Licochalcone A (LCA) is a characteristic compound of Glycyrrhiza inflata with anti-inflammatory, antioxidant and antitumor activities. However, G. inflata produces LCA in low quantities that does not meet the market demand. In this study, we found that DNA methylation inhibitor 5-azacitidine (5-azaC) successfully improved the LCA contents in G. inflata seedlings. Transcriptome analysis revealed a series of differentially expressed genes (DEGs), including transcription factors such as MYB, ERF, WRKY, and some structural genes related to flavonoid biosynthesis. However, whole genome bisulfite sequencing (BS-seq) results showed little effect of the 5-azaC treatment on the alteration of DNA methylation on these genes, indicating the possibility that 5-azaC acts as a stimulus, but not an epigenetic modulation factor to improve the LCA content in G. inflata. Additionally, we applied the 5-azaC treatment to field plants and hairy roots and successfully increased the LCA contents in both cases. This research demonstrates the feasibility of 5-azaC treatments in future applications to improve plant production of LCA.

Licochalcone A (LCA) 是胀果甘草 (Glycyrrhiza infata)的特征化合物，具有抗炎、抗氧化和抗肿瘤活性。然而，G. infata生产的 LCA 产量较低，无法满足市场需求。在本研究中，我们发现DNA甲基化抑制剂5-azacitidine (5-azaC)成功提高了G. infata幼苗的LCA含量。转录组分析揭示了一系列差异表达基因（DEG），包括MYB、ERF、WRKY等转录因子以及一些与类黄酮生物合成相关的结构基因。然而，全基因组亚硫酸氢盐测序（BS-seq）结果显示，5-azaC 处理对这些基因 DNA 甲基化的改变几乎没有影响，表明 5-azaC 可能作为刺激物，而不是表观遗传调节因子，提高膨胀棉的LCA含量。此外，我们对田间植物和毛状根进行了 5-azaC 处理，并成功提高了两种情况下的 LCA 含量。这项研究证明了 5-azaC 处理在未来应用中提高 LCA 植物产量的可行性。