Boron confers salt tolerance through facilitating BnaA2.HKT1-mediated root xylem Na+ unloading in rapeseed (Brassica napus L.)

Boron (B) is an important limiting factor for plant growth and yield in saline soils, but the underlying molecular mechanisms remain poorly understood. In this study, we found that appropriate B supply obviously complemented rapeseed (Brassica napus L.) growth under salinity accompanied by higher biomass production and less reactive oxygen species accumulation. Determination of Na⁺ content in shoots and roots indicated that B significantly repressed root-to-shoot Na⁺ translocation, and non-invasive micro-tests of root xylem sap demonstrated that B increased xylem Na⁺ unloading in the roots of rapeseed plants under salinity. Comparative transcriptomic profiling revealed that B strongly upregulated BnaHKT1s expression, especially BnaA2.HKT1, in rapeseed roots exposed to salinity. In situ hybridizations analysis showed that BnaA2.HKT1 was significantly induced in root stelar tissues by high B (HB) under salinity. Green fluorescent protein and yeast heterologous expression showed that BnaA2.HKT1 functioned as a plasma membrane-localized Na⁺ transporter. Knockout of BnaA2.HKT1 by CRISPR/Cas9 resulted in hypersensitive of rapeseed plants to salinity even under HB condition, with higher shoot Na⁺ accumulation and lower biomass production. By contrast, overexpression of BnaA2.HKT1 ameliorated salinity-induced growth inhibition under B deficiency and salinity. Overall, our results proposed that B functioned as a positive regulator for the rapeseed growth and seed production under salt stress through facilitating BnaA2.HKT1-mediated root xylem Na⁺ unloading. This study may also provide an alternative strategy for the improvement of crop growth and development in saline soils.