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Bioproduction and optimization of rosmarinic acid production in Solenostemon scutellarioides through media manipulation and conservation of high yielding clone via encapsulation.

Natural product communications (2013-11-28)
Ranabir Sahu, Saikat Dewanjee, Moumita Gangopadhyay
ABSTRACT

The present study describes the role of different exogenous phytohormones, polyamines and sucrose on growth and rosmarinic acid (RA) production in whole plant culture of Solenostemon scutellarioides. It was further aimed to conserve elite clones via synthetic seed technology. S. scutellarioides was treated either singly or in combination with different phytohormones. Cultures incubated with NAA (0.5 mg L(-1)) yielded the highest RA accumulation (g(-1FW)), but negatively affected the growth. So, overall RA content was insignificant. Cultures incubated with IBA, BAP and GA3 at low concentration significantly improved growth and RA bioaccumulation. In the combinatorial study, IBA+BAP+GA3 (0.5 mg L(-1) each) was found optimum for plant biomass and RA production (65.2% improvement of total RA). Amongst polyamines, putrescine (1 mg L(-1)) exhibited 20.4% improvement of total RA content. The intracellular RA accumulation (g(-1FW)) was significantly higher between 5 and 7% of sucrose concentrations. However, the total increase in RA content was inhibited due to deterioration of the culture with increasing sucrose concentration. Based on the effect of different treatments on growth and RA accumulation, a high yielding and stable plant line was selected for conservation via alginate encapsulation. Uniform shaped alginate coated synthetic seeds conserved up to 6 months exhibited high regeneration potential and RA content.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Sucrose, for molecular biology, ≥99.5% (GC)
Sigma-Aldrich
Sucrose, ≥99.5% (GC)
Sigma-Aldrich
Sucrose, BioXtra, ≥99.5% (GC)
Sigma-Aldrich
Sucrose, BioUltra, for molecular biology, ≥99.5% (HPLC)
Sigma-Aldrich
Sucrose, ≥99.5%
Sigma-Aldrich
Sucrose, puriss., meets analytical specification of Ph. Eur., BP, NF
Supelco
Sucrose, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Sucrose, ACS reagent
Sigma-Aldrich
Sucrose, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99.5% (GC)
Millipore
Sucrose, ACS reagent, suitable for microbiology, ≥99.0%
Sigma-Aldrich
Rosmarinic acid, ≥98% (HPLC), from Rosemarinus officinalis L.
Supelco
Sucrose, analytical standard
Sigma-Aldrich
Rosmarinic acid, 96%
Sigma-Aldrich
Sucrose, Grade II, suitable for plant cell culture
Sigma-Aldrich
Sucrose, meets USP testing specifications
Sucrose, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Sucrose, Grade I, suitable for plant cell culture
Supelco
Sucrose, analytical standard, for enzymatic assay kit SCA20
Rosmarinic acid, primary reference standard