Colanero Sara, Perata Pierdomenico, Gonzali Silvia
PlantLab, Institute of Life Sciences, Scuola Superiore Sant'Anna, Pisa, Italy.
Front Plant Sci. 2018 Jun 19;9:830. doi: 10.3389/fpls.2018.00830. eCollection 2018.
The anthocyanin biosynthetic pathway is well characterized in plants. However, in tomato ( L.) an exhaustive knowledge of its regulation is still lacking. Tomato mutants showing higher levels of anthocyanins in fruits or vegetative tissues, such as ) or (), have been extensively exploited in the attempt to clarify the process. Nevertheless, only candidate genes have been proposed as responsible for such phenotypes. The recessive mutation likely represents an allelic variant of a gene introgressed in tomato from wild species. We performed genome sequencing of plants followed by candidate gene analysis, and identified a mutated gene encoding an R3-MYB protein. When overexpressed, this protein abolished anthocyanin production in tomato seedlings and plants, by silencing key regulators and biosynthetic genes of the pathway. The functional analysis of the protein clearly showed that it can negatively interfere with the activation of the anthocyanin biosynthetic pathway mediated by the endogenous MYB-bHLH-WDR (MBW) complexes. In particular, this R3-MYB protein can directly bind the bHLH factors which are part of the MBW complexes, therefore acting as a competitive inhibitor. The R3-MYB protein here described is therefore involved in a feedback mechanism that dampens the production of anthocyanins once activated by endogenous or exogenous stimuli. The mutation causes the production of a truncated version of the R3-MYB factor that cannot retain the full potential to inhibit the MBW complexes, thus leading to a constitutively higher production of anthocyanins.
花青素生物合成途径在植物中已得到充分表征。然而,在番茄中,对其调控的详尽了解仍然缺乏。番茄果实或营养组织中花青素水平较高的突变体,如或,已被广泛用于试图阐明这一过程。尽管如此,仅提出了候选基因作为此类表型的原因。隐性突变可能代表了从野生种渗入番茄的一个基因的等位变异。我们对植株进行了基因组测序,随后进行候选基因分析,鉴定出一个编码R3-MYB蛋白的突变基因。当该蛋白过表达时,通过沉默该途径的关键调控因子和生物合成基因,消除了番茄幼苗和植株中的花青素生成。该蛋白的功能分析清楚地表明,它可以负向干扰由内源性MYB-bHLH-WDR(MBW)复合体介导的花青素生物合成途径的激活。特别是,这种R3-MYB蛋白可以直接结合作为MBW复合体一部分的bHLH因子,因此起到竞争性抑制剂的作用。因此,这里描述的R3-MYB蛋白参与了一种反馈机制,一旦受到内源性或外源性刺激激活,就会抑制花青素的生成。突变导致产生截短版本的R3-MYB因子,该因子不能完全保留抑制MBW复合体的潜力,从而导致花青素的组成型高产量。