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3 Biotech

, 8:146 | Cite as

An efficient protocol for in vitro direct shoot organogenesis of Sesamum indicum L. using cotyledon as explant

  • A. J. Debnath
  • G. Gangopadhyay
  • D. Basu
  • S. R. Sikdar
Original Article
  • 54 Downloads

Abstract

Establishment of a suitable regeneration protocol is a pre-requisite to carry out transformation study in Sesamum indicum L. (sesame). In this paper, different parameters of regeneration were standardised to develop an efficient protocol for in vitro plant regeneration via direct adventitious shoot organogenesis using de-embryonated cotyledons of sesame as explants. Among the various treatments of MS medium supplemented with 6-benzylaminopurine, thidiazuron and indole-3-acetic acid, maximum regeneration frequency (25.93  ±  2.21%) was obtained in BTI 4 medium (MS supplemented with 33.33 µM BAP with 2.85 µM IAA) within 6 weeks of culture. Regeneration frequency increased further (50.37 ± 2.49%) by fortifying BTI 4 with 29.43 µM silver nitrate (AG 3 medium). Pre-culture of cotyledon explants in AB 3 medium (AG 3 supplemented with 3.78 µM abscisic acid) for 14 days followed by sub-culture in AG 3 medium further improved the regeneration frequency (68.15 ± 2.68%). The highest rate of shoot regeneration (94.82 ± 1.34%) was obtained by pre-culturing 4-day-old cotyledon in a vertical position in AB 3 medium for 14 days and sub-culturing in AG 3 medium for 4 weeks. Regenerated shoots proliferated in MS medium supplemented with 4.44 μM BAP and 1.44 μM gibberelic acid (GA3). The highest frequency (65.33 ± 3.78%) of root induction was achieved by culturing the elongated shoots in MS medium supplemented with 2.69 μM α-naphthalene acetic acid (NAA) for 6 weeks. Rooted plants were acclimatised in soilrite and transferred to soil after 6–8 weeks. The rate of acclimatisation of plants was 76%.

Keywords

Adventitious shoot Direct organogenesis Explant orientation Regeneration Scanning electron microscopy Sesame 

Abbreviations

ABA

Abscisic acid

AgNO3

Silver nitrate

BAP

6-Benzylaminopurine

DMSO

Dimethyl sulfoxide

GA3

Gibberelic acid

GUS

β-glucuronidase

IAA

Indole-3-acetic acid

IBA

Indole-3-butyric acid

MS

Murashige and skoog

NAA

1-Naphthalene acetic acid

PGR

Plant growth regulator

SAAT

Sonication-assisted Agrobacterium-mediated transformation

SAM

Shoot apical meristem

SE

Standard error

SEM

Scanning electron microscopy

TDZ

1-Phenyl-3-(1, 2, 3-Thidiazol-5-yl)-urea (Thidiazuron)

Notes

Acknowledgement

This work was supported by National Agricultural Innovation Project, Indian Council of Agricultural Research (ICAR-NAIP) under Grant [Funding Number: NAIP/C4/C1090].

Author contribution statement

G. Gangopadhyay, D. Basu, and S. R. Sikdar conceived and designed the research. A. J. Debnath organized, conducted experiments, analysed data, and wrote the manuscript. All of the authors read and approved the final version of the manuscript.

Compliance with ethical standards

Conflict of interest

On behalf of all authors, the corresponding author states that there is no conflict of interest.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • A. J. Debnath
    • 1
  • G. Gangopadhyay
    • 2
  • D. Basu
    • 1
  • S. R. Sikdar
    • 1
  1. 1.Division of Plant BiologyBose InstituteKolkataIndia
  2. 2.Division of Plant BiologyBose InstituteKolkataIndia

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