Articles
EFFICIENT PLANT REGENERATION VIA SHOOT ORGANOGENESIS FROM EXPLANTS OF IN VITRO SEEDLINGS OF A RECALCITRANT WOODY SPECIES OF TEAK (TECTONA GRANDIS L.F.)
Article number
1083_4
Pages
53 – 60
Language
English
Abstract
In this study, adventitious shoot regeneration and rooting of shoots is reported from teak (Tectona grandis) seedlings.
A factorial experiment was conducted with 4 explants (cotyledon, leaf, node, internode), 2 culture conditions (light, dark), 2 basal media (MS, WPM), 5 levels each of Dicamba, 2, 4-Dichlorophenoxyacetic acid, α-naphthaleneacetic acid, Picloram (1, 4, 9, 13, 20 µM) and Thidiazuron (0.001, 0.1, 1, 4, 10 µM) for callus induction from explants derived from 4-week-old-seedlings grown on MS agar-solidified medium along with 0.1% activated charcoal without sucrose.
Callus induction varying in frequency was observed from all explant types at all TDZ levels on MS medium.
However, 100% callus induction was obtained with 1 or 4 µM TDZ from nodal explants in dark after 7 weeks of initial culture.
All NAA concentrations induced callus from nodal explants whereas 1 or 4 µM 2, 4-D was only effective for cotyledons.
The calluses were then shifted to MS medium containing N6-benzyleadenine (BA), Kinetin or TDZ (0.5, 1, 4, 6 or 8 µM) in combination with indole-3-acetic acid, indole-3-butyric acid or NAA (0.5 µM) resulting in 45 combinations for shoot regeneration.
Adventitious shoot regeneration (100%) with 42.25 shoots was obtained on MS medium supplemented with 8 µM TDZ after 17 weeks of culture.
The regenerating shoots were elongated (5.14 cm shoot length) on MS medium supplemented with BA (8 µM) + IBA (1 µM) for 30 days.
Such shoots were then rooted on half-strength MS medium with IBA + NAA (6+6, 8+8 or 10+10 µM). Rooting frequency of elongated shoots was 75.45% treated with 8+8 µM IBA and NAA with 1.65 mean number and 3.2 cm length after 35 days.
The plantlets were successfully acclimatized with 55% survival in soil.
A factorial experiment was conducted with 4 explants (cotyledon, leaf, node, internode), 2 culture conditions (light, dark), 2 basal media (MS, WPM), 5 levels each of Dicamba, 2, 4-Dichlorophenoxyacetic acid, α-naphthaleneacetic acid, Picloram (1, 4, 9, 13, 20 µM) and Thidiazuron (0.001, 0.1, 1, 4, 10 µM) for callus induction from explants derived from 4-week-old-seedlings grown on MS agar-solidified medium along with 0.1% activated charcoal without sucrose.
Callus induction varying in frequency was observed from all explant types at all TDZ levels on MS medium.
However, 100% callus induction was obtained with 1 or 4 µM TDZ from nodal explants in dark after 7 weeks of initial culture.
All NAA concentrations induced callus from nodal explants whereas 1 or 4 µM 2, 4-D was only effective for cotyledons.
The calluses were then shifted to MS medium containing N6-benzyleadenine (BA), Kinetin or TDZ (0.5, 1, 4, 6 or 8 µM) in combination with indole-3-acetic acid, indole-3-butyric acid or NAA (0.5 µM) resulting in 45 combinations for shoot regeneration.
Adventitious shoot regeneration (100%) with 42.25 shoots was obtained on MS medium supplemented with 8 µM TDZ after 17 weeks of culture.
The regenerating shoots were elongated (5.14 cm shoot length) on MS medium supplemented with BA (8 µM) + IBA (1 µM) for 30 days.
Such shoots were then rooted on half-strength MS medium with IBA + NAA (6+6, 8+8 or 10+10 µM). Rooting frequency of elongated shoots was 75.45% treated with 8+8 µM IBA and NAA with 1.65 mean number and 3.2 cm length after 35 days.
The plantlets were successfully acclimatized with 55% survival in soil.
Authors
M. Akram, F. Aftab
Keywords
callus, nodal explant, shoot organogenesis, Tectona grandis, TDZ, teak
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