Abstract
This work presents a rapid and reliable micropropagation method for a Lycaste hybrid using a field-grown axillary bud culture system. Intact buds (2–4 mm in length) were excised from a mature pseudobulb and were cultured in half-strength MS basal medium, which was supplemented with 0.5 mg l−1 benzyladenine (BA), 1.0 mg l−1 thidiazuron (TDZ) and 2% (w/v) sucrose. After 2 months, the calli exhibited vigorous growth and eventually turned green, forming protocorm-like bodies (PLBs) originating in the surface of each callus. The results of this work reveal that the combination of 0.5 mg l−1 BA and 1.0 mg l−1 TDZ treatments was highly effective in indirectly multiplying shoots from callus-PLB mixed explants, which yielded up to 400 shoots in the fourth time subcultures (within 24 weeks). Histological observations showed the apical meristem of adventitious bud is based on a longitudinal section of a callus sample. Histological and scanning electronic microscopy also indicated that PLBs derived from calli could be regarded as organogenesis but not somatic embryogenesis. Shoots with a length of around 2–3 cm generated in vitro were excised and cultured in MS medium supplemented with 0.5 mg l−1 IBA exhibited the best rooting response (78.3%), and an average of 1.8 roots per explant was produced within 4 weeks.
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Abbreviations
- BA:
-
Benzyladenine
- TDZ:
-
Thidiazuron
- PLBs:
-
Protocorm-like bodies
- MS:
-
Murashige and Skoog medium
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Acknowledgment
We thank the Agriculture and Food Agency, Council of Agriculture, Executive Yuan, Taiwan (ROC) for financially supporting this research under Contract No. 95AS-6.1.3-FD-Z2.
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Huang, CH., Chung, JP. Efficient indirect induction of protocorm-like bodies and shoot proliferation using field-grown axillary buds of a Lycaste hybrid. Plant Cell Tiss Organ Cult 106, 31–38 (2011). https://doi.org/10.1007/s11240-010-9890-6
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DOI: https://doi.org/10.1007/s11240-010-9890-6