Citric acid is an important organic acid with wide applications and diverse biological functionality. As the predominant organic acid in lemons, citric acid plays a crucial role in determining the flavor of citrus, especially in lemons. ATP citrate lyase (ACL, EC4.1.3.8) is the keg gene in citric acid metabolism. Several research studies on ACL only focused on high-sugar- and low-acid-content citrus varieties; however, the ACL mechanism in lemons with high acid and low sugar levels remains undetermined. In this study, a key candidate gene, ClACLB-1, for citrate cleavage was identified from the genome data of 'Xiangshui' lemon [Citrus limon (L.) Burm f.]. The putative protein coded by the gene ClACLB-1 is localized in the nuclear and cell membranes. The ClACLB-1 gene was expressed in all tissues, with the highest expression in male flowers and the lowest expression in mature fruits; the expression decreased during lemon fruit development. The overexpression of ClACLB-1 in transgenic tomatoes significantly increases the activity of citrate lyase, which subsequently reduces citric acid content. This study clarified the function of the ClACLB-1 gene in cleaving citric acid, provided new insights into the citric acid metabolism of citrus, and offered a theoretical reference for reducing acid and increasing sugar in citrus to improve fruit quality. It also helped to enhance the understanding of the metabolism and role of citrate in plants.
Keywords: ClACLB-1; citric acid; gene function; genome-wide identification; lemon.