Nemacheilid loaches of the genus Paraschistura has been widely distributed from the Tigris drainage in Turkey, east throughout Iran and Pakistan to the Indus River and the Hari, Murghab and Helmand endorheic basins in Afghanistan, Iran, Pakistan and Turkmenistan. Herein, Paraschistura specimens from seven stations in Hari river basin were captured using hand-net and COI sequences of Paraschistura cristata and Paraschistura turcmenica were presented for all recognized species. Moreover; 119 sequences from ten species of Paraschistura in Genbank were added in order to assess of the phylogenetic position of the Paraschistura specimens and compare resulting tree with other studies. Molecular data obtained from Maximum likelihood (ML) tree and Bayesian tree (BI) were congruent and clearly indicate two main clades in phylogenetic tree and the genus Paraschistura in the Hari River basin, are considered as P. cristata and P. turcmenica.

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