European Journal of Forest Research

, Volume 130, Issue 2, pp 135–144 | Cite as

An efficient in vitro process for recurrent production of cloned plants of Vitex negundo L

  • Naseem AhmadEmail author
  • Mohammad Anis
Original Paper


An efficient method for cultivation of Vitex negundo L. through axillary shoot (collected from micropropagated plants) proliferation has been successfully developed, which can be employed at a commercial scale. Axillary shoot induction was most successful using nodal explants for propagation on Murashige and Skoog (MS) medium supplemented with various concentrations of single cytokinin or in various combination with auxins. We obtained the maximum percentage (97.6 ± 1.45) response with highest number (16.4 ± 0.60) of shoots per explant on MS medium supplemented with 6-benzyladenine (BA) and α-naphthalene acetic acid (NAA) at concentrations of 5.0 and 0.5 μM, respectively. Shoot regeneration frequency was optimized by manipulating pH and using various media. MS medium and pH 5.8 was found to be the optimum for maximum regeneration. Nodal explants from in vitro regenerated microshoots too developed shoots, thus making the process recurrent. Shootlets with 4–5 nodes were utilized for in vitro rooting, and best response was evaluated on MS medium supplemented with 1.0 μM indole-3-butyric acid (IBA). The well-developed micropropagated plants were acclimatized (97%) successfully within 2 weeks in soilrite and planted ex vitro in normal garden soil, where they grew well without any morphological and genetic variations. The present regeneration process not only favoured the rapid multiplication but also expressed the regeneration capability of micropropagated plants.


Cyclic protocol Micronodes Micropropagated plant Microshoots Vitex negundo 



6-Benzyl adenine


Indole-3-acetic acid


Indole-3-butyric acid


2-Isopentenyl adenine.




Plant growth regulators


Murashige and Skoog (1962) medium


α-Naphthalene acetic acid





N. Ahmad is thankful to the Department of Science and Technology (DST), Govt. of India, New Delhi, for its award of a SERC Fast Track Young Scientist Scheme (SR/FT/LS-014/2009). The authors acknowledge the Department of Science and Technology (DST) and the University Grants Commission (UGC), Govt. of India, New Delhi, for rendering assistance to the Department under DST-FIST (2005) and UGC-DRS (2009) programmes, respectively.


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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  1. 1.Plant Biotechnology Laboratory, Department of BotanyAligarh Muslim UniversityAligarhUPIndia
  2. 2.Department of Plant Production, College of Food and Agriculture SciencesKing Saud UniversityRiyadhSaudi Arabia

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