Journal of Chemical Ecology

, Volume 41, Issue 5, pp 465–472 | Cite as

N-3-Methylbutanoyl-O-methylpropanoyl-L-serine Methyl Ester – Pheromone Component of Western Black Widow Females

  • Catherine ScottEmail author
  • Sean McCann
  • Regine Gries
  • Grigori Khaskin
  • Gerhard Gries


Chemical communication is common in spiders but few pheromones have been identified. Female widow spiders in the genus Latrodectus spin webs that disseminate an attractive sex pheromone, and a contact pheromone on the silk elicits courtship behavior by males. The methyl ester of N-3-methylbutanoyl-O-(S)-2-methylbutanoyl-L-serine is a contact pheromone of the Australian redback spider Latrodectus hasselti. We hypothesized that the contact pheromone of congeneric L. hesperus resembles that of L. hasselti. The silk of virgin L. hesperus females was extracted with methanol, and analyses by gas chromatography–mass spectrometry (GC/MS) provided evidence for the presence of N-3-methylbutanoyl-O-methylpropanoyl-L-serine methyl ester (MB-MP-S), a lower homologue of the L. hasselti contact pheromone. Behavioral responses of L. hesperus males to test stimuli were assayed on T-shaped rods with the end sections of the horizontal arm enveloped in filter paper. Males spent 40 % longer in contact with paper bearing female silk than with blank paper, and 39 % longer in contact with paper treated with silk extract than with solvent controls. Contact with silk and silk extract induced courtship behavior by 96 % and 80 % of males, respectively, indicating that there was a methanol-soluble courtship-eliciting contact pheromone on the silk. Males responded less strongly to synthetic MB-MP-S than to silk or silk extract. Paper impregnated with synthetic MB-MP-S (10 or 100 μg) induced courtship behavior in 3–16 % of males, and prompted males to stay 10–16 % longer than on control paper. Our data support the conclusion that MB-MP-S is part of a multi-component contact pheromone of L. hesperus.


Latrodectus Chemical communication Pheromone Amino acid Silk Courtship 



We thank the Tsawout First Nation for permission to do field work on their lands, the Capital Regional District for granting permits to collect spiders from Island View Beach; Maydianne Andrade, Bernard Roitberg, Tanya Stemberger, Samantha Vibert, and two anonymous reviewers for comments that improved the manuscript; Huimin Zhai for comments on the chemistry section; and Susan Chen and Elaine Wu for assistance with spider rearing and maintenance. Funding was provided by a Natural Sciences and Engineering Research Council of Canada (NSERC) – Discovery Grant and an NSERC – Industrial Research Chair to G.G., with Contech Enterprises Inc. and Global Forest Science as industrial sponsors.

Supplementary material

10886_2015_582_MOESM1_ESM.doc (154 kb)
ESM 1 General methods and instrumentation for syntheses, a representative synthesis of ester-amide mixtures, and synthesis of N-3-methylbutanoyl-O-methylpropanoyl-L-serine methyl ester (DOC 153 kb)

A Latrodectus hesperus male silk-wrapping a filter paper treated with methanol extract of a virgin female’s web, first at normal speed, then slowed to half-speed (MP4 12085 kb)


  1. Andrade M, Kasumovic M (2005) Terminal investment strategies and male mate choice: extreme tests of Bateman. Integr Comp Biol 45:838–47. doi: 10.1093/icb/45.5.838 CrossRefPubMedGoogle Scholar
  2. Arn H, Städler E, Rauscher S (1975) The electroantennographic detector—a selective and sensitive tool in the gas chromatographic analysis of insect pheromones. Z Naturforsch 30c:722–725Google Scholar
  3. Blumstein DT, Daniel JC, Evans CS (2012) Jwatcher. Available online:
  4. Chinta SP, Goller S, Lux J, Funke S, Uhl G, Schulz S (2010) The sex pheromone of the wasp spider Argiope bruennichi. Angew Chem Int Ed 49:2033–2036. doi: 10.1002/anie.200906311 CrossRefGoogle Scholar
  5. Gaskett AC (2007) Spider sex pheromones: emission, reception, structures, and functions. Biol Rev Camb Philos Soc 82:27–48. doi: 10.1111/j.1469-185X.2006.00002.x CrossRefPubMedGoogle Scholar
  6. Hebets EA, Chapman RF (2000) Electrophysiological studies of olfaction in the whip spider Phrynus parvulus (Arachnida, Amblypygi). J Insect Physiol 46:1441–1448. doi: 10.1016/S0022-1910(00)00068-8 CrossRefPubMedGoogle Scholar
  7. Jerhot E, Stoltz JA, Andrade MCB, Schulz S (2010) Acylated serine derivatives: a unique class of arthropod pheromones of the Australian redback spider, Latrodectus hasselti. Angew Chem Int Ed 49:2037–2040. doi: 10.1002/anie.200906312
  8. Kasumovic M, Andrade MCB (2004) Discrimination of airborne pheromones by mate-searching male western black widow spiders (Latrodectus hesperus): species- and population-specific responses. Can J Zool 82:1027–1034. doi: 10.1139/z04-081 CrossRefGoogle Scholar
  9. MacLeod EC, Andrade MCB (2014) Strong, convergent male mate choice along two preference axes in field populations of black widow spiders. Anim Behav 89:163–169. doi: 10.1016/j.anbehav.2013.12.023 CrossRefGoogle Scholar
  10. Papke MD, Riechert SE, Schulz S (2001) An airborne female pheromone associated with male attraction and courtship in a desert spider. Anim Behav 61:877–886. doi: 10.1006/anbe.2000.1675 CrossRefGoogle Scholar
  11. Prouvost O, Trabalon M, Papke M, Schulz S (1999) Contact sex signals on web and cuticle of Tegenaria atrica (Araneae, Agelenidae). Arch Insect Biochem Physiol 40:194–202. doi: 10.1002/(SICI)1520-6327(1999)40:4<194::AID-ARCH4>3.0.CO;2-P CrossRefGoogle Scholar
  12. R Core Team (2013) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, URL
  13. Ross K, Smith RL (1979) Aspects of the courtship behavior of the black widow spider, Latrodectus hesperus (Araneae: Theridiidae), with evidence for the existence of a contact sex pheromone. J Arachnol 7:69–77Google Scholar
  14. Schulz S (2013) Spider pheromones – a structural perspective. J Chem Ecol 39:1–14. doi: 10.1007/s10886-012-0231-6
  15. Schulz S, Toft S (1993) Identification of a sex pheromone from a spider. Science 260:1635–1637. doi: 10.1126/science.260.5114.1635 CrossRefPubMedGoogle Scholar
  16. Scott C, Vibert S, Gries G (2012) Evidence that web reduction by western black widow males functions in sexual communication. Can Entomol 144:672–678. doi: 10.4039/tce.2012.56 CrossRefGoogle Scholar
  17. Stoltz JA, McNeil JN, Andrade MCB (2007) Males assess chemical signals to discriminate just-mated females from virgins in redback spiders. Anim Behav 74:1669–1674. doi: 10.1016/j.anbehav.2007.03.011 CrossRefGoogle Scholar
  18. Tichy H, Gingl E, Ehn R, Papke M, Schulz S (2001) Female sex pheromone of a wandering spider (Cupiennius salei): identification and sensory reception. J Comp Physiol A 187:75–78. doi: 10.1007/s003590000175 CrossRefPubMedGoogle Scholar
  19. Trabalon M, Niogret J, Legrand-Frossi C (2005) Effect of 20-hydroxyecdysone on cannibalism, sexual behavior, and contact sex pheromone in the solitary female spider, Tegenaria atrica. Gen Comp Endocrinol 144:60–6. doi: 10.1016/j.ygcen.2005.04.011 CrossRefPubMedGoogle Scholar
  20. Uhl G, Elias DO (2011) Communication. In: Herberstein ME (ed) Spider behaviour: flexibility and versatility. Cambridge University Press, Cambridge, pp 127–189CrossRefGoogle Scholar
  21. Van den Dool H, Kratz P (1963) A generalization of retention index system including linear temperature programmed gas–liquid partition chromatography. J Chromatogr A 11:463–471. doi: 10.1016/S0021-9673(01)80947-X CrossRefGoogle Scholar
  22. Xiao Y, Zhang J, Li S (2010) Male-specific (Z)-9-tricosene stimulates female mating behaviour in the spider Pholcus beijingensis. Proc R Soc B Biol Sci 277:3009–3018. doi: 10.1098/rspb.2010.0763
  23. Xiao Y, Zhang J, Li S (2009) A two-component female-produced pheromone of the spider Pholcus beijingensis. J Chem Ecol 35:769–778. doi: 10.1007/s10886-009-9660-2 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Catherine Scott
    • 1
    Email author
  • Sean McCann
    • 1
  • Regine Gries
    • 1
  • Grigori Khaskin
    • 1
  • Gerhard Gries
    • 1
  1. 1.Department of Biological SciencesSimon Fraser UniversityBurnabyCanada

Personalised recommendations