Petridis Antonios, Döll Stefanie, Nichelmann Lars, Bilger Wolfgang, Mock Hans-Peter
Department of Physiology and Cell Biology, Leibniz Institute for Plant Genetics and Crop Plant Research, Corrensstraße 3, D-06466, Gatersleben, Germany.
Botanical Institute, University of Kiel, D-24098, Kiel, Germany.
New Phytol. 2016 Aug;211(3):912-25. doi: 10.1111/nph.13986. Epub 2016 Apr 29.
Flavonoid synthesis is predominantly regulated at the transcriptional level through the MYB-basic helix-loop-helix (bHLH)-WD40 (MBW) (MYB: transcription factor of the myeloblastosis protein family, WD40: tanscription factor with a short structural motif of 40 amino acids which terminates in an aspartic acid-tryptophan dipeptide) complex, and responds to both environmental and developmental stimuli. Although the developmental regulation of flavonoid accumulation in Arabidopsis thaliana has been examined in great detail, the response of the flavonoid synthesis pathway to abiotic stress (particularly low temperature) remains unclear. A screen of a Dissociation element (Ds) transposon-induced mutation collection identified two lines which exhibited an altered profile of phenylpropanoid accumulation following exposure to low-temperature stress. One of the mutated genes (BRASSINOSTEROID ENHANCED EXPRESSION1 (BEE1)) encoded a brassinosteroid enhanced expression transcription factor, while the other (G2-LIKE FLAVONOID REGULATOR (GFR)) encoded a G2-like flavonoid regulator. Phenylpropanoid-targeted analysis was performed using high-performance LC-MS, and gene expression analysis using quantitative reverse transcription-PCR. In both mutants, the accumulation of quercetins and scopolin was reduced under low-temperature growing conditions, whereas that of anthocyanin was increased. BEE1 and GFR were both shown to negatively regulate anthocyanin accumulation by inhibiting anthocyanin synthesis genes via the suppression of the bHLH (TRANSPARENT TESTA8 (TT8) and GLABROUS3 (GL3)) and/or the MYB (PRODUCTION OF ANTHOCYANIN PIGMENTS2 (PAP2)) components of the MBW complex. Our results provide new insight into the regulatory control of phenylpropanoid metabolism at low temperatures, and reveal that BEE1 and GFR act as important components of the signal transduction chain.
类黄酮的合成主要在转录水平上通过MYB-碱性螺旋-环-螺旋(bHLH)-WD40(MBW)复合物进行调控(MYB:成髓细胞瘤蛋白家族的转录因子,WD40:具有40个氨基酸短结构基序且以天冬氨酸-色氨酸二肽结尾的转录因子),并对环境和发育刺激作出响应。尽管已对拟南芥中类黄酮积累的发育调控进行了详细研究,但类黄酮合成途径对非生物胁迫(尤其是低温)的响应仍不清楚。对解离元件(Ds)转座子诱导的突变体库进行筛选,鉴定出两个品系,它们在暴露于低温胁迫后表现出苯丙烷类积累模式的改变。其中一个突变基因(油菜素类固醇增强表达1(BEE1))编码一种油菜素类固醇增强表达转录因子,另一个(G2类黄酮调节剂(GFR))编码一种G2类黄酮调节剂。使用高效液相色谱-质谱联用仪进行了苯丙烷类靶向分析,并使用定量逆转录-聚合酶链反应进行了基因表达分析。在两个突变体中,低温生长条件下槲皮素和东莨菪苷的积累减少,而花青素的积累增加。研究表明,BEE1和GFR均通过抑制MBW复合物的bHLH(透明种皮8(TT8)和无毛3(GL3))和/或MYB(花青素色素合成2(PAP2))成分来抑制花青素合成基因,从而负调控花青素积累。我们的研究结果为低温下苯丙烷类代谢的调控控制提供了新的见解,并揭示BEE1和GFR是信号转导链的重要组成部分。