Federal Scientific Center of the East Asia Terrestrial Biodiversity of the Far East Branch of Russian Academy of Sciences, Vladivostok 690022, Russia.
Federal Scientific Center of the East Asia Terrestrial Biodiversity of the Far East Branch of Russian Academy of Sciences, Vladivostok 690022, Russia.
J Biotechnol. 2021 Dec 10;342:102-113. doi: 10.1016/j.jbiotec.2021.10.012. Epub 2021 Nov 1.
Indole alkaloid camalexin has potential medicinal properties such as suppressing the viability of leukemic but not normal cells. Camalexin is not produced in plants and an external factor is required to activate its biosynthesis. In this work, we stimulated camalexin biosynthesis in Arabidopsis calli by blocking one of repressors of the jasmonate pathway, the jasmonate ZIM-domain protein 1 (JAZ1) by using amiRNA targeting JAZ1 gene transcripts. Inhibition of the JAZ1 gene led to an increase in camalexin content from trace amounts in control culture to 9 µg/g DW in the jaz1 line without affecting growth. In addition, JAZ1 silencing enhanced tolerance to cold stress with simultaneous increasing camalexin content up to 30 µg/g DW. Real-time quantitative PCR determination of marker gene expression showed that effects caused by the JAZ1 silencing might be realized through crosslinking JA, ROS, and abscisic acid signaling pathways. Thus, targeting the distal components of signaling pathways can be suggested as a tool for bioengineering of secondary metabolism, along with standard techniques for targeting biosynthetic genes or genes encoding transcription factors.
色胺生物碱卡马西平具有潜在的药用特性，例如抑制白血病细胞但不抑制正常细胞的活力。卡马西平在植物中不产生，需要外部因素来激活其生物合成。在这项工作中，我们通过使用针对 JAZ1 基因转录本的 amiRNA 阻断茉莉酸途径的一种抑制剂——茉莉酸 ZIM 结构域蛋白 1 (JAZ1)，刺激拟南芥愈伤组织中的卡马西平生物合成。JAZ1 基因的抑制导致卡马西平含量从对照培养物中的痕量增加到 jaz1 系中的 9 µg/g DW，而不影响生长。此外，JAZ1 沉默增强了对冷胁迫的耐受性，同时将卡马西平含量提高到 30 µg/g DW。标记基因表达的实时定量 PCR 测定表明，JAZ1 沉默引起的效应可能是通过交联 JA、ROS 和脱落酸信号通路实现的。因此，靶向信号通路的远端组分可以被建议作为生物工程次生代谢的一种工具，同时还可以使用靶向生物合成基因或转录因子编码基因的标准技术。
