Because of rapid heating, cooling, and solidification during metal additive manufacturing (AM), the resulting products exhibit strong anisotropy and are at risk of quality problems from metallurgical defects. The defects and anisotropy affect the fatigue resistance and material properties, including mechanical, electrical, and magnetic properties, which limit the applications of the additively manufactured components in the field of engineering. In this study, the anisotropy of laser power bed fusion 316L stainless steel components was first measured by conventional destructive approaches using metallographic methods, X-ray diffraction (XRD), and electron backscatter diffraction (EBSD). Then, anisotropy was also evaluated by ultrasonic nondestructive characterization using the wave speed, attenuation, and diffuse backscatter results. The results from the destructive and nondestructive methods were compared. The wave speed fluctuated in a small range, while the attenuation and diffuse backscatter results were varied depending on the build direction. Furthermore, a laser power bed fusion 316L stainless steel sample with a series of artificial defects along the build direction was investigated via laser ultrasonic testing, which is more commonly used for AM defect detection. The corresponding ultrasonic imaging was improved with the synthetic aperture focusing technique (SAFT), which was found to be in good agreement with the results from the digital radiograph (DR). The outcomes of this study provide additional information for anisotropy evaluation and defect detection for improving the quality of additively manufactured products.