Induction and Regeneration of Tobacco Callus using Benzyl Aminopurine (BAP) and Furfuryl Aminopurine (Kinetin) With Thin Cell Layer (TCL) Technique

Abstract Tobacco was a high-value crop that required effective and efficient cultivation methods to produce quality seedlings. This study aimed to enhance the efficiency of tobacco cultivation through tissue culture using the Thin Cell Layer (TCL) method. The study took place at the Tissue Culture Laboratory, Faculty of Agriculture, University of Jember, between January and March 2024. Leaf bud explants from the Broadleaf One-Sucker tobacco variety were cultivated on Murashige & Skoog (MS) medium supplemented with different concentrations of BAP (0, 2, 3, 4 ppm) and Kinetin (0, 3, 4 ppm). The study employed a Completely Randomized Design (CRD) and was analyzed using SPSS 25 and Microsoft Excel. Callus color analysis used the Munsell Color Chart. The findings revealed that 3 ppm BAP was the best concentration for inducing callus in tobacco, with the callus appearing 10.78 days after planting (DAP) and explants sprouting within 8.3 DAP, producing 81.3 shoots. Visual observations indicated that the combination of 3 ppm BAP and 4 ppm Kinetin (P2M2) produced green callus with a compact texture, suitable for shoot formation. The green color of the callus indicated chlorophyll content, which was important for photosynthesis and shoot growth.