Micro-structural responses of Pisonia grandis R. Br. (Nyctaginaceae) to growth regulators for in vitro regeneration: a study on an avenue tree in urban landscapes

Pisonia grandis R. Br. is a tree commonly found in urban coastal and tropical regions, valued for its ornamental beauty and utility in agroforestry and traditional medicine. To facilitate its propagation, particularly through in vitro techniques, this study aimed to refine a propagation system by inducing adventitious buds from node explants of mature tree. The study revealed that the Murashige and Skoog (MS) medium, supplemented with 30 g L⁻¹ sucrose and 2.0 mg L⁻¹ thidiazuron (TDZ), effectively stimulated bud break. Furthermore, a combination of 1.0 mg L⁻¹ TDZ and 0.5 mg L⁻¹ α-naphthalene acetic acid (NAA) yielded optimal shoot multiplication, resulting in an average of 16.0 shoots per explant with a length of 5.9 cm. In contrast, utilizing 1.0 mg L⁻¹ 6-benzylaminopurine (BAP) and 0.5 mg L⁻¹ NAA led to producing 10.0 shoots with a height of 3.7 cm. Incorporating TDZ significantly enhanced shoot numbers, size, and overall health of the shootlets. Microscopic analysis revealed that leaves and petioles derived from the TDZ and NAA combination exhibited desirable features, such as a thick cuticle, well-differentiated epidermis, mesophyll, vascular tissues, stomata, and improved vein density compared to those derived from BAP and NAA. The maximum root percentage (87.5%) and adventitious root formation (averaging 8.0 roots with a length of 6.2 cm) were observed on half-strength media supplemented with 1.5 mg L⁻¹ indole-3-butyric acid (IBA). Conversely, NAA fortification resulted in lower rooting percentages (77.8% response with an average of 4.7 roots measuring 4.0 cm in length). Subsequently, the rooted plantlets were successfully acclimatized using a mixture of soilrite®, cocopeat, and garden soil (1:1:1 v/v ratio), with a survival rate of 92.0% in the field. This study contributes valuable insights into the optimized use of growth regulators for the large-scale propagation of P. grandis and related species, ensuring the production of micro-morpho-structurally stable plantlets.