Abstract
Members of 14-3-3 protein family are involved in the proper operation of Fe acquisition mechanisms at physiological and gene expression levels in Arabidopsis thaliana. To more directly and effectively observe whether members of 14-3-3 non-epsilon group have a function in Fe-deficiency adaptation, three higher order quadruple KOs, kappa/lambda/phi/chi (klpc), kappa/lambda/upsilon/nu(klun), and upsilon/nu/phi/chi (unpc) were generated and applied for physiological analysis in this study. The mutant plants that combine kl with un (klun) or kl with pc (klpc) mutations showed a better Fe uptake than Wt plants at low medium Fe, while this phenotype was absent in unpc mutant. The higher Fe uptake by klun correlated with a higher Fe-deficiency induced expression of selected Fe-related genes. The dynamics of 14-3-3-client interactions analysis showed that a subset of 27 proteins differentially interacted with 14-3-3 in roots caused by Fe deficiency. Many of these Fe responsive proteins have a role in glycolysis and TCA cycle, the FoF1-synthase and in the cysteine/methionine synthesis. Also, the 14-3-3 interactome of the klun roots showed significant differences with that of Wt roots under Fe sufficient conditions, where most of these differential binding proteins showed enhanced binding in the klun mutant. Nevertheless, a clear explanation for the observed phenotypes awaits a more detailed analysis of the functional aspects of 14-3-3 binding to the target proteins identified in this study.
Protocol: optimising hydroponic growth systems for nutritional and physiological analysis of Arabidopsis thaliana and other plants
