Sex is generally determined by sex chromosomes in vertebrates, and sex chromosomes exhibit the most rapidly-evolving traits. Sex chromosome evolution has been revealed previously in numerous cases, but the association between sex chromosome origin and the reproduction mode transition from unisexual to sexual reproduction remains unclear. Here, we have isolated a male-specific sequence via analysis of amplified fragment length polymorphism from polyploid gibel carp (Carassius gibelio), a species that not only has the ability to reproduce unisexually but also contains males in wild populations. Subsequently, we have found through FISH analysis that males have several extra microchromosomes with repetitive sequences and transposable elements when compared to females. Moreover, we produced sex-reversed physiological females with a male-specific marker by using estradiol hormone treatment, and two gynogenetic families were established from them. In addition, the male incidence rates of two gynogenetic families were revealed to be closely associated with the extra microchromosome number of the sex-reversed physiological females. These results suggest that the extra microchromosomes in males might resemble a common feature of sex chromosomes and might play a significant role in male determination during the evolutionary trajectory of the reproduction mode transition from unisexual to sexual reproduction in the polyploid fish.KEYWORDS microchromosome; sex determination; sex chromosome; polyploid; gibel carp; genetics of sex S EX is a common phenomenon in nature and also one of the most important topics in life sciences, especially in evolutionary biology and genetics (Graves 2008). Most vertebrates are gonochoristic and reproduce by sexual reproduction. Different sex-determination systems, such as male heterogametic XX/XY sex chromosomes and female heterogametic ZZ/ZW sex chromosomes, as well as their numerous variants, have been revealed (Bachtrog et al. 2014;Mei and Gui 2015). Along with the rapid development of genomics and molecular genetic techniques, labile sex-determination systems and rapid sex chromosome turnovers have been noticed in both animals and plants recently (Bachtrog et al. 2014;Chen et al. 2014;Cortez et al. 2014;Graves 2014;Wei and Barbash 2015). Although how neo-sex chromosomes evolved (Roberts et al. 2009;Cortez et al. 2014;Vicoso and Bachtrog 2015) and how unisexual and sexual reproduction modes transformed (Jokela et al. 2009;Zhang et al. 2015) have been revealed, the association between sex chromosome origin and reproduction mode transition from unisexual to sexual reproduction remains unclear in vertebrates.The gibel carp, Carassius gibelio, has a wide geographic distribution in the Eurasian continent (Hanfling et al. 2005;Li and Gui 2008;Gui and Zhou 2010;Jakovlic and Gui 2011;Jiang et al. 2013) and diverse gynogenetic strains have been identified using biological traits and genetic markers (Zhou et al. 2000a;Yang and Gui 2004;Guo and Gui 2008). As a hexaploid with .150 chromosomes (...