Abstract
Male genital organs are among the fastest evolving morphological structures. However, large parts of the male’s genitalia are often hidden inside the female during mating. In several bushcricket species, males bear a pair of sclerotized genital appendices called titillators. By employing synchrotron-based in vivo X-ray cineradiography on mating couples, we were able to visualize titillator movement and spermatophore attachment inside the female. Titillators are inserted and retracted rhythmically. During insertion the titillator processes tap the soft and sensillae-covered dorsal side of the female’s flap-like genital fold, which covers the opening of the female’s genitalia, without tissue penetration. Titillators thus appear to be initially used for stimulation; later they may apply pressure that forces the female’s genital fold to stay open, thereby aiding mechanically in spermatophore transfer.
Highlights
From the ampullae into the female’s spermatheca[19]
The styli at the end of the male’s subgenital plate are placed around the female’s ovipositor; they seem to have no function in copulation and spermatophore transfer (Fig. 3 and Video S2)
The live videos show the three major functions of the titillators of M. roeselii: the titillators tapping on the membranous dorsal parts of the genital fold stimulate the females, open their genital fold completely and allow the male to transfer the spermatophore
Summary
From the ampullae into the female’s spermatheca[19]. If the spermatophore transfer is not adequately ensured, this migration cannot take place and the sperm will otherwise be consumed by the female, impairing the reproductive success of the donor of the nuptial gift. Polygamous bushcricket females use several indicators, such as mating calls and male body mass, to assess the male’s fitness[18,20]. Experimental shortening of titillators in M. roeselii reduced the male’s capacity to transfer the spermatophore and increased female resistance behaviours during copulation[15]. The titillators in this species are expected to be important in spermatophore attachment and/or for suppressing female resistance behaviour via stimulation[14,15]. Video recordings of mating pairs during the phase of copulation are not conclusive as the titillators are covered by the female’s and male’s genital tissues for most of their motion sequence[14]. The aim of this study is to understand the complex function of the male’s titillators in copulation and spermatophore transfer. Using synchrotron-based in vivo X-ray cineradiography[21,22,23,24], we followed the real-time genital movements of mating couples
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