Benzoquinones from Scent Glands of Phalangiid Harvestmen (Arachnida, Opiliones, Eupnoi): a Lesson from Rilaena Triangularis
Total Page：16
File Type：pdf, Size：1020Kb
Chemoecology DOI 10.1007/s00049-014-0177-y CHEMOECOLOGY RESEARCH PAPER Benzoquinones from scent glands of phalangiid harvestmen (Arachnida, Opiliones, Eupnoi): a lesson from Rilaena triangularis Gu¨nther Raspotnig • Miriam Schaider • Petra Fo¨ttinger • Verena Leutgeb • Christian Komposch Received: 22 May 2014 / Accepted: 13 October 2014 Ó The Author(s) 2014. This article is published with open access at Springerlink.com Abstract In case of disturbance, the phalangiine har- Introduction vestman Rilaena triangularis (Eupnoi, Phalangiidae) emits a directed jet from large prosomal scent (‘‘defensive’’) The scent glands of harvestmen are well known for the glands. The pungent-smelling secretion was analyzed by production of odoriferous secretions that serve in chemical gas chromatography–mass spectrometry and found to defense against predators (e.g., Eisner et al. 2004; Machado contain mainly 1,4-benzoquinone along with 1,4-naphtho- et al. 2005), but also in microbial protection (e.g., Fieser quinone and caprylic (=octanoic) acid. While various and Ardao 1956; Rocha et al. 2013) and chemical com- alkylated benzoquinones are characteristic for the scent munication (e.g., Holmberg 1986; Machado et al. 2002). gland secretions of many grassatorean Laniatores, this is Moreover, scent gland secretions represent a stunningly the first incidence of benzoquinone-based chemical defense diverse source of natural products, including rare naph- in palpatorean harvestmen. thoquinones and methyl ketones in Cyphophthalmi (Raspotnig et al. 2005, 2012; Jones et al. 2009), nitrogen- Keywords Scent glands Á Chemical defense Á containing compounds in travunioid Insidiatores (Ekpa Benzoquinone Á Naphthoquinone Á Caprylic acid Á et al. 1984; Raspotnig et al. 2011), and alkylated phenols in Palpatores Á Eupnoi some Grassatores (Shear et al. 2010a, b; Pomini et al. 2010). The best-known chemical class of scent gland- derived products, however, is certainly the benzoquinones which have been documented for many Grassatores of superfamily Gonyleptoidea (Hara et al. 2005; Caetano and Machado 2013; Raspotnig et al. 2014b). In detail, benzo- Handling Editor: Michael Heethoff. quinones were found to prevail the secretions of G. Raspotnig (&) Á M. Schaider Á P. Fo¨ttinger Á V. Leutgeb gonyleptoid families, Cosmetidae, Manaosbiidae, and Institute of Zoology, University of Graz, Universita¨tsplatz 2, Gonyleptidae (e.g., Hara et al. 2005;Fo¨ttinger et al. 2010), 8010 Graz, Austria but have not yet been reported from the secretions of any e-mail: [email protected] other major harvestman clade. Indeed, an assumed chem- G. Raspotnig ical dichotomy between the benzoquinone-rich exudates Research Unit of Osteology and Analytical Mass Spectrometry, from gonyleptoid Laniatores and the non-benzoquinone Medical University, University Children’s Hospital, chemistry of the remaining Opiliones represented an early Auenbruggerplatz 30, 8036 Graz, Austria paradigm of harvestman chemosystematics (e.g., Roach M. Schaider et al. 1980; Duffield et al. 1981). Department of Limnology and Bio-Oceanography, University of However, large gaps in the knowledge of opilionid scent Vienna, Althanstraße 14, 1090 Vienna, Austria gland chemistry still exist, and the largest gap certainly refers to the Palpatores—a species-rich group of ‘‘daddy- C. Komposch Institute of Animal Ecology and Landscape Planning, long legs’’ from which more than 2,000 representatives O¨ KOTEAM, Bergmanngasse 22, 8010 Graz, Austria have so far been described. The Palpatores can be classed 123 G. Raspotnig et al. into two large groups, often given the rank of suborders, compounds to the secretion chemistry of Eupnoi. We fur- namely the Dyspnoi (about 330 species) and the Eupnoi thermore provide evidence for the multiple independent (about 1,820 species) (Kury 2013). The phylogenetic evolution of benzoquinones in the scent glands of relationship between Dyspnoi and Eupnoi is still being Opiliones. discussed, either considering them as sister groups, resulting in monophyletic Palpatores, or regarding Dyspnoi as the sister group of Laniatores, leading to the so-called Materials and methods Dyspnolaniatores hypothesis (Machado et al. 2007). Any- way, apart from few studies on their scent gland Collection of specimens morphology (Juberthie et al. 1991; Holmberg 1970; Lopez et al. 1980; Clawson 1988; Schaider and Raspotnig 2009), In total, 183 individuals of Rilaena triangularis (Herbst, data on palpatorean scent glands, particularly on the 1799) were collected by hand at different localities in South- chemistry of secretions, are scarce. The secretion chemistry Eastern Austria (mainly Carinthia and Styria: Table 1). This of only 15 species (=less than 1 % of described Palpatores) species is widespread in Europe, ranging from Western and has so far been investigated. Indeed, scent glands in Pal- Central Europe to Southern Scandinavia, Russia and south- patores appear to be generally less developed and more wards to the northern parts of the Balkan Peninsula. Its heterogenous than in representatives of Cyphophthalmi and vertical distribution reaches from the lowlands to the montane Laniatores; they may even represent ‘‘cryptic’’ organs with level in the Alps. Habitats are forests, flood-plain forests and hidden or covered orifices such as in certain taxa of meadowsweet fens (Martens 1978; Komposch and Gruber Dyspnoi (Schaider and Raspotnig 2009). Also, the mode of 2004). Juveniles live in the soil layer whereas adults can be secretion discharge varies from emission of forceful jets or found on low vegetation in spring, and later on tree trunks up sprays to not observable or even no discharge at all (e.g., to treetops. We used adults of both sexes as well as juveniles Pabst 1953; Gnaspini and Hara 2007). These aberrant for our chemical study. conditions led to many methodological problems with respect to the extraction of glandular contents (Raspotnig Extraction of secretions et al. 2014a), resulting in decreasing attention on palpato- rean scent glands during the last decades. Scent gland secretions were obtained by three different However, initial chemical data indicate chemically methods: (1) directly by dabbing secretion on small pieces diverse secretions, in any case representing relevant data of filter paper (2 9 2 mm), immediately after emission for a consistent chemosystematic overall picture of the from ozopores (subsequent to mechanical irritation of Opiliones. Regarding the Dyspnoi, scent gland chemistry specimens by e.g., gentle squeezing). Secretion-loaded has eventually been studied on a few representatives of filter papers were extracted in hexane for 30 min; (2) by Nemastomatidae, demonstrating the presence of naphtho- whole body extraction of individuals in 150 ll of hexane or quinones, methoxy-naphthoquinones and anthraquinones in methylene chloride for 30 min as already described and Paranemastoma quadripunctatum (Raspotnig et al. 2010) standardized for other harvestmen (e.g., Raspotnig et al. and naphthoquinones along with highly volatile ketones in 2005, 2010); (3) by extraction of excised single glands in the species of Carinostoma (Raspotnig et al. 2014a). The methylene chloride for 30 h. little that is known on Eupnoi scent gland chemistry is still It should be mentioned beforehand that the variation in restricted to investigations from the 1970s and 1980s and secretion patterns as described in the results largely resul- refers to an extremely small taxonomic unit of nine North ted from the generally problematic methodological access American species of the sclerosomatid genus Leiobunum, to the Rilaena secretion. In detail, the handling of speci- one species of Hadrobunus (both genera in Leiobuninae) as mens during extract preparation greatly affected the well as one single representative of family Phalangiidae, resulting profiles. Thus, for a reliable evaluation of the full Phalangium opilio. While the secretions of leiobunine secretion profile of R. triangularis, we exclusively used Sclerosomatidae contained a variety of acyclic ethyl extracts prepared in a standardized manner as follows: ketones and biosynthetically related compounds (review in Individuals were handed with extreme care to avoid early Ekpa et al. 1985), P. opilio exclusively produced naph- emission of secretion beginning with the very gentle han- thoquinones (Wiemer et al. 1978). dling of specimens in the course of collecting in the field. Here, we report on the scent gland secretion of a second Individuals that could not be collected without disturbing/ phalangiid, Rilaena triangularis, that produces a mixture of stressing them were excluded from the investigation. Also, benzo- and naphthoquinones. Benzoquinones—so far to avoid effects of handling in the course of extract prep- considered characteristic of scent glands of gonyleptoid aration in the laboratory, we excluded individuals that Laniatores only—are introduced as a third class of could not be transferred into the solvent rapidly, i.e. 123 Benzoquinones from scent glands of phalangiid harvestmen Table 1 Collection of Rilaena triangularis Location (in Austria) Geographical co-ordinates (altitude) Date Individuals Carinthia, near Ferlach, Osce N 46°3103200,E14°1901000 (475 m) 24 February 2008 2 juv 22 March 2009 3 juv Carinthia, near Ferlach, Rauth N 46°3103200,E14°1903200 (453 m) 12 May 2012b 2 #,8$ N46°3102900,E14°1903800 (462 m) 9 June 2012b 6 #,6$ 24 June 2012 2 inda Styria, Graz, Berliner Ring N 47°0403700,E15°2904200 (398 m) 6 May 2009 3 $ 7 May 2009 3 #,4$ Styria,