AtERF13 and AtERF6 double knockout fine-tunes growth and the transcriptome to promote cadmium tolerance in Arabidopsis.

The toxic heavy metal cadmium (Cd) restricts plant growth. However, how plants fine-tune their growth to modulate Cd resistance has not been determined. Ethylene response factors (ERFs) are key regulators of Cd stress, and Arabidopsis thaliana ERF13 and ERF6 (AtERF13 and AtERF6) negatively regulate growth. We previously demonstrated that AtERF13 is a transcriptional activator that binds a Cd-responsive element. Herein, we report that Arabidopsis plants improve Cd tolerance by repressing AtERF13 and AtERF6. We found that AtERF13 and AtERF6 were strongly downregulated by Cd stress and that AtERF6 weakly bound Cd-responsive elements. Moreover, AtERF13 physically interacted with AtERF6. Importantly, AtERF13 and AtERF6 double knockout mutants, but not single mutants or overexpression lines, grew better, tolerated more Cd and had higher Cd contents than did the wild type. Comparative transcriptome analysis revealed that the double mutants regulate the defense response to cope with Cd toxicity. Accordingly, we propose that, upon Cd stress, Arabidopsis plants repress AtERF13 and AtERF6 to relieve their growth inhibition effects and adjust the transcriptome to adapt to Cd stress, leading to increased Cd tolerance. Our findings thereby provide deep mechanical insights into how dual-function transcription factors fine-tune growth and the transcriptome to promote Cd tolerance in plants.

IbInvInh2, a novel invertase inhibitor in sweet potato, regulates starch content through post-translational regulation of vacuolar invertase IbßFRUCT2.

As a key enzyme in the starch and sugar metabolic pathways in sweet potato (Ipomoea batatas (L.) Lam.), the vacuolar invertase (EC 3.2.1.26) IbßFRUCT2 is involved in partitioning and modulating the starch and sugar components of the storage root. However, the post-translational regulation of its invertase activity remains unclear. In this study, we identified three invertase inhibitors, IbInvInh1, IbInvInh2, and IbInvInh3, as potential interaction partners of IbßFRUCT2. All were found to act as vacuolar invertase inhibitors (VIFs) and belonged to the plant invertase/pectin methyl esterase inhibitor superfamily. Among the three VIFs, IbInvInh2 is a novel VIF in sweet potato and was confirmed to be an inhibitor of IbßFRUCT2. The N-terminal domain of IbßFRUCT2 and the Thr39 and Leu198 sites of IbInvInh2 were predicted to be engaged in their interactions. The transgenic expression of IbInvInh2 in Arabidopsis thaliana plants reduced the starch content of leaves, while its expression in the Ibßfruct2-expressing Arabidopsis plants increased the starch content of leaves, suggesting that the post-translational inhibition of IbßFRUCT2 activity by IbInvInh2 contributes to the regulation of the plant starch content. Taken together, our findings reveal a novel VIF in sweet potato and provide insights into the potential regulatory roles of the VIFs and invertase-VIF interaction in starch metabolism. These insights lay the foundation for using VIFs to improve the starch properties of crops.