Journal of Medicinal Plants Biotechnology

Abstract Active substances or secondary metabolites in medicinal plants are regularly produced in response to biotic and abiotic stresses and determine the ability of plants to adapt during growth and development. In order to increase the quantity and quality of the effective substance of the green cumin medicinal plant, with the scientific name Cuminum cyminum L from the Chetrian family, we need to identify the germplasm and useful genes in domestic and wild populations of this plant. In the current research, five selected genes in the biosynthesis pathway of flavonoids in cumin, obtained from previous research based on NGS method, which had the highest expression changes in drought stress, were validated. DNA from young leaves of selected ecotypes was extracted and quality measured with agarose gel. Considering the PCR test and the observation of a single band in all cases and the lack of difference in the length of the amplified fragment among the selected ecotypes; The length polymorphism of the fragments resulting from the amplification was rejected due to the presence of deletion and addition regions between different genotypes. This experiment could not confirm the placement of the two studied ecotypes of Sadouq and Rafsanjan in terms of seed yield, essential oil and extract content, in two different groups in terms of the studied genes. Also, he could not confirm the difference in the expression of two genes (with sequence numbers DN1196 and DN32640) in drought stress and normal irrigation in the previous research

Abstract Cumin is an annual and herbaceous plant belonging to the parsley family. In this research, five selected genes effective in the biosynthesis of flavonoids, which were identified by NGS method in cumin and had the most expression changes, were studied bioinformatically. Among these, two gene fragments, DN32640 and DN1196, which had the greatest evolutionary distance and were from two different families, were investigated for further identification. The sequence of DN1196 is from the GT6 family, a glycosyltransferase in the biosynthesis pathway of phenolic compounds, and the sequence of DN32640 is from the BHLH family, a transcription factor in the anthocyanin biosynthesis pathway. The coded protein of the sequence DN1196, with more than 85% similarity with a protein from the carrot plant, glycosylates it in the final step of flavonol synthesis by transferring a glucose molecule. In this process, two transcription factor proteins interact with this protein. The predicted secondary structure of the product of this gene was identified in accordance with the carrot protein sequence. The encoded protein of the DN32640 sequence, with more than 90% similarity to the carrot protein, is a transcription factor of the BHLH family and a component of the MBW complex. This transcription factor is involved in regulating the final stages of the path leading to the biosynthesis of anthocyanins and tannins. The secondary structure of this gene product was determined based on the identified protein sequence of carrot. Considering the importance of these two genes and the interaction of their product with other proteins, they can be used in genetic manipulations to produce more phenolic compounds, especially anthocyanins in cumin.