The behavioral functions of larval firefly lights have been studied, and several theories have been proposed. 1 Male flash patterns are thought to be affected by sexual selection and females have been shown to prefer certain light signal characteristics. 2 In addition to these behaviors, it is generally assumed that most non-luminous fireflies locate mates through the use of pheromones. The courtship patterns of Japanese fireflies seem to indicate numerous types of communication, which include pheromones as well as light signals. 3 Our analytical study of fireflies was based on the hypothesis that a certain chemical signal mediates a particular interaction between male and female fireflies. This hypothesis can be tested by collecting, analyzing and identifying these compounds. Using our previously reported method, pheromones were adsorbed by direct contact with glass surfaces. 4 We call this method direct contact extraction. Because this technique is nondestructive, it is possible to subject extracted hydrocarbons to gas chromatography-mass spectrometry (GC-MS) directly using a solvent. Here, we report on the GC-MS profiles of extracts from three Japanese firefly species (Luciola lateralis, Luciola cruciata, and Lucidina biplagiata) and three North American firefly species (Lucidota atra, Photuris lucicrescens, and Photuris cinctipennis). Several research aspects concerning present analytical methods are discussed: (1) distinguish firefly species using the GC-MS profile as finger prints, (2) the intraspecific differences of fireflies, which could be interpreted as a geographic variation, and (3) chemical-ecological strategy of luminous and non-luminous fireflies.
ExperimentalLuciola lateralis were collected in Yokohama, Kanagawa prefecture and Chino, Nagano prefecture. Luciola cruciata were collected in Yokohama, Kanagawa prefecture. Lucidina biplagiata were collected in Hayama, Kanagawa prefecture. Lucidota atra, Photuris lucicrescens, and Photuris cinctipennis were collected in Meramec state park Missouri, USA.Glass vials (50 ml) were washed in hexane and dried at 200˚C in an electric oven. Three fireflies were collected; if this was not possible, one or two fireflies were collected and placed into one clean glass vial at the field site. Fireflies were free to crawl inside the glass vials for 2 -3 h during the mating period. After the fireflies were released, 1 ml of dichloromethane was added to the glass vials, which were then shaken several times to collect any remaining compounds from the firefly without further condensation.Extracts (1 µl) were analyzed using a mass spectrometer (MS) (JMS-SX102A; JEOL, Tokyo, Japan) equipped with a gas chromatograph (GC) (HP 5890A; Hewlett Packard, Palt Alto, USA). An HP-5 capillary column (30 m length, 0.32 mm i.d., 0.25 mm film thickness; Hewlett Packard, Palt Alto, USA) was used to separate the products into the volatile components. The initial temperature was 50˚C, the final temperature was 280˚C and a linear temperature program of 20˚C/min was applied. The injector te...