Expression of a KDEL-tagged dengue virus protein in cell suspension cultures of <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> and <Emphasis Type="Italic">Morinda citrifolia</Emphasis> |
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Authors: | C A Martínez A M Giulietti J Rodríguez Talou |
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Institution: | (1) NRC-Plant Biotechnology Institute, 110 Gymnasium Place, Saskatoon, SK, S7NOW9, Canada;(2) Genome Prairie, 101-111 Research Drive, Saskatoon, SK, S7N3R2, Canada |
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Abstract: | Hypericum perforatum L. (St. John’s wort) produces a number of phytochemicals having medicinal, anti-microbial, anti-viral and anti-oxidative
properties. Plant extracts are generally used for treatment of mild to medium cases of depression. Plant regeneration can
be achieved in this species by in vitro culture of a variety of explants. However, there are no reports of regeneration from
petal explants. In this report plant regeneration from petal explants of St. John’s wort was evaluated. Petals of various
ages were cultured on agarized Murashige and Skoog 1962 (MS) medium supplemented with auxin and cytokinin (kinetin), maintained in the dark and callus and shoot regeneration determined
after 28 days. At an auxin to cytokinin ratio of 10:1, callus and shoot formation were induced by all levels of indole-3-acetic
acid (IAA), indole-3-butyric acid (IBA) and 1-naphthaleneacetic acid (NAA), while 2,4-dichlorophenoxyacetic acid (2,4-D) induced
only callus formation. The optimum level of auxin for shoot regeneration was 1.0 and 0.1 mg/l kinetin, where the regeneration
frequency was 100 percent for all three auxins. The highest number of shoots per explant (57.4 and 53.4) was obtained with
IAA and IBA, respectively. In the absence of auxin, kinetin levels of 0.1 and 0.25 mg/l induce callus and shoot formation
at low frequency but not at lower levels. Callus and shoot formation did not occur in the absence of growth regulators. Petal-derived
shoots were successfully rooted on half-strength MS medium without a requirement for exogenous auxin and flowering plants
were established under greenhouse conditions. From these results it can be concluded that auxin type is a critical factor
for plant regeneration from petal explants of Hypericum perforatum and there is no absolute requirement for high levels of cytokinin. |
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