Genome-wide identification and characterization of flavonol synthase (FLS) gene family in Brassica vegetables and their roles in response to biotic and abiotic stresses

Flavonol synthase (FLS) serves as a pivotal enzyme in the flavonol biosynthesis pathway, contributing to the abundance of flavonol compounds found in Brassica vegetables. Nevertheless, there are limited investigations on the identification and analysis of FLS gene family and their precise roles in responding to biotic and abiotic stresses remain elusive in Brassica vegetables. Herein, a total of 6 (Brassica rapa), 9 (Brassica oleracea), and 14 (Brassica napus) FLS genes were identified and distributed on 4, 5, and 8 chromosomes, respectively. Notably, while FLS genes exhibited considerable variation in sequence, exon-intron structure, and molecular weight, they displayed highly conserved domain and motif compositions. Of the six identified BrFLSs, namely BrFLS1, BrFLS2, BrFLS3.1, BrFLS3.2, BrFLS3.3, and BrFLS4, all were located in the nucleus and cell membrane except for BrFLS4 exclusively in the nucleus. Specifically, BrFLS1, BrFLS3.2, and BrFLS3.3 showed up-regulation after 6-BA, ABA, BR, and GA3 treatments, while BrFLS2 exhibited down-regulation. Intriguingly, BrFLS members displayed distinct expression patterns under diverse abiotic and biotic stresses. Employing the virus-induced gene silencing (VIGS) technology, we validated BrFLS1/BrFLS3.2, BrFLS1/BrFLS3.3, BrFLS2, BrFLS1, and BrFLS1/BrFLS2/ BrFLS3.1/BrFLS3.3 as vital candidate genes, contributing to the flavonol biosynthesis pathway under drought, salt, high temperature, low temperature, and anthracnose stresses in Brassica vegetables, respectively, and they enhanced their resistance to stresses by bolstering antioxidant capacity. Overall, our findings shed light on the biological roles of FLS gene family in the flavonoid biosynthesis pathway, offering valuable genetic resources for crop enhancement.