To investigate effects of heat treatments including grain size control in substrate aluminum on nanopore arrays in anodic alumina template, aluminum was heat treated at $500^{circ}C$ for 1h. The heat treated aluminum was anodized by two successive anodization processes in oxalic solution and the nanopore arrays in anodic alumina layer were studied using TEM and FE-SEM. The highly ordered porous alumina templates with 110 nm interpore distance and 40 nm pore diameter have been observed and the pore array of the anodic alumina has a uniform and closely-packed honeycomb structure. In the case of alumina template obtained from heat treated aluminum substrate, the well- ordered nanopore region in anodic alumina increased and became more homogeneous compared with that from non-heattreated one.