TITLE:
In Vitro Plant Regeneration of Dendrocalamus stocksii (Munro) M. Kumar, Remesh & Unnikrisnan, through Somatic Embryogenesis
AUTHORS:
P. V. Somashekar, T. S. Rathore, Tanzeem Fatima
KEYWORDS:
Dendrocalamus stocksii, Somatic Embryogenesis, 2, 4 D, BAP, Kinetin, NAA
JOURNAL NAME:
American Journal of Plant Sciences,
Vol.9 No.12,
November
26,
2018
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.