Elite Stevia rebaudiana clones for sustainable tropical highland cultivation: Agronomic traits and glycoside profiles

Bambang Heliyanto; Mala  Murianingrum; Sri Hartati; Rully Dyah Purwati; Parnidi Parnidi; Betalini Widhi Hapsari; Erwin Al Hafiidz; Andri Fadillah; Cece Suhara; Rina Laksmi Hendrati; Marjani Marjani; Suseno Amien

doi:10.17268/sci.agropecu.2026.023

Autores/as

Bambang Heliyanto National Reseach and Innovation Agency: Jakarta, Jakarta, Indonesia. https://orcid.org/0000-0002-4706-6651
Mala Murianingrum National Research and Innovation Agency: Jakarta, Jakarta, Indonesia. https://orcid.org/0000-0002-8677-1564
Sri Hartati National Research and Innovation Agency: Jakarta, Jakarta, Indonesia. https://orcid.org/0000-0003-4127-5564
Rully Dyah Purwati National Research and Innovation Agency: Jakarta, Jakarta, Indonesia. https://orcid.org/0000-0001-5172-7531
Parnidi Parnidi National Research and Innovation Agency: Jakarta, Jakarta, Indonesia. https://orcid.org/0000-0002-2879-4565
Betalini Widhi Hapsari Lembaga Ilmu Pengetahuan Indonesia: Jakarta, Jakarta, Indonesia. https://orcid.org/0000-0002-9785-2087
Erwin Al Hafiidz Research Center for Biotechnology, Indonesian Institute of Sciences, Bogor, West Java, Indonesia. https://orcid.org/0000-0002-2937-6509
Andri Fadillah National Research and Innovation Agency: Jakarta, Jakarta, Indonesia. https://orcid.org/0000-0002-6723-3058
Cece Suhara National Research and Innovation Agency: Jakarta, Jakarta, Indonesia. https://orcid.org/0000-0002-6086-069X
Rina Laksmi Hendrati Research Center for Biotechnology, Indonesian Institute of Sciences, Bogor, West Java, Indonesia. https://orcid.org/0000-0002-7267-5838
Marjani Marjani National Research and Innovation Agency: Jakarta, Jakarta, Indonesia. https://orcid.org/0000-0002-0182-9190
Suseno Amien Universitas Padjadjaran: Bandung, West Java, Indonesia. https://orcid.org/0000-0001-8324-9247

DOI:

https://doi.org/10.17268/sci.agropecu.2026.023

Palabras clave:

Stevia rebaudiana , tropical highland, sustainability, genotype evaluation, steviol glycosides, yield stability

Resumen

Stevia rebaudiana Bertoni (Bertoni) is a high-value source of non-caloric sweeteners, yet its genotype-specific performance under tropical highland conditions remains insufficiently documented. Eleven elite stevia clones grown in the tropical highlands of Indonesia were evaluated, integrating agronomic performance with the steviol glycoside profile. A random block design with four replicates was implemented across three harvests. Majalengka exhibited the greatest plant height (45.06 cm) and total biomass (21.32 g plant⁻¹), while Karang Anyar and Garut also exceeded 20 g plant⁻¹, indicating robust vegetative growth. Dry leaf yield ranged from 4.98 to 14.00 g plant⁻¹, with Majalengka, Karang Anyar, and TIA-001A among the highest-yielding genotypes. High-performance liquid chromatography (HPLC) identified Garut as the leading clone in total glycoside content (346.78 mg g⁻¹), with the highest concentrations of stevioside (75.91 mg g⁻¹, 7.59%) and rebaudioside A (26.40 mg g⁻¹, 2.64%). However, its Reb A/stevioside ratio (0.35) was lower than that of TIA-006 (0.74), TIA-003 (0.54), and Karang Anyar (0.52), which exhibited more favorable sweetness profiles. TIA-003 combined high glycoside content (286.58 mg g⁻¹, 28.66%) with environmental stability, despite moderate leaf yield. Pujon and TIA-002A showed consistent glycoside profiles suitable for herbal and blended extract applications, while Karang Anyar demonstrated dual potential for leaf yield and sweetener quality. These findings identify Garut, TIA-003, and Karang Anyar as priority genotypes for glycoside extraction and agro-industrial scaling, while Majalengka and TIA-001A offer biomass advantages for whole-plant utilization. These findings offer genotype-specific insights to guide breeding programs and facilitate agro-industrial deployment in tropical highland stevia systems.

Citas

Abouelela, M. B., Eid, M., Ali, F. M., et al. (2025). Optimizing a rapid tissue culture method for steviol glycoside production from Stevia rebaudiana to address Egypt’s sugar deficit. Sci Rep, 15, 25495. https://doi.org/10.1038/s41598-025-10491-3

Adabiyah, R., Ratnadewi, D., & Ermayanti, T. M. (2019). Growth evaluation of Stevia rebaudiana Bert. tetraploid in vitro and in the field for stevioside and Reb-A production. J. Biol. Indones, 15(2), 153–165.

Ahmadirad, S., Tavakoli, A., Mokhtassi-Bidgoli, A. et al. (2024). Optimizing Biomass and Steviol Glycoside Yield in Hydroponically Grown Stevia (Stevia rebaudiana Bertoni) with Ammonium Nitrate and 6-Benzylaminopurine Concentrations. Sugar Tech, 26, 595–607. https://doi.org/10.1007/s12355-024-01365-5

Alves, N. B., Balestre, M., Pennacchi, J. P., Fernandes, M. C. N., Castro, D. G., & Botelho, F. B. S. (2020). Genetic progress of upland rice (Oryza sativa L.) lines for disease resistance. Plant Breed., 139(5), 853–861. https://doi.org/10.1111/pbr.12829

Amarakoon, S. (2021). Stevia rebaudiana–A review on agricultural, chemical and industrial applications. J. Nat. Appl. Res., 1(1), 14–27.

Amien, S., Maulana, H., Ruswandi, D., & Nurjanah, S. (2021). Genetic gain and relationship of yield and yield attributes of mutant and cross-bred stevia (Stevia rebaudiana) genotypes. Biodiversitas, 22(8), 3119–3126. https://doi.org/10.13057/biodiv/d220805

Amien, S., Maulana, H., Ruswandi, D., & Nurjanah, S. (2022). Stevia (Stevia rebaudiana B.) genotypes assessment for leaf yield stability through genotype by environment interactions, AMMI, and GGE biplot analyses. SABRAO J. Breed. Genet., 54(4), 1001–1010.

Andrade, M. I., Naico, A., Ricardo, J., Eyzaguirre, R., Makunde, G. S., Ortiz, R., & Grüneberg, W. J. (2016). Genotype × environment interaction and selection for drought adaptation in sweet potato (Ipomoea batatas L.) in Mozambique. Euphytica, 209, 261–280. https://doi.org/10.1007/s10681-016-1684-4

Arumugam, B., Subramaniam, A., & Alagaraj, P. (2020). Stevia rebaudiana as a natural sweetener: a review. Cardiovasc. Hematol. Agents Med. Chem, 18(2), 94–103.

Benhmimou, A., Ibriz, M., Faiz, C. A., Gaboun, F., Douaik, A., Amchra, F. Z., & Khiraoui, A. (2017). Effects of planting density and harvesting time on productivity of natural sweetener plant (Stevia rebaudiana Bertoni) in Larache Region, Morocco. Int. J. Plant Res, 7(4), 83–89.

Bhasker, R., Prasad, S. V., & Sherlin, D. M. (2015). Pharmacological properties of Stevia rebaudiana Bertoni. J. Med. Plant Res., 9(3), 89–96.

Bundgaard Anker, C. C., Rafiq, S., & Jeppesen, P. B. (2019). Effect of steviol glycosides on human health with emphasis on type 2 diabetic biomarkers: A systematic review and meta-analysis of randomized controlled trials. Nutrients, 11(9), 1965.

Burgess, A. J., Retkute, R., Herman, T., & Murchie, E. H. (2017). Exploring relationships between canopy architecture, light distribution, and photosynthesis in contrasting rice genotypes using 3D canopy reconstruction. Front. Plant Sci., 8, 1316.

Castro, F. C. D., Faleiro, F. G., Amabile, R. F., Oliveira, J. D. S., Melo, J. V. P., & Santos, M. A. (2025). Selection and genetic characterization of Stevia genotypes based on the rebaudioside A and total steviol glycosides for Brazilian Savanna. Int. J. Agron., 2025(1), 6630489.

Chou, C.-T., Christabel, V., Le, M. A., Tsai, M.-L., & Wang, S.-T. (2025). Effects of Light Conditions on the Leaf Growth and Steviol Glycoside Yields of Hydroponically Cultivated Stevia Across Growth Stages. Horticulturae, 11(3), 316. https://doi.org/10.3390/horticulturae11030316

Cho, J., Park, H.-J., Yun, C.-I., & Kim, Y.-J. (2025). Validation and determination of steviol glycosides in processed food products. Food Sci. Biotechnol., 34(4), 1123–1135.

Duan, M., Zhang, X., Wei, Z., Zhang, B., & Liu, Y. (2024). Effect of maize canopy structure on light interception and radiation use efficiency at different canopy layers. Agronomy, 14(7), 1511. https://doi.org/10.3390/agronomy14071511

Gardana, C., Scaglianti, M., & Simonetti, P. (2010). Evaluation of steviol and its glycosides in Stevia rebaudiana leaves and commercial sweetener by ultra-high-performance liquid chromatography–mass spectrometry. J. Chromatogr. A, 1217(9), 1463–1470.

Hashem, M. M., Ashry, H. A., ElShahed, E. S. et al. (2024). Differential expression of steviol glycoside pathway genes in NaCl-treated stevia genotypes with varied SG contents. Discov. Plants, 1, 42. https://doi.org/10.1007/s44372-024-00045-8

Heliyanto, B., Djumali, Abdurrakhman, Syakir, M., Sugiyarta, E., & Cholid, M. (2020). Development of high yielding sugarcane varieties for rainfed areas: yield multilocation trial of promising sugarcane clones. IOP Conf. Ser.: Earth Environ. Sci, 1302, 012005. https://doi.org/10.1088/1755-1315/1302/1/012005

Heliyanto, B., Murianingrum, M., Hartati, S., Hapsari, B. W., Purwati, R. D., Suhara, C., & Amien, S. M. (2024). Observation on local high-yielding stevia (Stevia rebaudiana Bertoni L.) clone specific to Bandung District, West Java. IOP Conf. Ser.: Earth Environ. Sci., 1302, 012065. https://doi.org/10.1088/1755-1315/1302/1/012005

Huber, B. M., & Wehner, T. C. (2023). Heritability and genetic variance estimates for agronomic traits and glycoside yield in four elite stevia breeding populations. Crop Sci., 63(3), 1234–1245. https://doi.org/10.1002/csc2.20940

Joshi, J., Gautam, S., & Gautam, A. (2024). Optimization of parameters for extraction and isolation of steviol glycosides (SGs) from Stevia rebaudiana leaves using water. Indian Journal of Environmental Protection, 44(14), 1268–1276.

Li, R., Xu, C., Wu, Z., Xu, Y., Sun, S., Song, W., & Wu, C. (2025). Optimizing canopy-spacing configuration increases soybean yield under high planting density. Crop J., 13(1), 233–245. https://doi.org/10.1016/j.cj.2024.12.005

Libik-Konieczny, M., Capecka, E., Kąkol, E., Dziurka, M., Grabowska-Joachimiak, A., Sliwinska, E., & Pistelli, L. (2018). Growth, development and steviol glycosides content in relation to photosynthetic activity of several Stevia rebaudiana Bertoni strains cultivated under temperate climate conditions. Sci. Hortic., 234, 10–18. https://doi.org/10.1016/j.scienta.2018.02.015

Martini, R. E., Tavares, M. C. H., & Silva, J. A. G. (2015). Vegetative propagation of Stevia rebaudiana Bertoni using stem cuttings. Rev. Bras. Plantas Med., 17(2), 295–301.

Martono, Y., Riyanto, S., Rohman, A., & Martono, S. (2016). Improvement method of fast and isocratic RP-HPLC analysis of major diterpene glycoside from Stevia rebaudiana leaves. AIP Conf. Proc., 1755, 080001.

Myint, K. Z., Chen, J., Zhou, Z., Xia, Y., Lin, J., & Zhang, J. (2020). Structural dependence of antidiabetic effect of steviol glycosides and their metabolites on streptozotocin-induced diabetic mice. J. Sci. Food Agric., 100(10), 3841–3849. https://doi.org/10.1002/jsfa.10421

Pal, P. K., Kumar, R., Guleria, V., Mahajan, M., Prasad, R., Pathania, V., & Ahuja, P. S. (2015). Crop ecology and nutritional variability influence growth and secondary metabolites of Stevia rebaudiana Bertoni. BMC Plant Biology, 15, 1–16. https://doi.org/10.1186/s12870-015-0457-x

Parris, C. A., Shock, C. C., & Qian, M. (2016). Dry leaf and steviol glycoside productivity of Stevia rebaudiana in the Western United States. HortScience, 51(10), 1220–1227. https://doi.org/10.21273/hortsci11149-16

Ribeiro, M. M., Diamantino, T., Domingues, J., Montanari, Í., Alves, M. N., & Gonçalves, J. C. (2021). Stevia rebaudiana germplasm characterization using microsatellite markers and steviol glycosides quantification by HPLC. Mol. Biol. Rep., 48(3), 2573–2582. https://doi.org/10.1007/s11033-021-06308-x

Sowmya, H. H., Kamatar, M. Y., Shanthakumar, G., Brunda, S. M., Shadakshari, T. V., Showkath Babu, B. M., & Rajput, S. S. (2018). Stability analysis of maize hybrids using Eberhart and Russell model. Int. J. Curr. Microbiol. Appl. Sci., 7(7), 963–970.

Sinta, M. M., & Sumaryono. (2019). Growth, biomass production and steviol glycoside content of five stevia clones in Bogor, Indonesia. J. Agron. Indones., 47(1), 105–110.

Tao, R., & Cho, S. (2020). Consumer-based sensory characterization of steviol glycosides (Rebaudioside A, D, and M). Foods, 9(8), 1026. https://doi.org/10.3390/foods9081026

Tian, X., Zhong, F., & Xia, Y. (2022). Dynamic characteristics of sweetness and bitterness and their correlation with chemical structures for six steviol glycosides. Food Res. Int., 151, 110848. https://doi.org/10.1016/j.foodres.2021.110848

Uçar, E., Turgut, K., Özyiğit, Y., Özek, T., & Özek, G. (2018). The effect of different nitrogen levels on yield and quality of stevia (Stevia rebaudiana Bertoni). J. Plant Nutr., 41(9), 1130–1137. https://doi.org/10.1080/01904167.2018.1431673

Xu, X., Wei, Q., Guo, J., Zhang, J., Yang, Y., Wang, L., & Dong, C. (2024). Identification of geographic, climatic, and soil factors dominating Stevia rebaudiana yield and quality. Ind. Crops Prod., 214, 118556.

Zaman, M. M. (2015). Growth parameters and leaf biomass yield of stevia (Stevia rebaudiana Bertoni) as influenced by different soil types of Bangladesh. J. Bangladesh Agric. Univ., 13(1), 25–31. https://doi.org/10.22004/ag.econ.211173

Zhou, X., Gong, M., Lv, X., Liu, Y., Li, J., Du, G., Liu, L. (2021) Metabolic engineering for the synthesis of steviol glycosides: current status and future prospects. Appl Microbiol Biotechnol, 105(13), 5367-5381.