A biaxial nematic phase had been predicted with D2h symmetry, wherein the mesogen’s long and short transverse axes are simultaneously aligned along the two orthogonal, primary and secondary directors, n and m, respectively. The unique low-angle x-ray diffraction patterns in the nematic phases exhibited by three rigid bent-core mesogens clearly reveal their biaxiality. The results of x-ray diffraction can be readily reproduced by ab initio calculations that explicitly include the bent-core shape in the form factor and assume short-range positional correlations.