Effects of Different Carbon Sources on In Vitro Shoot Multiplication: Solid MS Medium Optimization and Bioreactor Validation for Aquatic Plant Bucephalandra pygmaea (Becc.) P. C. Boyce & S.Y. Wong

Year 2026, Volume: 15 Issue: 1, 288 - 299, 24.03.2026

Ibrahim Halil Hatipoglu , Meral Doğan , Yakup Mert Kul , Burak Akyüz

https://doi.org/10.17798/bitlisfen.1799714

https://izlik.org/JA68XU75LY

Abstract

Bucephalandra pygmaea (Becc.) P. C. Boyce & S.Y. Wong is an aquarium plant that is gaining importance in the ornamental plant sector due to its vibrant leaf colours and high adaptability to underwater conditions.
This study aimed to enhance the in vitro shoot propagation efficiency of B. pygmaea ‘Bukit Kelam’ through a two-stage experimental design. In the first stage (solid MS optimisation), three carbon sources—maltose, sucrose, and glucose—were tested at 10 and 20 g L⁻¹ concentrations to determine their effects on key growth parameters, including shoot multiplication rate, plant height, leaf number, leaf area, and plant width. According to the results, 10 g L⁻¹ maltose promoted the highest shoot proliferation and leaf expansion, whereas 20 g L⁻¹ glucose caused a pronounced reduction in all morphological traits.
In the second stage (bioreactor validation), the optimised 10 g L⁻¹ maltose medium was applied in both solid MS and temporary immersion (Plantform TIS) systems. Plants grown on solid MS displayed superior morphological performance (plant height, leaf area, and SPAD values), while bioreactor-cultured plants exhibited colour values, reflecting enhanced pigment intensity. Overall, 10 g L⁻¹ maltose was identified as the most effective carbon source for Bucephalandra micropropagation. The findings suggest that solid MS medium ensures compact growth and high chlorophyll content, whereas the bioreactor system favours visual colour quality—providing complementary strategies for sustainable large-scale propagation of this ornamental aquatic species.

Keywords

Bucephalandra , maltose , glucose , sucrose , SPAD , micropropagation

Ethical Statement

The study is complied with research and publication ethics.

Thanks

This research was not supported by any external funding and was conducted using the facilities of the Faculty of Agriculture, Department of Horticulture, Ondokuz Mayıs University, Samsun, Türkiye.

References

• N. Caraco, J. Cole, S. Findlay, and C. Wigand, "Vascular Plants as Engineers of Oxygen in Aquatic Systems", in BioScience, Vol. 56, Issue 3, 2006, pp. 219–225.
• C. L. Irkin, "Aquatic Miracles: Some Aquatic Plants and Their Uses", in International Journal of Scientific and Technological Research www.iiste.org ISSN 2422-8702 Vol. 4, No. 10, 2018, https://www.iiste.org/Journals/index.php/JSTR/article/view/45439 [Accessed: Sep. 30, 2025].
• L. Jham, A. Vaishnav, A. Patel, S. Kumar, D. K. Verma, S. Kumar, K. Rathod, K. Kumari, K. Kumari, K. Kanaujiya, and A. Kumar, “Aquatic Greenery: Managing Aquatic Vegetation and Harnessing Their Potential”, in Journal of Scientific Research and Reports Vol. 30 Issue 7, 2024, pp. 830-42.
• M. Jabade, and J. Kaur, “Systematic review of phytoremediation: efficacy of aquatic plants in wastewater treatment and pollutant removal”, in Nature Environment and Pollution Technology, Vol. 24, Issue 2, 2025, pp. 42-43.
• M. Doğan, “Farklı Sükroz Konsantrasyonlarını İçeren Kültür Ortamında Pogostemon erectus (Dalzell) Kuntze’un İn Vitro Sürgün Rejenerasyon Performansı“, in International Journal of Eastern Mediterranean Agricultural Research, Vol. 2, Issue 1, 2019, pp. 1-12.
• P. K. Rai, “Heavy metal phytoremediation from aquatic ecosystems with special reference to macrophytes”, in Critical Reviews in Environmental Science and Technology, Vol. 39, 2009, pp. 697-753.
• S. Y. Wong, and P. C. Boyce, ‘Studies on Schismatoglottideae (Araceae) of Borneo LVII: Bucephalandra filiformis — a new species from Maligan’ Sarawak, Malaysian Borneo’, in Aroideana, Vol. 39, Issue 2, 2016, pp. 56–60.
• R. Yunita, M. F. I. Nugraha, and E. G. Lestari, “Micropropagation of Bucephalandra sp”, in Bioscience Research, Vol. 17, Issue 2, 2020, pp. 1339–1343.
• I. Iliev, A. Gajdosová, G. Libiaková, and S. M. Jain, “Plant micropropagation” in Plant cell culture: Essential methods, M. R. Davey & P. Anthony (Eds.), John Wiley & Sons, Ltd., 2010.
• K. R. Goswami, P. Sharma, K. Singh, and G. Singh, ‘Micropropagation of seedless lemon (Citrus limon L. cv. Kaghzi Kalan) and assessment of genetic fidelity of micropropagated plants using RAPD markers’, in Physiology and Molecular Biology of Plants, Vol. 19, Issue 1, 2013, pp. 137–145.
• B. Akyüz, ‘Effect of different carbon sources and concentrations on in vitro propagation of chestnut’, in Plant Cell, Tissue and Organ Culture (PCTOC), Vol. 160, Issue 2, 2025, p. 25.
• M. I. Haque, P. K. Singh, S. Ghuge, A. Kumar, A. C. Rai, A. Kumar, and A. Modi, ‘A general introduction to and background of plant tissue culture: past, current, and future aspects’, in Advances in Plant Tissue Culture, Elsevier, 2022, pp. 1–30.
• N. Al Ghasheem, R. A. Al-Saeed, and L. T. Fadala, ‘İn vitro, effect of sucrose concentration and type of culture medium on fungal colonies infections on Murashige and Skoog medium’, in J Surv Fisheries Sci, Vol. 10, Issue 3, 2023, pp. 2667–2678.
• M. Cioloca, A. Tican, M. Popa, C. Badarau, A. Diaconu, and R. Draghici, ‘Effects of sucrose medium content and Sterilant Treatment on Microbial Contamination of Sweet potato cultures initiated in vitro’, in Annals Univ Craiova-Agriculture Montanology Cadastre Ser, Vol. 47, Issue 1, 2018, pp. 82–88.
• N. Vidal, C. Sánchez, and B. Cuenca, ‘Proliferation of Axillary Shoots of Chestnut in Temporary Immersion Systems’, in Micropropagation Methods in Temporary Immersion Systems; Methods in Molecular Biology, Ramírez-Mosqueda, M. A., Cruz-Cruz, C. A., Eds.; Springer: Humana, NY, USA, Volume 2759, 2024, pp. 167–181.
• S. Miranda, M. Malnoy, A. Aldrey, M. J. Cernadas, C. Sánchez, B. Christie, and N. Vidal, ‘Micropropagation of Apple Cultivars ‘Golden Delicious’ and ‘Royal Gala’ in Bioreactors’, in Plants, 14(17), 2025, pp. 27–40.
• H. D. Hwang, S. H. Kwon, H. N. Murthy, S. W. Yun, S. S. Pyo, and S.-Y. Park, ‘Temporary Immersion Bioreactor System as an Efficient Method for Mass Production of İn vitro Plants in Horticulture and Medicinal Plants’, in Agronomy, Vol. 12, Issue 2, 2022, p. 346.
• T. Murashige, and F. Skoog, ‘A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures’, in Physiol Plantarum, Vol. 15, 1962, pp. 473–497.
• W. S. Yeng, and P. C. Boyce, “Studies on Schismatoglottideae (Araceae) of Borneo XXXXI: Additional new species of Bucephalandra”, in Willdenowia, Vol. 44, Issue 3, 2014, pp. 414–421.
• P. C. Boyce, and S. Y. Wong, ‘Studies on Schismatoglottideae (Araceae) of Borneo XX: Beccari's «La Più piccola delle Aracee» (Microcasia pygmaea) re-collected and transferred to Bucephalandra Schott’, in Webbia, Vol. 67, 2012, pp. 139–146.
• N. M. Zalan, M. F. N. Hassim, and A. Ahmad, ‘Effect of Sucrose Concentration on Proliferation of Bucephalandra Sp. ‘Red-Mini’ Cultures’, in Universiti Malaysia Terengganu Journal of Undergraduate Research, Vol. 7, Issue 1, 2025, pp. 56–65.
• S. S. Bhojwani, and Razdan, Plant tissue culture: Theory and practice. Elsevier, 1986.
• M. Karatas, and M. Aasim, ‘Influence of carbon sources and cytokinin types on shoot proliferation of Bacopa monnieri L. Pennell.’, in Turkish Journal of Agriculture and Forestry, Vol. 38, Issue 4, 2014, pp. 561–568.
• M. Doğan, ‘Effect of pre-treatment of zeatin on multiple shoot regeneration from leaf explants of aquatic plant Limnophila aromatica (Lamk.) Merr.’, in Journal of Anatolian Environmental and Animal Sciences, Vol. 3, Issue 1, 2018, pp. 45–49.
• M. Doğan, ‘Effect of Pre-treatment of Zeatin on Multiple Shoot Regeneration from Leaf Explants of Aquatic Plant Bacopa monnieri (L.) Wettst.’, in Journal of Anatolian Environmental and Animal Sciences, Vol. 4, Issue 2, 2019, pp. 161–165.
• M. Yaseen, T. Ahmad, G. Sablok, A. Standardi, and I. A. Hafiz, ‘Role of carbon sources for in vitro plant growth and development’, in Molecular Biology Reports, 40(4), 2013, pp. 2837–2849.
• D. M. Braun, and T. L. Slewinski, ‘Genetic control of carbon partitioning in grasses: roles of sucrose transporters and tie-dyed loci in phloem loading’, in Plant Physiology, 149(1), 2009, pp. 71–81.
• M. Karatas, M. Aasim, M. Dogan, ‘Multiple shoot regeneration of Ceratophyllum demersum L. on agar solidified and liquid mediums’, in Fresenius Envir. Bull., Vol. 23, 2014, pp. 3–9.
• M. Dogan, ‘Impact of Growth Regulators on Rapid Clonal Propagation of Rotala rotundifolia (Buch-Ham. ex Roxb) in Liquid Culture Medium’, in Electronic Letters on Science and Engineering, Vol. 18, Issue 2, 2022, pp. 79–86.
• J. A. Silva, ‘The effect of carbon source on in vitro organogenesis of chrysanthemum thin cell layers’, in Bragantia, Vol. 63, 2004, pp. 165–177.
• M. Welander, and N. Pawlicki, ‘Carbon compounds and their influence on in vitro growth and organogenesis’, in Physiology, Growth and Development of Plants in Culture, Dordrecht: Springer Netherlands, 1994, pp. 83–93.