Optimization of Culture Media and Elicitation for Enhancing Steviol Glycosides in Stevia rebaudiana Callus Culture

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Published May 30, 2026
https://doi.org/10.55043/jaast.v10i2.548
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Vol. 10 No. 2 (2026): Journal of Applied Agricultural Science and Technology

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Zozy Aneloi Noli
Universitas Andalas
Betalini Widhi Hapsari
National Research and Innovation Agency (BRIN)
Rezka Sutrianti
Universitas Andalas
Erwin Al Hafiidz
National Research and Innovation Agency (BRIN)
Deritha Ellfy Rantau
National Research and Innovation Agency (BRIN)
Evan Maulana
National Research and Innovation Agency (BRIN)

Abstract

Stevia rebaudiana is a valuable medicinal plant known for producing steviol glycosides, which are widely used as natural sugar substitutes. This study aims to optimize culture media and elicitation treatments to enhance callus induction, growth, and steviol glycoside accumulation in S. rebaudiana callus culture. The experiment was conducted using a Completely Randomized Design (CRD) with four treatments: MS0, DKW0, MS + 2 mg/L 2,4-D + 0.5 mg/L BAP + 0.5 mg/L GA₃, and DKW + 2 mg/L 2,4-D + 0.5 mg/L BAP + 0.5 mg/L GA₃, each with five replicates. The results showed that the highest callus induction percentage (100%) and the fastest callus initiation time (6.33–6.40 days) were achieved on MS and DKW media supplemented with plant growth regulators (2 mg/L 2,4-D + 0.5 mg/L BAP + 0.5 mg/L GA₃). The highest fresh weight (0.158 g) and dry weight (0.011 g) were obtained on DKW medium supplemented with 2 mg/L 2,4-D + 0.5 mg/L BAP + 0.5 mg/L GA₃. The highest stevioside content (0.147 mg/g DW) was recorded in the 4 mg/L GA₃ treatment, whereas the highest rebaudioside A content (0.114 mg/g DW) was observed in the 0.5 mg/L SA treatment. The highest rebaudioside D content (3.256 mg/g DW) was obtained in the 0.25 mg/L SA treatment, while the highest dulcoside content (0.120 mg/g DW) was found in the control treatment. In addition, the highest total phenolic content (19.37 mg GAE/g DW) was observed in the 20 mg/L daminozide treatment. In conclusion, DKW medium supplemented with 2 mg/L 2,4-D + 0.5 mg/L BAP + 0.5 mg/L GA₃ was the most effective treatment for promoting callus growth, while elicitation treatments, particularly 4 mg/L GA3, significantly enhanced the accumulation of steviol glycosides in S. rebaudiana callus culture (0.147 mg/L).

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Author Biographies

Zozy Aneloi Noli, Universitas Andalas

Biology Department, Faculty of Mathematics and Natural Sciences, Universitas Andalas, Padang, Indonesia

Betalini Widhi Hapsari, National Research and Innovation Agency (BRIN)

Research Center for Genetic Engineering, National Research and Innovation Agency (BRIN), Bogor, Indonesia

Rezka Sutrianti, Universitas Andalas

Undergraduate Program in Biology, Faculty of Mathematics and Natural Sciences, Universitas Andalas, Padang, Indonesia

Erwin Al Hafiidz, National Research and Innovation Agency (BRIN)

Research Center of Applied Botany, National Research and Innovation Agency (BRIN), Bogor, Indonesia

Deritha Ellfy Rantau, National Research and Innovation Agency (BRIN)

Research Center of Applied Botany, National Research and Innovation Agency (BRIN), Bogor, Indonesia

Evan Maulana, National Research and Innovation Agency (BRIN)

Research Center of Applied Botany, National Research and Innovation Agency (BRIN), Bogor, Indonesia

How to Cite
1.
Noli ZA, Hapsari BW, Sutrianti R, Hafiidz EA, Rantau DE, Maulana E. Optimization of Culture Media and Elicitation for Enhancing Steviol Glycosides in Stevia rebaudiana Callus Culture. J. appl. agricultural sci. technol. [Internet]. 2026May30 [cited 2026May31];10(2):328-42. Available from: https://jaast.org/index.php/jaast/article/view/548

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