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Hematoxylin staining is an early indicator of Aluminum (Al) toxicity effects on the apices of young, developing roots grown in nutrient solution. In this work, the potential of this technique as a reliable and reproducible phenotypic index for Al tolerance in tropical maize genotypes was assessed, with its performance systematically compared to two other parameters widely used in breeding programs – relative seminal-root length (RSRL) and net seminal-root length (NSRL). Seeding roots from contrasting genotypes for Al sensitivity stained remarkably different after 24- and 48-h and 7-day exposures to 222 μM Al in nutrient solution, with the Al-dye complex being detected in both the outer (epidermis) and inner (cortex) portions of the roots from the sensitive cultivar. Hematoxylin staining was compared to the RSRL and NSRL parameters using 20 families from the third generation of selfing (S3) following the cross …