Reproductive isolation in Chara aspera populations

Possible reproductive isolation between freshwater and brackish water populations of the dioecious charophyte Chara aspera was studied by means of cross-fertilization experiments and AFLP (Amplified Fragment Length Polymorphism). Three Swedish freshwater populations and three (German and Swedish) Baltic Sea populations of C. aspera were sampled. Cross-fertilization experiments were performed in a full combination setup of all populations and with two different salinities (0 and 10 PSU). Both freshwater and brackish water females formed about 70% more gametangia at 0 than at 10 PSU. Male individuals collected from freshwater had higher fertility than brackish water males at both salinities. 57% of all gametangia of females from freshwater developed into oospores compared to only 8% of gametangia of brackish water females. 42% of all oospores were fertilized in crosses between habitats (freshwater–brackish water) compared to 36% in crosses within habitats, the difference was not significant.Oospore and bulbil germination was investigated using propagules from freshwater and brackish water populations and incubation salinities of 0, 5, 10 and 20 PSU. None of the oospores collected from brackish water germinated. Germination of oospores and bulbils from freshwater was higher at 0 and 5 PSU than at higher salinities. Only around 40% of bulbils from brackish water germinated at 20 PSU compared to around 70% at the other three salinities. Germination of all bulbils was delayed at 20 PSU compared to other salinities.Genetic similarities (Jaccard indices of AFLP data) were higher within than between populations, but comparisons within habitat (freshwater–freshwater and brackish water–brackish water) were not different from comparisons between habitats.Our results did not identify any reproductive isolation between freshwater and brackish water populations, but indicate low gene flow between the two habitats. Oospore and bulbil germination success were highest at salinities corresponding to the conditions of their original habitat, suggesting genetic adaptation to their environmental conditions and indicating that propagules transported from freshwater to brackish water or vice versa will hardly develop into fertile plants. Additionally, brackish water plants perform poorer in all aspects of sexual reproduction than freshwater plants. Possibly, successful dispersal of oospores is not subjected to high selective pressure within the Baltic Sea where new sites easily can be colonized by means of vegetative reproduction. We assume that these adaptations will favour speciation within C. aspera and support the idea of the geologically young Baltic Sea as a “cradle of plant evolution”.