Conservation, ex vitro direct regeneration, and genetic uniformity assessment of alginate-encapsulated nodal cuttings of Sphagneticola calendulacea (L.) Pruski

  • Suprabuddha Kundu
  • Umme Salma
  • Md. Nasim Ali
  • Nirmal Mandal
Original Article


A well-organized procedure was established for the conservation and distribution of Sphagneticola calendulacea (L.) Pruski [synonym Wedelia chinensis (Osbeck) Merrill] for the first time, using alginate-encapsulated nodal segments (NSs) as synthetic seeds. The ideal beads were obtained through a combination of 2.5% sodium alginate and 75 mM calcium chloride with 84.40 ± 2.20% rate of shoot emergence. The maximum regeneration (88.84 ± 2.24%) from synthetic seeds was achieved on liquid 1/2Murashige and Skoog (MS) medium in comparison to its other formulations. Furthermore, superior frequency (91.09 ± 2.24%) of complete plantlet (having both shoots and roots) formation was achieved when synthetic seeds were cultured on liquid 1/2MS (1.5% sucrose) fortified with 1.0 mg L−1 N6-benzyladenine plus 0.25 mg L−1 α-naphthalene acetic acid. Synthetic seeds could be effectively stored at low temperature (8 °C) up to 90 days with a survival rate of 52.38 ± 3.06%, whereas higher temperature (25 °C) did not support regeneration after 75 days of storage. The plantlets were successfully acclimatized to natural conditions with ~ 89% survival frequency. To by-pass the time-consuming in vitro culture step after encapsulation, synthetic seeds were directly regrown into complete plantlets ex vitro on sand, soil, and vermicompost (1:1:1; w/w). Regeneration rate of 42.22 ± 2.22% was attained when NSs were pretreated on 1/2MS medium containing 4.0 mg L−1 indole-3-acetic acid for 24 h in dark, prior to encapsulation. The random amplified polymorphic DNA and intersimple sequence repeat fingerprinting profiles demonstrated genetic uniformity amongst the regenerated plantlets, in vitro mother plant, as well as in vivo wild plant.


Encapsulation Germplasm exchange Sodium alginate Calcium chloride Genetic uniformity 





Calcium chloride


Indole-3-butyric acid


Laminar airflow


Murashige and Skoog medium


α-Naphthalene acetic acid


Nodal segment


Plant growth regulator


Sodium alginate



Authors acknowledge the laboratory as well as library assistance from the Bidhan Chandra Krishi Viswavidyalaya, West Bengal, India. We further are thankful to the anonymous reviewers and the editor of this article for their critical comments and suggestions on the manuscript. The authors declare that there are no conflicts of interest. This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.


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Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2018

Authors and Affiliations

  1. 1.Department of Agricultural Biotechnology, Faculty of AgricultureBidhan Chandra Krishi ViswavidyalayaNadiaIndia

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