Abstract
Oxymatrine is one of the most important biologically active compounds and is present in Sophora alopecuroides L. The present investigation focuses on the development of an efficient tissue culture method to induce callus and cell suspension culture of S. alopecuroides by studying the effect of jasmonic acid and nitric oxide on cell suspension culture. Callus induction efficiency is high in axenic leaf explants grown in MS medium supplemented with 1.0 mg/L Kinetin (Kin), 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D). The cell suspension culture was developed using the same callus induction medium without agar. The maximum cell number and dry weight of suspension culture were obtained by the 9th day of incubation. The synthesis of oxymatrine is higher in jasmonic acid and nitric oxide (200 µMJA and 50 µMNO) combination (11.91 µg/g) when compared to the non-elicited control (8.3 µg/g) of callus.
Highlights
Sophora is a perennial herb belonging to the Fabaceae family and is used in Mongolian traditional medicine to treat heart disease, bacterial infection, fever, and rheumatism [1]
The production of secondary metabolites is limited in the case of cell culture whereas the biosynthesis of the secondary metabolites in plants is due to its stress factors and its response [7]
No callus induction was found on MS medium basal MS medium supplemented with either kinetin or 2,4-D alone .with Friable yellowish calli along with higher callus growth (75-100%) was observed in MS solid medium provided with 0.5 mg/LKin, 1 mg/L 2,4-D and 1.0mg/LKin, 1 mg/L 2,4-D (Table 1)
Summary
Sophora is a perennial herb belonging to the Fabaceae family and is used in Mongolian traditional medicine to treat heart disease, bacterial infection, fever, and rheumatism [1]. Properties of callus tissue are one of the important factors to establish cell cultures. The production of secondary metabolites is limited in the case of cell culture whereas the biosynthesis of the secondary metabolites in plants is due to its stress factors and its response [7]. Synthesis and accumulation of secondary metabolites are due to the compounds called elicitors which are produced at the time of stress and enhance the accumulation of secondary metabolites [8]. Based on the previous study, the elicitors viz., Jasmonic acid (JA), and Nitric oxide (NO) are selected on the enhancing ability of the compound oxymatrine. The present investigation focuses on developing an efficient procedure for the establishment of callus and cell suspension cultures, to determine the oxymatrine production from these cultures with the influence of jasmonic acid and nitric oxide. Cell viability and culture biomass of S. alopecuroides is taken up in the present study
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