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

Abstract

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.

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Acknowledgments

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.

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Correspondence to Catherine Scott.

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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)

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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)

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Scott, C., McCann, S., Gries, R. et al. N-3-Methylbutanoyl-O-methylpropanoyl-L-serine Methyl Ester – Pheromone Component of Western Black Widow Females. J Chem Ecol 41, 465–472 (2015). https://doi.org/10.1007/s10886-015-0582-x

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Keywords

  • Latrodectus
  • Chemical communication
  • Pheromone
  • Amino acid
  • Silk
  • Courtship