Investigation of tropane alkaloid production and expression of pmt gene, in Atropa belladonna cell suspension culture treated with Piriformospora indica

Hyoscyamine and scopolamine are tropane alkaloids employed in medicine as anticholinergic agents and extracted from plants that belong to several species of Solanaceae because their chemical synthesis is complicated and expensive obtaining these compounds through cell suspension culture is a promising approach. The effect of Piriformospora indica, a plant growth-promoting root endophyte fungus, on tropane alkaloids production and expression of one rate-limiting enzyme gene, Putrescine N-methyltransferase (pmt) were studied in A. belladonna cell suspension culture. Based on our results the highest level of total alkaloids in MS + 1 mg.L-1 NAA (naphthalene acetic acid) + 1 mg.L-1 BA(6-Benzylaminopurin) medium (84.07 mg/g FW) was observed in treated suspension culture. High performance liquid chromatography (HPLC) analysis show that fungal treatment had a positive effect on production of hyoscyamine (17.60 mg/gFW) in cell suspension culture with NAA/BA hormones compared with control. Also, fungal treatment significantly increase scopolamine content (11.91 mg/gFW). The results determined by semi-quantitative RT-PCR indicates that pmt gene expression in cell suspension increased 1.7 fold compare to the tubulin gene as an internal control. These results for the first time indicated that, cell suspension culture of A. belladonna L. are susceptible to elicitation by P. indica. The current study provides an effective approach for commercially production of hyoscyamine and scopolamine by P. indica inoculation in A. belladonna cell suspension culture

کلیدواژه ها:

Atropa belladonna ، elicitor ، Hyoscyamine ، pmt gene ، scopolamine