Department of Plant Breeding and Biotechnology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Department of Agronomy and Plant Breeding, College of Agriculture and Natural Resources, University of Tehran, 14174, Karaj, Iran.
Appl Biochem Biotechnol. 2022 Jul;194(7):3228-3241. doi: 10.1007/s12010-022-03879-8. Epub 2022 Mar 29.
Biotechnology and nanotechnology are important tools for understanding biochemical pathways. They can be used efficiently for stimulating and increasing the production of secondary metabolites in medicinal plants. The present study aimed to identify the γ-terpinene synthase gene (CcTPS2) as an effective contributor to the biosynthetic pathway of monoterpenes. The effects of silver nanoparticles (AgNPs; 50 and 100 mg l) and time (24 and 48 h) were examined on secondary metabolites in cell suspension cultures of Carum carvi. This involved the identification, isolation, and sequencing of a partial sequence in the CcTPS2 gene of C. carvi. The genomic sequence of CcTPS2 comprised 292 bp which were organized into two exons (110 and 82 bp) and one intron (100 bp), while the cDNA was 192 bp. In the scale of nucleotides, the CcTPS2 gene showed 96% similarity with the TPS2 gene of Oliveria decumbens. We generated sequence data of the CcTPS2 gene for the first time in this species, thereby enabling further developments in understanding the molecular mechanisms responsible for terpene biosynthesis and other chemical derivatives in C. carvi. The results of GC/MS and GC/FID showed that AgNPs strongly affected the secondary metabolites in cell suspension cultures of C. carvi. According to the results, the AgNPs (50 mg l) increased p-cymene and carvone contents in comparison with the control. The exposure of plants to 100 mg l AgNPs induced the production of thymol and carvacrol. The results of real-time PCR revealed that the exposure of plants to 100 mg l AgNPs caused a significant upregulation of CcTPS2 expression for 24 h. These cell suspension cultures were elicited by AgNPs, the application of which proved as an effective method to improve the production of secondary metabolites in vitro.
生物技术和纳米技术是理解生化途径的重要工具。它们可以有效地用于刺激和增加药用植物中次生代谢产物的产生。本研究旨在鉴定 γ-松油烯合酶基因（CcTPS2）作为单萜生物合成途径的有效贡献者。研究了银纳米粒子（AgNPs；50 和 100 mg l）和时间（24 和 48 h）对 Carum carvi 细胞悬浮培养物中次生代谢产物的影响。这涉及鉴定、分离和测序 C. carvi 中 CcTPS2 基因的部分序列。CcTPS2 基因的基因组序列由 292 bp 组成，分为两个外显子（110 和 82 bp）和一个内含子（100 bp），而 cDNA 为 192 bp。在核苷酸的尺度上，CcTPS2 基因与 Oliveria decumbens 的 TPS2 基因具有 96%的相似性。我们首次在该物种中生成了 CcTPS2 基因的序列数据，从而为进一步了解萜烯生物合成和 C. carvi 中其他化学衍生物的分子机制提供了支持。GC/MS 和 GC/FID 的结果表明，AgNPs 强烈影响 C. carvi 细胞悬浮培养物中的次生代谢产物。结果表明，与对照相比，AgNPs（50 mg l）增加了对伞花烃和香芹酮的含量。暴露于 100 mg l AgNPs 的植物诱导了百里香酚和香芹酚的产生。实时 PCR 的结果表明，暴露于 100 mg l AgNPs 的植物在 24 h 内导致 CcTPS2 表达的显著上调。这些细胞悬浮培养物被 AgNPs 激发，AgNPs 的应用被证明是一种提高体外次生代谢产物产生的有效方法。
