A full-automatic sturgeon gonad tissue puncture sampling method comprises the following steps of preparation before puncture, puncture depth determination through B-mode ultrasound, slitting in an epidermal layer of a specimen for 2-3 mm, puncture position determination, sampling trigger and full-automatic emission gun storage. By adopting the full-automatic sturgeon gonad tissue puncture samplingmethod, sturgeon gonad tissue sampling can be achieved, and the sampling success rate can be also greatly improved.