Efficient plant regeneration from embryogenic cell suspension cultures of Euonymus alatus

Hyun-A Woo,Hyeran Kim,Seong Sub Ku,Won-Joong Jeong,Sung Ran Min,Hye Sun Cho,Hyun-Soon Kim,Eun Yee Jie,Sang Un Park,Suk Weon Kim

doi:10.1038/s41598-021-94597-4

Abstract

To establish an efficient plant regeneration system from cell suspension cultures of Euonymus alatus, embryogenic callus formation from immature embryos was investigated. The highest frequency of embryogenic callus formation reached 50% when the immature zygotic embryos were incubated on Murashige and Skoog (MS) medium supplemented with 1 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D). At higher concentrations of 2,4-D (over 2 mg/L), the frequency of embryogenic callus formation declined significantly. The total number of somatic embryos development was highest with the 3% (w/v) sucrose treatment, which was found to be the optimal concentration for somatic embryo formation. Activated charcoal (AC) and 6-benzyladenine (BA) significantly increased the frequency of plantlet conversion from somatic embryos, but gibberellic acid (GA3) had a negative effect on plantlet conversion and subsequent development from somatic embryos. Even though the cell suspension cultures were maintained for more than 1 year, cell aggregates from embryogenic cell suspension cultures were successfully converted into normal somatic embryos with two cotyledons. To our knowledge, this is the first successful report of a plant regeneration system of E. alatus via somatic embryogenesis. Thus, the embryogenic cell line and plant regeneration system established in this study can be applied to mass proliferation and production of pharmaceutical metabolite in E. alatus.

Highlights

To establish an efficient plant regeneration system from cell suspension cultures of Euonymus alatus, embryogenic callus formation from immature embryos was investigated
After 6 to 16 weeks of culture, the highest frequency of embryogenic callus formation reached 50% when the immature zygotic embryos were incubated on Murashige and Skoog (MS) medium supplemented with 1 mg/L 2,4-D
Under a lower concentration of 2,4-D, the frequency of embryogenic callus formation from immature embryo cultures was increased in a dosedependent manner

Summary

Introduction

To establish an efficient plant regeneration system from cell suspension cultures of Euonymus alatus, embryogenic callus formation from immature embryos was investigated. Even though the cell suspension cultures were maintained for more than 1 year, cell aggregates from embryogenic cell suspension cultures were successfully converted into normal somatic embryos with two cotyledons To our knowledge, this is the first successful report of a plant regeneration system of E. alatus via somatic embryogenesis. Plant cell and tissue culture can be utilized as a new means of producing high-value recombinant proteins or pharmaceutical metabolites[14,15,16] To achieve this goal, a plant cell culture system, an efficient plant regeneration system via organogenesis and somatic embryogenesis, and in vitro mass proliferation system must be established.

Methods

Results

Conclusion