Sains Malaysiana 54(4)(2025): 993-1003

http://doi.org/10.17576/jsm-2025-5404-03

 

Steviol Glycoside Production and Growth in Stevia rebaudiana Shoot Culture after Methyl Jasmonate Elicitation

(Penghasilan Steviol Glikosida dan Pertumbuhan Kultur Pucuk Stevia rebaudiana selepas Elisitasi Metil Jasmonat)

 

ANDIRA RAHMAWATI^1,2, IRIAWATI², RIZKITA RACHMI ESYANTI^2,* & ROOHAIDA OTHMAN³

 

¹Doctoral Programme, School of Life Sciences and Technology, Institut Teknologi Bandung, Jl. Ganesha 10, 40132 Bandung, Indonesia

²School of Life Sciences and Technology, Institut Teknologi Bandung, Jl. Ganesha 10, 40132 Bandung, Indonesia

³Department of Biological Sciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia

 

Diserahkan: 24 Mei 2024/ Diterima: 23 Disember 2024

 

Abstract

Steviol glycosides are a group of compounds derived from the Stevia rebaudiana metabolites which are commonly used as sugar replacements. Methyl jasmonate (MeJA) elicitation is one strategy to induce high steviol glycoside content. MeJA can enhance the synthesis of secondary metabolites, however, it can also negatively impact the growth of Stevia. The objective of this study was to establish the concentration and exposure duration of MeJA elicitation to obtain the highest level of steviol glycoside production while achieving the best growth for Stevia shoot cultures. The shoot cultures were elicited using MeJA at concentrations of 10 μM, 50 μM, and 100 μM for 24, 48, and 72 h. The cultures were then transferred into the medium without MeJA for 7 days, and then growth characteristics were measured. Analysis of stevioside and rebaudioside A levels was performed using HPLC from samples harvested directly after treatment and 7 days post-elicitation. MeJA was found to suppress growth in terms of height, adventitious shoot formation, and leaf size across all concentrations. However, the number of leaves, fresh weight, number of axillary buds, and number of nodes showed no significant difference. The optimal condition for stevioside synthesis was achieved at 50 μM MeJA for 48 h (0.73 mg/g), while the optimal condition for rebaudioside A synthesis was achieved with 100 μM MeJA 24 h treatment (0.63 mg/g). Analysis of the plant growth and secondary metabolite production suggested that the treatment using 50 μM MeJA for 48 h was found to be the best conditions which gave high secondary metabolite production while plant growth was also high. This finding can serve as a fundamental approach for enhancing the production of steviol glycosides in the industry.

Keywords: Growth; in vitro culture; rebaudioside A; Stevia; stevioside

 

Abstrak

Steviol glikosida adalah sekumpulan sebatian terbitan daripada metabolit Stevia rebaudiana yang biasa digunakan sebagai pengganti gula. Elisitasi menggunakan metil jasmonat (MeJA) merupakan satu strategi yang boleh dilakukan untuk mengaruh kandungan steviol glikosida yang tinggi. MeJA boleh meningkatkan sintesis metabolit sekunder, namun begitu, ia juga boleh menjejaskan pertumbuhan Stevia. Objektif kajian ini adalah untuk menentukan kepekatan dan tempoh pendedahan elisitasi MeJA untuk memperoleh tahap penghasilan steviol glikosida paling tinggi di samping mencapai pertumbuhan terbaik bagi kultur pucuk Stevia. Kultur pucuk telah dielisitasi menggunakan MeJA pada kepekatan 10 μM, 50 μM dan 100 μM selama 24, 48 dan 72 jam. Kultur kemudiannya dipindahkan ke dalam medium tanpa MeJA selama 7 hari, seterusnya ciri pertumbuhan telah diukur. Analisis tahap steviosida dan rebaudiosida A dilakukan menggunakan HPLC daripada sampel yang diambil sejurus selepas rawatan MeJA dan 7 hari selepas elisitasi. MeJA didapati merencat pertumbuhan dari segi ketinggian, pembentukan pucuk adventitius dan saiz daun pada semua kepekatan. Walau bagaimanapun, bilangan daun, berat segar, bilangan tunas aksilari dan bilangan nod tidak menunjukkan perbezaan signifikan. Keadaan yang optimum bagi sintesis steviosida dicapai pada 50 μM MeJA selama 48 jam (0.73 mg/g), manakala keadaan yang optimum bagi sintesis rebaudiosida A pula dicapai dengan rawatan 100 μM MeJA selama 24 jam (0.63 mg/g). Analisis pertumbuhan tumbuhan dan penghasilan metabolit sekunder mencadangkan bahawa rawatan menggunakan 50 μM MeJA selama 48 jam didapati merupakan keadaan terbaik yang memberikan penghasilan metabolit sekunder yang tinggi di samping pertumbuhan tumbuhan yang juga tinggi. Penemuan ini boleh bertindak sebagai pendekatan asas dalam meningkatkan penghasilan steviol glikosida dalam industri.

Kata kunci: Kultur in vitro; pertumbuhan; rebaudiosida A; Stevia; steviosida

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*Pengarang untuk surat-menyurat; email: rizkita@itb.ac.id