Simultaneous triple staining for detecting cell-type specific spatio-temporal distribution of cell wall materials in monocot roots

Abstract

Summary Anatomical and histochemical imaging of grass root systems relies on tissue sectioning and cell wall staining dyes because molecular reporter lines are limited for most organisms. Distinct staining dyes require variable incubation time and concentration across different tissues and organisms. As a result, staining with multiple dyes becomes time consuming or challenging. Here, we report a rapid method to perform simultaneous triple staining on a glass slide. The entire protocol requires ∼4 hours and a smaller volume of stain than traditional methods. We tested this method using the roots of two economically important crops, Triticum aestivum (wheat) and Zea mays (maize), as proof of concept. We have also demonstrated the presence of exodermis in wheat roots. Additionally, we identified the formation of polar lignin caps in maize exodermis using our simultaneous triple staining method. This method empowers a quantitative approach to cell biology by elucidating cell-type specific spatio-temporal distribution of cell wall materials in monocot root systems.

Plant phenotyping relevance

単子葉植物の根における細胞壁物質の細胞型別・時空間分布を定量的に可視化する同時三重染色法の開発が中心であり、植物状態の取得・解析手法に該当する。

Code and data availability

The paper describes the STAR triple-staining protocol for wheat and maize root imaging with ImageJ/Fiji quantification, but no public phenotype dataset, image collection, analysis code, or trained model is deposited. Supplemental figures/movies are referenced but no public repository or URL for them is given; vendor/ma

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