Journal of Plant Science and Biotechnology

ABSTRACT

Stevia rebaudiana is a high-value medicinal and natural sweetener plant with increasing commercial demand in Bangladesh; however, conventional propagation is limited by low seed viability and inconsistent field establishment. This study aimed to develop an efficient, reproducible, and region-specific direct in vitro regeneration protocol coupled with a climate-adaptive acclimatization strategy suitable for Bangladeshi conditions. Nodal explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of kinetin and 6-benzylaminopurine (BAP) for shoot multiplication, while rooting was induced using different concentrations of naphthaleneacetic acid (NAA) and indole-3-butyric acid (IBA). Acclimatization was performed using six different substrate combinations. The optimized regeneration system resulted in a significantly higher shoot induction frequency (94.33%), number of shoots per explant (8.67 ± 0.58), shoot length (6.42 ± 0.34 cm), and rooting percentage (92.67%), while minimizing callus formation. A Bangladesh-specific acclimatization protocol was developed using cocopeat-vermicompost-garden soil mixture (1:1:1), gradual humidity reduction, and incremental exposure to natural light and ambient temperature, yielding 91.33% survival after 21 days of acclimatization under nursery conditions. The integrated regeneration and acclimatization protocol developed in this study provides a scalable and cost-effective approach suitable for commercial Stevia production in Bangladesh. This study provides a comprehensive optimization of direct regeneration and acclimatization of S. rebaudiana aligned with the subtropical microclimatic conditions of Bangladesh, supporting future research in phytopharmaceutical and biotechnological applications. 

Keywords: Direct Organogenesis; Micropropagation; Phyto-Hormone; Hardening; Subtropical Climate