State Key Laboratory of Plant Cell and Chromosome Engineering, National Center for Plant Gene Research, Institute of Genetics and Developmental Biology/Innovative Academy of Seed Design, Chinese Academy of Sciences, Beijing, China.
University of Chinese Academy of Sciences, Beijing, China.
Plant Biotechnol J. 2021 Sep;19(9):1863-1877. doi: 10.1111/pbi.13604. Epub 2021 Jun 5.
Seed storage proteins (SSPs) are determinants of wheat end-product quality. SSP synthesis is mainly regulated at the transcriptional level. Few transcriptional regulators of SSP synthesis have been identified in wheat and this study aims to identify novel SSP gene regulators. Here, the R2R3 MYB transcription factor TuODORANT1 from Triticum urartu was found to be preferentially expressed in the developing endosperm during grain filling. In common wheat (Triticum aestivum) overexpressing TuODORANT1, the transcription levels of all the SSP genes tested by RNA-Seq analysis were reduced by 49.71% throughout grain filling, which contributed to 13.38%-35.60% declines in the total SSP levels of mature grains. In in vitro assays, TuODORANT1 inhibited both the promoter activities and the transcription of SSP genes by 1- to 13-fold. The electrophoretic mobility shift assay (EMSA) and ChIP-qPCR analysis demonstrated that TuODORANT1 bound to the cis-elements 5'-T/CAACCA-3' and 5'-T/CAACT/AG-3' in SSP gene promoters both in vitro and in vivo. Similarly, the homolog TaODORANT1 in common wheat hindered both the promoter activities and the transcription of SSP genes by 1- to 112-fold in vitro. Knockdown of TaODORANT1 in common wheat led to 14.73%-232.78% increases in the transcription of the tested SSP genes, which contributed to 11.43%-19.35% elevation in the total SSP levels. Our data show that both TuODORANT1 and TaODORANT1 are repressors of SSP synthesis.
种子贮藏蛋白(SSPs)是小麦终产品质量的决定因素。SSP 的合成主要受转录水平调控。在小麦中,仅鉴定到少数 SSP 合成的转录调控因子,本研究旨在鉴定新的 SSP 基因调控因子。本研究发现,来自节节麦(Triticum urartu)的 R2R3 MYB 转录因子 TuODORANT1 在灌浆过程中,于发育中的胚乳中特异性表达。在过表达 TuODORANT1 的普通小麦中,通过 RNA-Seq 分析测试的所有 SSP 基因的转录水平在整个灌浆过程中降低了 49.71%,导致成熟谷物中总 SSP 水平下降 13.38%-35.60%。在体外测定中,TuODORANT1 抑制 SSP 基因启动子活性和转录,抑制倍数为 1-13 倍。电泳迁移率变动分析(EMSA)和 ChIP-qPCR 分析表明,TuODORANT1 在体内和体外均结合到 SSP 基因启动子中的顺式元件 5'-T/CAACCA-3'和 5'-T/CAACT/AG-3'。同样,普通小麦中的同源物 TaODORANT1 在体外抑制 SSP 基因启动子活性和转录,抑制倍数为 1-112 倍。在普通小麦中敲低 TaODORANT1 导致测试的 SSP 基因的转录增加 14.73%-232.78%,导致总 SSP 水平升高 11.43%-19.35%。我们的数据表明,TuODORANT1 和 TaODORANT1 均为 SSP 合成的抑制剂。