GHCYP706A7 governs anthocyanin biosynthesis to mitigate ROS under alkali stress in cotton.

key messageFlavonoid 3'-hydroxylase synthesis gene-GHCYP706A7, enhanced cotton resistance to alkali stress by scavenging ROS to regulate anthocyanin synthesis. Anthocyanins are a class of flavonoids that play a significant role in mediating plant responses to adverse environmental conditions. Flavonoid 3'-hydroxylase (F3'H), a member of the cytochrome P-450 (CYP) family, is a pivotal enzyme involved in the biosynthesis of anthocyanins. The present study identified 398 CYPs in the Gossypium hirsutum genome, of which GHCYP706A7 was responsible for F3'H synthesis and its ability to respond to alkaline stress. GHCYP706A7 suppression through virus-induced gene silencing (VIGS) diminished tolerance to alkali stress in cotton, evidenced by significantly reduced anthocyanin synthesis, markedly decreased antioxidant capacity, notable increases in reactive oxygen species, severe cellular damage, and observably decreased stomatal opening. The cumulative effects of these physiological disruptions ultimately manifest in cotton wilting and fresh weight decline. These findings lay a foundation for further investigations into the role of CYPs in regulating anthocyanin synthesis and responding to alkali stress.