Enhancing anthocyanin production by altering competition for substrate between flavonol synthase and dihydroflavonol 4-reductase

Abstract

Flavonoids, in particular the anthocyanins,are responsible for flower colour in manyspecies. The dihydroflavonols represent abranch point in flavonoid biosynthesis,being the intermediates for production ofboth the coloured anthocyanins, through theaction of the enzyme dihydroflavonol4-reductase (DFR), and the colourlessflavonols, produced by flavonol synthase(FLS). In this study the white-flowered,flavonol accumulating Mitchell line ofpetunia was used as a model to examine theinteraction between DFR and FLS enzymeactivities and possibilities forredirecting flavonoid biosynthesis awayfrom production of flavonols and towardsanthocyanins. Introduction of a 35SCaMV-DFR sense transgene construct causedthe production of anthocyanins, resultingin a pink-flowered phenotype. Furthermore,inhibition of FLS production throughintroduction of an FLS antisense RNAconstruct also led to anthocyaninproduction and a pink-flowered phenotype. A combination of both transgenes gave thehighest level of anthocyanin formation. Anthocyanins were produced in the DFR-senseand FLS-antisense transgenic lines in spiteof the greatly reduced levels of geneexpression in the Mitchell line for threeenzymes late in anthocyanin biosynthesis,anthocyanindin synthase, UDP-glucose:flavonoid 3-O-glucosyltransferase andUDP-rhamnose: anthocyanidin-3-glucosiderhamnosyltransferase. Thus, the level ofgene activity required for visibleanthocyanin formation is much lower thanthe high levels normally induced duringpetal development. Altering the balancebetween the DFR and FLS enzyme activities,using genetic modification, may be a usefulstrategy for introducing or increasinganthocyanin production in target ornamentalspecies.