Callus and cell suspension culture of bamboo plant, Phyllostachys nigra

Abstract

A modified 1/2MS medium supplemented with 3 µM 2,4-D was effective for callus induction from bamboo shoots of Phyllostachys nigra Munro var. Henonis. In the first phase (phase 1), some parts of the explants enlarged and gave rise to whitish-yellow calli after 2–3 weeks of culture. During maintenance subcultures, almost all explants and the initially formed calli turned brown and these calli gradually lost their proliferation capacity (phase 2). Removal of the necrotic potions of explants, and frequent subcultures at phase 2 was essential. Secondary proliferated calli were subsequently produced on the surface of brown tissues (phase 3). These calli could be maintained on both solid and liquid media. The liquid suspension cells had a blue to pale blue autofluorescence in the cell walls. These cells fluoresced strongly when stained with Calcofluor White M2R and Aniline Blue, indicating the presence of callose (β-1,3-glucan) in a cellulosic wall. Endogenous free amino acids analyses indicated that glutamine, γ-aminobutyric acid, and alanine were the major amino acids in callus tissues whereas asparagine and tyrosine were abundant in the regenerated bamboo shoots.