Key Laboratory of Urban Agriculture (South), Ministry of Agriculture, Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, People's Republic of China.
College of Pharmaceutical Science, Zhejiang Chinese Medical University, Hangzhou, People's Republic of China.
Biotechnol Appl Biochem. 2021 Apr;68(2):338-344. doi: 10.1002/bab.1931. Epub 2020 Jun 29.
Artemisinin, an effective antimalarial compound, is isolated from the medicinal plant Artemisia annua L. However, because of the low content of artemisinin in A. annua, the demand of artemisinin exceeds supply. Previous studies show that the artemisinin biosynthesis is promoted by light in A. annua. Cryptochrome1 (CRY1) is involved in many processes in the light response. In this study, AaCRY1 was cloned from A. annua. Overexpressing AaCRY1 in Arabidopsis thaliana cry1 mutant resulted in blue-light-dependent short hypocotyl phenotype and short coleoptile under blue light. Yeast two-hybrid and subcellular colocalization showed that AaCRY1 interacted with AtCOP1 (ubiquitin E3 ligase CONSTITUTIVE PHOTOMORPHOGENIC1). Overexpression of AaCRY1 in transgenic A. annua increased the artemisinin content. When AaCRY1 was overexpressed in A. annua driven by the CYP71AV1 (cytochrome P450 dependent amorpha-4,11-diene 12-hydroxylase) promoter, the artemisinin content was 1.6 times higher than that of the control. Furthermore, we expressed the C terminal of AaCRY1(CCT) involved a GUS-CCT fusion protein in A. annua. The results showed that the artemisinin content was increased to 1.7- to 2.4-fold in GUS-CCT transgenic A. annua plants. These results demonstrate that overexpression of GUS-CCT is an effective strategy to increase artemisinin production in A. annua.
青蒿素是一种有效的抗疟化合物，从药用植物青蒿中分离得到。然而，由于青蒿中青蒿素的含量低，青蒿素的需求超过了供应。先前的研究表明，青蒿素的生物合成在青蒿中受光的促进。隐花色素 1（CRY1）参与了光响应中的许多过程。本研究从青蒿中克隆了 AaCRY1。在拟南芥 cry1 突变体中过表达 AaCRY1 导致蓝光依赖性短下胚轴表型和蓝光下短的子叶。酵母双杂交和亚细胞共定位表明 AaCRY1 与 AtCOP1（泛素 E3 连接酶 CONSTITUTIVE PHOTOMORPHOGENIC1）相互作用。在转基因青蒿中过表达 AaCRY1 增加了青蒿素的含量。当 AaCRY1 由 CYP71AV1（细胞色素 P450 依赖的 amorpha-4,11-diene 12-羟化酶）启动子时，青蒿素的含量比对照高出 1.6 倍。此外，我们在青蒿中表达了 AaCRY1 的 C 端（CCT），该基因与 GUS-CCT 融合蛋白有关。结果表明，GUS-CCT 转基因青蒿素的含量增加了 1.7 至 2.4 倍。这些结果表明，过表达 GUS-CCT 是提高青蒿素产量的有效策略。
