Accelerated germination of aged recalcitrant seeds by K⁺-rich bulk oxygen nanobubbles

Bulk nanobubbles, measuring less than 200 nm in water, have shown their salient properties in promoting growth in various species of plants and orthodox seeds, and as potential drug-delivery carriers in medicine. Studies of recalcitrant seeds have reported markedly increased germination rates with gibberellin treatment; however, neither the mechanism promoting germination nor the implication for food safety is well elucidated. In our study, recalcitrant wasabi (Eutrema japonicum) seeds treated with bulk oxygen nanobubbles (BONB) containing K⁺, Na⁺, and Cl^- (BONB-KNaCl) showed significantly accelerated germination. As germination progressed, 99% of K⁺ ions in the BONB-KNaCl medium were absorbed by the seeds, whereas Ca²⁺ ions were released. These results suggest that the germination mechanism involves the action of K⁺ channels for migration of K⁺ ions down their concentration gradient and Ca²⁺ pumps for the movement of Ca²⁺ ions, the first potential discovery in germination promotion in recalcitrant seeds using nutrient solutions with BONB-KNaCl.