Visualization and quantitative analysis of endosperm cavities in maize kernels via X-ray micro-computed tomography

Endosperm cavities within maize kernels influence quality traits such as kernel plumpness and hardness, serving as a key phenotypic indicator for assessing maize yield and quality. Research on endosperm cavities remains relatively scarce due to the small size of maize kernels and limitations in technical approaches. This study employed X-ray micro-computed tomography (μCT) three-dimensional reconstruction technology to extract morphological parameters and spatial configurations of endosperm cavities in multiple maize varieties, enabling visualisation and quantification of endosperm cavities within maize kernels. Endosperm cavities exhibit spatial heterogeneity within the kernels: embryo-adjacent cavities (EACs) are distributed in a conical pattern around the embryo, whereas internal endosperm cavities (IECs) are located in the floury endosperm at the tip region of the kernel and exhibit a boat-shaped morphology. The volume ratio of EACs to IECs is approximately 5:1. A coordinate system was established with the kernel length axis perpendicular to the horizontal plane, revealing the spatial positions of IECs (x = 3.5 mm, y = 2.1 mm, z = 1.1 mm) and EACs (x = 2.5 mm, y = 2.3 mm, z = 7.1 mm). Significant differences in endosperm cavity characteristics were observed among the different varieties. The average volume of the endosperm cavities was 4.1 mm 3 , with kernel porosities ranging from 0.4% to 3.3%. These parameters exhibited highly significant positive correlations with kernel volume, kernel thickness, cavity surface density, etc. Although manual sectioning methods cannot capture the 3D features of endosperm cavities, their operational simplicity and rapid data extraction allow them to reflect, to some extent, the characteristics of endosperm cavities across different maize varieties, as confirmed by this study. This study elucidates the morphology and spatial distribution of endosperm cavities, revealing significant varietal differences in cavity characteristics that correlate with grain morphological traits. These findings lay the groundwork for research into maize grain digital characterisation and the relationship between grain structure and function.