Influence of water availability on gender determination of gametophytes in a diploid-polyploid complex of a xerophytic fern genus

Environmental sex determination (ESD) is present in several animal and plant lineages. Diverse factors such as temperature, light or water availability have been described as sex determinants in these organisms. Among plants, ferns frequently display ESD. This work compares the effect of different levels of water availability in two diploid species of the xerophytic fern genus Cheilanthes and in their derived tetraploid, and if they are sensitive to antheridio- gen (i.e. maleness-inducing pheromone). Different watering regimes were applied to isolated gametophyte cultures of the three study species. Gametophyte survival, size, gender and sporophyte production were assessed after 13, 18 and 23 weeks of culture. Cultures combining spores and adult gametophytes were established to test the effect of anther- idiogen. Isolated gametophytes had an asexual to female to bisexual sequence that did not depend upon the degree of soil moisture. Both gender expression and growth reduction in response to water scarcity of the allotetraploid were more similar to those of one of the diploid parents. In all watering regimes, survival was higher in the allotetraploid, suggesting hybrid vigour, whereas automixis rate was similar in the three species and reached � 50 % at high mois- ture. This breeding system can ensure reproduction in the absence of males. In the three species, female gameto- phytes produced antheridiogens that enhanced maleness. This promotes a mixed mating system that could be favourable for ferns growing in xeric habitats.