Development of shoot initiation protocol for in vitro regeneration in bamboo through nodal segments (Bambusa balcooa Roxb.)

Abstract

Bamboo propagation is typically carried out using two primary methods: one involves the use of offsets, culm, or side branch cuttings, while the other relies on micropropagation, also known as clonal propagation, which is most effectively achieved through tissue culture techniques. In vitro propagation is particularly useful to meet the growing demand for bamboo species, given their diverse applications. This study focused on developing a sterilization and initiation protocol for in vitro regeneration of Bambusa balcooa Roxb. The experiment was conducted using nodal explants collected from the Research Farm at the College of Agriculture, Pune, and the tissue culture work was carried out in the Biotechnology Laboratory at the same institution. Nodal explants, when cultured on MS (Murashige and Skoog) media, proved to be ideal for in vitro regeneration. The sterilization protocol for the nodal explants, which included treating them with Tween 20 (10 min), 1% Bavistin (7 min), 0.1% HgCl2 (5 min), 5% sodium hypochlorite (5 min), and 70% ethanol (1 min), followed by three rinses in distilled water between each treatment, resulted in the lowest contamination rate of 13.3%. Various plant growth hormones were tested in MS media to assess their effects on shoot initiation. Among the treatments, BAP (3 mg/l) yielded the best results, with 83.3% of the explants showing shoot initiation as early as 8 days after inoculation.