onstruction of a phylogenetic tree by categorizing the studys 21 species allowed phylogenetic relationships between each of them to be established in order to test alternative hypotheses for the diversification that occurred over time. Examination of these data alongside geological-scale changes that have affected aquatic ecosystems with time brought out evidence that marine incursions played a fundamental role in the appearance then the distribution of piranha species. Five million years ago, the Atlantic Ocean advanced, its waters finding their way far onto the Amazon flood plain. The saline water invaded the lowland expanse of the great river and penetrated its tributaries situated below 100 metres of altitude, provoking the disappearance of many species of freshwater fish. Some of these would nevertheless have succeeded in finding refuge at high altitude, in particular in rivers that flowed on the Guianan and Brazilian shields.
DNA analysis confirmed this hypothesis and showed that the piranha populations present in the Amazon flood plain but situated 100 metres above sea-level have been in existence for no more than 3 million years. Hitherto, certain specialists had suggested that the present-day piranha species had arisen in the lower sections of the great rivers of South America. The scientists thought that from centres of speciation, piranhas would subsequently have dispersed to colonize the more upstream reaches of the river system. However, the results of the study give sustenance to another scenario.
According to that new hypothesis, during the marine incursion phase some piranha populations would have survived in the upstream parts of the network. Such populations would have differentiated into speciesfollowing the fragmentation of their zone of distribution, but probably also in response to ecological constraints specific to the basin where they were kept in isolation from each other. Once the ocean had regressed again, 3 million years ago
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