Journal of Nanomedicine & Nanotechnology

High frequency plant regeneration and enhanced production of shikonin from hairy root cultures in Arnebia hispidissima

Joint Event on 23^rd International Conference on Nanomaterials science & Nanoengineering & Technology & International Conference and Exhibition on Pharmaceutical Nanotechnology and Nanomedicine

April 18-19, 2018 | Las Vegas, USA

Minakshi Pal, Ashok Chaudhury and Ashok k Dhawan

Guru Jambheshwar University of Science and Technology, India
CCSHAU, India

Accepted Abstracts: J Nanomed Nanotechnol

Abstract:

Arnebia belonging to family Boraginaceae, a group of hispid herbs, is a source of shikonin, a naphthoquinone red pigment. A rapid and efficient protocol for micro propagation from such as shoot tip, nodal, leaf and internodal tissue has been developed. Highest plant regeneration was observed from shoot tip (93±0.00%) and nodal segment (60.0±0.13%) cultured on MS media supplemented with plant growth regulators (KIN 0.5mgl-1, BAP 0.25mgl-1, IAA 0.1mgl-1) and CH (100mgl-1). Proliferating shoots were efficiently rooted on MS medium supplemented with 2.0mgl-1 IBA upon transfer to soil. Highest response for induction of callus for shikonin production of 80.0% from nodal segments was observed on media combination MS+mgl-1 BAP 0.25+1.0mgl-1 IAA. UV-visible spectroscopic analysis of fresh callus showed induction and accumulation of 0.50mgg-1 of shikonin content at the end of the 50 days of culture. Additionally, a method for Agrobacterium rhizogenes-mediated genetic transformation of Arnebia hispidissima for hairy root cultures was also optimized for enhancing the shikonin production to meet its ever-increasing demand in pharmaceutical industry. Among the various tissues employed, leaf explants showed maximum (70.7%) response followed by shoot tips (52%), nodal segments (38.7%) and internodal segments (9.3%). The presence of Ri plasmid rolB gene in the transformed hairy root cultures was confirmed by PCR analysis using forward (FrolB) and reverse (RrolB) primers of rolB gene resulting in the amplification of ~0.8kb fragments. Medium composition has been optimized for in vitro induction of shikonin in hairy root cultures of Arnebia hispidissima. The shikonin content in transformed hairy root was estimated spectrophotometrically using authentic sample of shikonin. The total content of shikonin was 0.85mgg-1 fresh weight of tissue at the end of day 50 of the culture period. These results will help to design strategies for bridging the gap between ever increasing demand and supply of raw products necessary for obtaining shikonin for cosmetic, dyeing, food, medicinal, and pharmaceutical industries. minakshiarnav@gmail.com