High frequency in vitro propagation of <Emphasis Type="Italic">Trichopus zeylanicus</Emphasis> subsp. <Emphasis Type="Italic">travancoricus</Emphasis> using branch–petiole explants |
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Authors: | Kottackal Poulose Martin A K Pradeep Joseph Madassery |
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Institution: | (1) Department of Biotechnology, University of Calicut, Calicut, Kerala, 673 635, India;(2) Department of Botany, University of Calicut, Calicut, Kerala, 673 635, India;(3) Present address: Department of Plant Sciences, University of California, One Shields Ave., Davis, CA 95616, USA |
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Abstract: | Trichopus zeylanicus subsp. travancoricus (known as Arogyapacha), an endangered ethnomedicinal plant of the Western Ghats of South India, serves as the major source
of the commercial drug Jeevani. The present study established a long-term high frequency in vitro propagation protocol for Arogyapacha. Callus obtained from
the branch–petiole explants cultured on Murashige and Skoog (MS) medium with 4.5 μM 2,4-dichlorophenoxyacetic acid upon subculture
to medium with different concentrations of 6-benzyladenine (BA) either alone or in combination with an auxin favoured shoot
morphogenesis. Medium with 13.3 μM BA alone facilitated high frequency shoot bud (mean of 93.2) formation. Medium with lower
concentrations of BA (4.4, 6.6 and 8.8 μM) alone or in combination with lower concentration of α-naphthaleneacetic acid (NAA)
or indole-3-butyric acid (IBA) favoured better shoot growth than 13.3 μM BA containing medium, but with reduced number of
shoot buds. Subsequent cultures on medium with lower concentrations of BA and also on MS basal media facilitated shoot formation
as well as growth of shoots. The shoot regeneration potential showed no decline up to 5 years. Culture of the in vitro-derived
whole branch–leaf explants on MS basal medium developed shoots directly from the node. On medium with 19.6 μM IBA, the whole
branch–leaf explants induced nodular callus from the node, which developed shoots later. Subsequent cultures on medium with
BA exhibited high frequency shoot formation. The transfer of shoots after 10–15 days culture on half-strength MS medium containing
2.7 μM NAA to half-strength basal medium induced a mean of 11.3 roots. Field survival of plantlets relied on the soil mix:
a 1:4 ratio of sand and red-soil exhibited the highest plantlets survival (86.6%). RAPD profile of the source plant and plants
regenerated from calli after 4 years showed no polymorphism. The established plantlets with morpho-floral features similar
to that of the source plants flowered normally and set fruits. |
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