IN VITRO PROPAGATION AND SECONDARY METABOLITE PRODUCTION IN Podophyllum hexandrum ROYLE (INDIAN MAYAPPLE)
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Date
2021-12
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UHF,NAUNI
Abstract
ABSTRACT
The present study reports an optimized protocol for high frequency in vitro propagation
through seeds and rhizome buds, molecular analysis and podophyllotoxin production in Podophyllum
hexandrum. Maximum percent uncontamination of seeds (89%) was achieved on treatment with 0.1%
HgCl2 for 3 minutes and highest percent surviving rhizome buds (82.62%) were obtained after surface
sterilization with 0.1% HgCl2+1.0% NaOCl for 4 minutes with maximum in vitro establishment
(95.97%) on MS medium fortified with 0.2mg/l BA and 0.1mg/l GA3. Maximum in vitro
establishment through seeds (89.00%) and buds (90%) was achieved during spring, followed by 68
and 79% in winter season. The proliferated shoots from seeds showed highest multiplication (1:11) on
MS medium fortified with 2.0mg/l BA + 0.2mg/l NAA+ 0.7mg/l CaCl2. Addition of CaCl2 (0.7mg/l)
into the medium was done to prevent shoot chlorosis. For shoot regenerating from rhizome buds
highest multiplication rate of 1:5 was achieved on MS medium fortified with 2.0mg/l BA
+0.2mg/l NAA. Molecular analysis was done using SCoT and CBDP markers. SCoT markers showed
71% polymorphism in samples from different altitudes in comparison to 50.90% polymorphism
depicted by CBDP markers. However, both the makers showed 100% monomorphism among mother
plants and their tissue culture raised progeny. In vivo leaves and petioles as well as in vitro roots and
leaves were used as explants for callus induction. Highest callus induction was observed under dark
incubation in in vivo leaves (81.07%), petioles (87.19%), in vitro roots (90.67%) and leaves
(89.20%).HPLC analysis revealed maximum (0.267%) production of podophyllotoxin from callus
initiated from in vitro roots procured from experimental material of district Kinnaur followed by
0.258% from in vitro root callus of Lahaul – Spiti. For enhancing podophyllotoxin yield callus
induced from in vitro roots of P. hexandrum plants of district Kullu were subjected to elicitation by
incorporating different concentrations (0.5-1.5 mM) of elicitors (methyl jasmonate and chitin) in
production medium. Highest podophyllotoxin content (0.33%) was observed after elicitation with
1mM methyl jasmonate followed by its higher concentration of 1.5 mM (0.31%).