Abstract
BackgroundThe flavonoid pathway is a long-standing and important tool for plant genetics, biochemistry, and molecular biology. Numerous flavonoid mutants have been identified in Arabidopsis over the past several decades in a variety of ecotypes. Here we present an analysis of Arabidopsis lines of ecotype Columbia carrying T-DNA insertions in genes encoding enzymes of the central flavonoid pathway. We also provide a comprehensive summary of various mutant alleles for these structural genes that have been described in the literature to date in a wide variety of ecotypes.FindingsThe confirmed knockout lines present easily-scorable phenotypes due to altered pigmentation of the seed coat (or testa). Knockouts for seven alleles for six flavonoid biosynthetic genes were confirmed by PCR and characterized by UPLC for altered flavonol content.ConclusionSeven mutant lines for six genes of the central flavonoid pathway were characterized in ecotype, Columbia. These lines represent a useful resource for integrating biochemical and physiological studies with genomic, transcriptomic, and proteomic data, much of which has been, and continues to be, generated in the Columbia background.
Highlights
The flavonoid pathway is a long-standing and important tool for plant genetics, biochemistry, and molecular biology
Confirmation of homozygous tt alleles T-DNA insertion lines in ecotype Col-0 were obtained from the Arabidopsis Biological Resource Center (ABRC, Columbus, OH) for genes encoding six of the eight enzymes of the central flavonoid pathway: chalcone synthase (CHS, SALK_020583), chalcone isomerase (CHI, SALK_034145 and CS300857 from the GABI-Kat project), flavanone 3-hydroxylase (F3H, SALK_113904), flavonoid 30-hydroxylase (F30H, SALK_053394), anthocyanidin synthase (ANS, SALK_073183), and anthocyanidin reductase (ANR, SALK_040250)
Note that a mutant allele for dihydroflavonol reductase (DFR) was recently identified in the Col-0 background that was not included in this study; no stable mutant allele has yet been identified in this ecotype for flavonol synthase 1 (FLS1)
Summary
The flavonoid pathway is a long-standing and important tool for plant genetics, biochemistry, and molecular biology. We describe the confirmation and preliminary characterization of mutant alleles for genes encoding flavonoid enzymes in Arabidopsis ecotype Columbia-0 (Col-0) that are available as part of the SALK collection of T-DNA insertion lines [19]. These lines represent a useful set of tools for analyzing the organization of flavonoid biosynthetic enzymes and their end products, as well the cellular, physiological and ecological roles of flavonoids. We present a compilation of mutant alleles for flavonoid structural gene that have been described in the literature to date in a variety of different ecotypes
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