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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
  • 114 Downloads

Abstract

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.

Keywords

Colchicine Cytology HPTLC Sphagneticola calendulacea Tetraploid Wedelolactone 

Abbreviations

MS

Murashige and Skoog

TDZ

Thidiazuron

NAA

α-Naphthalene acetic acid

IAA

Indole-3-acetic acid

ST

Shoot tip

NS

Nodal segment

RT

Root tip

HPLC

High-performance liquid chromatography

HPTLC

High-performance thin-layer chromatography

Notes

Acknowledgements

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.

References

  1. Abdoli M, Moieni A, Badi HN (2013) Morphological, physiological, cytological and phytochemical studies in diploid and colchicine-induced tetraploid plants of Echinacea purpurea (L.). Acta Physiol Plant 35:2075–2083CrossRefGoogle Scholar
  2. Agarwala B, Azam FMS, Khatun MA, Rahman F, Rahmatullah M (2010) Simultaneous shoot regeneration and rhizogenesis of Wedelia chinensis for in vitro clonal propagation. Am Eurasian J Sustain Agric 4:65–69Google Scholar
  3. 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
  4. Arnon DI (1949) Copper enzymes in isolated chloroplasts polyphenoloxidase in Beta vulgaris. Plant Physiol 24:1–15CrossRefGoogle Scholar
  5. Ashraf M, Orooj A (2006) Salt stress effects on growth, ion accumulation and seed oil concentration in an arid zone traditional medicinal plant ajwain (Trachyspermum ammi [L.] Sprague). J Arid Environ 64:209–220CrossRefGoogle Scholar
  6. Bouvier L, Pillon FR, Lespinasse Y (1994) Oryzalin as an efficient agent for chromosome doubling of haploid apple shoots in vitro. Plant Breed 113:343–346CrossRefGoogle Scholar
  7. Chen Z, Sun X, Shen S, Zhang H, Ma X, Liu J, Kuang S, Yu Q (2013) Wedelolactone a naturally occurring coumestan, enhances interferon-G signaling through inhibiting STAT1 protein dephosphorylation. J Biol Chem 288:14417–14427CrossRefGoogle Scholar
  8. Chung MY, Kim CY, Min JS, Lee DJ, Naing AH, Chung JD, Kim CK (2014) In vitro induction of tetraploids in an interspecific hybrid of Calanthe (Calanthe discolor × Calanthe sieboldii) through colchicine and oryzalin treatments. Plant Biotechnol Rep 8:251–257CrossRefGoogle Scholar
  9. De Oliveira VM, Forni-Martins ER, Magalhães PM, Alves MN (2004) Chromosomal and morphological studies of diploid and polyploid cytotypes of Stevia rebaudiana (Bertoni) (Eupatorieae, Asteraceae). Genet Mol Biol 27:215–222CrossRefGoogle Scholar
  10. Dhooghe E, Van Laere K, Eeckhaut T, Leus L, Van Huylenbroeck J (2011) Mitotic chromosome doubling of plant tissues in vitro. Plant Cell Tissue Organ Cult 104:359–373CrossRefGoogle Scholar
  11. Doyle JJ, Doyle JL (1990) A rapid total DNA preparartion procedure from fresh plant tissue. Focus 12:13–15Google Scholar
  12. Duncan DB (1955) Multiple range and multiple F test. Biometrics 11:1–42CrossRefGoogle Scholar
  13. Elyazid DMA, El-Shereif AR (2014) In vitro induction of polyploidy in Citrus reticulata Blanco. Am J Plant Sci 5:1679–1685CrossRefGoogle Scholar
  14. Galbraith DW, Harkins KR, Maddox JM, Ayres NM, Sharma DP, Firoozabady E (1983) Rapid flow cytometric analysis of the cell cycle in intact plant tissues. Science 220:1049–1051CrossRefGoogle Scholar
  15. Gantait S, Mandal N, Bhattacharyya S, Das PK (2011) Induction and identification of tetraploids using in vitro colchicine treatment of Gerbera jamesonii Bolus cv Sciella. Plant Cell Tissue Organ Cult 106:485–493CrossRefGoogle Scholar
  16. Hamill SD, Smith MK, Dodd WA (1992) In vitro induction of banana autotetraploids by colchicine treatment of micropropagated diploids. Aust J Bot 40:887–896CrossRefGoogle Scholar
  17. Hannweg K, Sippel A, Bertling I (2013) A simple and effective method for the micropropagation and in vitro induction of polyploidy and the effect on floral characteristics of the South African iris, Crocosmia aurea. S Afr J Bot 88:367–372CrossRefGoogle Scholar
  18. Hegde SN, Rameshsing CN, Vasundhara M (2015) Characterization of Stevia rebaudiana Bertoni polyploids for growth and quality. Med Plants Int J Phytomed Relat Ind 7:188–195CrossRefGoogle Scholar
  19. Iannicelli J, Elechosa MA, Juárez MA, Martínez A, Bugallo V, Bandoni AL, van Baren CM (2016) Effect of polyploidization in the production of essential oils in Lippia integrifolia. Ind Crops Prod 81:20–29CrossRefGoogle Scholar
  20. Javadian N, Karimzadeh G, Sharifi M, Moieni A, Behmanesh M (2017) In vitro polyploidy induction: changes in morphology, podophyllotoxin biosynthesis, and expression of the related genes in Linum album (Linaceae). Planta 245:1165–1178CrossRefGoogle Scholar
  21. Koul S, Pandurangan A, Khosa RL (2012) Wedelia chinenis (Asteraceae)—an overview. Asian Pac J Trop Biomed 2:S1169–S1175CrossRefGoogle Scholar
  22. Kundu S, Salma U, Ali MN, Mandal N (2017) Factors influencing large scale micropropagation of Sphagneticola calendulacea (L.) Pruski and clonality assessment using RAPD and ISSR markers. In Vitro Cell Dev Biol Plant 53:167–177CrossRefGoogle Scholar
  23. Lavania UC (2005) Genomic and ploidy manipulation for enhanced production of phyto-pharmaceuticals. Plant Genet Resour 3(2):170–177CrossRefGoogle Scholar
  24. Lin FM, Chen LR, Lin EH, Ke FC, Chen HY, Tsai MJ, Hsiao PW (2007) Compounds from Wedelia chinensis synergistically suppress androgen activity and growth in prostate cancer cells. Carcinogenesis 28:2521–2529CrossRefGoogle Scholar
  25. Madani H, Hosseini B, Dehghan E, Rezaei-chiyaneh E (2015) Enhanced production of scopolamine in induced autotetraploid plants of Hyoscyamus reticulatus L. Acta Physiol Plant 37(3):55CrossRefGoogle Scholar
  26. Meena AK, Rao MM, Meena RP, Panda P, Renu (2011) Pharmacological and phytochemical evidences for the plants of Wedelia genus—a review. Asian J Pharm Res 1:7–12Google Scholar
  27. Mehra PN, Remanandan P (1974) Cytological investigations on the Indian compositae. II. Astereae, Heliantheae, Helenieae and Anthemideae. Caryologia 27:255–284CrossRefGoogle Scholar
  28. Mishra BK, Pathak S, Sharma A, Trivedi PK, Shukla S (2010) Modulated gene expression in newly synthesized auto-tetraploid of Papaver somniferum L. S Afr J Bot 76:447–452CrossRefGoogle Scholar
  29. Moghbel N, Borujeni MK, Bernard F (2015) Colchicine effect on the DNA content and stomata size of Glycyrrhiza glabra var. glandulifera and Carthamus tinctorius L. cultured in vitro. J Genet Eng Biotechnol 13:1–6CrossRefGoogle Scholar
  30. Murashige T, Nakano R (1966) Tissue culture as a potential tool in obtaining polyploid plants. J Hered 57:114–118CrossRefGoogle Scholar
  31. Murashige T, Skoog FA (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  32. Niu L, Tao YB, Chen MS, Fu Q, Dong Y, He H, Xu ZF (2016) Identification and characterization of tetraploid and octoploid Jatropha curcas induced by colchicine. Caryologia 69:58–66CrossRefGoogle Scholar
  33. 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 130:543–551CrossRefGoogle Scholar
  34. Salma U, Kundu S, Mandal N (2017) Artificial polyploidy in medicinal plants: advancement in the last two decades and impending prospects. J Crop Sci Biotechnol 20:9–19CrossRefGoogle Scholar
  35. Salma U, Kundu S, Ali MN, Mandal N (2018) Augmentation of wedelolactone through in vitro tetraploid induction in Eclipta alba (L.) Hassk. Plant Cell Tissue Organ Cult 133:289–298CrossRefGoogle Scholar
  36. Shao JZ, Chen CL, Deng XX (2003) In vitro induction of tetraploid in pomegranate (Punica granatum). Plant Cell Tissue Organ Cult 75:241–246CrossRefGoogle Scholar
  37. Talebi SF, Saharkhiz MJ, Kermani MJ, Sharafi Y, Raouf Fard F (2017) Effect of different antimitotic agents on polyploid induction of anise hyssop (Agastache foeniculum L.). Caryologia 70:1–10CrossRefGoogle Scholar
  38. Tavan M, Mirjalili MH, Karimzadeh G (2015) In vitro polyploidy induction: changes in morphological, anatomical and phytochemical characteristics of Thymus persicus (Lamiaceae). Plant Cell Tissue Organ Cult 122:573–583CrossRefGoogle Scholar
  39. Widoretno W (2016) In vitro induction and characterization of tetraploid Patchouli (Pogostemon cablin Benth.) plant. Plant Cell Tissue Organ Cult 125(2):261–267CrossRefGoogle Scholar
  40. Yan HJ, Xiong Y, Zhang HY, He ML (2016) In vitro induction and morphological characteristics of octoploid plants in Pogostemon cablin. Breed Sci 66:169–174CrossRefGoogle Scholar
  41. Zahedi AA, Hosseini B, Fattahi M, Dehghan E, Parastar H, Madani H (2014) Overproduction of valuable methoxylated flavones in induced tetraploid plants of Dracocephalum kotschyi Boiss. Bot Stud 55:22CrossRefGoogle Scholar
  42. Zhang Z, Dai H, Xiao M, Liu X (2008) In vitro induction of tetraploids in Phlox subulata L. Euphytica 159:59–65CrossRefGoogle Scholar

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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