Adenine nucleotide translocase (ANT) is a constitutive component located in the inner membrane of mitochondria involved in the ADP/ATP exchange, which also has been known to be a major component of the mitochondrial permeability transition pore (PTP) complex and contributes to mitochondria-mediated apoptosis in yeast and mammalian.However, little has been known as for the function of ANT in fungi.Herein, in this study isolated an ANT of Puccinia striiformis (Pst), designated as PsANT.PsANT contains three typical conserved repeat mitochondria carrier protein domains, which are conserved among animals, Caenorhabditis elegan and fungi.Immuno-cytochemical localization confirmed the mitochondria localization of PsANT in normal Pst invading hypha cells or cells under collapse status,implying its roles in energy supply for the pathogen growth and cell death modulation for senescent cells.Transient expression of PsANT verified a pro-cell death role of PsANT in tobacco leaves as well as in wheat leaves.Further mutation analyses revealed a cumulative effect of the three conserved motifs in cell death induction.qRT-PCR analyses revealed an in-planta induced expression of PsANT during infection.Silencing of PsANT during the compatible wheat-Pst interaction by host induced gene silencing (HIGS) led to fewer branches and haustoria mother cells at the early stage but recovered Pst growth at the late stage.Our results suggested that PsANT may modulate the Pst growth via supplying energy, but not essential for the Pst pathogenicity.