Agrobacterium rhizogenes mediated transformation, initiation and multiplication of hairy roots in Spilanthes acmella Murr.

Authors

  • Hajera Sana Department of Botany, Veeranari Chakali Ilamma Women’s University, Koti, Hyderabad, Telangana State, India Author https://orcid.org/0000-0003-2604-9841
  • A. Sabitha Rani Department of Botany, University College of Sciences, Osmania University, Hyderabad- 500007, Telangana State, India Author

DOI:

https://doi.org/10.21276/pt.2025.v2.i1.9

Keywords:

Spilanthes acmella, Agrobacterium rhizogenes, hairy roots, transformation, PCR

Abstract

Spilanthes acmella is an important medicinal plant that belongs to the family Asteraceae. Traditionally, this plant has been used for years to cure toothaches, stammering, stomatitis and many other ailments. This plant is an important source of various medicinally important secondary metabolites like phenolics, coumarin, spilanthol, scopoletin, triterpenoids etc. The present study was carried out with the objective to transform this plant with Agrobacterium rhizogenes to initiate the hairy roots formation. This study demonstrated the transformation, initiation and multiplication of hairy roots from nodal segments and leaf explants taken from field grown and in vitro developed S. acmella plants. The hairy roots produced from different explants were white, slender, showed negative geotropism and lateral branching. PCR analysis of hairy root was performed that confirmed the bacterial transformation. Among the field grown and in vitro grown plants, the explants from the in vitro grown plants gave a high percentage of root induction. Nodal segments from in vitro grown plant gave 90% hairy root induction and leaf segments from in vitro grown plants with petiole gave the highest rate of root induction which was 92%. Nodal segments from field grown plants gave 85% and the leaves from field grown plants gave 80% of hairy root induction. This study offers great potential to establish the protocol for hairy root induction which can be used as an alternative source for the continuous production of this plant’s important secondary metabolites and active biocompounds.

 

Author Biographies

  • Hajera Sana, Department of Botany, Veeranari Chakali Ilamma Women’s University, Koti, Hyderabad, Telangana State, India

    Lecturer, Department of Botany

  • A. Sabitha Rani, Department of Botany, University College of Sciences, Osmania University, Hyderabad- 500007, Telangana State, India

    Professor, Department of Botany

References

Nelofar GN, Tareq A, Wani MS, Abid W, Syed Naseer S. Spilanthes acmella an endangered medicinal plant - its Traditional, Phytochemical and Therapeutic properties – An overview. Int J Adv Res. 2016; 4(1): 627 – 639.

Guillon S, Trémouillaux-Guiller J, Pati PK, Rideau M, Gantet P. Hairy root research: recent scenario and exciting prospects. Curr Opin Plant Biol. 2006; 9(3):341-6. https://doi.org/10.1016/j.pbi.2006.03.008

Ono NN, Tian L. The multiplicity of hairy root cultures: prolific possibilities. Plant Sci. 2011; 180(3):439-46. https://doi.org/10.1016/j.plantsci.2010.11.012

Murashige T. and Skoog F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant. 15: 473-497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x

Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979; 7(6):1513-23. https://doi.org/10.1093/nar/7.6.1513

Doyle JJ, Doyle JL. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull. 1987; 19:11-15

. Shanks JV, Morgan M. (1999). Plant ‘hairy root’ culture. Curr Opin Biotechnol. 1999; 10(2):151-155. https://doi.org/10.1016/s0958-1669(99)80026-3

Dixit AK, Vaidya S, Devaliya R. Agrobacterium rhizogenes MTCC 532 induced hairy hoot development in Tephrosia purpurea (L) and its effect on production of Tephrosin. J Sci Res. 2012; 4(3):741-748

Commisso M, Guarino F, Marchi L, Muto A, Piro A, Sheela C, Sharma HP, Ramesh C, Jha S. Agrobacterium rhizogenes mediated genetic transformation of Spilanthes paniculata (DC.) Jansen. Int. J Plant Sc. 2008; 3(2): 471-476.

Yogananth N, Basu MJ. TLC method for the determination of plumbagin in hairy root culture of Plumbago rosea L. Global J Biotech and Biochem. 2009; 4 (1):66–69.

Ercan AG, Taskin KM. Agrobacterium rhizogenes-mediated hairy root formation in some Rubia tinctorum L. population growth in Turkey. Turk J Bot. 1999; 23:373–377.

Karthikeyan A, Palanivel S, Parvathy S, Bhakyaraj R. Hairy root induction from hypocotyl segments of groundnut (Arachis hypogaea L.) Afr J Biotechnol. 2007; 6(15):1817–1820. https://doi.org/10.5897/AJB2007.000-2268

Doma M, Abhayankar G, Reddy VD, Kavi Kishor PB. Carbohydrate and elicitor enhanced withanolide (withaferin A and withanolide A) accumulation in hairy root cultures of Withania somnifera (L.). Indian J Exp Biol. 2012; 50(7):484-90

Chaudhury A, Pal M. Induction of Shikonin production in hairy root cultures of Arnebia hispidissima via Agrobacterium rhizogenes-mediated genetic transformation. J Crop Sci Biotechnol. 2010; 13: 99–106. https://doi.org/10.1007/s12892-010-0007-x

Brijwal L, Tamta S. Agrobacterium rhizogenes mediated hairy root induction in endangered Berberis aristata DC. SpringerPlus; 2015; 4: 443. https://doi.org/10.1186/s40064-015-1222-1

Murthy HN, Dijkstra C, Anthony P, White DA, Davey MR, Power JB, Hahn EJ, Paek KY. Establishment of Withania somnifera hairy root cultures for the production of withanolide A. J Integr Plant Biol. 2008; 50(8):975-81. https://doi.org/10.1111/j.1744-7909.2008.00680.x

Downloads

Published

2025-04-29 — Updated on 2025-05-02

Versions

Data Availability Statement

All data generated or analyzed during this study are included in this published article.

How to Cite

1.
Sana H, A. SR. Agrobacterium rhizogenes mediated transformation, initiation and multiplication of hairy roots in Spilanthes acmella Murr. phytoTalks. 2025;2(1):297-303. doi:10.21276/pt.2025.v2.i1.9