Exogenous Trimethylamine N-Oxide (TMAO) Improves Apple Rootstock Drought Tolerance Through Physiological Modulation

Drought stress represents a major constraint on global apple production, with the widely used semi-dwarfing rootstock ‘M.26’ being particularly vulnerable to water deficit. Although the osmolyte trimethylamine N-oxide (TMAO) has been shown to improve abiotic stress tolerance in the model plant Arabidopsis, its potential role in enhancing drought resilience in woody fruit trees remains largely unexplored. Under prolonged moderate drought stress, exogenous TMAO application significantly promoted plant growth, mitigating the drought-induced suppression of plant height by 5.3–12.2% compared to untreated drought-stressed controls and alleviating the decline in above-ground biomass. This improvement was underpinned by a substantial alleviation of root growth inhibition, with TMAO restoring total root length and biomass from 37% in the control to only 6.1–9.5%. TMAO also fine-tuned the root-to-shoot ratio to favor resource allocation to roots. Consequently, TMAO-treated plants maintained superior leaf water status, exhibiting higher relative water content (drought-induced reduction limited to ~17.5% with TMAO versus 26.3% in the control). Physiologically, TMAO alleviated the drought-induced stomatal limitation of photosynthesis, sustaining higher net photosynthetic rate, stomatal conductance, and transpiration rate. Crucially, under severe drought stress, TMAO pretreatment markedly enhanced ‘M.26’ survival rates from approximately 39% in the untreated control to 60–68%, representing a relative increase of approximately 74%. Collectively, this study demonstrates that exogenous application TMAO significantly enhances drought tolerance in apple rootstock ‘M.26’, highlighting its potential as an effective and environmentally safe plant growth regulator for more sustainable cultivation of fruit trees under irregular/erratic irrigation conditions.