Effectiveness of Sterilization Methods of Coffee Leaf Explants (Coffea canephora Var. Milo Pace) To Decrease Contamination and Browning In Vitro
Keywords:
coffea, milo pace, NaCl, sterilization, tissue cultureAbstract
Coffee is one of the leading plantation commodities that are in demand by the public. Robusta coffee var. Milo Pace is Jember's flagship local coffee that was only released at the end of 2023. Seedling propagation continues to be pursued to preserve the coffee. In vitro culture is one of the effective plant propagation methods for production in large quantities. However, explant sterilization is a crucial stage in in vitro culture. Reports related to in vitro culture of robusta milo pace have never been reported before. Therefore, optimization of sterilization methods is the initial stage in supporting the success of in vitro culture of robusta milo pace coffee. The purpose of this study was to obtain the optimal method of sterilization of leaf explants of robusta milo pace coffee. This study used a one-factor randomized design, namely the method of sterilization of coffee leaf explants consisting of 12 methods. The sterilants used were detergent, bactericide, fungicide, NaOCl, H2O2, alcohol and distilled water with different time and concentration. The results showed that sterilization of explants by method XII with 2 g/L detergent for 20 minutes, 2 g/L bactericide and 2 g/L fungicide for 60 minutes, 1.05% NaOCl for 15 minutes, 0.525% NaOCl 10 minutes and 1.5% H2O2 can suppress 20% contamination such as fungi and bacteria with an average contaminant appearance time of 11 days after inoculation. The percentage of browning was 20% with an average appearance of 8.8 days after inoculation. This method maintains the percentage of live explants up to 60% characterized by fresh green explants.
References
Asmono, S. L., Wardana, R., & Rahmawati. (2021). Optimization of Sterilization Methods in Vitro of Arabica (Coffea Arabica L.) and Robusta (Coffea Canephora Var. Robusta Chev.) Leaf Exsplans. Jurnal Ilmiah Inovasi, 21(3), 140–145.
Felek, W., Mekibib, F., & Admassu, B. (2015). Optimization of explants surface sterilization condition for field grown peach (Prunus persica L. Batsch. Cv. Garnem) intended for in vitro culture. African Journal of Biotechnology, 14(8), 657–660. https://doi.org/10.5897/ajb2014.14266
Handayani, E, Irsyadi, M. B., Alawiyah, R. L. M. N., & Aris, I. (2022). Effect of Explants Sterilization and Plant Growth Regulators on Embryo Culture of Kepel (Stelechocarpus burahol). IOP Conference Series: Earth and Environmental Science, 985(012016). https://doi.org/10.1088/1755-1315/985/1/012016
Handayani, Etty, Irsyadi, M. B., Aris, I., Alawiyah, R. L. M. N., Kusumaningtyas, N., Permatasari, F., & Rineksane, innaka ageng. (2021). Optimasi sterilisasi endosperma kepel (Stelechocarpus burahol [Bl] Hook f. & Th) secara in vitro. Bio-Edu:Jurnal Pendidikan Biologi, 6(2), 113–121. https://doi.org/10.32938/jbe.v6i2.1179
Harum, S. (2022). Analisis Produksi Kopi Di Indonesia Tahun 2015-2020 Menggunakan Metode Cobb-Douglass. Jurnal Ilmiah Ekonomi Pembangunan, 1(2), 102–109.
Irsyadi, M. B. (2021). Factors That Effect of the Optimal Plantlet Growth from Tissue Culture on the Acclimatization Stage. Proceeding International Conference on Science and Engineering, 100–104.
Kumparannews. (2023). Jember Patenkan Varietas Kopi Robusta Milo Pace. Https://Kumparan.Com/Kumparannews/.
Natasha, K., & Restiani, R. (2019). Optimasi sterilisasi eksplan pada kultur in vitro ginseng jawa (Talium paniculatum). Symposium of Biology Education, 87–95.
Purwandhini, A. S., Pudjiastutik, W., & Suhaeriyah, N. E. (2023). Analisis Perwilayahan Komoditas Kopi. Jurnal Sosial Ekonomi Pertanian, 19(3), 167–178.
Rismayani, & Hamzah, F. (2010). Pengaruh Pemberian Chlorox (NaOCl) Pada Sterilisasi Permukaan Untuk Perkembangan Bibit Aglaonema (Donna carmen) Secara In Vitro. Prosiding Seminar Ilmiah Dan Pertemuan Tahunan PGJ Dan PEJ XX.
Setiani, N. A., Nurwinda, F., & Astriany, D. (2018). Pengaruh Desinfektan dan Lama Perendaman pada Sterilisasi Eksplan Daun Sukun (Artocarpus altilis (Parkinson ex. F.A Zorn) Fosberg). Biotropika: Journal of Tropical Biology, 6(3), 78–82. https://doi.org/10.21776/ub.biotropika.2018.006.03.01
Shofiyani, A., & Damajanti, N. (2017). Pengembangan Metode Sterilisasi pada Berbagai Eksplan Guna Meningkatkan Keberhasilan Kultur Kalus Kencur (Kaemferia galangal L). Agritech: Jurnal Fakultas Pertanian Universitas Muhammadiyah Purwokerto, 17(1), 55–64. https://doi.org/10.30595/agritech.v17i1.1345
Singh, V., Tyagi, A., Clrauhan, P. K., Kumari, P., & Kaushal, S. (2011). Idenrification And Prevention Of Bacterial Contimination On Explant Used In Plant Tissue Culture Labs. International Journal of Pharmacy and Pharmaceutical Sciences, 3(4), 160–163. https://innovareacademics.in/journal/ijpps/Vol3Issue4/2557.pdf
Solin, N. W. N. M., & Siregar, L. A. M. (2025). Optimization of Sterilization Methods for In Vitro Gambir (Uncaria gambir (Hunter) Roxb) Leaf Explants from Pakpak Bharat. PROSIDING Semnas Integrasi Pertanian Dan Peternakan, 3(1), 58–68.
Suratman, A. P., & Mulyani, S. (2013). Keefektifan Penggunaan Bahan Sterilisasi Dalam Pengendalian Kontaminasi Eksplan Pada Perbanyakan Tanaman Sirsak (Annona muricata L.) Secara In vitro. Publikasi Hasil Penelitian Hibah Bersaing Dikti, 1–8.
Wulandari, A. S., & Nasution, S. S. (2014). The Effect of Sterilant Material on the Success of Initiation Pulownia Explant (Paulownia elongata SY Hu) by In Vitro. Jurnal Silvikultur Tropika, 5(1), 1–6.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2025 Journal of Soilscape and Agriculture
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
