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

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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.

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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

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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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Authors and Affiliations

  1. Department of Biotechnology, Bharathidasan University, Tiruchirappalli, Tamil Nadu, 620 024, India

    Chandrasekaran Ajithan, Venkatachalam Vasudevan, Dorairaj Sathish, Selvam Sathish & Markandan Manickavasagam

  2. Division of Biochemistry, ICAR- Indian Agricultural Research Institute, New Delhi, 110 012, India

    Veda Krishnan

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  1. Chandrasekaran Ajithan
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  2. Venkatachalam Vasudevan
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Correspondence to Markandan Manickavasagam.

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Communicated by Sergio J. Ochatt.

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Ajithan, C., Vasudevan, V., Sathish, D. et al. The influential role of polyamines on the in vitro regeneration of pea (Pisum sativum L.) and genetic fidelity assessment by SCoT and RAPD markers. Plant Cell Tiss Organ Cult 139, 547–561 (2019). https://doi.org/10.1007/s11240-019-01699-z

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