In vitro cloning of Bambusa pallida Munro through axillary shoot proliferation and evaluation of genetic fidelity by random amplified polymorphic DNA markers

Abstract

Multiple shoots emerged from the nodal shoot segments of the field-grown candidate plus clump explants of Bambusa pallida Munro when cultured on Murashige and Skoog (MS) liquid medium with additives (ascorbic acid 50 mg/L + citric acid 25 mg/L + cysteine 25 mg/L) and combined use of a-naphthalene acetic acid (NAA) 1.34 pM + thiodiozuron 1.125 pM in a 2-week period. Further shoot multiplication was achieved in MS liquid medium with additives + NAA 1.34 pM + 6-benzylaminop- urine 4.4 pM at 25±2°C and 33.78 pmol photons m-2 s-1 light illumination for a 12-h photoperiod. These shoots were rooted within four weeks in MS/2 basal salt medium with addi-tives + 2% sucrose +1% glucose, and 0.6% agar by pulse treatment of shoots with indole 3 butyric acid 0.5 mg/mL for 30 min prior to inoculation. Rooted plants were successfully hardened in the mist chamber. Survival rate during hardening was more than 95%. Micropropagated plants achieved a height of 25-30 cm with 3-4 tillers (shoots) with miniature rhizome in a 4-month period. Genetic stability was observed in the micropropagated plants.