Chloroplast proteome analysis of Nicotiana tabacum overexpressing TERF1 under drought stress condition

Chloroplast is indispensable for plant response to environmental stresses, growth and development, whose function is regulated by different plant hormones. The chloroplast proteome is encoded by chloroplast genome and nuclear genome, which play essential roles in plant photosynthesis, metabolism and other biological processes. Ethylene response factors (ERFs) are key transcription factors in activating the ethylene signaling pathway and plant response to abiotic stress. But we know little about how ethylene regulates plastid function under drought stress condition. In this study we utilized tobacco overexpressing tomato ethylene responsive factor 1 (TERF1), an ERF transcription factor isolated from tomato, to investigate its effects on the plastid proteome under drought stress condition by method of iTRAQ technology.

Results show that TERF1 represses the genes encoding the photosynthetic apparatus at both transcriptional and translational level, but the genes involved in carbon fixation are significantly induced by TERF1. TERF1 regulates multiple retrograde signaling pathways, providing a new mechanism for regulating nuclear gene expression. TERF1 also regulates plant utilization of phosphorus (Pi) and nitrogen (N). We find that several metabolic and signaling pathways related with Pi are significantly repressed and gene expression analysis shows that TERF1 significantly represses the Pi transport from root to shoot. However, the N metabolism is upregulated by TERF1 as shown by the activation of different amino acids biosynthesis pathways due to the induction of glutamine synthetase and stabilization of nitrate reductase although the root-to-shoot N transport is also reduced. TERF1 also regulates other core metabolic pathways and secondary metabolic pathways that are important for plant growth, development and response to environmental stresses. Gene set linkage analysis was applied for the upregulated proteins by TERF1, showing some new potential for regulating plant response to drought stress by TERF1.

Our research reveals effects of ethylene signaling on plastid proteome related with two key biological processes, including photosynthesis and nutrition utilization. We also provide a new mechanism to regulate nuclear gene expression by ERF1 transcription factor through retrograde signals in chloroplast. These results can enrich our knowledge about ERF1 transcription factor and function of ethylene signaling pathway.