Augmentation of wedelolactone through in vitro tetraploid induction in Eclipta alba (L.) Hassk.
- 207 Downloads
A complete protocol for the in vitro induction of Eclipta alba tetraploids has been optimized to enhance the wedelolactone content, an anti-cancerous compound. The effects of different concentrations of colchicine (0, 0.01, 0.05, 0.1, 0.2 and 0.3%; w/v) along with treatment durations (12, 24, 36 and 48 h) were investigated on shoot tip (ST) and nodal segment (NS). The treated explants were then incubated on Murashige and Skoog (MS) medium having 1.5 mg L−1 N6-benzylaminopurine and 0.5 mg L−1 α-napthalene acetic acid for shoot regeneration and afterward root was induced on 1.0 mg L−1 indole-3-acetic acid enriched ½MS medium. The tetraploids of E. alba were proficiently induced by the treatment of 0.1% colchicine for 24 h. The highest tetraploid induction efficiency was obtained from ST (30.56%) in comparison to the NS (22.22%). Analysis by spectrophotometry and flow cytometry showed that colchicine treated plants contained higher quantity of DNA than diploid plants. Cytological studies demonstrated doubled the chromosome number in tetraploids (2n = 4x = 44) than diploids (2n = 2x = 22). The ploidy level enhancement lead to alteration of other traits, like increased plant height, stem diameter, leaf size, stomatal size and chlorophyll content. As determined through high performance thin-layer chromatography, the ultimate achievement of this technique is the higher accumulation of wedelolactone in tetraploid plants (300.32 µg g−1 dry weight) in evaluation to in vitro diploid (131.31 µg g−1 dry weight) and in vivo diploid mother plants (93.26 µg g−1 dry weight), thus improving the pharmaceutical value of E. alba.
KeywordsColchicine Chromosome counting Eclipta alba Tetraploid Wedelolactone
High performance thin-layer chromatography
Murashige and Skoog (1962)
α-Napthalene acetic acid
Authors acknowledge the laboratory as well as library assistance from the Bidhan Chandra Krishi Viswavidyalaya, West Bengal, India.
US, SK, MNA and NM conceived the idea; US and SK performed the experiments; US and SK surveyed the literature and wrote the manuscript; AKH assisted in the phytochemical assessment; MNA and NM edited the manuscript. All the authors approved the final version of the manuscript prior to submission.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflicts of interest.
- Amiri S, Kazemitabaar S, Ranjbar G, Azadbakht M (2010) The effect of trifluralin and colchicine treatments on morphological characteristics of jimsonweed (Datura Stramonium L.). Trakia J Sci 8:47–61Google Scholar
- Gomes SSL, Saldanha CW, Neves CS, Trevizani M, Raposo NRB, Notini MM, de Oliveira Santos M, Campos JMS, Otoni WC, Viccini LF (2014) Karyotype, genome size, and in vitro chromosome doubling of Pfaffia glomerata (Spreng.) Pedersen. Plant Cell Tissue Organ Cult 118:45–56Google Scholar
- Li W, Sixiang Z, Zhijian G (2002) In vitro culture of tetraploids of Aloe vera L. Acta Hortic Sin 29:176–178Google Scholar
- Majdi M, Karimzadeh G, Malboobi MA, Omidbaigi R, Mirzaghaderi G (2010) Induction of tetraploidy to feverfew (Tanacetum parthenium Schulz-Bip.): morphological, physiological, cytological, and phytochemical changes. HortScience 45:16–21Google Scholar
- Nilanthi D, Yang YS (2013) In vitro induction of octaploid from colchicine-treated tetraploid petiole explants of purple coneflower (Echinacea purpurea L.). Trop Agric Res Ext 16:25–30Google Scholar
- Noori SAS, Norouzi M, Karimzadeh G, Shirkool K, Niazian M (2017) Effect of colchicine induced polyploidy on morphological characteristics and essential oil composition of ajowan (Trachyspermum ammi L.). Plant Cell Tissue Organ Cult 1–9Google Scholar
- Omidbaigi R, Mirzaee M, Hassani ME, Sedghi Moghadam M (2010) Induction and identification of polyploidy in basil (Ocimum basilicum L.) medicinal plant by colchicine treatment. Int J Plant Prod 4:87–98Google Scholar
- Pôças ES, Lopes DV, da Silva AJ, Pimenta PH, Leitão FB, Netto CD, Buarque CD, Brito FV, Costa PR, Noël F (2006) Structure-activity relationship of wedelolactone analogues: structural requirements for inhibition of Na, K-ATPase and binding to the central benzodiazepine receptor. Bioorg Med Chem 14:7962–7966CrossRefPubMedGoogle Scholar
- Salma U, Kundu S, Ali MN, Mandal N (2017a) An efficient micropropagation protocol for Eclipta alba (L.) Hassk.: an endangered, medicinally important plant. J Crop Weed 13:49–54Google Scholar
- Sharma A, Bhansali S, Kumar A (2013) In vitro callus induction and shoot regeneration in Eclipta alba (l.) Hassk. Life Sci 3:43–46Google Scholar