Topological Dirac semimetal is a newly discovered class of materials and has attracted intense attentions. This material can be viewed as a three-dimensional (3D) analogue of graphene and has linear energy dispersion in bulk, leading to a range of exotic transport properties. Here we report direct quantum transport evidence of 3D Dirac semimetal phase of layered material ZrTe5 by angular dependent magnetoresistance measurements under high magnetic fields up to 31 Tesla. We observed very clear

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... gative longitudinal magnetoresistance induced by chiral anomaly under the condition of the magnetic field aligned only along the current direction. Pronounced Shubnikov-de Hass (SdH) quantum oscillations in both longitudinal magnetoresistance and transverse Hall resistance were observed, revealing anisotropic light cyclotron masses and high mobility of the system. In particular, a nontrivial π-Berry phase in the SdH gives clear evidence for 3D Dirac semimetal phase. Furthermore, we observed clear Landau Level splitting under high magnetic field, suggesting possible splitting of Dirac point into Weyl points due to broken time reversal symmetry. Our results indicate that ZrTe5 is an ideal platform to study 3D massless Dirac and Weyl fermions in a layered compound.