Journal of Crop Science and Biotechnology

, Volume 19, Issue 2, pp 157–165 | Cite as

Direct shoot organogenesis from rhizomes of medicinal zingiber Alpinia calcarata Rosc. and evaluation of genetic stability by RAPD and ISSR markers

  • Sudipta Shekhar Das Bhowmik
  • Adreeja Basu
  • Lingaraj SahooEmail author
Research Article


A simple and efficient protocol for direct in vitro shoot multiplication and plant regeneration was established for an important aromatic medicinal plant, Alpinia calcarata. Preinduction of rhizome segments in medium containing 8.8 μM 6-benzylamino purine (BAP) rescued the buds from dormancy in 60% of the cultures. An average of 6.2 shoots were produced from rhizomatous bud explants on Murashige and Skoog (MS) medium supplemented with 5 μM BAP, 10 μM kinetin, and 2.5 μM α-Naphthalene acetic acid (NAA). The mother cultures retained their morphogenetic potential upto four subcultures and a maximum of 1.77-fold increase in shoot multiplication was recorded after the 3rd subculture. Rooting was simultaneously induced during subculture on shoot multiplication medium eliminating an additional step for rooting induction. Rooted plantlets were successfully acclimatized in pots in the greenhouse and subsequently established in the experimental garden without any visible symptoms of wilting and necrosis. The genetic fidelity of regenerated plants was evaluated by adapting to two PCR-based DNA marker techniques, Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeats (ISSR) which detected no variability in the in vitro multiplied plantlets of A. calcarata. This efficient method of clonal multiplication may be useful for commercial scale multiplication, and in situ and ex situ conservation of elite germplasm of A. calcarata.


Alpinia calcarata genetic uniformity micropropagation rhizome 


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Copyright information

© Korean Society of Crop Science and Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Sudipta Shekhar Das Bhowmik
    • 1
    • 2
  • Adreeja Basu
    • 3
  • Lingaraj Sahoo
    • 1
    Email author
  1. 1.Department of Biosciences and BioengineeringIndian Institute of Technology GuwahatiGuwahatiIndia
  2. 2.Center for Tropical Crops and Biocommodities, Queensland University of TechnologyGardens Point CampusBrisbaneAustralia
  3. 3.Center for EnergyIndian Institute of Technology GuwahatiGuwahatiIndia

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