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Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 139, Issue 3, pp 547–561 | Cite as

The influential role of polyamines on the in vitro regeneration of pea (Pisum sativum L.) and genetic fidelity assessment by SCoT and RAPD markers

  • Chandrasekaran Ajithan
  • Venkatachalam Vasudevan
  • Dorairaj Sathish
  • Selvam Sathish
  • Veda Krishnan
  • Markandan ManickavasagamEmail author
Original Article

Abstract

A proficient and reliable in vitro plant regeneration protocol was established for pea by utilizing cotyledonary node as an explants, which excised from 3-days old imbibed seeds. The present research work explains the positive role of polyamines (PAs) accompanied by plant growth regulators (PGRs) on the multiple shoot induction and rooting in pea plant regeneration. The highest multiple shoots (65.1 shoots/explant) was attained from the cotyledonary node explant on Murashige and Skoog (MS) medium accompanied with 20 mg l−1 spermidine (SPD) along with 1.5 mg l−1 6-benzyladenine (BA). Moreover, the highest elongation of multiplied shoots (10 cm length/shoot) was noted on MS medium enriched with 1 mg l−1 of gibberellic acid (GA3). The elongated shoots produced the highest number of roots (33.66 roots/shoot) on MS medium augmented with 30 mg l−1 putrescine (PUT) along with 0.6 mg l−1 1-naphthaleneacetic acid (NAA). Rooted plants were hardened and acclimatized in the greenhouse with an existence rate of 98%. The standardized procedure with the use of PAs has enhanced the multiple shoot induction to threefold higher than the plants raised from PGRs treatments. The regenerated pea plants revealed the fivefold enhanced photosynthetic pigment, twofold enhanced antioxidant profile and the substantial growth in chloroplast count have been achieved in optimized SPD and BA supplemented MS medium compared to control plant. Start Codon Targeted polymorphism and Random Amplified Polymorphic DNA molecular markers recognized the genetic purity of the in vitro regenerated pea plants for their true type of nature.

Key message

An enhanced and consistence in vitro regeneration protocol was established for pea, by the application of polyamines along with PGRs, which significantly improved the multiple shooting, shoot elongation, rooting, total chlorophyll and antioxidant profile of regenerated plants, further not hampering their genetic fidelity.

Keywords

Antioxidant Chlorophyll Chloroplast Genetic fidelity Molecular marker Pea Polyamines Regeneration 

Abbreviations

PAs

Polyamines

PGRs

Plant growth regulators

BA

6-Benzyladenine

TDZ

Thidiazuron

KIN

Kinetin

GA3

Gibberellic acid

NAA

1-Naphthaleneacetic acid

IAA

Indole-3-acetic acid

IBA

Indole-3-butyric acid

SPD

Spermidine

SPM

Spermine

PUT

Putrescine

SCoT

Start Codon Targeted polymorphism

RAPD

Random Amplified Polymorphic DNA

Notes

Acknowledgements

The first author is thankful to the University Grants Commission, India for the award of Rajiv Gandhi National Fellowship (F1-17.1/2016-17-SC-TA,-3517/SA-III/Website-Date: 01.09.2016).

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© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of BiotechnologyBharathidasan UniversityTiruchirappalliIndia
  2. 2.Division of BiochemistryICAR- Indian Agricultural Research InstituteNew DelhiIndia

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