Sugar Tech

, Volume 20, Issue 6, pp 734–744 | Cite as

Differential pH-Induced Biosynthesis of Steviol Glycosides and Biochemical Parameters in Submerge Root Cultures of Stevia rebaudiana (Bert.)

  • Naveed AhmadEmail author
  • Abdur Rab
  • Nisar AhmadEmail author
  • Hina Fazal
Research Article


The major objective of the current study was to investigate the potential effects of differential pH levels on culture development and production of steviol glycosides and other polyphenolics content in submerge root cultures of Stevia rebaudiana. In vitro grown cultures require an optimum pH level for rapid growth and uniform production of secondary metabolites. Herein, varying media pH levels (5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9 and 6.0) significantly influenced fresh and dry biomasses of adventitious root cultures. Among tested levels, adventitious root cultures favored high media pH (6.0) for accumulation of maximum fresh biomass (112.86 g l−1) and dry biomass (8.29 g l−1). In this study, we observed that lower pH level (5.1) strongly supported the production of steviosides (79.48 mg/g-DW) and rebaudioside-A (13.10 mg/g-DW) contents but reduced the polyphenolics content in adventitious cultures. However, dulcoside contents (2.57 mg/g-DW) in adventitious root cultures were found in higher quantities at pH level 5.8. Similarly, maximum total phenolics (70.06 mg/g-DW) and flavonoids (50.19 mg/g-DW) were observed on media having 5.8-pH level. The same pH level also improved DPPH-radical scavenging activities (92.67%). This study will offer an approach to enhance medicinal products in the in vitro tissues, rather than to over-exploit the wild plants and ultimately putting them on the brink of being endangered. Furthermore, the consistent production of secondary metabolites in these cultures could be scaled-up on bioreactor level, which will ultimately affect the society by potential introduction of cost-effective and biologically stable medicinal drugs.


Stevia Adventitious roots pH levels Steviol glycosides Polyphenolics 


Authors’ Contribution

NA performed the major experiment and developed adventitious root cultures of Stevia and finally investigated the phenolic and flavonoid content. AR designed the experiment and thoroughly checked the whole manuscript. NA wrote the manuscript and performed antioxidant activity and HPLC analysis for Steviol glycosides. HF provided plant materials.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest regarding this manuscript.


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© Society for Sugar Research & Promotion 2018

Authors and Affiliations

  1. 1.Department of HorticultureThe University of Agriculture, PeshawarPeshawarPakistan
  2. 2.Center for Biotechnology and Microbiology (CB&M)University of SwatSwatPakistan
  3. 3.Pakistan Council of Scientific and Industrial Research (PCSIR)PeshawarPakistan

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