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Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 117, Issue 3, pp 323–334 | Cite as

Plant regeneration from protoplasts in Indian local Coriandrum sativum L.: scanning electron microscopy and histological evidences for somatic embryogenesis

  • A. MujibEmail author
  • Dipti Tonk
  • Muzamil Ali
Original Paper

Abstract

Coriandrum sativum L. is an annual herb belonging to the family Umbelliferae. It is used as a spice plant in Indian subcontinent and it has several medicinal applications as well. In this present article, an efficient plant regeneration protocol from protoplasts via somatic embryogenesis was established and is reported. This is the first ever protoplast isolation study in Indian local coriander in which plant regeneration was achieved. Hypocotyl-derived embryogenic callus was used as a source of protoplast. The embryogenic callus suspension was prepared by transferring tissues onto rotary-agitated liquid Murashige and Skoog, added with 1.0 mg l−1 2,4-Dichlorophenoxyacetic acid (2,4-D) and 1.0 mg l−1 KIN (6-furfurylaminopurine). The suspension was digested with enzymatic solutions and a combination of cellulase (2.0 %), pectinase (1.0 %), macerozyme (0.02 %) and driselase (0.50 %) induced maximum yield of protoplasts (34.25 × 105). In 1.0 mg l−1 2,4-D + 1.0 mg l−1 KIN containing medium, protoplasts divided well and formed maximum number of microcolonies (14.30/test tube). The protoplast callus (PC) biomass grew well in solid medium. The protoplast embryogenic callus was rich in protein, proline and sugar compared to non-embryogenic PC. The protoplast originated callus later differentiated into somatic embryos. The somatic embryo morphology, scanning electron microscopy and histology of embryo origin and development were investigated and discussed in details in this present communication. In 1.0 mg l−1 2,4-D + 0.5 mg l−1 BA (6-Benzyladenine), maximum number of embryos were formed on microcallus (26.6/callus mass). The embryo matured and germinated into plantlets at a low to moderate rate, highest (31.3 %) embryo germination was observed in 1.0 mg l−1 BA + 0.5 mg l−1 α-Naphthalene acetic acid added medium. The entire process of regeneration took about 4–5 months’ time for recovering plantlets from protoplasts.

Keywords

Coriandrum sativum Protoplasts Enzymatic digestion Micro-colony Somatic embryos Scanning electron microscopy Histology 

Abbreviations

2,4-D

2,4-Dichlorophenoxyacetic acid

BA

6-Benzyladenine

KIN

6-Furfurylaminopurine

MS

Murashige and Skoog

NAA

α-Naphthalene acetic acid

PGR

Plant growth regulator

SEM

Scanning electron microscopy

Notes

Acknowledgments

The authors are thankful to University Grant Commission (UGC) and Department of Botany, Hamdard University (Jamia Hamdard) for receiving facilities of various kinds. The first author also acknowledges the help, rendered by M.Sc and Ph.D students.

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Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  1. 1.Cellular Differentiation and Molecular Genetics Section, Department of BotanyHamdard UniversityNew DelhiIndia

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