Identification of BoFAR3a Reveals the Genetic Basis of a Glossy Green Trait in Broccoli

Mutants with a bright green appearance due to wax synthesis or deposition defects have been reported in various plants such as <i>Arabidopsis thaliana</i>, corn, and rice, but they are relatively rare in broccoli (a brassicaceae crop). Here, we describe SY03, a natural mutant of broccoli with a glossy green phenotype owing to epidermal wax deficiency. Genetic analysis indicated that the leaf luster trait of SY03 was controlled by a single recessive gene. By using the F₂ generation and combining bulked segregant analysis and molecular marker techniques, the candidate gene <i>BoFAR3a,</i> homologous to the Arabidopsis <i>FAR</i> gene, was identified within a 96.678 kb interval of chromosome C01. The A→G point mutation in exon 1 of the <i>BoFAR3a</i> coding sequence substitutes the canonical ATG start codon with GTG, which is predicted to abrogate or severely reduce translation initiation. RT-qPCR indicated that the expression levels of <i>BoFAR3a</i> were significantly decreased in the leaves of the glossy green phenotype mutant. Heterologous expression of <i>BoFAR3a</i> in <i>A. thaliana</i> restored the phenotype of <i>A. thaliana</i> mutant FAR3. The discovery of <i>BoFAR3a</i> is of great significance for breeding lustrous and commercially appealing broccoli varieties. This study systematically analyzed the molecular basis of the lustrous green phenotype in broccoli, providing new insights into the epidermal waxy regulatory network of cruciferous crops. In the future, the wax synthesis pathway can be precisely improved through gene editing technology, achieving a coordinated enhancement of the appearance quality and stress resistance of broccoli.