Axillary shoots: Initiation to acclimatization on micropropagation of Philodendron ‘Super Atom’

Budi Winarto; Fitri Rachmawati; Sri Rianawati; Fitrahtunnisa Fitrahtunnisa; Joko Pramono; Sigid Handoko

doi:10.17268/sci.agropecu.2026.041

Autores/as

Budi Winarto Reasearch Center for Horticulture; Research Organization of Agriculture and Food, National Research and Innovation Agency, The Republic of Indonesia https://orcid.org/0009-0000-6603-1959
Fitri Rachmawati Research Center for Horticulture, Research Organization of Agriculture and Food, National Research and Innovation Agency, The Republic of Indonesia https://orcid.org/0000-0003-4532-903X
Sri Rianawati Research Center for Horticulture, Research Organization of Agriculture and Food, National Research and Innovation Agency, The Republic of Indonesia https://orcid.org/0000-0001-8019-5386
Fitrahtunnisa Fitrahtunnisa Research Center for Horticulture, Research Organization of Agriculture and Food, National Research and Innovation Agency, The Republic of Indonesia https://orcid.org/0000-0002-7672-0907
Joko Pramono Research Center for Horticulture, Research Organization of Agriculture and Food, National Research and Innovation Agency, The Republic of Indonesia https://orcid.org/0000-0003-2793-2142
Sigid Handoko Research Center for Horticulture, Research Organization of Agriculture and Food, National Research and Innovation Agency, The Republic of Indonesia https://orcid.org/0000-0002-2655-8630

DOI:

https://doi.org/10.17268/sci.agropecu.2026.041

Palabras clave:

axillary shoot, acclimatization, in vitro culture, plant growth regulator, V, Lacy tree philodendron

Resumen

Philodendron ‘Super Atom’ is among Indonesia's most favoured ornamental foliage plants, yet traditional propagation methods pose challenges. Establishing an in vitro propagation protocol for this plant was crucial to sustainably produce high-quality planting materials to meet consumer demand. All experiments were conducted using a completely randomized design (CRD) with four replicates. High axillary shoots with regeneration issues were used as explant sources, initiated on Murashige and Skoog (MS) medium containing 1 mg L^-1 thidiazuron (TDZ) and 3 mg L^-1 N6-benzylaminopurine (BAP). A total of 4.8 shoots per explant were regenerated on MS medium supplemented with 0.3 mg L^-1 BAP and 0.005 mg L^-1 IAA. A slight enhancement in axillary shoot proliferation was noted on MS medium with 0.5 mg L^-1 BAP, 0.5 mg L^-1 TDZ, and 0.025 mg L^-1 α-naphthalene acetic acid (NAA), yielding 6.7 shoots explant^-1, and 0.2 mg L^-1 BAP and 0.01 mg L^-1 indole-3-butyric acid (IBA), yielding 6.9 shoots explant^-1. A significant improvement was achieved on MS medium fortified with 0.2 mg L^-1 BAP and 0.005 mg L^-1 indole-3-acetic acid (IAA), resulting in 13.6 shoots explant^-1. Rooted shoots were readily observed in the proliferation medium. A 100% survival rate of plantlets was successfully acclimatized using a mixture of burned rice husks, hyacinth organic manure, and cocopeat (1:2:1, v/v/v), as well as burned rice husks and hyacinth organic manure (1:1, v/v). The findings of this study can serve as a model for developing and establishing new in vitro mass propagation protocols for other Philodendron species with appropriate modifications.

Citas

Agustina, M., Maisura, M., & Handayani, R. S. (2020). The Effect of Different Seed Cutting Treatments and Concentrations of BAP for the Successful In Vitro Micrografting of Mangosteen (Garcinia mangostana L.). Journal of Tropical Horticulture, 3(1), 1. https://doi.org/10.33089/jthort.v3i1.37

Akramian, M., Khaleghi, A., & Salehi Arjmand, H. (2024). Optimization of plant growth regulators for in vitro mass propagation of Philodendron cv. Birkin through shoot tip culture. Greenhouse Plant Production Journal, 1(1), 55–62. https://doi.org/10.61186/gppj.1.1.55

Alawaadh, A. A., Dewir, Y. H., Alwihibi, M. S., Aldubai, A. A., El-Hendawy, S., & Naidoo, Y. (2020). Micropropagation of Lacy Tree Philodendron (Philodendron bipinnatifidum Schott ex Endl.). HortScience, 55(3), 294–299. https://doi.org/10.21273/HORTSCI14612-19

Alwahibi, M. S., Alawaadh, A. A., Dewir, Y. H., Soliman, D. A., & Seliem, M. K. (2022). Assessment of genetic fidelity of lacy tree philodendron (Philodendron bipinnatifidum Schott ex Endl.) micro propagated plants. Bionatura, 7(1), 1–5. https://doi.org/10.21931/RB/2022.07.01.10

Anbazhakan, R., Parthibhan, S., Rajasekar, C., Muthukumar, M., & Gao, J. (2025). In vitro micropropagation of Theriophonum sivaganganum (Ramam. and Sebastine) Bogner—an endemic plant of South Tamil Nadu, India. In Vitro Cellular and Developmental Biology - Plant, 61(1), 239–247. https://doi.org/10.1007/s11627-025-10515-7

Ayala, P. G., Acevedo, R. M., Luna, C. V., Rivarola, M., Acuña, C., Marcucci Poltri, S., González, A. M., & Sansberro, P. A. (2022). Transcriptome Dynamics of Rooting Zone and Leaves during In Vitro Adventitious Root Formation in Eucalyptus nitens. Plants, 11(23), 1–25. https://doi.org/10.3390/plants11233301

Bhat, A. Y., & Shahzad, A. (2026). Optimizing efficient and reproducible in vitro regeneration via shoot organogenesis and somatic embryogenesis from leaf and nodal explants of Phlomis bracteosa Royle ex Benth.—a highly valued medicinal herb of the Himalayas. In Vitro Cellular and Developmental Biology - Plant, 0123456789. https://doi.org/10.1007/s11627-025-10622-5

CAPOTESCU, M., PETCU, F., UNGUR, R., ALDA, S., & DANCI, M. (2025). Effect of growth regulator combinations on In Vitro shoot proliferation and root formation in Fragaria vesca and Fragaria × ananassa. Journal of Horticulture, Forestry and Biotechnology, 29(2), 485–490. https://doi.org/10.59463/zr2cwk24

Chen, S., Xiong, Y., Yu, X., Pang, J., Zhang, T., Wu, K., Ren, H., Jian, S., Teixeira da Silva, J. A., Xiong, Y., Zeng, S., & Ma, G. (2020). Adventitious shoot organogenesis from leaf explants of Portulaca pilosa L. Scientific Reports, 10(1), 1–8. https://doi.org/10.1038/s41598-020-60651-w

Chiewchan, N., Saetiew, K., & Teerarak, M. (2023). The effect of BA on inducing shoots of Philodendron erubescent “Pink Princes” in vitro. International Journal of Agricultural Technology, 19(6), 2385–2398.

del Rosario Cárdenas-Aquino, M., Sarria-Guzmán, Y., & Martínez-Antonio, A. (2022). Review: Isoprenoid and aromatic cytokinins in shoot branching. Plant Science, 319(February). https://doi.org/10.1016/j.plantsci.2022.111240

Dewir, Y. H., Habib, M. M., AlQarawi, A. A. A., Alshahrani, T. S., Alaizari, A. A., Malik, J. A., Alwahibi, M. S., & Murthy, H. N. (2023). Mycorrhization Enhances Vegetative Growth, Leaf Gas Exchange, and Root Development of Micropropagated Philodendron bipinnatifidum Schott ex Endl. Plantlets during Acclimatization. Horticulturae, 9, 276. https://doi.org/10.3390/horticulturae9020276

Güney, M. (2023). Optimization of in vitro micropropagation protocol for Eminium rauwolffii var. rauwolffii: an ornamental plant with prominent pharmaceutical value. Genetic Resources and Crop Evolution, 70(8), 2439–2449. https://doi.org/10.1007/s10722-023-01572-1

Kang, I., & Sivanesan, I. (2025). Micropropagation of Philodendron ‘White Knight’ via Shoot Regeneration from Petiole Explants. Plants, 14(11), 1–12. https://doi.org/10.3390/plants14111714

Karthik, G., Ravindra Kumar, K., Pradesh, A., Aruna Kumari, I. K., Dorajee Rao, A., Raju, D., Aruna Kumari, K., & Gouri sankar, T. (2023). The Pharma Innovation Journal 2023; 12(9): 52-58 Optimization of in vitro culture establishment in Aglaonema sp. by employing shoot tips and nodal segments as explants. The Pharma Innovation Journal, 12(9), 52–58. www.thepharmajournal.com

Khamrit, R., & Jongrungklang, N. (2024). Determining Optimal Mutation Induction of Philodendron billietiae Using Gamma Radiation and In Vitro Tissue Culture Techniques. Horticulturae, 10(11), 1–12. https://doi.org/10.3390/horticulturae10111164

Kharrazi, M., Moradian, M., Moghaddam, Z. S., Khadem, A., & Sharifi, A. (2023). Micropropagation and ex vitro rooting of three ZZ plant (Zamioculcas zamiifolia Engl.) cultivars. In Vitro Cellular and Developmental Biology - Plant, 59(1), 129–139. https://doi.org/10.1007/s11627-022-10323-3

Klanrit, P., Kitwetcharoen, H., Thanonkeo, P., & Thanonkeo, S. (2023). In Vitro Propagation of Philodendron erubescens ‘Pink Princess’ and Ex Vitro Acclimatization of the Plantlets. Horticulturae, 9(6), 1–10. https://doi.org/10.3390/horticulturae9060688

Kuathan, N., & Judsri, W. (2023). Effect of Micropropagation of Philodendron Pink Princess ( Philodendron erubescens ) by Tissue Culture Technique. วารสารเกษตรและอาหาร มรวอ., 2(1), 16–21.

Lakho, M. A., Jatoi, M. A., Solangi, N., Abul-Soad, A. A., Qazi, M. A., & Abdi, G. (2023). Optimizing in vitro nutrient and ex vitro soil mediums-driven responses for multiplication, rooting, and acclimatization of pineapple. Scientific Reports, 13(1), 1–10. https://doi.org/10.1038/s41598-023-28359-9

Lamboro, A., Han, X., Yang, S., Li, X., Yao, D., Song, B., & Zhang, J. (2022). Combination of 6-Benzylaminopurine and Thidiazuron Promotes Highly Efficient Shoot Regeneration from Cotyledonary Node of Mature Peanut (Arachis hypogaea L.) Cultivars. Phyton-International Journal of Experimental Botany, 91(12), 2619–2631. https://doi.org/10.32604/PHYTON.2022.021404

Li, Q., Xing, X., Gong, Y., Li, Z., Huang, M., & Jiang, Y. (2026). Effects of plant growth regulator combinations on the growth of ginger (Zingiber officinale Roscoe) plantlets in vitro. Kuwait Journal of Science, 53(2). https://doi.org/10.1016/j.kjs.2026.100539

Maikaeo, L., Puripunyavanich, V., Limtiyayotin, M., Orpong, P., & Kongpeng, C. (2024). Micropropagation and gamma irradiation mutagenesis in Philodendron billietiae. Thai Journal of Agricultural Science, 57(1), 11–19.

Martini, A. N., Vlachou, G., & Papafotiou, M. (2022). Effect of Explant Origin and Medium Plant Growth Regulators on In Vitro Shoot Proliferation and Rooting of Salvia tomentosa, a Native Sage of the Northeastern Mediterranean Basin. Agronomy, 12(8). https://doi.org/10.3390/agronomy12081889

Mhimdi, M., & Pérez-Pérez, J. M. (2020). Understanding of Adventitious Root Formation: What Can We Learn From Comparative Genetics? Frontiers in Plant Science, 11(October), 1–10. https://doi.org/10.3389/fpls.2020.582020

Mohammed, M., Munir, M., & Ghazzawy, H. S. (2023). Design and Evaluation of a Smart Ex Vitro Acclimatization System for Tissue Culture Plantlets. Agronomy, 13(1). https://doi.org/10.3390/agronomy13010078

Mongkolsawat, W., Punjansing, T., & Loma-in, P. (2023). Effects of BA , TDZ , and NAA on growth of philodendron ‘Birkin’ in vitro. PSRU Journal of Science and Technology, 8(1), 27–36.

Murashige, T., & Skoog, F. (1962). A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures. Physiologia Plantarum, 15(3), 473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x

Nautiyal, A., Ramlal, A., Agnihotri, A., & Rashid, A. (2023). Stress-induced somatic embryogenesis on seedlings of Azadirachta indica A. Juss. by thidiazuron and its inhibition by ethylene modulators. Plant Cell, Tissue and Organ Culture, 153(2), 357–366. https://doi.org/10.1007/s11240-023-02473-y

Nautiyal, A., Rashid, A., & Agnihotri, A. (2022). Induction of multiple shoots in Oryza sativa: roles of thidiazuron, 6-benzylaminopurine, decapitation, flooding, and Ethrel® treatments. In Vitro Cellular and Developmental Biology - Plant, 58(6), 1126–1137. https://doi.org/10.1007/s11627-022-10316-2

Nishesh, S., Nishant, K., Jerin, J., Sonika, K., & Shivani, J. (2023). Strategies for successful acclimatization and hardening of in vitro regenerated plants: Challenges and innovations in micropropagation techniques. Plant Science Today, 10(x), 90–97. https://doi.org/10.14719/pst.2376

Pasternak, T. P., & Steinmacher, D. (2024). Plant Growth Regulation in Cell and Tissue Culture In Vitro. Plants, 13(2), 1–24. https://doi.org/10.3390/plants13020327

Pourhassan, A., Kaviani, B., Kulus, D., Miler, N., & Negahdar, N. (2023). A Complete Micropropagation Protocol for Black-Leaved Zamioculcas zamiifolia (Lodd.) Engl. ‘Dowon.’ Horticulturae, 9(4), 1–10. https://doi.org/10.3390/horticulturae9040422

Raju, R. I., Hashi, A. K., Jazib, A., & Hossain, M. T. (2022). Micropropagation of Alocasia amazonica through Indirect Shoot Organogenesis. Plant Tissue Culture and Biotechnology, 32(1), 13–20. https://doi.org/10.3329/ptcb.v32i1.60468

Sakulsathaporn, A., & Mapanao, R. (2025). An ex vitro rooting-based clonal propagation system with ISSR confirmed genetic fidelity for conservation of Cryptocoryne crispatula var. yunnanensis (Araceae) from the Mekong River Basin. Plant Cell, Tissue and Organ Culture, 161(3), 1–11. https://doi.org/10.1007/s11240-025-03112-4

Sangchanjiradet, S., Kaewprom, C., Dasri, K., Sutthiphapa, N., & Rattana, K. (2024). In vitro propagation of Philodendron erubescens ’White princess’ and Philodendron verrucosum l. Mathieu ex schott by temporary immersion bioreactor system (TIBS). PSRU Journal of Science and Technology, 9(3), 94–108.

Seliem, M. K., El-Mahrouk, M. E., El-Banna, A. N., Hafez, Y. M., & Dewir, Y. H. (2021). Micropropagation of Philodendron selloum: Influence of copper sulfate on endophytic bacterial contamination, antioxidant enzyme activity, electrolyte leakage, and plant survival. South African Journal of Botany, 139, 230–240. https://doi.org/10.1016/j.sajb.2021.01.024

Shintiavira, H., Pramanik, D., Daniyanti, R. D., Pertiwi, M. D., & Rachmawati, F. (2024). Effects of explants and culture medium compositions on quality of chrysanthemum ‘Jayanti Agrihorti’ rooted cuttings. Ilmu Pertanian (Agricultural Science), 9(1), 51–56.

Sjahril, R., Haring, F., Riadi, M., Rahim, M. D., Khan, R. S., Amir, A., & R., T. A. (2016). Performance of NAA, 2iP, BAP and TDZ on Callus Multiplication, Shoots Initiation and Growth for Efficient Plant Regeneration System in Chrysanthemum (Chrysanthemum morifolium Ramat.). International Journal of Agriculture System, 4(1), 52–61. http://pasca.unhas.ac.id/ojs/index.php/ijas/article/view/241

Sosnowski, J., Truba, M., & Vasileva, V. (2023). The Impact of Auxin and Cytokinin on the Growth and Development of Selected Crops. Agriculture (Switzerland), 13(3), 1–14. https://doi.org/10.3390/agriculture13030724

Surve, S. P. (2022). Micropropagation Studies In Philodendron (Philodendron Bipinnatifidum). International Journal of Scientific Research in Science and Technology, 9(15), 126–134.

Sushmarani, Y. S., Venkatesha Murthy, P., & Deeksha Raj, N. (2021). Influence of BAP with TDZ growth regulators on in vitro regeneration in chrysanthemum (Dendranthema grandiflora T.) cv. Marigold. Journal of Pharmacognosy and Phytochemistry, 10(2), 1171–1176.

Talukdar, M., Swain, D. K., & Bhadoria, P. B. S. (2022). Effect of IAA and BAP application in varying concentration on seed yield and oil quality of Guizotia abyssinica (L.f.) Cass. Annals of Agricultural Sciences, 67(1), 15–23. https://doi.org/10.1016/j.aoas.2022.02.002

Vy, B. K., Klanrit, P., Thanonkeo, S., & Thanonkeo, P. (2025). Development of an Efficient Micropropagation Protocol for Philodendron erubescens ‘Pink Princess’ Using a Temporary Immersion System and Assessment of Genetic Fidelity. Horticulturae, 11(9), 1–18. https://doi.org/10.3390/horticulturae11091085

Wu, Q., Yang, H., Sun, Y., Hu, J., & Zou, L. (2021). Organogenesis and high-frequency plant regeneration in Caryopteris terniflora Maxim. using thidiazuron. In Vitro Cellular and Developmental Biology - Plant, 57(1), 39–47. https://doi.org/10.1007/s11627-020-10114-8

Zhang, Y., Zhang, T., Zheng, Y., Wang, J., Luo, C., Li, Y., & Liu, X. (2025). Physiological and Transcriptomic Analyses Reveal Regulatory Mechanisms of Adventitious Root Formation in In Vitro Culture of Cinnamomum camphora. International Journal of Molecular Sciences, 26(15), 1–22. https://doi.org/10.3390/ijms26157264

Zhi, L., & Hu, X. (2023). Adventitious Root Regeneration: Molecular Basis and Influencing Factors. Phyton-International Journal of Experimental Botany, 92(10), 2825–2840. https://doi.org/10.32604/phyton.2023.030912