Involvement of carbon catabolite repression on regulation of endopolygalacturonase gene expression in citrus fruit

Abstract

 The Acpg1 gene of Alternaria citri encodes an extracellular endopolygalacturonase that is important for virulence in citrus fruits. Expression of Acpg1 is regulated by substrate induction and carbon catabolite repression. A green fluorescent protein (GFP) gene was employed as a reporter gene to define 813 bases upstream of the translation start site comprising the Acpg1 promoter. This upstream sequence contains five putative binding sequences of catabolite repressive element A (CreA), a cis-acting zinc finger repressor involved in carbon catabolite repression. We constructed each CreA-binding site-deleted Acpg1 promoters with GFP reporter gene and transformed them to A. citri. The construct PGPDL4 deleted from −401 to −813 showed both substrate induction and catabolite repression, whereas PGPDL5 additionally deleted from −1 to −84, including one putative CreA-binding site, resulted in a loss of catabolite repression function. Green fluorescence of PGPDL4 was induced by pectin in the peel but was repressed completely in the juice sac area of citrus fruit. However, green fluorescence of PGPDL5 was induced in both the peel and juice sac area, indicating that repression of Acpg1 in the juice sac area is likely accomplished by carbon catabolite repression.