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Sains Malaysiana 54(4)(2025): 993-1003
http://doi.org/10.17576/jsm-2025-5404-03
Steviol Glycoside Production and Growth in Stevia rebaudianaShoot Culture after Methyl Jasmonate Elicitation
(Penghasilan Steviol Glikosida dan Pertumbuhan Kultur Pucuk Stevia rebaudiana selepas Elisitasi Metil Jasmonat)
ANDIRA RAHMAWATI1,2, IRIAWATI2,
RIZKITA RACHMI ESYANTI2,* & 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
Received:
24 May 2024/Accepted: 23 December 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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*Corresponding
author; email: rizkita@itb.ac.id
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