Effect of total nitrogen content and NH4+/NO3- ratio on biomass accumulation and secondary metabolite production in cell culture of S. nemorosa

Document Type: Research paper

Authors

1 Department of Horticulture Science, Faculty of Agriculture, University of Mohaghegh Ardabili, Ardabil, Iran

2 Department of Horticulture Science, Faculty of Agriculture, University of Mohaghegh Ardabili, Ardabil, Iran.

3 Department of horticultural science, Faculty of agriculture and natural resources, University of Mohaghegh Ardabili, Ardabil, Iran

10.30479/ijgpb.2020.12321.1258

Abstract

Salvia nemorosa is an important native medicinal plant in Iran, which considered a rich source of phenolic and flavonoids compounds. The present research was conducted with respect to the optimization of medium for biomass accumulation and secondary metabolite production in cell suspension culture of Woodland Sage. We examined the effects of total nitrogen content (0, 15, 30, 60, 90, and 120 mM) and NH4+/NO3- ratio (0:60, 10:50, 20:40, 30:30, 40:20, 50:10, and 60:0) in the Murashige and Skoog Media on biomass growth, total phenolic, flavonoid and Rosmarinic acid content. The maximum accumulation of fresh biomass (294.80 g/l) and total phenolic content (76.61 mg GAE/g DW) was obtained in the medium supplemented with 90 mM nitrogen. The highest Rosmarinic acid content (16.41 and 16.16 mg/g DW) was recorded in the medium containing 30 and 60 mM nitrogen. Increasing total nitrogen more than 30 mM resulted in a decline in Rosmarinic acid production. The ammonium to nitrate ratio also affected the biomass growth and secondary products accumulation. The highest fresh biomass accumulation (267.74 mg/l) total phenolic content (87.30 mg GAE/g DW) and Rosmarinic acid content (18.43 mg/ DW) was recorded in 10:50 ratio of NH4+/NO3-. Increasing in NH4+ level or complete elimination of it from culture medium reduced the rosmarinic acid and total phenolic content of S. nemorosa. Our findings revealed that Woodland Sage cell suspension needs both nitrogen forms, but ammonium is required at low concentration and nitrate at high levels. The results of the current study are beneficial for medium optimization in establishment large scale and bioreactor assist cell culture of Woodland Sage for the production Phenolic acids.

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