In Vitro Method of High-Frequency Plant Regeneration Through Internodal Callus of Ruta graveolens L.

  • Malik Aabid Hussain
  • Varsha Nitin Nathar


Ruta graveolens L. is of considerable interest worldwide because of its medicinal properties. In vitro culture is a useful tool for both multiplication and study of important secondary metabolites. It was intended to develop an effective in vitro indirect propagation protocol for Ruta graveolens L. from internodal explants. The explants excised from 10-week-old plants were placed on Murashige and Skoog (MS) medium supplemented with various concentrations and combinations of auxins, indole-3-butyric acid (IBA), 2,4-dichlorophenoxy acetic acid (2,4-D), and naphthalene acetic acid (NAA), and cytokinins (benzyl amino purine (BAP) and kinetin) singly and in combinations for callus induction. The callus response was maximum (90.00 ± 1.66) in 2,4-D (1.5 mg/l) + NAA (1.5 mg/l). The friable callus obtained was subcultured on MS full-strength medium fortified with different concentrations of plant growth regulators. The highest percent response (81.22 ± 0.57) per culture for shoot bud regeneration was noted for the concentration of BAP (1.5 mg/l) with IBA (1.0 mg/l). The same concentration effectively increased the number of shoots per culture. Different concentrations of indole-3-butyric acid (IBA) and indole acetic acid (IAA) were used in half-strength MS medium for in vitro rooting of regenerated shoots. The maximum percentage of rooting (86.10 ± 0.50) and the highest number of root formations (8.11 ± 0.19) per shoot were observed on the medium containing 0.50 mg/l of IBA. Plantlets with well-developed root and shoot systems were successfully acclimatized (85%) and established in earthen pots.


Callus culture Organogenesis Regeneration Ruta graveolens L. 


  1. Ahmad N, Faisal M, Anis M, Aref IM (2010) In vitro callus induction and plant regeneration from leaf explants of Ruta graveolens L. S Afr J Bot 76:597–600CrossRefGoogle Scholar
  2. Al-Ajloun ZI, Abbas S, Shatnawi M, Al-Makhadmeh I (2015) In vitro propagation, callus induction, and evaluation of active compounds on Ruta graveolens. J Food Agric Environ 13(2):101–106Google Scholar
  3. Fadlalla K, Watson A, Yehualaeshet T, Turner T, Samuel T (2011) Ruta graveolens extract induces DNA damage pathways and blocks akt activation to inhibit cancer cell proliferation and survival. Anticancer Res 31:233–241PubMedPubMedCentralGoogle Scholar
  4. Faisal M, Anis M (2003) Rapid mass propagation of Tylophora indica Merrill via leaf callus culture. Plant Cell Tissue Org Cult 75:125–129CrossRefGoogle Scholar
  5. Faisal M, Ahmad N, Anis M (2005) In vitro regeneration and mass propagation of Ruta graveolens L. - multipurpose shrub. HortScience 40(5):1478–1480CrossRefGoogle Scholar
  6. Feo V, Simone F, Senatore F (2002) Eight peak index of mass spectra, potential allelochemicals from the essential oil of Ruta graveolens. Phytochemistry 61:573–578CrossRefGoogle Scholar
  7. Ghirardini MP, Carli M, del Vecchio N, Rovati A, Cova O, Valigi F, Agnetti G, Gurudeeban S, Satyavani K, Ramanathan T, Balasubramanian T (2007) Effect of antioxidant and anti-aggregating properties of micro-propagated plantlets of Ruta graveolens. Afr J Biotechnol 11(6):1497–1504Google Scholar
  8. Kataki MS, Kakoti BB, Bhuyan B, Rajkumari A, Rajak P (2014) Garden rue inhibits the arachidonic acid pathway, scavenges free radicals, and elevates FRAP: role in inflammation. Chin J Nat Med 12:172–179PubMedGoogle Scholar
  9. Massot B, Milesi S, Gontier E, Bourgaud F, Guckert A (2000) Optimized culture conditions for the production of furanocoumarins by micropropagated shoots of Ruta graveolens. Plant Cell Tissue Organ Cult 62:11–19CrossRefGoogle Scholar
  10. Ratheesh M, Sindhu G, Helen A (2013) Anti-inflammatory effect of quinoline alkaloid skimmianine isolated from Ruta graveolens L. Inflamm Res 62:367–376CrossRefGoogle Scholar
  11. Sallal AJ, Alkofahi A (1996) Inhibition of the hemolytic activities of snake and scorpion venoms in vitro with plant extracts. Biomed Lett 53:211–215Google Scholar
  12. Shabana MM, El-Alfy TS, El-Tantawy ME, Ibrahim AI, Ibrahim GF (2002) Tissue culture and evaluation of some active constituents of Ruta graveolens L. Arab J Biotechnol 5(1):45–56Google Scholar

Copyright information

© Springer Nature Singapore Pte Ltd. 2020

Authors and Affiliations

  • Malik Aabid Hussain
    • 1
  • Varsha Nitin Nathar
    • 1
  1. 1.Plant Tissue Culture Laboratory, Department of BotanySant Gadge Baba Amravati UniversityAmravatiIndia

Personalised recommendations