Integrating Cytochrome P450-Mediated Herbicide Tolerance into Anthocyanin-Rich Maize Through Conventional Breeding

Meeting the rising demand for staple grains now requires cultivars that combine high yield, enhanced nutritional value, and strong chemical resilience. Blue-kernel landraces from central Mexico are rich in anthocyanins yet remain highly susceptible to post-emergence herbicides, whereas modern hybrids detoxify these compounds through cytochrome P450 (CYP450) enzymes. We crossed the anthocyanin-rich variety Polimaize with a CYP450-tolerant hybrid and evaluated the two parents and their F₁ segregants (designated “White” and “Yellow”) under greenhouse applications of mesotrione (75 g a.i. ha⁻¹), nicosulfuron (30 g a.i. ha⁻¹), and their mixture. Across 160 plants, the hybrid retained 95% of control dry matter and showed ≤7% foliar injury under all treatments, whereas Polimaize lost 28% biomass and exhibited 36% injury after nicosulfuron. The Yellow class matched hybrid performance while maintaining a blue pericarp and a β-carotene-rich endosperm, demonstrating that nutritional and agronomic traits can be stacked. The White class displayed heterosis-driven compensatory growth, exceeding its untreated biomass by 60% with nicosulfuron and by 82% with the mixture despite transient bleaching. Chlorophyll and carotenoid fluorescence revealed rapid, zeaxanthin-linked photoprotection in all tolerant genotypes, consistent with accelerated CYP450-mediated detoxification. These findings show that broad-spectrum herbicide tolerance can be introgressed into pigment-rich germplasm through conventional breeding, providing a non-transgenic path to herbicide-ready, anthocyanin-rich maize. The strategy preserves local biodiversity while delivering cultivars suited to intensive, weed-competitive agriculture and offers a template for integrating metabolic resilience into other native crops.