Reproductive isolation among geographically and temporally isolated marine Brachionus strains

Using a polyclonal antibody against the mate recognition pheromone (MRP) of Brachionus rotundiformis Koshiki strain, we investigated the behavioral reproductive isolation and the similarity of MRP among geographically and temporally isolated B. rotundiformis strains. Males of the Koshiki strain did not discriminate in mating attempts among females of the Koshiki strain and those of conspecific allopatric strains from Hamana, Fiji, Thailand and Spain. Likewise, Koshiki males attempted mating with statistically indistinguishable frequency with Koshiki females and B. plicatilis strains. However, copulation was not consummated between Koshiki males and B. plicatilis females. The amount of anti-MRP binding to three allopatric B. rotundiformis strains was similar to that of the Koshiki strain, but binding to Hamana and the B. plicatilis strain was significantly lower. Four temporally separated B. rotundiformis populations were hatched from resting eggs collected from 0, 5, 10 and 15 cm depth in the sediment of Kai-ike pond in Koshiki island, Japan. Sediment age was determined using the ²¹⁰Pb method, allowing us to estimate that resting eggs from 15 cm depth were produced 65 years ago. Results of mating assays and anti-MRP binding showed that no behavioral reproductive isolation exists among the four temporally isolated Koshiki strains. B. rotundiformis appears to be reproductively isolated from B. plicatilis, but heterospecific matings are still attempted between B. plicatilis and B. rotundiformis, suggesting that the MRP remains sufficiently similar to elicit circling behavior.