Analysis of T-DNA alleles of flavonoid biosynthesis genes in Arabidopsis ecotype Columbia

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.

Contrasting cadmium resistance strategies in two metallicolous populations of Arabidopsis halleri

“…Arabidopsis thaliana (thale cress) wild‐type (WT) and transgenic lines were in the Columbia‐0 (Col‐0) background. The T‐DNA insertion mutants of FLAVANONE 3‐HYDROXYLASE ( F3H ) (SALK_113904) and FLAVONOL SYNTHASE 1 ( FLS1 ) (SALK_076420) were kindly provided by Prof. Brenda Winkel (Department of Biological Science, VirginiaTech, Blacksburg, VA, USA; Bowerman et al ., ), transparent testa 8 ( tt8 ) (SALK_082999) seeds were provided by Prof. Song Susheng (Tsinghua‐Peking Center for Life Sciences, MOE Key Laboratory of Bioinformatics, Tsinghua University, Beijing, China; Qi et al ., ) and cyclic nucleotide gate channel 12 ( cngc12 ) knockout (ko) mutants were provided by Prof. Keiko Yoshioka (Department of Cell & Systems Biology, University of Toronto, Toronto, Canada; DeFalco et al ., ). Finally, T‐DNA insertion mutants of VACUOLAR MEMBRANE ATPAse 10 ( VMA10 ) (CS473188) and CYCLIC NUCLEOTIDE GATE CHANNEL 3 ( CNGC3 ) (SALK_056832; Gobert et al ., ) were obtained from GABI‐kat (Center for Biotechnology and Department of Biology, Bielefeld University, Bielefeld, Germany) or Nottingham Arabidopsis Stock Centre (NASC, University of Nottingham, Nottingham, UK).…”

Arabidopsis ROOT HAIR DEFECTIVE3 is involved in nitrogen starvation‐induced anthocyanin accumulation

“…Arabidopsis anthocyanin biosynthesis mutants tt4 –13 (SALK_020583), tt5 – 2 (GK‐176H03), tt6 – 3 (SALK_113904), and tt7 – 5 (SALK_053394) in the Col‐0 background were used, and are described in Bowerman et al (). Arabidopsis ethylene mutants were obtained from Dr. Brad Binder at the University of Tennessee, USA.…”

Use of the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique to analyse the Anthocyanidin Synthase (ANS) gene Locus in Zimbabwean sorghum landraces with different seed proanthocyanidin profiles

“…It is important to identify such mutations and establish their exact effects on the flavonoid pathway and on flavonoid profiles. Such studies have been carried out in other plants like arabidopsis, alfalfa and barley (Misyura et al, 2012;Abrahams et al, 2002;Bowerman et al, 2012;Buer and Djordjevic, 2009;von Wettstein 2007). However, similar studies are yet to be carried out in sorghum, hence this study is aimed at attempting to establish a link between sequence polymorphisms at the ANS gene locus and differences in proanthocyanidin profiles in sorghum.…”

“…Our interest is to find out if any sequence polymorphisms at this locus may explain observed differences in grain proanthocyanidin profiles. A number of flavonoid ANS gene mutants in Arabidopsis, namely tt 11, tt17 and tt 18 (tds-4) have been identified and characterised in Arabidopsis and alfalfa, both at the biochemical and molecular level (Misyura et al, 2012;Bowerman et al, 2012;Abrahams et al, 2002;Buer and Djordjevic, 2009). All these mutants were characterised by altered flavonoid profiles and deposition patterns in the grain.…”