<i>In Vitro</i> Plant Regeneration of <i>Dendrocalamus stocksii</i> (Munro) M. Kumar, Remesh & Unnikrisnan, through Somatic Embryogenesis

Abstract

Dendrocalamus stocksii is fast cultivating economically important forest crop species. National Mission of Bamboo Application (NMBA) of India has been identified in 15 industrially important bamboo species. Traditionally it was propagated through by offset cuttings and rhizome splitting which was not meeting the demand, culm cuttings needed mass material to propagate and rooting percentage mixed. Plant regeneration through somatic embryogenesis was achieved in callus cultures derived from the callus initiated through type of explants viz. leaf, leaf sheath, shoot tip, nodal shoot segments, and inter node segments from aseptic cultures. Explants were cultured on Murashige & Skoog basal media supplemented with 2,4 Dichloro diphenyle ethane 0.44 μM/L with additives (Ascorbic acid 8.8 μM/L, citric acid 4.8 μM/L Cysteine 3.02 μM/L and Glutamine 14.6 μM/L) with 3% sucrose and Agar agar 0.6%. Cultures were incubated in the dark at 25°C ± 1°C. Out of five types of explants nodal shoot induced callus > 80% followed by leaf sheath (60%) and no callus was induced in leaf. Various nutrient media viz. Murashige and Skoog (MS), Woody Plant Media (WP), Gamborg media (B5) and Heller’s (HE) media fortified with 2,4 D (0.2 - 1.10 μM/L) and Kinetin 0.10 μM/L were tested for high frequency callus induction. Among four nutrient media tested MS media fortified with 2,4 D (0.55 - 1.1 μM/L) 100% callus induction. Calli multiplication was carried out with various concentrations of PGR’s with 10% coconut milk. Out of these MS media 2,4 D 0.55 μM /L and 10% coconut milk concentration were found best for high frequency (80%) calli multiplication. Various combinations of α-naphthalene acetic acid (NAA) with N6-benzyiaminopurine (BAP) and kinetin were tested for embryo germination, out of which MS media supplemented with NAA 0.55 μM /L and BAP 0.22 μM /L were showed high frequency (80%) germination. Germinated plantlets carefully transferred to polybags containing potting mixture of sand soil and compost in the ratio of 40:10:50 with 10 Kg/m3 + 250 gm/m3 fungicide. Plantlets were kept 4 weeks under poly tunnel inside mist chamber followed by two weeks outside poly tunnel in mist chamber. Plants are lifted to the canopy condition directed to a week before subjected to them in the institute division nursery.