The high-frequency ultrasonic transducers with larger bandwidths yield excellent imaging performance in the biomedical field. However, achieving perfect acoustic impedance matching from the piezo-element to the target medium in the operating frequency spectrum is still a challenge. Conventional matching layers are mostly fabricated by only one or two uniform materials which are limited by their acoustic property. We propose a novel composite matching layer with gradient acoustic impedance based on a 1-3 gradient composite structure and multilevel matching theory. The proposed gradient-composite matching layer applied for ultrasonic transducer provides efficient impedance matching and ultrawide bandwidth which can significantly improve the quality of biomedical imaging. The active aperture size of the matching layer is 5× 5 mm2, and the overall thickness for five equivalent layers is 115 [Formula: see text]. The -6-dB bandwidth and the center frequency obtained by the ultrasonic transducer equipped with the 1-3 gradient composite matching layer are 141.7% and 22.3 MHz, respectively. The exceedingly good imaging performance of the fabricated ultrasonic transducer was demonstrated by the tungsten wire phantom and study on the biological tissues of a zebrafish and porcine eyeball. The theoretical and experimental results provide a novel train of thought for improving the quality of biomedical ultrasonic imaging.