Hyperoliids often call in mixed choruses with closely related species without an apparent spatial segregation (Lötters et al., 2004; Schiøtz, 1999), and multimodal signals might have evolved to avoid mismating and to facilitate navigation towards a conspecific mate in dense vegetation.
Since direct observations of mating in the wild are not possible, we assayed the frequency of outcrossing in individual trials in which two or three asci were placed in contact with one another, and in mass mating assays of dense sporulated populations using natural strains.
300 µl of each of two cultures with different antibiotic resistances were combined, washed, resuspended in 500µl of water, and plated on 60mm SOE plates, which created a dense lawn of asci on the mating plate.
By increasing a male's conspicuousness and its contrast to females, the blue colouration can act as a new type of visual signal directed at other males evolved to promote swift mate recognition and mate finding in dense aggregations.
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Smaller or less dense breeding aggregations may attract fewer females, thereby reducing mating probability for males attending smaller choruses, and may have subsequent negative population impacts.
In rodents, a dense, hard copulatory plug forms in the vagina following mating as a consequence of the protein crosslinking activity of transglutaminase IV, which is generated in the prostate gland and induces coagulation of seminal vesicle proteins (Dean, 2013).
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Extra-group mating has been observed during intergroup encounters (Cant et al. 2002) but such mating is difficult to observe as it is often surreptitious and occurs in dense bushes.
During early generations, organisms are forced to mate with partners which may be a considerable distance away, and therefore clusters will be less dense (more diverse).
