Cell and Tissue Journal

Abstract Aim: Papaver somniferum remains the sole commercial source for several pharmaceutical benzophenanthridine alkaloids from benzylisoquinoline branch alkaloids, which includes the narcotic analgesics codeine and the semi-synthetic drugs such as; oxycodone, buprenorphine and naltrexone. Although, most of the biosynthetic pathways genes of this alkaloid have been identified, the post-transcriptional regulation of these alkaloids pathway has not been completely determined. Virus induced gene silencing (VIGS) is a method for fast functional genomics, and in this study, this technique was used to investigate the silencing one of the most important genes in this alkaloid pathway.
Materials and Methods: In the current research VIGS technique was used for systematic reduction in the level of genes expression involves in benzylisoquinoline alkaloids pathway. For silencing of codeinone reductase gene, pTRV vector was used for cloning. The specific silencing of COR gene was evaluated in 2-3 weeks old leaves of Papaver somniferum using agro-infiltration method.
Results: Result of cloning was confirmed by using of different molecular methods such as enzyme digestion and also PCR. PCR technique was used for confirmation of transgenic plants in some transformed plants. Using of semi-quantitative PCR and real-time PCR showed that the level of reduction in transcription of COR gene was about 89 percent.
Conclusion: The obtained results confirmed that by application of RNA interference method, the level of COR gene expression was significantly reduced compared with control.

Abstract Aim: The aim of this study was to investigate the expression of key genes, 1-deoxy-D-xylulose-5-phosphate reductoisomerase )DXR( and gamma-terpinene synthase (GTS), involve in thymol and carvacrol biosynthesis pathway in Summer savory (Satureja hortensis). This species is one of the important medicinal plants of the Lamiaceae family, and consider as an important source of the mentioned compounds.
Material and Methods: Plants were treated with salicylic acid, methyl jasmonate and UV-B rays. RNAs were extracted from control and treated plants and cDNAs were synthesized. Primers were designed for gene isolation and expression studies. Transcript expression analyses for the DXR and GTS were performed using semi-quantitative RT-PCR method.
Results: A partial segment for DXR and GTS genes was sequenced. The relative gene expression analyses showed differential expression of both genes at transcript level in different tissues (roots, stems, leaves and inflorescence), with higher levels of expression in leaf and inflorescence. The expression of both genes under the effect of abiotic elicitors including salicylic acid, methyl jasmonate and UV-B rays exhibited significant alteration.
Conclusion: Under controlled conditions, using abiotic elicitors such as: salicylic acid, methyl jasmonate and UV-B radiation could elevate the level of gene expression and possibly increase the production of secondary metabolite such as: thymol and carvacrol.