Zeng Meijuan, Zhong Yongjia, Guo Zhiying, Yang Huiyong, Zhu Haisheng, Zheng Liling, Diao Yong
School of Biomedical Sciences, Huaqiao University, Quanzhou, China.
Crops Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou, China.
Front Plant Sci. 2022 Jul 1;13:919071. doi: 10.3389/fpls.2022.919071. eCollection 2022.
(Nees) Bremek () is an important medicinal plant. Its effective substances including indigo and indirubin are metabolites in indoleacetate metabolic pathway. Based on a previous transcriptome sequencing analysis, a WRKY transcription factor, BcWRKY1, in was identified, showing significant correlation with effective substances from . In this study, was cloned by reverse transcription-polymerase chain reaction (RT-PCR). Further analysis showed that the gene was 916 bp in length, containing three exons and two introns. The open reading frame (ORF) of was 534 bp in length and encoded a WRKY domain-containing protein with 177 amino acids residues. Subcellular localization showed that BcWRKY1 protein was mainly localized in the nucleus. It could bind to the W-box motif and its role in transcriptional activation was confirmed in yeast. The function of BcWRKY1 was investigated by overexpressing in . Metabolic profiles in wild type and transgenic were analyzed with LC-MS. Results showed that the metabolic profile was significantly changed in compared with wild type. Furthermore, indole-related metabolites were significantly increased in transgenic , and the metabolic pathway analysis showed that flavonoid biosynthesis was significantly enriched. Overexpression of significantly changed flavonoid and indole metabolism and indole-related metabolites were significantly upregulated. We postulated that the BcWRKY1 transcription factor might be involved in the regulation of effective substances metabolism in .
（菘蓝）板蓝是一种重要的药用植物。其有效物质包括靛蓝和靛玉红，是吲哚乙酸代谢途径中的代谢产物。基于之前的转录组测序分析，在板蓝中鉴定出一个WRKY转录因子BcWRKY1，它与板蓝中的有效物质显示出显著相关性。在本研究中，通过逆转录-聚合酶链反应（RT-PCR）克隆了BcWRKY1。进一步分析表明，BcWRKY1基因长度为916 bp，包含三个外显子和两个内含子。BcWRKY1的开放阅读框（ORF）长度为534 bp，编码一个含有177个氨基酸残基的WRKY结构域蛋白。亚细胞定位显示BcWRKY1蛋白主要定位于细胞核。它可以与W-box基序结合，并在酵母中证实了其转录激活作用。通过在板蓝中过表达BcWRKY1来研究其功能。用液相色谱-质谱联用（LC-MS）分析野生型和BcWRKY1转基因板蓝的代谢谱。结果表明，与野生型相比，BcWRKY1转基因板蓝的代谢谱发生了显著变化。此外，BcWRKY1转基因板蓝中吲哚相关代谢产物显著增加，代谢途径分析表明黄酮类生物合成显著富集。BcWRKY1的过表达显著改变了黄酮类和吲哚代谢，吲哚相关代谢产物显著上调。我们推测BcWRKY1转录因子可能参与了板蓝中有效物质代谢的调控。
