STUDIES ON CRYOPRESERVATION AND REGENERATION OF IN VITRO RAISED CULTURES OF PICRORHIZA KURROA
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Date
2020-09-28
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CSKHPKV. Palampur
Abstract
Picrorhiza kurroa is an endangered Himalayan perennial herb and important ingredient of
traditional medicinal systems of Asian region. It’s medicinal value primarily depends upon
presence of picrosides in its roots. These bioactives are used for preparation of hepatoprotective
phytopharmaceuticals. This herb is uprooted from wild for extraction of bioactive ingredients,
which is a major reason for its depletion from natural habitat. Considering the high demand and
low supply of P. kurroa population, efforts for its conservation through biotechnological tools
is urgently required. In current research, cryopreservation protocol of high picroside producing
cell lines of Picrorhiza kurroa were developed. P. kurroa shoot cultures incepted with 0.50
mg/L of Kn. Highest in vitro average shoot number was observed at 0.50 mg/L Kn strengthened
MS medium with 3.18 shoots per shoot tip whereas 4.29 cm shoot length with 11.46 average
number of leaves recorded on MS basal medium. Highest callus induction (100%) was found
on MS medium augmented with 0.0125 mg/L IBA + 0.125 mg/L TDZ from leaf and root
explants. Calli multiplied on the same medium with 6.89 fold increase in fresh weight. Leaf and
root explants of in vivo and in vitro grown plants were analysed for identification of high
metabolite producing cell lines. Among in vivo leaf and root explants, highest P-I content (5.65
%) was found in leaves whereas highest P-II content (8.37 %) was found in roots. In case of in
vitro leaves and roots, leaves contained higher P-I (1.160 %) & P-II (0.280 %) content. Callus
derived from leaves also showed highest total picroside content i.e 0.2 per cent, therefore, leaf
derived callus was used for cryopreservation. Among all the cryopreservation methods used i.e
encapsulation-vitrification, vitrification and desiccation, only encapsulation-vitrification
showed 60 per cent regeneration at 15 minutes of dehydration time with PVS2 solution.
Encapsulation-vitrification protocol would be useful for ex-situ conservation of P. kurroa callus
cultures.