Timing of male sex pheromone biosynthesis in a butterfly - different dynamics under direct or diapause development.

The life history traits and behavior of the butterfly Pieris napi are well-known, as the species is often used as a model organism for evolutionary and ecological studies. The species has two or more generations per year in the major part of its temperate distribution, and as different selection pressures affect the different generations, both behavioral and physiological seasonal polyphenisms have been shown previously. Here, we explored the dynamics of male sex pheromone production. The two generations are shown to have significantly different scent compositions early in life; the direct developers--who have shorter time for pupal development--need the first 24 hr of adult life after eclosion to synthesize the sex pheromone citral (geranial and neral 1:1)--whereas the diapausing individuals who have spent several months in the pupal stage eclose with adult scent composition. Resource allocation and biosynthesis also were studied in greater detail by feeding butterflies (13)C labeled glucose either in the larval or adult stage, and recording incorporation into geranial, neral, and other volatiles produced. Results demonstrate that the pheromone synthesized by newly eclosed adult males is based on materials ingested in the larval stage, and that adult butterflies are able to synthesize the pheromone components geranial and neral and the related alcohols also from adult intake of glucose. In summary, our study shows that time-stress changes the timing in biosynthesis of the complete pheromone between generations, and underpins the importance of understanding resource allocation and the physiological basis of life history traits.

Nonvolatile chemical cues affect host-plant ranking by gravid Polygonia c-album females.

In a multiple-choice test, the preference of egg-laying Polygonia c-album (comma butterfly) females was studied for oviposition on plants bearing surrogate leaves treated with crude methanol extracts obtained from leaves of seven host-plant species: Humulus lupulus, Urtica dioica, Ulmus glabra, Salix caprea, Ribes nigrum, Corylus avellana, and Betula pubescens. The ranking order of surrogate leaves treated with host-plant extracts corresponded well to that reported on natural foliage, except R. nigrum. Thus, host-plant choice in P. c-album seems to be highly dependent on chemical cues. Moreover, after two subsequent fractionations using reversed-phase chromatography the nonvolatile chemical cues residing in the most polar water-soluble fractions evidently provided sufficient information for egg-laying females to discriminate and rank between the samples of more and less preferred plants, since the ranking in these assays was similar to that for natural foliage or whole methanol extracts, while the physical traits of the surrogate leaves remained uniform.

Anti-aphrodisiac pheromone, a renewable signal in adult butterflies.

The male butterfly Pieris napi produces the anti-aphrodisiac pheromone methyl salicylate (MeS) and transfers it to the female during mating. After mating she releases MeS, when courted by conspecific males, which decreases her attractiveness and the duration of male harassment, thus increasing her time available for egg-laying. In previous studies we have shown that males produced MeS from the amino acid L-phenylalanine (L-Phe) acquired during larval stage. In this study we show that adult males of P. napi can utilize L-Phe and aromatic flower volatiles as building blocks for production of anti-aphrodisiac pheromone and transfer it to females during mating. We demonstrate this by feeding butterflies with stable isotope labelled molecules mixed in sugar solutions, and, to mimic the natural conditions, we fed male butterflies with floral nectar of Bunias orientalis plants treated with labelled L-Phe. The volatiles from butterflies and plants were collected and identified by solid phase micro extraction, gas chromatography and mass spectrometry techniques. Since P. napi is polygamous, males would gain from restoring the titre of MeS after mating and the use of aromatic precursors for production of MeS could be considered as an advantageous trait which could enable butterflies to relocate L-Phe for other needs.