SlPHL1 is involved in low phosphate stress promoting anthocyanin biosynthesis by directly upregulation of genes SlF3H, SlF3′H, and SlLDOX in tomato

Phosphate (Pi) deficiency is a common stress that limits plant growth and development. Plants exhibit a variety of Pi starvation responses (PSRs), including anthocyanin accumulation. The transcription factors of the PHOSPHATE STARVATION RESPONSE (PHR) family, such as AtPHR1 in Arabidopsis, play central roles in the regulation of Pi starvation signaling. Solanum lycopersicum PHR1-like 1 (SlPHL1) is a recently identified PHR involved in PSR regulation in tomato, but the detailed mechanism of its participation in Pi starvation-inducing anthocyanin accumulation remains unclear. Here we found that overexpression of SlPHL1 in tomato increases the expression of genes associated with anthocyanin biosynthesis, thereby promoting anthocyanin biosynthesis, but silencing SlPHL1 with Virus Induced Gene Silencing (VIGS) attenuated low phosphate (LP) stress-induced anthocyanin accumulation and expression of the biosynthesis-related genes. Notably, SlPHL1 is able to bind the promoters of genes Flavanone 3-Hydroxylase (SlF3H), Flavanone 3′-Hydroxylase (SlF3′H), and Leucoanthocyanidin Dioxygenase (SlLDOX) by yeast one-hybrid (Y1H) analysis. Furthermore, Electrophoretic Mobility Shift Assay (EMSA) and transient transcript expression assay showed that PHR1 binding t (sequence (P1BS) motifs located on the promoters of these three genes are critical for SlPHL1 binding and enhancing the gene transcription. Additionally, allogenic overexpression of SlPHL1 could promote anthocyanin biosynthesis in Arabidopsis under LP conditions through the similar mechanism to AtPHR1, suggesting that SlPHL1 might be functionally conserved with AtPHR1 in this process. Taken together, SlPHL1 positively regulates LP-induced anthocyanin accumulation by directly promoting the transcription of SlF3H, SlF3′H and SlLDOX. These findings will contribute to understanding the molecular mechanism of PSR in tomato.