VEGETOS: An International Journal of Plant Research & Biotechnology
(Society For Plant Research)

Research Articles

A SOCIETY FOR PLANT RESEARCH PUBLICATION


Volume: 34, Issue: 1, March 2021


Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Website:www.vegetosindia.org
Pub Email: contact@vegetosindia.org
Views: 178

Doi: 10.1007/s42535-020-00166-x
Doi Link: https://doi.org/10.1007/s42535-020-00166-x
First Page: 144
Last Page: 152
Published: 01 January, 2021

TDZ induced plant regeneration from immature cotyledons of Pterocarpus marsupium Roxb. and validation of genetic homogeneity using ISSR markers


Abstract:

Pterocarpus marsupium Roxb. (Indian kino) is a threatened leguminous tree species well known for its therapeutic properties. A genetically stable plant regeneration system in P.marsupium has been developed using immature cotyledon (IC) explants excised from 12-days old in vitro grown seedlings. ICs were cultured on Murashige and Skoog (MS) media augmented with various concentrations of plant growth regulators to induce multiple shoots. Among the various concentrations and combinations used, MS medium augmented with thidiazuron (TDZ; 4.45 µM) and naphthalene acetic acid (NAA; 1.07 µM) showed the highest regeneration frequency (95.83%) with a maximum number of adventitious shoot buds (25.4 ± 0.55) with 22.7 shoots per explant. The harmful effects of extended exposure to TDZ at long-term regimen was prevented by transferring the organogenic cultures to MS medium fortified with 6- benzylaminopurine (BAP) for further multiplication and elongation. The highest number of shoots (36.0 ± 0.74) and shoot length (5.8 ± 0.10) were observed on 4.44 µM BAP augmented media after the third subculture. The elongated shoots were treated for 24 h with 19.60 µM indole-3-butyric acid (IBA) solution and rooted on half strength MS medium amended with 2.85 µM IBA. The plantlets were acclimatized successfully in the greenhouse with a 90% survival rate. Genetic homogeneity of regenerants were analyzed by inter simple sequence repeat (ISSR) markers. The comparative amplified products across the in vitro developed plantlets and the mother plant showed good genetic consistency. The protocol mentioned herein would be beneficial for raising genetically uniform plantlets and commercial cultivation of P. marsupium.

Vegetos

Keywords:


Genetic homogeneity, Immature cotyledon, Indian kino, ISSR, Woody tree


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Acknowledgements :



Radhika Tippani is thankful to Jawaharlal Nehru Memorial Fund, New Delhi, India for providing fellowship (Ref: SU-A/180/2011-12/404). We gratefully acknowledge Prof. A. Sadanandam, Department of Biotechnology, Kakatiya University, Warangal, Telangana, India to provide ISSR primers.


Author Information:



Radhika Tippani
Department of Biotechnology, Kakatiya University, Warangal, India
tippanira@gmail.ocm




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