Thin Cell Layer Cultures for In Vitro Shoot Regeneration and Proliferation of Kaempferia parviflora
Keywords:
micropropagation, thin cell layer, Kaempferia parviflora, plant growth regulatorAbstract
Kaempferia parviflora contains bioactive compounds with beneficial effects on health, including the management of obesity, diabetes, and cardiorespiratory disorders, as well as anticancer properties. However, in vitro propagation of this species remains challenging due to the limited availability of rhizomes as explants, which is caused by the prolonged dormancy period and low rate of rhizome splitting. Therefore, this study aimed to evaluate the suitability of thin cell layer (TCL) cultures as an alternative explant source for the shoot regeneration of K. parviflora. TCLs of different thicknesses (1.0, 2.0, 3.0, 4.0 & 5.0 mm) were assessed to determine the optimum explant size using in vitro aerial shoots. The effects of various concentrations of BAP (1.0–5.0 mg/L), kinetin (0.5–4.0 mg/L), TDZ (0.5–4.0 mg/L), NAA (0.5–4.0 mg/L), and 2,4-D (0.2–1.0 mg/L) applied individually were tested for shoot regeneration. Subsequently, the effect of 1.0 mg/L BAP in combination with 2,4-D or TDZ (0–0.8 mg/L) was evaluated. TCLs of 5.0 mm thickness produced the highest shoot regeneration rate (46.33 ± 11.64%). Among the cytokinins, BAP at 1.0 mg/L induced the highest percentage of TCL-forming shoots (44.4 ± 15.7%) with an average of 2.38 ± 0.75 shoots per explant. Supplementing BAP with TDZ further enhanced shoot regeneration, with the combination of 1.0 mg/L BAP and 0.6 mg/L TDZ producing the highest regeneration rate (70.83 ± 8.33%). After 5 weeks of acclimatisation, 90% of plantlets survived in a soil and burnt rice husk mixture (1:1). These findings demonstrate that TCLs are effective explants for the in vitro regeneration of K. parviflora, reducing dependency on rhizomes and supporting large-scale propagation of this medicinal species.
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