Development of efficient micropropagation protocol through axillary shoot proliferation for Bambusa vulgaris ‘wamin’ and Bambusa bambos and assessment of clonal fidelity of the micropropagated plants through Random Amplified Polymorphic DNA markers

Abstract

Micropropagation protocols were described for two important bamboo species (Bambusa vulgaris ‘wamin’ and Bambusa bambos) through axillary shoot proliferation. Explants were surface sterilized by sequential application of antifungal (Carbandazim-50%), various antibacterial (Cefotaxime, Kanamycin and Streptocycline) agents and mercuric chloride. The axenic cultures established using B. vulgaris and B. bambos were 78% and 94%, respectively. Initiation and bud break were obtained on Murashige and Skoog (1962) medium supplemented with 3% sucrose. B. vulgaris shoots were best multiplied (6.20 ± 0.32 shoots/explant) on MS containing 8.87 μM 6-benzylaminopurine (BAP) + 0.54 μM naphthaleneacetic acid (NAA) while B. bambos shoots were best multiplied (7.10 ± 0.26 shoots/explant) on MS containing 8.87 μM BAP. Shoot clumps, containing three to four shoots, were rooted on half strength MS supplemented with varying concentrations of indole-3-butyric acid (IBA). B. vulgaris shoots were best rooted (4.4 ± 0.18 roots/clump) on ½ MS supplemented with 21.48 μM IBA while B. bambos shoots were best rooted (2.5 ± 0.12 roots/clump) on ½ MS supplemented with 10.74 μM IBA. Plantlets regenerated in this manner were primary hardened in the greenhouse and acclimatized in the polyhouse under a shade net. The overall survival rates recorded were 85% and 80% for B. vulgaris and B. bambos, respectively. The clonal fidelity of in vitro regenerated plantlets was assessed using 10 random amplified polymorphic DNA (RAPD) markers. All the bands generated using the 10 RAPD decamers were monomorphic and produced more than six scorable bands within the band size of 240–2,505 base pairs.