Three-dimensional nano-imaging reveals subtle changes in xylem structure in CAD-deficient sorghum

Abstract

Lignin plays a central role in the formation and function of secondary cell walls in vascular plants. However, the structural consequences of lignin modification for cell wall properties and cellular function in grasses remain poorly understood. Here, we investigated how cinnamyl alcohol dehydrogenase (CAD) deficiency alters vascular cell architecture in Sorghum bicolor, using the brown midrib-6 (bmr6) mutant as a model system. Biochemical and histochemical analyses confirmed altered lignin chemistry in bmr6, including increased incorporation of hydroxycinnamaldehyde residues and reduced tricin levels. We applied ptychographic X-ray computed tomography (PXCT) to quantify the cell wall geometry, in three dimensions, at nanometer-scale resolution. PXCT enabled measurements of wall thickness distribution and lumen shape along tracheary elements. Analyses revealed no significant differences in wall thickness between wild-type and bmr6 plants. However, three-dimensional morphometric descriptors indicated reduced lumen convexity in bmr6, suggesting localized modifications not detectable by conventional two-dimensional imaging. Water flow numerical simulations through PXCT-derived images indicated reduced vessel permeability and simulated hydraulic conductivity in bmr6, suggesting that subtle geometric changes may influence performance. These findings highlight the value of three-dimensional imaging for resolving cell wall organization and provide new insight into the architectural resilience of grass xylem in response to targeted lignin modification. HighlightThree-dimensional X-ray nano-imaging reveals alterations in the cell wall architecture that affect simulated hydraulic performance under reduced CAD activity in sorghum.

Plant phenotyping relevance

植物の木部細胞壁形状をナノスケール3D画像から定量化するPXCT手法が研究の中心であり、壁厚・内腔形状・形態記述子を抽出しているため、植物表現型計測の実質的な適用に該当する。

Code and data availability

The supplied blocks describe PXCT imaging, segmentation, morphometrics, and lattice Boltzmann water-flow simulations of sorghum xylem, but contain no public data repository, deposited image/tomogram dataset, or author analysis code with an availability statement or URL. OpenLB is cited as a generic third-party library,

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