ROS-induced ATP synthase mRNA degradation and metabolism dysfunction reveals the mechanism of artificial deteriorated cotton seeds

Seed aging is a complex biological process, the deterioration of oil crop seeds, in particular, has caused yield decline and economic loss to a large extent. However, research on the aging mechanism of oilseeds, such as cotton seeds, is still unclear. In this study, the physiological and biochemical changes in artificially aged cotton seeds were examined to further reveal the mechanism of seed aging and deterioration. Cotton seeds of "Xinluzao 74" were treated by artificial aging treatment method with high temperature (45°C) and high relative humidity (100%) for 1, 2, 3, 4, or 5 days, respectively, and untreated cotton seeds were used as control (CK). Our results showed that the germination rate, seed embryo viability, dehydrogenase activity, the activities of dehydrogenase antioxidant enzymes (SOD and POD), seed respiration rate, ATP content, ATP synthase activity and ATP synthase subunit mRNA integrity all showed a significant downward trend with the aging treatment time (P < 0.05), whereas the ROS generation (H2O2 content and ‧O2- production rate), relative conductivity, MDA content of seeds increased significantly, and the ultrastructure of cell membrane, mitochondria and chromatin of seed embryo was seriously damaged. Correlation analysis showed that there was a strong negative correlation between germination rate, SOD and POD activities, and respiration rate with H2O2 content (P < 0.05). This study reveals that excessive ROS, particularly H₂O₂, causes oxidative damage to ATP synthase subunit mRNAs, leading to impaired mitochondrial respiration and reduced seed vigor. These findings provide new molecular evidence linking oxidative RNA damage with seed aging, which could inform seed quality evaluation and storage strategies.