Abstract
Previous work has shown that male sorghum plant bugs, Stenotus rubrovittatus (Matsumura) (Heteroptera: Miridae), are more attracted to young virgin females than to old virgin females and are not attracted to mated females. Therefore, we examined the effects of age and mating status on female sex pheromone levels. The pheromone components, hexyl butyrate, (E)-hex-2-en-1-yl butyrate, and (E)-4-oxohex-2-enal, were collected from females in two different ways. First, the compounds were extracted from whole bodies of each female and quantified by gas chromatography-mass spectrometry (GC-MS). Pheromone levels extracted from virgin female bodies decreased with age. In females that had just mated, pheromone levels did not differ from those of virgin females, and pheromone levels in mated females remained relatively constant up to 15 d after mating. This absence of change in pheromone levels extracted from whole bodies of mated females is not congruent with previous reports of lack of male attraction to these females. In a second method for pheromone recovery (adsorption on a PDMS-coated stir bar), GC-MS analysis showed that mated females released lower amounts of pheromone components compared to those emitted by virgin females of the same age (4 d). As was the case with whole body extracts, young virgin females (3 d) released higher amounts of pheromone components than did old virgin females (8 d). The results suggest that male response in S. rubrovittatus is dictated more by the quantities of pheromone components released into the volatile headspace by females than by the quantities present in the body.
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Acknowledgments
We thank Dr. T. Watanabe of National Agricultural Research Center (NARC) for suggestions and encouragement, Dr. T. Mitsunaga of NARC for advice on statistical analyses, and two anonymous reviewers for their valuable comments. This study was partly supported by a Grant-in-Aid for Research Projects Utilizing Advanced Technologies in Agriculture, Forestry and Fisheries from the Ministry of Agriculture, Forestry, and Fisheries of Japan.
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Supplemental Fig. 1
Standard linear calibration curves for each pheromone component from female Stenotus rubrovittatus. The y-axis indicates the peak area ratio of each component to the internal standard (heptadecane, R t = 10.04 min), while the x-axis indicates the quantity of each component. The amounts of hexyl butyrate (R t = 7.65 min), (E)-hex-2-en-1-yl butyrate (R t = 8.19 min), and (E)-4-oxohex-2-enal (R t = 9.31 min) in the extracts from female bodies were quantified by gas chromatography-mass spectrometry in selected ion monitoring (SIM) mode. Two ions were monitored for each component, m/z 129 and 89 for hexyl butyrate, m/z 170 and 100 for (E)-hex-2-en-1-yl butyrate, m/z 112 and 83 for (E)-4-oxohex-2-enal, and m/z 240 and 99 for heptadecane. Quantification was based on the former ions of the ion pairs for each component, whereas the latter ions served as reference or qualifier ions for diagnosis and quantitative analysis. The presence of a qualifier ion in the correct amount relative to the target ion gives evidence of correct target compound identification (http://www.chem.agilent.com/Library/Support/Documents/a05214.pdf, accessed April 18, 2010). (DOC 128 kb)
Supplemental Fig. 2
Mass spectra of (a) hexyl butyrate, (b) (E)-hex-2-en-1-yl butyrate, (c) (E)-4-oxohex-2-enal, and (d) heptadecane (internal standard) demonstrating that neither the analytes nor the standard had co-eluting peaks representing chemical compounds that would confound the quantitative analysis. The mass spectra were recorded from whole-body extracts of 5-d virgin female Stenotus rubrovittatus. (DOC 83 kb)
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Oku, K., Yasuda, T. Effects of Age and Mating on Female Sex Attractant Pheromone Levels in the Sorghum Plant Bug, Stenotus rubrovittatus (Matsumura). J Chem Ecol 36, 548–552 (2010). https://doi.org/10.1007/s10886-010-9795-1
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DOI: https://doi.org/10.1007/s10886-010-9795-1