Citrus CitERF6 Contributes to Citric Acid Degradation via Upregulation of CitAclα1, Encoding ATP-Citrate Lyase Subunit α.

Citric acid is the most abundant organic acid in citrus fruit, and the acetyl-CoA pathway potentially plays an important role in citric acid degradation, which occurs during fruit ripening. Analysis of transcripts during fruit development of key genes in the acetyl-CoA pathway and transient overexpression assay in citrus leaves indicated that CitAclα1 could be a potential target gene involved in citrate degradation. In order to understand more about CitAclα1, 23 transcription factors coexpressed with CitAclα1 in citrus fruit were identified by RNA-seq. Using dual-luciferase assays, CitERF6 was shown to trans-activate the promoter of CitAclα1 and electrophoretic mobility shift assays (EMSAs) showed that CitERF6 directly bound to a 5'-CAACA-3' motif in the CitAclα1 promoter. Furthermore, citric acid content was significantly reduced when CitERF6 was overexpressed in transgenic tobacco leaves. Taken together, these results indicate an important role for CitERF6 in transcriptional regulation of CitAclα1 and control of citrate degradation.