Salinity is one of the major abiotic stresses that limits agricultural productivity worldwide. Many proteins with defined functions in salt stress adaptation are controlled through interactions with members of the 14-3-3 family. In the present study, we generated three 14-3-3 quadruple knockout mutants (qKOs: klpc, klun, and unpc) to study the role of six non-epsilon group 14-3-3 proteins for salt stress adaptation. The relative growth inhibition under 100 mM of NaCl stress was the same for wild-type (Wt) and qKOs, but the accumulation of Na⁺ in the shoots of klpc was significantly lower than that in Wt. This difference correlated with the higher expression of the HKT1 gene in klpc. Considering the regulatory role of 14-3-3 proteins in metabolism and the effect of salt stress on metabolite accumulation, we analyzed the effect of a 24-h salt treatment on the root metabolome of nutrient solution-grown genotypes. The results indicated that the klpc mutant had metabolome responses that were different from those of Wt. Notably, the reducing sugars, glucose and fructose, were lower in klpc under control and salt stress. On the other hand, their phosphorylated forms, glucose-6P and fructose-6P, were lower under salt stress as compared to Wt. This study provided insight into the functions of the 14-3-3 proteins from non-epsilon group members. In summary, it was found that these proteins control ion homeostasis and metabolite composition under salt stress conditions and non-stressed conditions. The analyses of single, double, and triple mutants that modify subsets from the most effective qKO mutant (klpc) may also reveal the potential redundancy for the observed phenotypes.

盐度是限制全球农业生产力的主要非生物胁迫之一。许多在盐胁迫适应中具有明确功能的蛋白质是通过与 14-3-3 家族成员的相互作用来控制的。在本研究中，我们生成了三个 14-3-3 四重敲除突变体（qKOs：klpc, 昆仑， 和 解压器) 研究六种非 epsilon 组 14-3-3 蛋白对盐胁迫适应的作用。在 100 mM NaCl 胁迫下的相对生长抑制对于野生型 (Wt) 和 qKOs 是相同的，但 Na ⁺在芽中的积累韩国石油公司显着低于 Wt。这种差异与高表达相关HKT1 基因输入 韩国石油公司. 考虑到 14-3-3 蛋白在代谢中的调节作用以及盐胁迫对代谢物积累的影响，我们分析了 24 小时盐处理对营养液生长基因型根部代谢组的影响。结果表明，韩国石油公司突变体具有与 Wt 不同的代谢组反应。值得注意的是，还原糖、葡萄糖和果糖，在韩国石油公司在控制和盐胁迫下。另一方面，与 Wt 相比，它们的磷酸化形式葡萄糖-6P 和果糖-6P 在盐胁迫下较低。这项研究提供了对来自非 epsilon 组成员的 14-3-3 蛋白功能的深入了解。总之，发现这些蛋白质在盐胁迫条件和非胁迫条件下控制离子稳态和代谢物组成。对修改最有效 qKO 突变体子集的单突变体、双突变体和三重突变体的分析（韩国石油公司) 也可能揭示观察到的表型的潜在冗余。