Several recent studies have demonstrated the great potential for exploiting semiochemicals in ecology and conservation studies. The cerambycid beetle Rosalia alpina represents one of the flagship species of saproxylic insect biodiversity in Europe. In recent years its populations appear to have declined substantially, and its range has shrunk considerably as a result of forest management and urbanization. Here, we collected volatile chemicals released by males and females of R. alpina. Analyses of the resulting extracts revealed the presence of a single male-specific compound, identified as a novel alkylated pyrone structure. In field bioassays in Slovenia, traps baited with the synthesized pyrone captured both sexes of R. alpina, indicating that the pyrone functions as an aggregation pheromone. Our results represent the first example of a new structural class of pheromones within the Cerambycidae, and demonstrate that pheromone-baited traps can provide a useful tool for sampling R. alpina. This tool could be particularly useful in the ongoing development of conservation strategies for the iconic but endangered Alpine longicorn.

Institute of Organic Chemistry and Biochemistry ASCR Prague Czech Republic

National Institute of Biology Department of Organisms and Ecosystem Research Ljubljana Slovenia

University of California Department of Entomology Riverside California United States of America

Zobrazit více v PubMed

Thomson DR, Gut LJ, Jenkins JW. Pheromones for insect control In Hall FR, Menn JJ editors. Methods in biotechnology: Biopesticides: use and delivery. Humana Press; 1998. pp 385–412.

Gitau CW, Bashford R, Carnegie AJ, Gurr GM. A review of semiochemicals associated with bark beetle (Coleoptera: Curculionidae: Scolytinae) pests of coniferous trees: A focus on beetle interactions with other pests and their associates. Forest Ecol Manag. 2013; 297: 1–14.

Hoover K, Keena M, Nehme M, Wang S, Meng P, Zhang A. Sex-specific trail pheromone mediates complex mate finding behavior in PubMed DOI

Hansen L, Xu T, Wickham J, Chen Y, Hao D, Hanks LM, et al. Identification of a male-produced pheromone component of the citrus longhorned beetle, PubMed DOI PMC

Welzel KF, Choe DH. Development of a pheromone-assisted baiting technique for Argentine ants (Hymenoptera: Formicidae). J Econ Entomol. 2016; 109: 1303–1309. PubMed

Larsson MC, Svensson GP. Pheromone monitoring of rare and threatened insects: Exploiting a pheromone-kairomone system to estimate prey and predator abundance. Cons Biol. 2009; 23: 1516–1525. PubMed

Kadej M, Zając K, Ruta R, Gutowski JM, Tarnawski D, Smolis A, et al. Sex pheromones as a tool to overcome the Wallacean shortfall in conservation biology: a case of

Burman J, Westerberg L, Ostrow S, Ryrholm N, Bergman KO, Winde I, et al. Revealing hidden species distribution with pheromones: The case of

CECC. Directive 92/43/EEC of 21 May 1992 on the Conservation of Natural Habitats and of Wild Fauna and Flora. 1992. http://eur-lex.europa.eu/.

McDonald LL. Sampling rare populations In: Thompson WL editor. Sampling Rare or Elusive Species: Concepts, Designs, and Techniques for Estimating Population Parameters. Island Press; 2004. pp. 11–42.

Larsson MC. Pheromones and other semiochemicals for monitoring rare and endangered species. J Chem Ecol. 2016; 42: 853–868. doi: 10.1007/s10886-016-0753-4 PubMed DOI PMC

Larsson MC, Hedin J, Svensson GP, Tolasch T, Francke W. Characteristic odor of PubMed

Tolasch T, von Fragstein M, Steidle JLM. Sex pheromone of PubMed DOI

Tolasch T, Konig C, von Fragstein M, Steidle JLM. Identification of the sex pheromone of PubMed DOI

Millar JG, McElfresh JS, Romero C, Vila M, Mari-Mena N, Lopez-Vaamonde C. Identification of the sex pheromone of a protected species, the Spanish moon moth PubMed DOI PMC

Levi-Zada A, Ben-Yehuda S, Dunkelblum E, Gindin G, Fefer D, Protasov A, et al. Identification and field bioassays of the sex pheromone of the yellow-legged clearwing

Ray AM, Barbour JD, McElfresh JS, Moreira JA, Swift I, Wright IM, et al. 2,3-Hexanediols as sex attractants and a female-produced sex pheromone for cerambycid beetles in the genus PubMed DOI PMC

Gago R, Allison JD, McElfresh JS, Haynes KF, McKenney J, Guerrero A, et al. A tetraene aldehyde as the major sex pheromone component of the promethea moth ( PubMed DOI

Yan Q, Kanegae A, Miyachi T, Naka H, Tatsuta H, Ando T. Female sex pheromones of two Japanese saturniid species, PubMed DOI

Konig C, Szallies A, Steidle JLM, Tolasch T. Sex pheromone of the rare click beetle PubMed DOI

Millar JG, Haynes KF, Dossey AT, McElfresh JS, Allison JD. 2016. Sex attractant pheromone of the luna moth, PubMed DOI

Larsson MC, Svensson GP. Monitoring spatiotemporal variation in abundance and dispersal by a pheromone-kairomone system in the threatened saproxylic beetles

Gouix N, Brustel H. Emergence trap, a new method to survey

Chiari S, Zauli A, Mazziotta A, Luiselli L, Audisio P, Carpaneto GM. Surveying an endangered saproxylic beetle,

Musa N, Andersson K, Burman J, Andersson F, Hedenström E, Jansson N, et al. Using sex pheromone and a multi-scale approach to predict the distribution of a rare saproxylic beetle. PLoS ONE. 2013; 8: e66149 doi: 10.1371/journal.pone.0066149 PubMed DOI PMC

Andersson K, Bergman KO, Andersson F, Hedenström E, Jansson N, Burman J, et al. High-accuracy sampling of saproxylic diversity indicators at regional scales with pheromones: the case of

Ray AM, Arnold RA, Swift I, Schapker PA, McCann S, Marshall CJ, et al. ( PubMed DOI PMC

Zauli A, Chiari S, Hedenstrom E, Svensson GP, Carpaneto GM. Using odour traps for population monitoring and dispersal analysis of the threatened saproxylic beetles

Zauli A, Carpaneto GM, Chiari S, Mancini E, Nyabuga FN, De Zan LR, et al. Assessing the taxonomic status of

Harvey DJ, Harvey H, Harvey RP, Kadej M, Hedenström E, Gange AC, et al. Use of novel attraction compounds increases monitoring success of a rare beetle,

Mari-Mena N, Lopez-Vaamonde C, Naveira H, Auger-Rozenberg MA, Vila M. Phylogeography of the Spanish moon moth PubMed DOI PMC

Nieto A, Alexander KNA. European Red List of Saproxylic Beetles. 2010. Publications Office of the European Union, Luxembourg.

Siitonen J. Threatened saproxylic species In: Stokland J.N., Siitonen J, Jonsson BG editors. Biodiversity in Dead Wood. Cambridge University Press; 2012; pp. 356–379.

Lachat T, Ecker K, Duelli P, Wermelinger B. Population trends of

Ciach M, Michalcewicz J. Correlation between selected biometric traits of adult

Čížek L, Schlaghamerský J, Bořucký J, Hauck D, Helešic J. Range expansion of an endangered beetle: Alpine longhorn

Annex II. The European Council Directive on the Conservation of Habitats, Flora and Fauna 92/43/EEC (92/43/EEC)-»The Habitat Directive«.

The IUCN Red List of Threatened Species. Version 2016–3. Downloaded on the 21st of December 2016. <http://www.iucnredlist.org>

Ciach M, Michalcewicz J, Fluda M. The first report on development of

Drag L, Hauck D, Pokluda P, Zimmermann K, Cizek L. Demography and dispersal ability of a threatened saproxylic beetle: a mark-recapture study of the Rosalia longicorn ( PubMed PMC

Bosso L, Rebelob H, Garonnac AP, Russo D. Modelling geographic distribution and detecting conservation gaps in Italy for the threatened beetle

Michalcewicz J, Ciach M. Current distribution of the Rosalia longicorn

Russo D, Cistrone L, Garonna AP. Habitat selection by the highly endangered long-horned beetle

Castro A, de Murguia LM, Fernandez J, Casis A, Molino F. Size and quality of wood used by

Castro A, Fernandez J. Tree selection by the endangered beetle

Vrezec A, Ambrožič Š, Kapla A. An overview of sampling methods test for monitoring schemes of saproxylic beetles in the scope of Natura 2000 in Slovenia In Jurc M editor. Saproxylic beetles in Europe: monitoring, biology and conservation. Silva Slovenica; 2014. pp. 73–89.

Russo D, Di Febbraro M, Cistrone L, Jones G, Smeraldo S, Garonna AP, et al. Protecting one, protecting both? Scale-dependent ecological differences in two species using dead trees, the Rosalia longicorn beetle and the barbastelle bat. J Zool. 2015; 297: 165–175.

Jeppsson T, Lindhe A, Gärdenfors U, Forslund P. The use of historical collections to estimate population trends: A case study using Swedish longhorn beetles (Coleoptera: Cerambycidae). Biol Conserv. 2010; 143: 1940–1950.

Pardo I, Pata MP, Gomez D, Garcıa MB. A novel method to handle the effect of uneven sampling effort in biodiversity databases. PLoS ONE. 2013; 8: e52786 doi: 10.1371/journal.pone.0052786 PubMed DOI PMC

Millar J, Hanks L. Chemical ecology of Cerambycids In: Wang Q editor. Cerambycidae of the World: Biology and Pest Management. CRC Press/Taylor & Francis; 2017. pp. 161–208.

Evans HF, Moraal LG, Pajares JA. Biology, ecology and economic importance of Buprestidae and Cerambycidae In Lieutier F, Day KR, Battisti A, Grégoire JC, Evans HF editors. Bark and Wood Boring Insects in Living Trees in Europe, a Synthesis. Kluwer; 2004. pp. 447–474.

Hanks LM, Millar JG. Sex and aggregation-sex pheromones of cerambycid beetles: Basic science and practical applications. J Chem Ecol. 2016; 42: 631–654. doi: 10.1007/s10886-016-0733-8 PubMed DOI

Ray AM, Millar JG, McElfresh JS, Swift IP, Barbour JD, Hanks LM. Male-produced aggregation pheromone of the cerambycid beetle PubMed DOI

Vrezec A, Kapla A. Naravovarstveno vrednotenje favne hroščev (Coleoptera) Krajinskega parka Boč-Donačka gora v občini Rogaška Slatina: kvantitativna varstveno-favnistična analiza. Varstvo narave. 2007; 20: 61–82.

Katritzky AR, Zhang Y, Singh SK. Efficient conversion of carboxylic acids into

Zhang Z, Kitamura Y, Myers AG. An efficient directed Claisen reaction allows for rapid construction of 5,6-disubstituted-1,3-dioxin-4-ones. Synthesis. 2015; 47: 2709–2712.

Vrezec A, de Groot M, Kobler A, Ambrožič Š, Kapla A. Ecological characteristics of habitats and potential distribution of selected qualification species of beetles (Coleoptera) in the scope of Natura 2000 network in Slovenia: the first modelling approach. Gozdarski vestnik. 2014; 72: 452–471.

Vrezec A. Fenološka ocena pojavljanja imagov štirih vrst varstveno pomembnih saproksilnih hroščev v Sloveniji:

Hammer Ø, Harper DAT, Ryan PD. PAST: Paleontological statistics software package for education and data analysis. Palaeontol Electron. 2001; 4: 1–9.

Sokal RR, Rohlf FJ. Biometry: The Principles and Practice of Statistics in Biological Research. 3rd ed USA: W.H. Freeman and Company; 1995.

Zar JH. Biostatistical Analysis. 5th ed New Jersey: Prentice Hall /Pearson; 2010.

Bach T, Kirsch S. Synthesis of 6-substituted-4-hydroxy-2-pyrones from aldehydes by addition of an acetoacetate equivalent, Dess-Martin oxidation and subsequent cyclization. Synlett. 2001: 1974–1976.

Acocella MR, De Rosa M, Massa A, Palombi L, Villano R, Scettri A. Silicon tetrachloride in organic synthesis: new applications for the vinylogous aldol reaction. Tetrahedron. 2005; 61: 4091–4097.

Prantz K, Mulzer J. 2010. Synthesis of ( PubMed DOI

Dong Y, Nakagawa-Goto K, Lai C-Y, Morris-Natschke SL, Bastow KF, Lee K-H. Antitumor agents 287. Substituted 4-amino-2H-pyran-2-one (APO) analogs reveal a new scaffold from neo-tanshinlactone with PubMed DOI PMC

Rentsch A, Kalesse M. The total synthesis of corallopyronin A and myxopyronin B. Angew Chem Int Ed. 2012; 51: 11381–11384. PubMed

Zhao Y-M, Maimone TJ. Short, enantioselective total synthesis of chatancin. Angew Chem Int Ed. 2015; 54: 1223–1226. PubMed PMC

Seiple IB, Zhang Z, Jakubec P, Langlois-Mercier A, Wright PM, Hog DT, et al. A platform for the discovery of new macrolide antibiotics. Nature 2016; 533: 338–345. doi: 10.1038/nature17967 PubMed DOI PMC

Katritzky AR, Wang Z, Wang M, Hall CD, Suzuki K. Facile syntheses of 2,2-dimethyl-6-(2-oxoalkyl)-1,3-dioxin-4-ones and the corresponding 6-substituted-4-hydroxy-2-pyrones. J Org Chem. 2005; 70: 4854–4856 doi: 10.1021/jo050307m PubMed DOI

Fuse S, Ikebe A, Oosumi K, Karasawa T, Matsumura K, Izumikawa M, et al. Asymmetric total synthesis of PubMed DOI

Sato M, Sakaki J, Sugita Y, Yasuda S, Sakoda H, Kaneko C. Two lactone formation reactions from 1,3-dioxin-4-ones having hydroxyalkyl group at the 6-position: Difference in ring opening and closure. Tetrahedron. 1991; 47: 5689–5708.

Boeckman RK Jr, Pruitt JR. A new, highly efficient selective methodology for formation of medium-ring and macrocyclic lactones via intramolecular ketene trapping: An application to a convergent synthesis of (-)-kromycin. J Am Chem Soc. 1989; 111: 8286–8288.

Shimamura H, Sunazuka T, Izuhara T, Hirose T, Shiomi K, Omura S. Total synthesis and biological evaluation of verticipyrone and analogues. Org Lett. 2007; 9: 65–67. doi: 10.1021/ol0626140 PubMed DOI

Song D, McDonald R, West FG. Facial diastereoselectivity in [4+4]-photocycloadditions of pyran-2-ones: Surprising effect of an adjacent chiral center. Org Lett. 2006; 8: 4075–4078. doi: 10.1021/ol061576h PubMed DOI

Sun CL, Fürstner A. Formal ring-opening/cross-coupling reactions of 2-pyrones: Iron-catalyzed entry into stereodefined dienyl carboxylates. Angew Chem Int Ed. 2013; 52: 13071–13075. PubMed

Khatri BB, Sieburth S McN. 2015. Enyne-2-pyrone [4 + 4]-photocycloaddition: Sesquiterpene synthesis and a low-temperature Cope rearrangement. Org Lett. 2015; 17: 4360–4363. doi: 10.1021/acs.orglett.5b02207 PubMed DOI

Fairlamb IJS, Lu FJ, Schmidt JP. Palladium-catalysed alkynylations of 2-pyrone (pyran-2-one) halides. Synthesis. 2003; 16: 2564–2570.

Menu F. Strategies of emergence in the chestnut weevil PubMed DOI

Stoks R. Male-biased sex ratios in mature damselfly populations: real or artefact? Ecol Entomol. 2001; 26: 181–187.

Larsson MC, Hedin J, Svensson GP, Tolasch T, Francke W. Characteristic odor of PubMed

Cardé RT. Defining attraction and aggregation pheromones: teleological versus functional perspectives. J Chem Ecol. 2015; 40: 519–520. PubMed

Prikryl ZB, Turcani M, Horak J. Sharing the same space: foraging behaviour of saproxylic beetles in relation to dietary components of morphologically similar larvae. Ecol Entomol. 2012; 37: 117–123.

Schäberle TF. Biosynthesis of α-pyrones. Beilstein J Org Chem. 2016; 12: 571–588. doi: 10.3762/bjoc.12.56 PubMed DOI PMC

Charlton RE, Webster FX, Zhang A, Schal C, Liang D, Sreng I, et al. Sex pheromone for the brownbanded cockroach is an unusual dialkyl-substituted alpha-pyrone. Proc Nat Acad Sci USA. 1993; 90: 10202–10205. PubMed PMC

Brachmann AO, Brameyer S, Kresovic D, Hitkova I, Kopp Y, Manske C et al. Pyrones as bacterial signaling molecules. Nat Cheml Biol. 2013; 9: 573–578. PubMed

Hanks LM, Millar JG. Field bioassays of cerambycid pheromones reveal widespread parsimony of pheromone structures, enhancement by host plant volatiles, and antagonism by components from heterospecifics. Chemoecology. 2013; 23: 21–44.

Graham EER, Mitchell F, Reagel PF, Barbour JD, Millar JG, Hanks LM. Treating panel traps with a fluoropolymer enhances their efficiency in capturing cerambycid beetles. J Econ Entomol. 2010; 103: 641–647. PubMed

Graham EET, Poland TM. Efficacy of fluon conditioning for capturing cerambycid beetles in different trap designs and persistence on panel traps over time. J Econ Entomol. 2012; 105: 395–401. PubMed

Allison JD, Johnson CW, Meeker JR, Strom RL, Butler SM. Effect of aerosol surface lubricants on the abundance and richness of selected forest insects captured in multiple-funnel and panel traps. J Econ Entomol. 2011; 104: 1258–1264. PubMed

Allison JD, Bhandari BD, McKenney JL, Millar JG. Design factors that influence the performance of flight intercept traps for the capture of longhorned beetles (Coleoptera: Cerambycidae) from the subfamilies Lamiinae and Cerambycinae. PlosONE 2014; 9: e93203. PubMed PMC

Drag L, Hauck D, Pokluda P, Zimmermann K, Cizek L. Demography and dispersal ability of a threatened saproxylic beetle: A mark-recapture study of the Rosalia Longicorn ( PubMed DOI PMC

Ginzel MD, Hanks LM. Role of host plant volatiles in PubMed

Silk PJ, Sweeney J, Wu J, Price J, Gutowski JM, Kettela EG. Evidence for a male-produced pheromone in PubMed DOI

Teale SA, Wickham JD, Zhang F, Su J, Chen Y, Xiao W, et al. Male-produced aggregation pheromone of PubMed

Collignon RM, Swift IP, Zou Y, McElfresh JS, Hanks LM, Millar JG. The influence of host plant volatiles on the attraction of longhorn beetles to pheromones. J Chem Ecol. 2016; 42: 215–229. doi: 10.1007/s10886-016-0679-x PubMed DOI

Miller DR. Ethanol and (−)-α-pinene: attractant kairomones for some large wood-boring beetles in southeastern USA. J Chem Ecol. 2006; 32: 779–794. doi: 10.1007/s10886-006-9037-8 PubMed DOI