A two-step procedure for adventitious shoot regeneration on excised leaves of lowbush blueberry |
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Authors: | S C Debnath |
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Institution: | (1) Atlantic Cool Climate Crop Research Centre, Agriculture and Agri-Food Canada, P.O. Box 39088, 308 Brookfield Road, St. John’s, Newfoundland and Labrador, Canada, A1E 5Y7 |
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Abstract: | An efficient system to regenerate shoots on excised leaves of greenhouse-grown wild lowbush blueberry (Vaccinium angustifolium Ait.) was developed in vitro. The effect of thidiazuron (TDZ) on adventitious bud and shoot formation from apical, medial, and basal segments of the leaves
was tested. Leaf cultures produced multiple buds and shoots with or without an intermediary callus phase on 2.3–4.5 μM TDZ
within 6 wk of culture initiation. The greatest shoot regeneration came from young expanding basal leaf segments positioned
with the adaxial side touching the culture medium and maintained for 2 wk in darkness. Callus development and shoot regeneration
depended not only on the polarity of the explants but also on the genotype of the clone that supplied the explant material.
TDZ-initiated cultures were transferred to medium containing 2.3–4.6 μM zeatin and produced usable shoots after one additional
subculture. Elongated shoots were dipped in 39.4 mM indole-3-butyric acid powder and planted on a peat:perlite soilless medium
at a ratio of 3:2 (v/v), which yielded an 80–90% rooting efficiency. The plantlets were acclimatized and eventually established in the greenhouse
with 75–85% survival. |
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Keywords: | Organogenesis Vaccinium angustifolium Cytokinins In vitro culture Ex vitro rooting |
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