In Vitro Culture of Vanilla Planifolia: Evaluation of Explant Type, with BAP–GA₃ Effects on Nodal Segments

Leow, Ke Zheng (2026) In Vitro Culture of Vanilla Planifolia: Evaluation of Explant Type, with BAP–GA₃ Effects on Nodal Segments. Final Year Project (Bachelor), Tunku Abdul Rahman University of Management and Technology.

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Abstract

Vanilla planifolia pods are the natural source of vanilla flavoring essence. To meet the high demand for vanilla flavor, in vitro culture of vanilla plantlets is promising as a replacement for conventional propagation using stem cuttings. Besides, it also aids in studies of DNA sequencing and gene modification. Nevertheless, microbial contamination is the most intractable issue in plant tissue culture, particularly when using field-grown plants as sources of explants. An effortless and effective surface sterilization protocol ensures the mass production of genetically identical and healthy plantlets. In the study, the effectiveness of Plant Preservative Mixture (PPM™, Plant Cell Technology) on minimizing microbial contamination was evaluated and integrated into the current surface sterilization protocol. The viability of different types of explants, such as nodal segments, leaves, and aerial roots, was tested using the modified protocol by treating them with bleach solution (Clorox®) and alcohol (70% ethanol), followed with PPM™. Furthermore, the concentration of BAP (6-benzylaminopurine) and KIN (kinetin) and their joint action with GA3 (gibberellic acid) on shoot multiplication of nodal segments were assessed to enhance the multiplication of healthy shoots. Results showed that the nodal explant viability reached 86% while using PPM™. Due to the unfavorable cultural environment, it was hard to maintain uniformity and thus not successful in getting scientific data. Yet, it was feasible to conduct shoot regeneration using nodal segments of vanilla orchids and the application of BAP and KIN. By experimenting with a Completely Randomized Design (CRD) and replicated with ten explants, results revealed statistically highly significant (p < 0.01) differences from the different hormone treatment and concentration, whereby 2 mg/L BAP proved to be the best, providing the highest average length (0.8 cm) of shoots after a week. Meanwhile, no regeneration was observed on leaf tissues or aerial roots, which might be attributed to harsh surface sterilization or a slow regeneration rate before the date. The findings indicate that in vitro culture of Vanilla planifolia has the potential to provide a huge number of vanilla plantlets, provided that the cultural environment meets the minimal requirements.