Articles
Shoot induction from node and basal tissue from an elongated stem of Paphiopedilum ‘Delrosi’
Article number
1262_22
Pages
167 – 172
Language
English
Abstract
Micropropagation of ladys slipper orchids is difficult and has been tested using various parts of the plant.
This experiment was aimed to culture Paphiopedilum Delrosi from the node and basal tissue via in vitro propagation.
The shoot was cultured on modified Hyponex medium in the dark for 16 weeks.
After removing the elongated shoot from a plant with 2 leaves, the node or basal tissue of the stem were used to develop as an explant.
The explant was individually cultured for 16 weeks on modified Hyponex medium supplemented with 0.45 µM thidiazuron (TDZ) alone or in combination with 4.52 µM 2,4-D or with 4.65 µM kinetin (Kn) in combination with 4.52 µM 2,4-D and containing 20 g L‑1 of sucrose, glucose or maltose.
For growth of nodes, medium containing 20 g L‑1 sucrose with or without 4.65 µM Kn and 4.52 µM 2,4-D gave the best results, with 80-90% survival of explants, 90-100% new shoot formation with 1 new shoot and 374.80-419.22 mg total fresh weight.
For growth of basal tissue, there was no significant difference among the percentage survival of explants (80-100%), new shoot induction (60-100%), number of new shoots (0.9-1.6) and number of leaves (3.5-4.1) obtained from media containing the combination of growth regulator and carbon source.
However, modified Hyponex medium containing 20 g L‑1 maltose with or without 0.45 µM TDZ provided 100% new shoot formation, with 1.5-1.6 new shoots.
When comparing the node and basal tissue, the results showed that both parts were able to regenerate to the shoot stage.
However, the shoot induction rate was low.
This experiment was aimed to culture Paphiopedilum Delrosi from the node and basal tissue via in vitro propagation.
The shoot was cultured on modified Hyponex medium in the dark for 16 weeks.
After removing the elongated shoot from a plant with 2 leaves, the node or basal tissue of the stem were used to develop as an explant.
The explant was individually cultured for 16 weeks on modified Hyponex medium supplemented with 0.45 µM thidiazuron (TDZ) alone or in combination with 4.52 µM 2,4-D or with 4.65 µM kinetin (Kn) in combination with 4.52 µM 2,4-D and containing 20 g L‑1 of sucrose, glucose or maltose.
For growth of nodes, medium containing 20 g L‑1 sucrose with or without 4.65 µM Kn and 4.52 µM 2,4-D gave the best results, with 80-90% survival of explants, 90-100% new shoot formation with 1 new shoot and 374.80-419.22 mg total fresh weight.
For growth of basal tissue, there was no significant difference among the percentage survival of explants (80-100%), new shoot induction (60-100%), number of new shoots (0.9-1.6) and number of leaves (3.5-4.1) obtained from media containing the combination of growth regulator and carbon source.
However, modified Hyponex medium containing 20 g L‑1 maltose with or without 0.45 µM TDZ provided 100% new shoot formation, with 1.5-1.6 new shoots.
When comparing the node and basal tissue, the results showed that both parts were able to regenerate to the shoot stage.
However, the shoot induction rate was low.
Publication
Authors
K. Obsuwan, C. Thepsithar, A. Thongpakdee, E. Nisayan
Keywords
lady’s slipper orchids, etiolated stem without shoot, carbon source, TDZ, Kn, 2,4-D
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