Physiology and Molecular Biology of Plants

, Volume 21, Issue 3, pp 407–415 | Cite as

A high-frequency in vitro multiplication, micromorphological studies and ex vitro rooting of Cadaba fruticosa (L.) Druce (Bahuguni): a multipurpose endangered medicinal shrub

  • Deepika Lodha
  • Ashok Kumar Patel
  • N. S. Shekhawat
Research Article


An efficient and reproducible in vitro propagation protocol has been established for Cadaba fruticosa (L.) Druce. Surface-sterilized nodal stem segments of mature plant were used as explants for culture establishment. Multiple shoots were optimally differentiated from the nodal stem explants through bud breaking on Murashige and Skoog (1962) medium containing 3.0 mg l−1 benzyladenine (BA). The effect of different plant growth regulators and minerals were studied on different stages of micropropagation procedure (i.e., explant establishment, shoot multiplication/growth and ex vitro rooting). Additionally, for enhancing shoot multiplication during subculture, MS medium was modified (MMS) with higher levels of magnesium, potassium and sulphate ions. Out of these, MMS3 medium containing 0.25 mg l−1 each of BA and Kin (N6-furfuryladenine), with 0.1 mg l−1 NAA (α-naphthalene acetic acid) was found the best for shoot multiplication (42.45 ± 3.82 per culture vessel). The in vitro regenerated shoots were rooted under ex vitro conditions on treating the shoot base with 500 mg l−1 of IBA (indole-3 butyric acid) for 3 min on sterile Soilrite®. The ex vitro rooted plants were hardened in the greenhouse and transferred to the field with ≈85 % survival rate. There were not any visual differences between wild and micropropagated plants in the field, although the later underwent significant changes during acclimatization. Micromorphological changes on leaf surface characters from in vitro to acclimatized plantlets were studied in terms of development of glandular trichomes, changes in vein spacing and vein structure in order to understand the nature of plant responses towards environmental conditions. The method developed and defined can be applied for commercial cultivation, which may be important for extraction of bioactive compounds and may facilitate conservation of this multipurpose endangered medicinal shrub.


Cadaba fruticosa Cadabicine Endangered plant Ex vitro rooting Micropropagation Modified MS medium 





Indole-3-butyric acid


N6–(2-isopentenyl) adenine


Kinetin (N6-furfuryladenine)


Murashige and Skoog (1962) medium


Modified MS medium


α-naphthalene acetic acid


Sodium hydroxide


Naphthoxyacetic acid


Photon Flux Density


Plant Growth Regulators


Relative Humidity



DL gratefully acknowledges the financial support from Council of Scientific and Industrial Research (CSIR), New Delhi in the form of Junior and Senior Research Fellowship (JRF-SRF). AKP is thankful to the University Grant Commission (UGC), New Delhi, for providing Special Assistance Program (SAP) in the form of Centre for Advanced Study (CAS) to the Department of Botany, Jai Narain Vyas University, Jodhpur (INDIA).

Author’s contribution

D. Lodha designed and performed the experiments and wrote the first draft of the manuscript. A. K. Patel helped in data analysis and organized it in figures and tables. N. S. Shekhawat guided the research and edited the final version of the manuscript.


  1. Abbas H, Qaiser M (2010) In vitro conservation of Cadaba heterotricha stocks, an endangered species in Pakistan. Pak J Bot 42:1553–1559Google Scholar
  2. Abbas H, Qaiser M, Alam J (2010) Conservation status of Cadaba heterotricha Stocks (Capparaceae): an endangered species in Pakistan. Pak J Bot 42:35–46Google Scholar
  3. Aluri RJS (1990) Studies on pollination in India: a review. Proc Natl Acad Sci India 56:375–388Google Scholar
  4. Amudha M, Rani S (2014) Assessing the bioactive constituents of Cadaba fruticosa (L.) Druce through GC-MS. Int J Pharm Pharm Sci 6:383–385Google Scholar
  5. Anonymous (2005) The wealth of India- publication and information directorate, Council of Scientific and Industrial Research, Volume-I A, New DelhiGoogle Scholar
  6. Aranda-Peres AN, Martinelli AP (2009) Adjustment of mineral elements in the culture medium for the micropropagation of three Vriesea bromeliads from the Brazilian Atlantic Forest: the importance of calcium. Hortic Sci 44:106–112Google Scholar
  7. Aziz-Ur-Rehman (1990) Studies in chemical constituents of Cadaba fruticosa. Pakistan Research Repository, PhD thesis, University of Karachi, KarachiGoogle Scholar
  8. Bhandari MM (1990) Flora of the Indian Desert. In: MPS Reports, Jodhpur, IndiaGoogle Scholar
  9. Compton ME (1994) Statistical methods suitable for the analysis of plant tissue culture data. Plant Cell Tissue Organ Cult 37:217–242Google Scholar
  10. Deora NS, Shekhawat NS (1995) Micropropagation of Capparis decidua (Forsk.) Edgew.-a tree of arid horticulture. Plant Cell Rep 15:278–281PubMedGoogle Scholar
  11. ENVIS (2014) The Environmental Information System, Ministry of Environment & Forests, India (; accessed on 13th April 2015)
  12. Gashi B, Abdullai K, Sota V, Kongjika E (2015) Micropropagation and in vitro conservation of the rare and threatened plants Ramonda serbica and Ramonda nathaliae. Physiol Mol Biol Plant 21:123–136CrossRefGoogle Scholar
  13. Gaspar TH, Kevers C, Faivre-Rampant O, Crevecoeur M, Penel C, Greppin H, Dommes J (2003) Changing concepts in plant hormone action. In Vitro Cell Dev Biol Plant 39:85–106CrossRefGoogle Scholar
  14. Gunasekaran M, Balasubramanian P (2012) Ethnomedicinal uses of Sthalavrikshas (temple trees) in Tamil Nadu, Southern India. Ethnobot Res Appl 10:253–268Google Scholar
  15. Hermans C, Johnson GN, Strasser RJ, Verbruggen N (2004) Physiological characterisation of magnesium deficiency in sugar beet: acclimation to low magnesium differentially affects photosystems I and II. Planta 220:344–355PubMedCrossRefGoogle Scholar
  16. Lodha D, Patel AK, Rai MK, Shekhawat NS (2014a) In vitro plantlet regeneration and assessment of alkaloid contents from callus cultures of Ephedra foliata (Unth phog), a source of anti-asthmatic drugs. Acta Physiol Plant 36:3071–3079CrossRefGoogle Scholar
  17. Lodha D, Rathore N, Kataria V, Shekhawat NS (2014b) In vitro propagation of female Ephedra foliata Boiss. & Kotschy ex Boiss.: an endemic and threatened Gymnosperm of the Thar Desert. Physiol Mol Biol Plant 20:375–383CrossRefGoogle Scholar
  18. Ludwig-Muller J (2000) Indole-3-butyric acid in plant growth and development. Plant Growth Regul 32:219–230CrossRefGoogle Scholar
  19. Mendonça D, Luna S, Bettencourt S, Lopes MS, Monteiro L, Neves JD, Monjardino P, Machado AC (2015) In vitro propagation of Picconia azorica (Tutin) Knobl. (Oleaceae) an Azorean endangered endemic plant species. Acta Physiol Plant 37:47CrossRefGoogle Scholar
  20. MOE (2012) The national red list 2012 of Sri Lanka; conservation status of the fauna and flora. Ministry of Environment, ColomboGoogle Scholar
  21. Murashige T, Skoog FA (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  22. Mythreyi R, Sasikala E, Geetha A, Madhavan V (2008) Antipyretic activity of leaves of Cadaba fruticosa (L.) Druce. Pharmacol Online 3:136–142Google Scholar
  23. Nas MN, Read PE (2004) A hypothesis for the development of a defined tissue culture medium of higher plants and micropropagation of hazelnuts. Sci Hortic 101:189–200CrossRefGoogle Scholar
  24. Osório ML, Gonçalves S, Coelho N, Osório J, Romano A (2013) Morphological, physiological and oxidative stress markers during acclimatization and field transfer of micropropagated Tuberaria major plants. Plant Cell Tissue Organ Cult 115:85–97CrossRefGoogle Scholar
  25. Patel AK, Phulwaria M, Rai MK, Gupta AK, Shekhawat S, Shekhawat NS (2014) In vitro propagation and ex vitro rooting of Caralluma edulis (Edgew.) Benth. & Hook. f.: an endemic and endangered edible plant species of the Thar Desert. Sci Hortic 165:175–180CrossRefGoogle Scholar
  26. Pospóšilová J, Tichá I, Kadleček P, Haisel D, Plzáková Š (1999) Acclimatization of micropropagated plants to ex vitro conditions. Biol Plant 42:481–497CrossRefGoogle Scholar
  27. Preece J (1995) Can nutrient salts partially substitute for plant growth regulators? Plant Tissue Cult Biotechnol 1:26–37Google Scholar
  28. Preece JE (2010) Acclimatization of plantlets from in vitro to the ambient environment. Encycl Ind Biotechnol 1–9Google Scholar
  29. Ramage CM, Williams RR (2002) Mineral nutrition and plant morphogenesis. In Vitro Cell Dev Biol Plant 38:116–124CrossRefGoogle Scholar
  30. Rathore JS, Rathore MS, Shekhawat NS (2005) Micropropagation of Maerua oblongifolia- a liana of arid areas. Phytomorphology 55:241–247Google Scholar
  31. Rathore NS, Rathore N, Shekhawat NS (2013) In vitro propagation and micromorphological studies of Cleome gynandra: a C4 model plant closely related to Arabidopsis thaliana. Acta Physiol Plant 35:2691–2698CrossRefGoogle Scholar
  32. Sánchez MC, San-José MC, Ferro E, Ballester A, Vieitez AM (1997) Improving micropropagation conditions for adult-phase shoots of chestnut. J Hortic Sci 72:433–443Google Scholar
  33. Shashikanth J, Mohan CH, Reddy PR (2014) A potent folklore medicinal plant: Cadaba fruticosa (L.) Druce. Res Rev J Bot Sci 3:30–33Google Scholar
  34. Vengadesan G, Pijut PM (2009) In vitro propagation of northern red oak (Quercus rubra L.). In Vitro Cell Dev Biol Plant 45:474–482CrossRefGoogle Scholar
  35. Vibha JB, Shekhawat NS, Mehandru P, Dinesh R (2014) Rapid multiplication of Dalbergia sissoo Roxb.: a timber yielding tree legume through axillary shoot proliferation and ex vitro rooting. Physiol Mol Biol Plant 20:81–87CrossRefGoogle Scholar
  36. Viqar Uddin A, Kaniz F, Azia-ur-Rahman A, Shoib A (1987) Cadabacine and cadabacine diacetate from Crataeva nurvala and Cadaba farinosa. J Nat Prod 50:1186Google Scholar
  37. Wada S, Niedz RP, DeNoma J, Reed BM (2013) Mesos components (CaCl2, MgSO4, KH2PO4) are critical for improving pear micropropagation. In Vitro Cell Dev Biol Plant 49:356–365CrossRefGoogle Scholar
  38. Zhang A, Wang H, Shao Q, Xu M, Zhang W, Li M (2015) Large scale in vitro propagation of Anoectochilus roxburghii for commercial application: pharmaceutically important and ornamental plant. Ind Crop Prod 70:158–162Google Scholar

Copyright information

© Prof. H.S. Srivastava Foundation for Science and Society 2015

Authors and Affiliations

  • Deepika Lodha
    • 1
  • Ashok Kumar Patel
    • 1
  • N. S. Shekhawat
    • 1
  1. 1.Biotechnology Unit, Department of Botany (UGC – Centre for Advanced Study)Jai Narain Vyas UniversityJodhpurIndia

Personalised recommendations