Abstract

BackgroundSomatic embryogenesis is one of the most popular in vitro regeneration methods for mass micropropagation. In the present study, somatic embryogenesis via zygotic embryos was studied in Asparagus racemosus Willd. Since the callus quality plays an important role in plantlet development, therefore, compact embryogenic callus was selected for further embryogenesis.ResultsSomatic embryos were induced by zygotic embryos germinating on callus induction medium (MS media with 1.54 mg L−1 2,4-D and 0.43 mg L−1 kinetin) in dark. Thereafter, the compact embryogenic callus differentiated up on MS media with 0.1 mg L−1 NAA and 0.5 mg L−1 kinetin supplemented with various concentrations of ancymidol. An addition of 0.75 mg L−1 ancymidol resulted in significant enhancement of somatic embryo formation and no malformed embryos were observed. Scanning electron micrographs and thin sections confirmed the structures as somatic embryos. Furthermore, these embryos were transferred to the same medium supplemented with glutamine, casein hydrolysate, and 3% sucrose for conversion into green shoots. These shoots could be multiplied in vitro using BAP-supplemented medium.ConclusionsAn effective method for conservation of an overexploited threatened medicinally important species Asparagus racemosus has been developed. This is the first report of the formation of somatic embryos from zygotic embryos in A. racemosus. Ancymidol in combination with kinetin and NAA was found to be most efficient for somatic embryo maturation and germination. The established protocol would certainly advance the efficient somatic embryo induction, maturation, and germination which could be utilized for large-scale propagation of Asparagus species.

Highlights

  • Asparagus racemosus Willd. is an important medicinal plant native to the Indian subcontinent

  • We report a method for induction of embryogenic callus and differentiation of somatic embryos from zygotic embryos

  • Developed somatic embryos from the somatic embryo induction (SEI) medium were transferred for germination to Murashige and Skoog (MS) with 0.1 mg L−1 αnaphthaleneacetic acid (NAA), 0.5 mg L−1 kinetin, and 0.75 mg L−1 ancymidol supplemented with either (i) 600 mg L−1 glutamine and 400 mg L−1 casein hydrolysate, or (ii) 800 mg L−1 glutamine and 500 mg L−1 casein hydrolysate

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Summary

Introduction

Asparagus racemosus Willd. (common name Shatavar; Asparagaceae) is an important medicinal plant native to the Indian subcontinent. The tuberous roots of A. racemosus are rich in the saponins, Shatavarin I to IV having rejuvenating and phytoestrogenic properties. These saponins are extensively used in hormone replacement therapy instead of synthetic estrogens that are neither safe nor effective (Barrett-Connor, 1998). A. racemosus roots are extensively collected from the wild causing mass destruction of the germplasm. This overexploitation has put considerable survival pressure on this plant making it endangered in the region of its natural existence (Kala et al 2006; Chaudhary and Dantu, 2011). Somatic embryogenesis via zygotic embryos was studied in Asparagus racemosus Willd. Since the callus quality plays an important role in plantlet development, compact embryogenic callus was selected for further embryogenesis

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