Cyprinodontiforms are a diverse group of approximately 900 pantropical and temperate fishes, mostly found in freshwater. Whereas the vast majority of fishes lay eggs (i.e. are oviparous), this group is unusual in that four groups of cyprinodont fishes give birth to living young (i.e. are viviparous). A molecular phylogenetic hypothesis was based on partial DNA sequences of the tyrosine kinase gene X-src. The study included the major lineages of fishes of the suborder Cyprinodontoidei, order Cyprinodontiformes. Our phylogeny agrees with some but not all of the conclusions of a previous morphological cladistic analysis (Parenti (Bull. Am. Mus. nat. Hist. 168, 335 (1981)). The differences are: (i) the Profundulidae are the sister group to the Goodeidae, not the sister group to all other cyprinodontoids; (ii) Fundulidae are the sister group to the Profundulidae and Goodeidae; (iii) Cubanichthys and the Cyprinodontinae might not be sister taxa; (iv) Cubanichthys, and not the Profundulidae, might be the most basal member of the cyprinodontoids; and (v) the Anablepinae and Poeciliinae might be sister groups. The molecular phylogeny was used to reconstruct the evolution of major life-history traits such as internal fertilization, copulatory organs, livebearing and placentas. Internal fertilization, modifications of the male's anal fin to form a copulatory organ, and viviparity probably evolved independently three times in cyprinodontiform fishes: in the subfamilies Goodeinae, Anablepinae and Poeciliinae (sensu Parenti 1981). The evolution of bundled sperm, spermatozeugmata, is probably not a prerequisite for internal fertilization because at least one species with internal fertilization has free spermatozoa. Livebearing (viviparity), which takes the form of ovoviviparity (where embryos are nourished by their yolk sac only), evolved only in the subfamily Poeciliinae. Advanced forms of viviparity, in which the mother provides additional nourishment to the embryos through placenta-like structures, apparently evolved at least three times from egg-laying ancestors: in the subfamilies Goodeinae, Anablepinae, and more than once in the Poeciliinae.
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