Cryopreservation studies in Callus culture of Arnebia euchroma
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Date
2022-05-09
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palampur
Abstract
Arnebia euchroma commonly known as Ratanjot belongs to Boraginaceae family. Arnebia species
is known for rich source of shikonin and their derivatives have antibacterial, antifungal, anti-HIV,
anti-inflammatory and several other medicinal properties. The genus Arnebia has been over exploited as a result of these characteristics, and it is now classified as a endangered species. These
resources must be protected in order to be used on a large scale in the future, and this valuable
treasure must be preserved. Callus was induced from the leaves with (75%) induction when
cultured on MS medium augmented with IBA (1.02 mg/l) and BAP (2.25 mg/l) and multiplied on
same medium. Leaf derived callus were used for the cryopreservation Therefore, a protocol is
optimized for cryopreservation of callus using vitrification, encapsulation vitrification and
desiccation technique. Callus was precultured on sucrose enriched medium containing 3% (w/v)
and 13.7% (w/v) sucrose. Among all these techniques of cryopreservation highest survival rate
(65%) was found in desiccation when precultured on 13.7% sucrose followed by 90 minutes
desiccation time. Callus used for the cryopreservation was not revived after cryogenic treatment
and necrosed after eight weeks of culture in encapsulation vitrification method. Hence, this method
of cryopreservation was not found suitable for preserving the callus tissues of A. euchroma. Non
cryopreserved callus showed the highest total Deoxyshikonin content i.e. 369.28 (µg/gm) and
301.31 (µg/gm) shikonin content was observed and Cryopreserved callus showed the highest
Deoxyshikonin content i.e. 319.23 (µg/gm) and total shikonin content of 296 (µg/gm). In UPLC
chromatography only deoxyshikonin content i.e. 65.05 (µg/gm) was observed in non cryopreserved
callus and 39.07(µg/gm) content was found in cryopreserved callus.