Zhejiang Provincial TCM Key Laboratory of Chinese Medicine Resource Innovation and Transformation, Zhejiang International Science and Technology Cooperation Base for Active Ingredients of Medicinal and Edible Plants and Health, Jinhua Academy, School of Pharmaceutical Sciences, Academy of Chinese Medical Sciences, Zhejiang Chinese Medical University, Hangzhou, 310053, China.
State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-Di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
BMC Genomics. 2024 Aug 9;25(1):776. doi: 10.1186/s12864-024-10683-7.
Transcription factors (TFs) of plant-specific SHORT INTERNODES (SHI) family play a significant role in regulating development and metabolism in plants. In Artemisia annua, various TFs from different families have been discovered to regulate the accumulation of artemisinin. However, specific members of the SHI family in A. annua (AaSHIs) have not been identified to regulate the biosynthesis of artemisinin.
We found five AaSHI genes (AaSHI1 to AaSHI5) in the A. annua genome. The expression levels of AaSHI1, AaSHI2, AaSHI3 and AaSHI4 genes were higher in trichomes and young leaves, also induced by light and decreased when the plants were subjected to dark treatment. The expression pattern of these four AaSHI genes was consistent with the expression pattern of four structural genes of artemisinin biosynthesis and their specific regulatory factors. Dual-luciferase reporter assays, yeast one-hybrid assays, and transient transformation in A. annua provided the evidence that AaSHI1 could directly bind to the promoters of structural genes AaADS and AaCYP71AV1, and positively regulate their expressions. This study has presented candidate genes, with AaSHI1 in particular, that can be considered for the metabolic engineering of artemisinin biosynthesis in A. annua.
Overall, a genome-wide analysis of the AaSHI TF family of A. annua was conducted. Five AaSHIs were identified in A. annua genome. Among the identified AaSHIs, AaSHI1 was found to be localized to the nucleus and activate the expression of structural genes of artemisinin biosynthesis including AaADS and AaCYP71AV1. These results indicated that AaSHI1 had positive roles in modulating artemisinin biosynthesis, providing candidate genes for obtaining high-quality new A. annua germplasms.
植物特异性短节间(SHORT INTERNODES,SHI)家族的转录因子(Transcription Factors,TFs)在植物的发育和代谢调控中发挥着重要作用。在青蒿中,已发现来自不同家族的各种 TF 调节青蒿素的积累。然而,青蒿中 SHI 家族的特定成员(AaSHIs)尚未被鉴定为调节青蒿素生物合成。
我们在青蒿基因组中发现了五个 AaSHI 基因(AaSHI1 至 AaSHI5)。AaSHI1、AaSHI2、AaSHI3 和 AaSHI4 基因的表达水平在毛状体和幼叶中较高,也受到光照诱导,而在黑暗处理下则降低。这四个 AaSHI 基因的表达模式与青蒿素生物合成的四个结构基因及其特定调节因子的表达模式一致。双荧光素酶报告基因检测、酵母单杂交检测和青蒿瞬时转化提供的证据表明,AaSHI1 可以直接结合到结构基因 AaADS 和 AaCYP71AV1 的启动子上,并正向调节它们的表达。本研究提出了候选基因,特别是 AaSHI1,可以考虑用于青蒿素生物合成的代谢工程。
总之,对青蒿 AaSHI TF 家族进行了全基因组分析。在青蒿基因组中鉴定出五个 AaSHIs。在所鉴定的 AaSHIs 中,AaSHI1 被发现定位于细胞核,并激活青蒿素生物合成的结构基因,包括 AaADS 和 AaCYP71AV1 的表达。这些结果表明,AaSHI1 在调节青蒿素生物合成中具有积极作用,为获得高质量的青蒿新种质提供了候选基因。