Mechanisms of reaction of hematoxylin with aluminium-treated wheat roots

Hematoxylin stain is used for localization of aluminium in plant root tissue and is the basis of a rapid assay of relative At tolerance among wheat cultivars. In the present study, mechanisms by which hematoxylin might selectively stain Al-sensitive wheat roots have been examined. The results are consistent with the idea that in Al-sensitive cultivars, hematoxylin forms complexes with Al that precipitate with phosphate as AlPO₄ in intercellular spaces: (1) Al and P are co-localized in the cell wall region of the outer cortex of Al-stressed roots by using x-ray microanalysis: (2) the molybdenum blue histochemical stain for extracellular phosphate reveals areas of stain that parallel those observed with hematoxylin; (3) in vitro, the presence of phosphate in an Al-hematoxylin reaction mixture causes formation of precipitate when the P/Al ratio exceeds 1. 0. I suggest that selective hematoxylin staining of Al-sensitive wheat cultivars is the result of direct damage by Al to root cells, leading to leakage of phosphorus into the cell wall region. Cultivars whose roots are damaged by Al in this way are likely to be judged Al-sensitive when other criteria such as growth or crop yield are used.