Genomic and transcriptomic characterisation of the f3h/fls1/ans Arabidopsis thaliana mutant

The flavonoid biosynthesis related 2-oxoglutarate-dependent dioxygenases (2-ODDs) flavanone 3-hydroxylase (F3H), flavonol synthase (FLS), and anthocyanidin synthase (ANS) frequently exhibit overlapping multifunctionalities. As the analysis of such multifunctionalities is often limited to in vitro enzyme assays that do not necessarily reflect the functionalities of flavonoid 2-ODDs in planta, a flavonoid 2-ODD mutant collection was generated followed by detailed phenotyping. The f3h/fls1/ans A. thaliana mutant was generated to promote future in planta studies of 2-ODD multifunctionalities and to investigate the accumulation of flavonoids in the absence of the three major flavonoid 2-ODDs. Genomic ONT long-reads were generated for the construction of a de novo genome assembly of the f3h/fls1/ans mutant to analyse the T-DNA and transposon insertions. Six-days-old seedlings of the wildtypes Columbia-0 and Nössen-0, as well as the f3h/fls1/ans mutant were used for RNA-Seq analysis to compare the gene expression. For each genotype four biological replicates were used and subjected to Illumina sequencing.