, Volume 32, Issue 2, pp 181–189 | Cite as

Cloning of mature pomegranate (Punica granatum) cv. Jalore seedless via in vitro shoot production and ex vitro rooting

  • Rachana Modi DineshEmail author
  • Ashok Kumar Patel
  • J. B. Vibha
  • Smita Shekhawat
  • N. S. Shekhawat
Research Articles


A novel approach of in vitro shoot amplification and ex vitro rooting for cloning of mature plants of Punica granatum cv. Jalore seedless/soft-seeded has been defined. Surface-sterilized nodal shoots were cultured for axillary meristem activation, bud breaking and shoot amplification. Multiple shoots differentiated by bud breaking from 82.8% of the explants on Murashige and Skoog (Physiol Plant 15:473–497, 1962) MS medium with 13.32 µM 6-benzylaminopurine (BAP). These were amplified by subculturing of fresh shoots and by repeated transfer of mother explants on MS medium + BAP or Kinetin (Kin)/Furfurylaminopurine (FAP) in combination with Indole acetic acid (IAA) or α-naphthalene acetic acid (NAA). MS medium with BAP 2.22 µM + IAA 0.57 µM was found to be most suitable for shoot (14.2 ± 1.03 shoots per culture vessel) multiplication. On half-strength MS medium with 9.84 µM of Indole butyric acid (IBA) and 16.65 µM of activated charcoal (AC), 72.9% of the micro-shoots rooted. Alternatively, base(s) of the micro-shoots treated with 1476 µM of IBA for 300 s. and transplanted on autoclaved soilrite (moistened with one-fourth strength of MS macro-salts) in glass bottles (420 ml; 70 mm diameter × 130 mm height). More than 85% of the IBA-treated shoots rooted ex vitro in a greenhouse. Ex vitro rooting of cloned shoots is a new approach for propagation of pomegranate. The process described is different and superior to all the described tissue culture methods for cloning of pomegranate. This is faster, cost-effective and saves resources enabling acclimatization/hardening with ease while minimizing microbial contamination, thus ensuring quick field transfer of hardened plantlets. This can be applied for mass and clonal propagation of selected genotypes and also for long-term conservation of germplasm of P. granatum.


Ex vitro rooting Horticultural fruit plant Micropropagation Lythraceae P. granatum cv. Jalore seedless 



Activated charcoal




Duncan’s multiple range test


Indole-3-acetic acid


Indole-3-butyric acid




Murashige and Skoog (1962)


α-Naphthalene acetic acid


Photon flux density


Plant growth regulators


Relative humidity



RD acknowledges the University Grants Commission (UGC), Government of India for startup Grant [F-30-16/2014(BSR)]. RD and AKP express their gratitude to the UGC for awarding the Special Assistance Programme (SAP) in the form of Centre of Advanced Study (CAS) to the Department of Botany, Jai Narain Vyas University, Jodhpur [F.5-1/2013(SAP-II)].

Author’s contribution

RMD designed, conducted the experiments and wrote the first draft of the manuscript. AKP and SS assisted in analyzing the data and organized them in tables and figures. VJB looked after the rooting and hardening experiments. NSS conceptualized, supervised the research and prepared the final draft of the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they do not have any conflicts of interest.


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Copyright information

© Society for Plant Research 2019

Authors and Affiliations

  • Rachana Modi Dinesh
    • 1
    Email author
  • Ashok Kumar Patel
    • 1
  • J. B. Vibha
    • 1
  • Smita Shekhawat
    • 1
  • N. S. Shekhawat
    • 1
  1. 1.Biotechnology Unit, Department of Botany, UGC–Centre of Advanced Study (CAS)Jai Narain Vyas UniversityJodhpurIndia

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