Somatic embryogenesis and plantlet regeneration in American ginseng (Panax quinquefolium L.) |
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Authors: | A Tirajoh T S Kyung Z K Punja |
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Institution: | (1) Department of Biological Sciences, Simon Fraser University, V5A 1S6 Burnaby, British Columbia, Canada |
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Abstract: | Summary Somatic embryogenesis in American ginseng (Panax quinquefolium L.) was investigated from three explant sources (root, leaf and epicotyl) with Murashige and Skoog (MS) medium containing
different growth regulators. Mature roots and leaves obtained from 3- to 5-yr-old field-grown plants, and seedling leaves
and epicotyls from plantlets grownin vitro, were evaluated. From root and epicotyl explants, callus development was optimal with 3,6-dichloro-o-anisic acid (dicamba)
(9.0 μM) and kinetin (KN) (5.0 μM) as the growth regulators. When these calluses were transferred after 3 mo. to dicamba alone (9.0 μM), somatic embryo formation was observed at an average frequency of 15.6% in root explants after an additional 3 mo., and
2% in epicotyl explants after an additional 6 mo. No plantlets were recovered because the embryos germinated to form shoots
with no roots. From leaf explants, callus growth was optimal with α-naphthaleneacetic acid (NAA) at 10.0 μM and 2,4-dichlorophenoxyacetic acid (2,4-D) at 9.0 μM. Somatic embryos developed on this medium, with the highest frequency (40%) obtained after 3 mo. from seedling-leaf explants.
Calluses on mature leaves formed somatic embryos after 7 mo. with NAA/2,4-D at an average frequency of 30%. Transfer of these
somatic embryos to 6-benzyladenine/gibberellic acid (4.4/2.9 μM) promoted shoot development but no roots were observed. Up to 100% of germination was observed within 6 wk on half-strength
MS salts containing activated charcoal (1%) and on NAA/2,4-D (5.0/4.5 μM) with charcoal (1%). On the latter medium, somatic embryos enlarged and frequently gave rise to new somatic embryos after
a brief callusing phase. The embryos germinated through a two-stage process, involving the elongation of the root followed
by the formation of a shoot. The highest recovery of ginseng plantlets from germinated embryos was 61.0%. Following transfer
to potting medium and maintenance under conditions of high humidity and low light intensity, the plantlets elongated and developed
new leaves. A high percentage (50%) of these plants have been acclimatized to soil. |
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Keywords: | medicinal plant propagation tissue culture |
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