Skip to main content

Advertisement

SpringerLink
Log in

Improvement of shoot morphogenesis in vitro and assessment of changes of the activity of antioxidant enzymes during acclimation of micropropagated plants of Desert Teak

  • Original Paper
  • Published:
Acta Physiologiae Plantarum Aims and scope Submit manuscript

Abstract

The aim of the study was to develop an improved and rapid regeneration system via axillary shoot proliferation for Tecomella undulata, an important endangered medicinal tree in India. The Murashige and Skoog (MS) medium augmented with different concentrations (0.1, 0.3, 0.5, 0.7, 1.0, 2.5, 5.0, 7.5 and 10.0 μM) of Thidiazuron (TDZ) was tested for the ability to induce axillary shoot development from cotyledonary node explants excised from 7-day-old sterile seedlings. Among the tried concentrations of TDZ, 0.7 μM showed optimum response in inducing maximum number (25.00 ± 2.30) of shoots and shoot length (4.06 ± 0.58 cm) after 3 weeks of incubation. The regenerated shoots when subcultured on MS medium lacking TDZ gave twofold shoot multiplication rate with maximum number (43.00 ± 2.86) of shoots per explant and longer shoot length (7.40 ± 0.34 cm) during the fourth subculture passage. Attempts were also made to study the morphogenetic effect of medium pH and sucrose concentration on axillary shoot induction and proliferation. The highest efficiency of shoot regeneration was recorded in MS medium supplemented with 0.7 μM TDZ and 3% sucrose at pH 5.8 after 3 weeks of culture. Different concentrations of Indole-3-butyric acid (IBA) were tested to determine the optimum conditions for ex vitro rooting of microshoots. The best result was accomplished with IBA (200 μM) pulse treatment given to the basal end of the microshoots for 30 min followed by their transfer in plastic cups containing soilrite and eventually established in natural soil with 80% survival rate. During acclimatization of plants, catalase (CAT) activity increased reaching maximum at 28th day after transplantation, while superoxide dismutase (SOD) activity increased reaching a maximum in the 21 days. Likewise, changes in the glutathione reductase (GR) and Ascorbate peroxidase (APX) were also detected. The observed changes reflect the plant’s capacity to develop antioxidant mechanisms during acclimatization which determine the ability to survive in oxidative stress. The standardized protocol for mass propagation of Tecomella undulata should eliminate the dependence on natural stands of plants for pharmaceutical purposes, and will also serve as a means of conservation as the species is highly overexploited.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

Abbreviations

APX:

Ascorbate peroxidase

BA:

6-Benzyladenine

CAT:

Catalase

CN:

Cotyledonary node

DDW:

Double distilled water

GR:

Glutathione reductase

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

MS:

Murashige and Skoog medium

SOD:

Superoxide dismutase

TDZ:

Thidiazuron

References

  • Aebi H (1984) Catalse in vitro methods. Methods Enzymol 105:121–126

    Article  PubMed  CAS  Google Scholar 

  • Ahmad N, Anis M (2007) Rapid clonal multiplication of a woody tree, Vitex negundo L. through axillary shoots proliferation. Agrofor Syst 71:195–200

    Article  Google Scholar 

  • Arya HC, Shekhawat NS (1986) Clonal multiplication of tree species in Thar Desert through tissue culture. Forest Ecol Manage 16:201–208

    Article  CAS  Google Scholar 

  • Arya S, Toky OP, Harris SM, Harris PJC (1992) Tecomella undulata (Rohida): a valuable tree of the Thar desert. Int Tree Crops J 7:141–147

    Article  Google Scholar 

  • Batkova P, Pospisilova J, Synkova H (2008) Production of reactive species and development of antioxidative systems during in vitro and ex vitro transfer. Biol Plant 52:413–422

    Article  CAS  Google Scholar 

  • Bhansali R (1993) Bud culture for shoot multiplication and plantlet formation of Tecomella undulata (Rohida), a woody tree of the arid zone. Trop Sci 3:31–38

    Google Scholar 

  • Bhau BS, Negi MS, Jindal SK, Singh M, Lakshmikumaran M (2007) Assessing genetic diversity of Tecomella undulata (Sm.)—an endangered tree species using amplified fragment length polymorphisms-based molecular markers. Curr Sci 93:67–72

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye-binding. Ann Biochem 72:248–254

    Article  CAS  Google Scholar 

  • Ceaser SA, Maxwell SL, Prasad KB, Karthigan M, Ignacimuthu S (2010) Highly efficient shoot regeneration of Bacopa monnieri (L.) using a two step culture procedure and assessment of genetic integrity of micropropagated plants by RAPD. Acta Physiol Plant 32:443–452

    Article  Google Scholar 

  • Corredoira E, Ballester A, Vieitez AM (2008) Thidiazuron induced high frequency plant regeneration from leaf explants of Paulownia tomentosa mature trees. Plant Cell Tiss Org Cult 95:197–208

    Article  CAS  Google Scholar 

  • Faisal M, Ahmad N, Anis M (2005) Shoot multiplication in Rauvolfia tetraphylla L. using thidiazuron. Plant Cell Tiss Org Cult 80:187–190

    Google Scholar 

  • Faisal M, Anis M (2009) Changes in photosynthetic activity, pigment composition, electrolyte leakage, lipid peroxidation and antioxidant enzymes during ex vitro establishment of micropropagated Rauvolfia tetraphylla plantlets. Plant Cell Tiss Org Cult 99:125–132

    Article  CAS  Google Scholar 

  • Faisal M, Anis M (2010) Effect of light irradiations on photosynthetic machinery and antioxidative enzymes during ex vitro acclimatization of Tylophora indica plantlets. J Plant Interact 5:21–27

    Article  CAS  Google Scholar 

  • Faisal M, Siddique I, Anis M (2006) In vitro rapid regeneration of plantlets from nodal segments of Mucuna pruriens. A valuable medicinal plant. Ann Appl Biol 148:1–6

    Article  CAS  Google Scholar 

  • Feng JC, Yu XM, Shang XL, Li JD, Wu YX (2010) Factors affecting efficiency of shoot regeneration in Ziziphus jujube Mill. ‘Huizao’. Plant Cell Tiss Org Cult 101:111–117

    Article  CAS  Google Scholar 

  • Giri CC, Shyankumar B, Anjaneyulu C (2004) Progress in tissue culture, genetic transformation and applications of biotechnology to trees: an overview. Trees 18:115–135

    Google Scholar 

  • Hazarika BN (2003) Acclimatization of tissue cultured plants. Curr Sci 85:1704–1712

    CAS  Google Scholar 

  • Husain MK, Anis M, Shahzad A (2007) In vitro propagation of Indian Kino (Pterocarpus marsupium Roxb.) using thidiazuron. In Vitro Cell Dev Biol Plant 43:59–64

    CAS  Google Scholar 

  • Huettemen CA, Preece JE (1993) Thidiazuron: a potent cytokinin for woody plant tissue culture. Plant Cell Tiss Org Cult 33:105–119

    Article  Google Scholar 

  • Jahan AA, Anis M (2009) In vitro rapid multiplication and propagation of Cardiospermum halicacabum L. through axillary bud culture. Acta Physiol Plant 31:133–138

    Article  CAS  Google Scholar 

  • Jones MPA, Yi Z, Murch SJ, Saxena PK (2007) Thidiazuron- induced regeneration of Echinacea purpurea L: micropropagation in solid and liquid culture systems. Plant Cell Rep 26:13–19

    Article  PubMed  CAS  Google Scholar 

  • Karuppanapandian T, Moon JC, Kim C, Manoharan K, Kim W (2011) Reactive oxygen species in plants: their generation, signal transduction and scavenging mechanisms. Aust J Crop Sci 5:709–725

    CAS  Google Scholar 

  • Kyle ME, Farber JL (1991) Biochemical mechanism of toxic cell injury. In: Haschek MW, Rousseavx CG (eds) Toxicological pathology. Academic Press, San Diego, pp 81–85

    Google Scholar 

  • Ludwig-Muller J (2000) Indole-3-butyric acid in plant growth and development. Plant Growth Regul 32:219–230

    Article  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Naik PM, Manohar SH, Praveen N, Murthy HN (2010) Effects of sucrose and pH levels on in vitro shoot regeneration from leaf explants of Bacopa monnieri and accumulation of bacoside A in regenerated shoots. Plant Cell Tiss Org Cult 100:235–239

    Article  Google Scholar 

  • Nair LG, Seeni S (2003) In vitro multiplication of Calophyllum apetalum (Clusiaceae), an endemic medicinal tree of the western ghats. Plant Cell Tiss Org Cult 75:169–174

    Article  CAS  Google Scholar 

  • Nandwani D, Mathur N, Ramawat KG (1995) In vitro shoot multiplication from cotyledonary node explants of Tecomella undulata. Gartenbauwisseschaft 60:65–68

    CAS  Google Scholar 

  • Nandwani D, Sharma R, Ramawat KG (1996) High frequency regeneration in callus culture of tree Tecomella undulata. Gartenbauwisseschaft 61:147–150

    CAS  Google Scholar 

  • Parveen S, Shahzad A (2011) A micropropagation protocol for Cassia angustifolia Vahl. from root explants. Acta Physiol Plant 33:789–796

    Article  Google Scholar 

  • Raghu AV, Geetha SP, Martin G, Balachandran I, Ravindran PN, Mohanan KV (2007) An improved micropropagation protocol for Bael—a vulnerable medicinal tree. Res J Bot 2:186–194

    Article  CAS  Google Scholar 

  • Rathore TS, Singh RP, Shekhawat NS (1991) Clonal propagation of desert tree (Tecomella undulata) through tissue culture. Plant Sci 79:217–222

    Article  CAS  Google Scholar 

  • Scandalios G (1990) Response of plant antioxidant defense genes to environmental stress. Adv Genet 28:1–41

    Article  PubMed  CAS  Google Scholar 

  • Sha Valli Khan PS, Hausman JF, Rao KR (1999) Clonal multiplication of Syzygium alternifolium (Wight.) Walp through mature nodal segments. Silvae Genet 48:45–50

    Google Scholar 

  • Siddique I, Anis M (2007) High frequency shoot regeneration and plantlet formation in Cassia angustifolia (Vahl.) using thidiazuron. Med Aromat Plant Sci Biotechnol 1:282–284

    Google Scholar 

  • Siddique I, Anis M, Jahan AA (2006) Rapid multiplication of Nyctanthes arbor-tristis L. through in vitro axillary shoot proliferation. World J Agricult Sci 2:188–192

    Google Scholar 

  • Taylor JLS, Van Staden J (2001) The effect of nitrogen and sucrose concentration on the growth of Eucomis autumnalis (Mill.) Chitt. plantlets in vitro and on subsequent anti-inflammatory activity in extracts prepared from the plantlets. Plant Growth Regul 34:49–56

    Article  CAS  Google Scholar 

  • Willekins H, Inze D, Van Montagu M, Van Camp W (1995) Catalases in plants. Mol Breed 1:207–228

    Article  Google Scholar 

  • Xu J, Yuzhen W, Zhang Y, Chai T (2008) Rapid in vitro multiplication and ex vitro rooting of Malus zumi (Matsumura) Rehd. Acta Physiol Plant 30:129–232

    Article  CAS  Google Scholar 

Download references

Acknowledgments

Authors gratefully acknowledge the Department of Science and Technology, and the University Grant Commission, Govt. of India, New Delhi for providing research support under DST-FIST (2005) and UGC-SAP DRS-I (2009) Programs, respectively.

Author information

Authors and Affiliations

  1. Plant Biotechnology Laboratory, Department of Botany, Aligarh Muslim University, Aligarh, 202 002, India

    Ankita Varshney & Mohammad Anis

  2. Department of Plant Production, College of Food & Agricultural Sciences, King Saud University, P.O. Box 2460, Riyadh, 11451, Saudi Arabia

    Mohammad Anis

Authors
  1. Ankita Varshney
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mohammad Anis
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mohammad Anis.

Additional information

Communicated by B. Borkowska.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Varshney, A., Anis, M. Improvement of shoot morphogenesis in vitro and assessment of changes of the activity of antioxidant enzymes during acclimation of micropropagated plants of Desert Teak. Acta Physiol Plant 34, 859–867 (2012). https://doi.org/10.1007/s11738-011-0883-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11738-011-0883-9

Keywords

Search

Navigation