Abstract

In this study, calli of Medicago sativa L. cv. Elçi (alfalfa Elçi) were inoculated in cell suspension culture and analyzed for aggregate assay, cell viability test, total phenolic content assay, DPPH free radical scavenging activity and formononetin assay by means of High-Performance Liquid Chromatography (HPLC). Hypocotyl, cotyledon and apical meristem explants were taken from 15-day-old aseptic seedlings and germinated in MS medium. 10 g calli were grown for each explant and then transferred into cell suspension culture. The highest cell viability rate, which was 75%, and the highest DPPH free radical scavenging activity with 51.36% was measured in 1000 mL cell suspension culture, while the highest total phenolic content, i.e. 40.2 mg/g, was quantified in 250 mL cell suspension culture. In accordance with the findings of the study, the production of formononetin was higher in the calli derived from cell suspension cultures than in herb samples of M. sativa. Moreover, in 1000 mL cell suspension culture, 4.99 mg/g of formononetin concentration was quantified, which scored the highest. In large-scale cell suspension cultures of M. sativa, it was possible to increase the production of formononetin production. Hence, due to its medicinal significance, a method has been tested to obtain higher amounts of this compound.

Highlights

  • Medicago sativa L. is an important feed plant in all over the world with its ability to adapt to different climates and its high feed efficiency and quality

  • Elçi were inoculated in cell suspension culture and analyzed for aggregate assay, cell viability test, total phenolic content assay, DPPH free radical scavenging activity and formononetin assay by means of High-Performance Liquid Chromatography (HPLC)

  • It was aimed to inoculate M. sativa calli in cell suspension culture. It was analyzed for aggregate assay, cell viability test, total phenolic content assay, DPPH free radical scavenging activity and formononetin assay by means of HPLC

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Summary

INTRODUCTION

Medicago sativa L. (alfalfa) is an important feed plant in all over the world with its ability to adapt to different climates and its high feed efficiency and quality. It was aimed to inoculate M. sativa calli in cell suspension culture. It was analyzed for aggregate assay, cell viability test, total phenolic content assay, DPPH free radical scavenging activity and formononetin assay by means of HPLC. Commercial sodium hypochlorite solution for 5 minutes They were rinsed 3 times in autoclaved distilled water and germinated in hormone-free Murashige and Skoog (MS) medium (Murashige & Skoog, 1962). Agar-free MS medium + 1.5 mg/L Kinetin, 1.5 mg/L NAA, 0.7 mg/L 2,4 D, 20 g/L sucrose were used for cell suspension cultures. The calli that was transferred to the cell suspension cultures were shaken at 180 rpm on the shaker for 20 days.

HPLC-UV Analyses
Statistical Analyses
RESULTS and DISCUSSION
Aggregate Measurement Results
Total Phenolic Compound Measurement Results
HPLC-UV Measurement Results
CONCLUSION
Declaration of Conflicting Interests and Ethics
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