Acta Physiologiae Plantarum

, 40:215 | Cite as

In vitro tetraploidization for the augmentation of wedelolactone in Sphagneticola calendulacea (L.) Pruski

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


A practical and reliable method for in vitro tetraploidization of Sphagneticola calendulacea (L.) Pruski [synonym Wedelia chinensis (Osbeck) Merrill] has been established to enhance the production of wedelolactone. Shoot tip and nodal explants from in vitro-grown culture (2n = 50) were exposed to the antimitotic chemical, i.e., colchicine, at various concentrations (0, 0.025, 0.05, 0.1, 0.3, and 0.5%; w/v) for 12, 24, 36, 48, and 60 h. The treated explants were then incubated and proliferated on Murashige and Skoog (MS) medium fortified with 0.2 mg l−1 thidiazuron and 0.05 mg l−1 naphthalene acetic acid, followed by root induction in 1.0 mg l−1 indole-3 acetic acid enriched ½MS medium. Treatment of shoot tips with 0.05% colchicine for 24 h supported the maximum rate of survival (63.33%) of explants as well as tetraploid induction (42.93%). Morphological, stomatal, and cytological characteristics along with the secondary metabolite content of the in vitro tetraploids were compared to that of diploids. The recovered tetraploid plants possessed superior plant height, stem diameter, leaf size, root number, and increased length and width of stomata but decreased stomatal frequency. The tetraploid plants demonstrated twice the chromosome number (2n = 4x = 100) than the diploids as confirmed through cytology, spectrophotometry and flow cytometry. High-performance thin-layer chromatography showed a significant enhancement in the wedelolactone content of tetraploid plants (541.48 µg g−1 of dried sample) in comparison to diploid plants (325.43 µg g−1 of dried sample), signifying the prospective of this technique for the trade value improvement.


Colchicine Cytology HPTLC Sphagneticola calendulacea Tetraploid Wedelolactone 



Murashige and Skoog




α-Naphthalene acetic acid


Indole-3-acetic acid


Shoot tip


Nodal segment


Root tip


High-performance liquid chromatography


High-performance thin-layer chromatography



Authors acknowledge the laboratory as well as library assistance from the Bidhan Chandra Krishi Viswavidyalaya, West Bengal, India.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.


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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 BiotechnologyFaculty of Agriculture, Bidhan Chandra Krishi ViswavidyalayaNadiaIndia

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