Tissue culture of multipurpose bamboo (Bambusa tulda Roxb) : Explant collection and aseptic culture establishment for mass clonal propagation of elite germplasm

Bambusa tulda Roxb., an economically important multipurpose bamboo, plays a vital role in rural livelihoods and sustainable development. However, its large-scale propagation is constrained by irregular flowering, limited seed availability, and the slow nature of conventional vegetative methods. In vitro propagation offers an effective alternative for rapid multiplication of elite, disease-free planting material, although the establishment of aseptic cultures is often hindered by microbial contamination and phenolic exudation. The present study was conducted to standardize an efficient protocol for in vitro propagation of B. tulda in relation to explant selection, culture conditions and growth regulators. Among different plant ages and collection times, explants collected during early morning (6:00–8:00 am) from 4-year-old plants exhibited maximum bud break (100%) with minimum contamination (10%), highlighting the importance of physiological status and collection timing. Among plant growth regulators, MS liquid medium supplemented with BAP (1.0 mg/l) showed superior response with 100% bud break, highest shoot number (8.33 ± 0.55), and maximum shoot length (9.28 ± 0.27 cm), outperforming kinetin and TDZ treatments. Nutrient media evaluation revealed that full-strength MS liquid medium was most effective for shoot proliferation compared to B5, WPM, and Heller’s medium. For shoot multiplication, BAP (1.0 mg/l) alone produced the highest number of shoots (22.11 ± 0.48), while among gelling agents, 0.25% clerigel yielded maximum shoot proliferation.In rooting experiments, MS medium supplemented with NAA (5.0 mg/l) resulted in the highest rooting (98%) with well-developed roots. During acclimatization, cocopeat: vermicompost (2:1) resulted maximum survival (92%), particularly during monsoon and spring seasons.The protocol developed in this study provides a reliable and efficient system for large-scale clonal propagation of B. tulda. These findings contribute to overcoming key challenges in bamboo tissue culture and have significant implications for commercial production, conservation of elite germplasm, and sustainable utilization of bamboo resources.