were displacement of species within the altisquamis group. likewise used in the classification of beetles (Jacob 1979; However, when the complete hydrocarbon blend was Page et al. 1990), Drosophila species from the Drosophila utilised major discrepancies in the dendrograms oc- virilis group (Bartelt et al. 1986), and termites (Bagn~res curred. CatagIyphis velox proved to be very similar to C. et al. 1990; Kaib et al. 1991). The concordance between bombycinus, whereas C. ftoricola clustered with the other chemical and morphological data that generally occurs two species of the altisquamis group. Based on the makes these chemicals a good tool in taxonomy. Some geographical distribution and paleontological data (Tin- precautions however should be taken in chemotaxo- aut 1993) it is assumed that C. floricola recently invaded nomic analyses, in particular with respect to the biolog- the Iberian Peninsula. Based on the chemical findings we ical role of the secretions utilized. The use of defensive postulate that chemical character displacement occurred secretions is relatively reliable since they need not be in C. floricola as a result of its sympatry with C. velox species specific, and therefore are not subjected to after the former colonized the Iberian Peninsula. We selective pressures to diversify. Any changes in their further discuss the possible reason for the different chemistry may thus be the consequence of speciation, dendrograms obtained when only the dimethylalkanes provided they remain pungent. are considered and its implication for the communicative On the other hand, secretions that are used for role of the postpharyngeal gland secretion in these ants. communication may be subjected to continuous selec- tive pressures to diversify, as they have to be species Key words, postpharyngeal gland - phylogeny - hydro- specific, and often must also exhibit individual compo- carbons - chemotaxonomy - Hymenoptera - sitions. Moreover, any changes in the chemical compo- Cataglyphis sition of pheromones may result in the formation of a behavioural barrier, leading to a quick fixation within the population. The selection for chemical diversity is accentuated in closely related sympatric species. This Introduction may result in two sympatric species which might be phylogenetically related, but exhibit disparate secretion- The extensive identification of exocrine products in ary compositions, while their more distant relatives, insects has opened the way for their use in taxonomy, being allopatric, retained the ancestral composition. In the present paper we have used the hydrocarbon con- * To whom correspondence should be addressed stituents of the postpharyngeal gland for the classifica- 164 A. Dahbi et al. CNEMOECOLO~¥ tion and the assessment of the phylogenetic relationship The character states used in the present work are the following: of several Cataglyphis species. l. Mandible with an apical tooth and a straight masticatory border without teeth (0), mandible pointed and only with an apical tooth The study of hydrocarbons in ants has focused (1). primarily on their role in intra- and interspecific com- 2. Discoidal cell vestigial (0), or absent (1). munication. Several lines of evidence suggest that these 3. Shape of sagitta ventral face straight or lightly concave (0), substances are involved in nestmate recognition lightly convex (1) or sharply convex (2). 4. Ventral face of sagitta with teeth (0) or without teeth (1). (Bonavita-Cougourdan et al. 1987; Morel et al. 1988; 5. Teeth in more than half sagitta ventral face (0) or occupying less Henderson et al. 1990; Nowbahari et al. 1990). More than half (1). recently the role of the glandular secretion in modifying 6. Sagitta with antero-dorsal process absent (0) or present (1). the aggressive behaviour of ants was established in the 7. Volsella curved (0) or straight (1). formicine ant Cataglyphis niger (Soroker et aI. 1994), 8. Basal process in the stipe absent (0), or present (1). 9. Proportion of the maxillary palps of males: 5 similar to 6 (0), 5 and in the myrmicine species Manica rubida (Hefetz et smaller than 6 (1). al. 1996). Using the glandular hydrocarbons, it was 10. Polymorphic workers (0), monomorphic workers (1). further demonstrated that the gland serves as a storage 11. Small process in the basis of the volsella (0 = absent, 1 = present). organ, and that the unified colonial odor is achieved by 12. Process in the basis of the volsella rounded (0) or pointed (1). 13. Volsella smooth (0) or with edges in the middle (1). constant exchanges of secretions through trophallaxis 14. Edges of the volsella small (0) or well developed (1). and mutual grooming, constituting therefore a "Gestalt- organ" (Soroker et aI. 1994; Vienne et al. 1995). Chemical studies Cataglyphis is a genus with a wide geographic distri- bution including various ecosystems ranging from For the chemical analyses, two colonies per species were collected and deserts to temperate regions around the Mediterranean individuals were immediately stored at -20°C until dissection. The basin. This distribution has resulted in a large generic postpharyngeal glands from about thirty ants of each species were diversity which complicates the study of their phy- dissected in chilled distilled water and immediately immersed in logeny. Taxonomic studies of this genus using essen- pentane for extraction. Chemical analysis of the samples was con- ducted primarily by coupled gas chromatography/mass spectrometry tially morphological and anatomical traits (Santschi (Fisons MD800 quadrupole). The samples were run on a 30 m x 0.32 1929; Wehner 1983; Agosti 1990; Wehner et al. 1994) mm DB-5 fused silica capillary column that was temperature pro- have proved insufficient leaving unresolved problems. grammed from 100°C to 300°C at 3°C/min with an initial hold of The few chemotaxonomic studies performed (Hefetz & 3 min. The various compounds were identified according to retention indices and to known fragmentation patterns, which enable structure Lenoir 1992; Keegans et al. 1992) relied mostly on assignments even of complex mixtures of branched hydrocarbons Dufour's gland secretion, and aimed at separating dif- (Doolittle et al. 1995 and references cited therein) as well as by ferent species within this genus. comparing their spectra to published spectra (McLafferty & Stauffer Six endemic species of Cataglyphis exist in the 1989; Bagn~res et al. 1991). Position of the double bonds was Iberian Peninsula representing the highest diversity of elucidated by alkylthiolation using dimethyl disulfide (DMDS) (Vin- centi 1987). For alkylthiolation, 10 gl of the extract were mixed with this genus in Europe. However, only a few studies 50 gl of carbon disulfide, 50 ~tl DMDS, and 5 gI of iodine solution pertaining to their phylogenetic relationships have been (60 mg in 1 ml ether) and heated to 60°C for 12h. The excess of iodine published (Tinaut & Plaza 1989; Tinaut 1990a, b, was destroyed with 50-100 i~1 aqueous sodium thiosulfate solution 1993). In this study we present