Autophagy, a bulk degradation system conserved among most eukaryotes, is also involved in responses to viral infection in plant. In our previous study, a new host factor P3IP was identified to interact with RSV (rice stripe virus) p3 and mediate its autophagic degradation to limit the viral infection. Here, we further discovered that P3IP of Nicotiana benthamiana (NbP3IP) participated in regulation of autophagy. Overexpression of NbP3IP induced autophagy and down-regulation of NbP3IP reduced autophagy. Combined the functions of autophagy-mediated plant defense against plant virus and regulation autophagy, we indicate that P3IP participates in the regulation of autophagy.