The accurate assessment of peanut seed plumpness is crucial for optimizing peanut production and quality. The current method is mainly manual and visual inspection, which is very time-consuming and causes seed deterioration. A novel imaging technique is used to enhance the detection of peanut seed fullness using a non-destructive soft X-ray, which is suitable for the analysis of the surface or a thin layer of a material. The overall grayscale of the peanut is similar to the background, and the edge of the peanut seed is blurred. The inaccuracy of peanut overall and peanut seed segmentation leads to low accuracy of seed plumpness detection. To improve accuracy in detecting the fullness of peanut seeds, a seed plumpness detection method based on level set and multi-threshold segmentation was proposed for peanut images. Firstly, the level set algorithm is used to extract the overall contour of peanuts. Secondly, the obtained binary image is processed by morphology to obtain the peanut pods (the peanut overall). Then, the multi-threshold OTSU algorithm is used for threshold segmentation. The threshold is selected to extract the peanut seed part. Finally, morphology is used to complete the cavity to achieve the segmentation of the peanut seed. Compared with optimization algorithms, in the segmentation of the peanut pods, average random index (RI), global consistency error (GCE) and variation of information (VI) were increased by 10.12% and decreased by 0.53% and 24.11%, respectively. Compared with existing algorithms, in the segmentation of the peanut seed, the average RI, VI and GCE were increased by 18.32% and decreased by 9.14% and 6.11%, respectively. The proposed method is stable, accurate and can meet the requirements of peanut image plumpness detection. It provides a feasible technical means and reference for scientific experimental breeding and testing grading service pricing.