Ion Homeostasis and Metabolome Analysis of Arabidopsis 14-3-3 Quadruple Mutants to Salt Stress.

Jing Gao,Alexander Erban,Paula J M Van Kleeff,Mark H De Boer,Dirk K Hincha,Albertus H De Boer,Joachim Kopka

doi:10.3389/fpls.2021.697324

Abstract

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.

Highlights

Salinity is one of the major abiotic stresses that limits agricultural productivity worldwide
The phosphorylation and interaction between SOS2-like protein kinase 5 (PKS5) and SOS2 could increase the interaction between SOS2 and Arabidopsis 14-3-3 kappa and lambda, which further repress SOS2 activity (Yang et al, 2019). These findings suggest that 14-3-3 and SOS3 proteins coordinately activate/inactivate the downstream protein kinases SOS2 and PKS5 to regulate cellular Na+ homeostasis
Our results demonstrated that 14-3-3 proteins are involved in multiple salt stress-related metabolic

Summary

INTRODUCTION

Salinity is one of the major abiotic stresses that limits agricultural productivity worldwide. Proteins from the 14-3-3 family are a class of molecular chaperones that have important functions in the adaptation of plants to salt and drought stress (Chen et al, 2006; Kaeodee et al, 2011) These functions are explained by the interaction of 14-3-3s with proteins that control ion homeostasis (Bunney et al, 2002; De Boer et al, 2013), ABA signaling (Sirichandra et al, 2010; Sun et al, 2015), and metabolism (Chung et al, 1999; Shin et al, 2011). 14-3-3 Functions in Ion Homeostasis and signaling pathways, with both redundancy and specificity between the 14-3-3 proteins

MATERIALS AND METHODS

RESULTS

DISCUSSION

Findings

DATA AVAILABILITY STATEMENT