Differential effects of thiamine, BAP, and thidiazuron combinations on shoot multiplication and phenolic-flavonoid accumulation in pineapple in vitro cultures

Dinda Septia Rachman, Erni Suminar, Anne Nuraini, Fiky Yulianto Wicaksono, Suseno Amien, Syariful Mubarok, Hermanto Hermanto

Abstract


The demand for high-quality pineapple (Ananas comosus (L.) Merr.) planting materials continues to increase, while conventional propagation methods are limited in multiplication rate, uniformity, and production efficiency. In vitro culture offers an alternative approach for rapid and large-scale seedling production. However, information on the effects of thiamine and cytokinin combinations on both shoot multiplication and phenolic-flavonoid accumulation in pineapple in vitro remains limited. This study evaluated ten defined combinations of thiamine and cytokinin for shoot multiplication and total phenolic-flavonoid contents of pineapple explants. The experiment employed a completely randomized design with 10 treatment combinations of thiamine (0.1 and 0.4 mg/L), cytokinin TDZ (0.01 and 0.1 mg/L) and BAP (1 and 2 mg/L) with three replications. The combination of 0.1 mg/L thiamine and 1 mg/L BAP produced the highest number of shoots (3.33 shoots) and leaves (17.56 leaves). In contrast, 0.4 mg/L thiamine combined with 0.1 mg/L resulted in the earliest shoot emergence (54.67 day after subculture) and the highest total phenolic content (13.62 mg GAE/g DW) and total flavonoid content (15.41 mg QE/g DW). No visible browning symptoms were observed in this treatment. These findings demonstrate that the optimal culture conditions differed according to the intended objective, 0.1 mg/L thiamine with 1 mg/L BAP favored shoot proliferation and leaf development, whereas 0.4 mg/L thiamine with 0.1 mg/L TDZ favored phenolic and flavonoid accumulation.

Keywords


Ananas comosus (L.) Merr; browning symptoms; cytokinin; secondary metabolite; shoot proliferation

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References


Ajadi A, Ehirim BO, Ishaq MN, Isah A, Isong A, Ghali A, Kester O, Ejizu AN. 2018. Regeneration of industrial sugarcane using in-vitro plant apical meristem. Electronic Journal of Plant Breeding, 9(4): 1342. https://doi.org/10.5958/0975-928X.2018.00167.9

Andrian R, Agustiansyah A, Junaidi A, Lestari DI. 2022. Aplikasi pengukuran luas daun tanaman menggunakan pengolahan citra digital berbasis android. Jurnal Agrotropika, 21(2): 115. https://doi.org/10.23960/ja.v21i2.6096

Aulia MI, Rustikawati E, Inoriah. 2020. Respon temu putih dan temu mangga dengan pemberian BA dan 2,4-D secara in vitro. Gema Agro, 25(2): 92-102.

Azmi R, Handriatni A. 2018. Pengaruh macam zat pengatur tumbuh alami terhadap pertumbuhan setek beberapa klon kopi robusta (Coffea canephora). Biofarm: Jurnal Ilmiah Pertanian, 14(2). https://doi.org/10.31941/biofarm.v14i2.794

Badan Pusat Statistik. 2024. Produksi Tanaman Buah-buahan, 2023. Badan Pusat Statistik Indonesia.

Baskara DR, Wijayani A, Rina. 2018. Kombinasi zat penghambat pencoklatan dan sukrosa terhadap pertumbuhan planlet pisang mas kirana (Musa acuminata C.) secara in vitro. Jurnal Agrivet, 24(1).

Bruce BB, Boateng ID. 2025. Pineapple by-products: A critical review of their bioactive compounds as eco-friendly pesticides in pest management. Food Chemistry: X, 28: 102567. https://doi.org/10.1016/j.fochx.2025.102567

Casas-Rodríguez AD, González-Martínez DW, Contreras CSA, Ascacio-Valdés JA, Dávila-Medina MD, Chávez-González ML, Sepúlveda-Torre L. 2025. Potential of pineapple waste: Bioactive compounds and their biological significance in human health. Journal of BioProcess and Chemical Technology, 17(35).

Chandran H, Meena M, Barupal T, Sharma K. 2020. Plant tissue culture as a perpetual source for production of industrially important bioactive compounds. Biotechnology Reports, 26: e00450. https://doi.org/10.1016/j.btre.2020.e00450

Cryssanti AD, Wijayani AMP, Wahyurini E. 2019. Induksi planlet kantong semar (Nepenthes ampullaria Jack) pada berbagai konsentrasi thiamin dan benzyl amino purine secara in vitro. AGRIVET, 25: 78-87.

Dahniar N, Elvavina P. 2022. Kombinasi BAP dan NAA untuk media perbanyakan nanas varietas Smooth Cayenne, Toboali in vitro. Agrotechnology Research Journal, 6(1): 21. https://doi.org/10.20961/agrotechresj.v6i1.55629

D’Amelia V, Aversano R, Chiaiese P, Carputo D. 2018. The antioxidant properties of plant flavonoids: their exploitation by molecular plant breeding. Phytochemistry Reviews, 17(3): 611-625. https://doi.org/10.1007/s11101-018-9568-y

Dar SA, Nawchoo IA, Tyub S, Kamili AN. 2021. Effect of plant growth regulators on in vitro induction and maintenance of callus from leaf and root explants of Atropa acuminata Royle ex Lindl. Biotechnology Reports, 32: e00688. https://doi.org/10.1016/j.btre.2021.e00688

de Ancos B, Sánchez-Moreno C, González-Aguilar GA. 2017. Pineapple composition and nutrition. In: Handbook of Pineapple Technology, 221-239. Wiley. https://doi.org/10.1002/9781118967355.ch12

Dinani ET, Shukla MR, Turi CE, Sullivan JA, Saxena PK. 2018. Thidiazuron: modulator of morphogenesis in vitro. In: Thidiazuron: From Urea Derivative to Plant Growth Regulator, 1-36. Springer Singapore. https://doi.org/10.1007/978-981-10-8004-3_1

Erland LAE, Giebelhaus RT, Victor JMR, Murch SJ, Saxena PK. 2020. The morphoregulatory role of thidiazuron: metabolomics-guided hypothesis generation for mechanisms of activity. Biomolecules, 10(9): 1253. https://doi.org/10.3390/biom100912531

Fauzi NI, Herawati IE, Hadisoebroto G. 2023. Kadar fenolik total, kadar flavonoid, dan aktivitas antioksidan dari ekstrak kulit buah nanas (Ananas comosus (L.) Merr.) varietas Pemalang. Jurnal Mandala Pharmacon Indonesia, 9(2): 492-500. https://doi.org/10.35311/jmpi.v9i2.413

Fitramala E, Khaerunnisa E, Djuita NRDR, Sunarso H, Ratnadewi D. 2017. Kultur in vitro pisang (Musa paradisiaca L.) cv. Kepok Merah untuk mikropropagasi cepat. E-Journal Menara Perkebunan, 84(2). https://doi.org/10.22302/iribb.jur.mp.v84i2.221

Fitzpatrick TB, Chapman LM. 2020. The importance of thiamine (vitamin B1) in plant health: from crop yield to biofortification. Journal of Biological Chemistry, 295(34): 12002-12013. https://doi.org/10.1074/jbc.REV120.010918

Levinsh G. 2023. Water content of plant tissues: so simple that almost forgotten? Plants, 12(6): 1238. https://doi.org/10.3390/plants12061238

Li Y, Kong D, Fu Y, Sussman MR, Wu H. 2020. The effect of developmental and environmental factors on secondary metabolites in medicinal plants. Plant Physiology and Biochemistry, 148: 80-89. https://doi.org/10.1016/j.plaphy.2020.01.006

Liu Z, Farkas P, Wang K, Kohli M, Fitzpatrick TB. 2022. B vitamin supply in plants and humans: the importance of vitamer homeostasis. The Plant Journal, 111(3): 662-682. https://doi.org/10.1111/tpj.15859

López-Hidalgo C, Meijón M, Lamelas L, Valledor L. 2021. The rainbow protocol: a sequential method for quantifying pigments, sugars, free amino acids, phenolics, flavonoids and MDA from a small amount of sample. Plant, Cell & Environment, 44(6): 1977-1986. https://doi.org/10.1111/pce.14007

Lukman, Fridayanti N, Budi S, Junaidi, Dahlan. 2023. Respon pertumbuhan bibit cabutan tanaman penghasil gaharu (Aquilaria sp.) akibat pemberian vitamin B1 pada media yang berbeda. Jurnal Ilmiah Mahasiswa Agroekoteknologi, 2(2): 50-55. https://doi.org/10.29103/jimatek.v2i2.12556

Lutfiani I, Lestari A, Widyodaru N, Suhesti S. 2022. Pengaruh pemberian berbagai konsentrasi NAA (Naphthalene Acetic Acid) dan BAP (Benzyl Amino Purine) terhadap multiplikasi tunas tanaman tebu (Saccharum officinarum L.). Jurnal Agro Industri, 1(7): 49-57. https://doi.org/10.33661/jai.v7i1.6111

Mawaddah Y, Erawati DN, Donianto M, Ryana WM, Ikanafi’ah A. 2021. Peran sitokinin terhadap kemampuan eksplan pada penggandaan tunas vanili (Vanilla planifolia Andrews.). Agriprima: Journal of Applied Agricultural Sciences, 5(2): 169-179. https://doi.org/10.25047/agriprima.v5i2.441

Megersa HG. 2017. Propagation methods of selected horticultural crops by specialized organs: review. Journal of Horticulture, 4(2). https://doi.org/10.4172/2376-0354.1000198

Mohd Ali M, Hashim N, Abd Aziz S, Lasekan O. 2020. Pineapple (Ananas comosus): a comprehensive review of nutritional values, volatile compounds, health benefits, and potential food products. Food Research International, 137: 109675. https://doi.org/10.1016/j.foodres.2020.109675

Muhammad DRA, Lemarcq V, Alderweireldt E, Vanoverberghe P, Praseptiangga D, Juvinal JG, Dewettinck K. 2020. Antioxidant activity and quality attributes of white chocolate incorporated with Cinnamomum burmannii Blume essential oil. Journal of Food Science and Technology, 57(5): 1731-1739. https://doi.org/10.1007/s13197-019-04206-6

Munir, Aini F, Jariah S. 2016. Pengaruh kadar thiamine (vitamin B1) terhadap pertumbuhan jamur tiram putih (Pleurotus ostreatus). Jurnal Biota, 2(2).

Naheed R, Zahid M, Aqeel M, Maqsood MF, Kanwal H, Khalid N, Hashem M, Alamri S, Noman A. 2022. Mediation of growth and metabolism of Pisum sativum in salt stress potentially be credited to thiamine. Journal of Soil Science and Plant Nutrition, 22(3): 2897-2910. https://doi.org/10.1007/s42729-022-00854-4

Numba S, Syam N, Imaniar M. 2024. Increasing flavonoid compounds through the use of elicitors in callus culture Catharanthus roseus L. Jurnal Penelitian Pertanian Terapan, 24(3): 366-374. https://doi.org/10.25181/jppt.v24i3.3406

Nurreni A, Susiyanti S, Hilal S, Sulistyorini E. 2025. The effect of vitamin B1 and nano zinc on drip irrigation systems for macakal mangosteen (Garcinia mangostana). Jurnal Biologi Tropis, 25(4a): 583-593. https://doi.org/10.29303/jbt.v25i4a.10858

Oberhuber W. 2017. Soil water availability and evaporative demand affect seasonal growth dynamics and use of stored water in co-occurring saplings and mature conifers under drought. Trees, 31(2): 467-478. https://doi.org/10.1007/s00468-016-1468-4

Patil VC, Waghmare RB, Mane RS. 2021. Challenges and prospects in pineapple propagation: a critical review. International Journal of Horticulture, 11(2): 22-28.

Purita SY, Rahmi N, Ardiarini, Basuki N. 2017. Pengaruh zat pengatur tumbuh jenis BAP terhadap pertumbuhan planlet sub kultur jaringan tanaman nanas (Ananas comosus L. Merr.). Jurnal Produksi Tanaman, 5(7): 1207-1212.

Pusat Data dan Sistem Informasi Pertanian. 2022. Analisis kinerja perdagangan nanas.

Putri ND, Setiari N, Nurchayati Y, Suedy SWA. 2024. The effect of thidiazuron treatment on the growth of Vanda limbata Blume in vivo. Makara Journal of Science, 28(4): 330-336. https://doi.org/10.7454/mss.v28i4.2207

Raisya E, Sobarna DS, Nuraini A, Mubarok S, Suminar E, Akutsu M. 2020. Multiplikasi in vitro stroberi kultivar Tochiotome dengan penambahan jenis dan konsentrasi sitokinin untuk perbanyakan bibit. Kultivasi, 19(3). https://doi.org/10.24198/kultivasi.v19i3.26932

Rani DEP, Elhany NA, Faisyah APN. 2023. Pengaruh konsentrasi thidiazuron (TDZ) terhadap multiplikasi tunas anggrek Grammatophyllum sp. secara in vitro. AGRIBIOS, 21(2): 165. https://doi.org/10.36841/agribios.v21i2.3465

Saepudin A, Amilin A, Undang U, Sudartini T. 2023. Kultur in vitro pisang Cavendish (Musa acuminata L.) pada media dengan konsentrasi berbeda ekstrak jambu batu dan benzyl amino purine. Paspalum: Jurnal Ilmiah Pertanian, 11(1): 87. https://doi.org/10.35138/paspalum.v11i1.481

Setiadi A, Hanum F, Indrayanti R, Hadipoentyanti E. 2025. In vitro shoot induction of vanilla (Vanilla planifolia Andrews.) through the application of kinetin, NAA, and TDZ. IOP Conference Series: Earth and Environmental Science, 1476(1): 012051. https://doi.org/10.1088/1755-1315/1476/1/012051

Setiawati T, Ayalla A, Nurzaman M, Kusumaningtyas VA, Bari I. 2020. Analisis metabolit sekunder kultur pucuk, kalus, dan tanaman lapang Chrysanthemum morifolium Ramat. Jurnal Ilmu Dasar, 21(1): 1. https://doi.org/10.19184/jid.v21i1.8665

Shoja HM, Shishavani HK. 2021. Effects of different hormonal treatments on growth parameters and secondary metabolite production in organ culture of Hyssopus officinalis L. BioTechnologia, 102(1): 33-41. https://doi.org/10.5114/bta.2021.103760

Srilestari R, Suwardi D. 2019. Penambahan thiamin dan pupuk daun pada tahap aklimatisasi pisang abaka (Musa textilis Nee.). AGRIVET, 25.

Subki A, Abidin AAZ, Yusof ZNB. 2018. The role of thiamine in plants and current perspectives in crop improvement. In: B Group Vitamins - Current Uses and Perspectives. InTech. https://doi.org/10.5772/intechopen.79350

Taha R, El Gohary A, Zaied N, Hassan S. 2022. Improving micropropagation efficiency and biochemical content of pineapple (Ananas comosus) using stevia extract. Egyptian Journal of Chemistry, 0(0): 0-0. https://doi.org/10.21608/ejchem.2022.122900.5500

Tuzzahra A, Mustakim A. 2026. Kajian fitokimia dan aktivitas pencernaan buah nanas (Ananas comosus) dalam mendukung nafsu makan. Jurnal Ilmiah Ilmu Kesehatan & Medis (MURAKES), 2(1): 37-46.

Wang F, Li Y, Pang Y, Hu J, Kang X, Qian C. 2025. Thidiazuron enhances strawberry shoot multiplication by regulating hormone signal transduction pathways. International Journal of Molecular Sciences, 26(9): 4060. https://doi.org/10.3390/ijms26094060

Yudha EP, Rachmadina V. 2023. Daya saing ekspor komoditas nanas Indonesia, Thailand dan Filipina di negara tujuan ekspor utama. Jurnal Universitas Padjadjaran.

Ziraluo YPB. 2021. Metode perbanyakan tanaman ubi jalar ungu (Ipomoea batatas Poiret) dengan teknik kultur jaringan atau stek planlet. Jurnal Inovasi Pertanian, 2(3).




DOI: https://doi.org/10.24198/kultivasi.v25i2.70924

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