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Background  Micro-computed tomography (μCT) bring a new opportunity to accurately quantify micro phenotypic traits of maize stem, also provide comparable benchmark to evaluate its dynamic development at the different growth stages. The progressive accumulation of stem biomass brings manifest structure changes of maize stem and vascular bundles, which are closely related with maize varietal characteristics and growth stages. Thus, micro-phenotyping (μPhenotyping) of maize stems is not only valuable to evaluate bio-mechanics and water-transport performance of maize, but also yield growth-based traits for quantitative traits loci (QTL) and functional genes location in molecular breeding.    Result  In this study, maize stems of 20 maize cultivars and two growth stages were imaged using μCT scanning technology. According to the observable differences of maize stems from the elongation and tasseling stages, function zones of maize stem were firstly defined to describe the substance accumulation of maize stems. And then a set of image-based μPhenotyping pipelines were implemented to quantify maize stem and vascular bundles at the two stages. The coefficient of determination (R 2 ) of counting vascular bundles was higher than 0.95. Based on the uniform contour representation, intensity-related, geometry-related and distribution-related traits of vascular bundles were respectively evaluated in function zones and structure layers. And growth-related traits of the slice, epidermis, periphery and inner zones were also used to describe the dynamic growth of maize stem. Statistical analysis demonstrated the presented method was suitable to the phenotyping analysis of maize stem for multiple growth stages.    Conclusions  The novel descriptors of function zones provide effective phenotypic references to quantify the differences between growth stages; and the detection and identification of vascular bundles based on function zones are more robust to determine the adaptive image analysis pipeline. Developing robust and effective image-based phenotyping method to assess the traits of stem and vascular bundles, is highly relevant for understanding the relationship between maize phenomics and genomics.

Phenotyping analysis of maize stem using micro-computed tomography at the elongation and tasseling stages