Plant Growth Regulation

, Volume 47, Issue 2–3, pp 121–127 | Cite as

Gradual Depletion of 2,4-D in the Culture Medium for Indirect Shoot Regeneration from Leaf Explants of Camellia sinensis (L.) O. Kuntze

  • Indra Sandal
  • Ajay Kumar
  • Amita Bhattacharya
  • Madhu Sharma
  • Adarsh Shanker
  • Paramvir Singh Ahuja


Adventitious shoot regeneration via callus phase from in vitro leaf explants is reported for the first time in tea. Callus was obtained on Murashige and Skoog medium supplemented with varied concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) (2.5, 5.0, 7.5 and 10.0 mg/l). Rhizogenesis was observed at all concentrations of 2,4-D. Adventitious shoot buds developed indirectly on leaf explants after prolonged culture for 16 weeks on medium supplemented with 10.0 mg/l 2,4-D. GC analysis of the medium and the tissues at different stages of development showed that specific levels of 2,4-D in the tissue were responsible for morphogenesis. Shoot buds developed on rhizogenic calli, only when 2,4-D declined to undetectable or negligible concentrations in the tissue probably due to detoxification and metabolism. Alternatively, shoot buds could also be evoked when rhizogenic calli were transferred to medium supplemented with low concentration of 2,4-D (1.5 mg/l). The adventitious nature of the shoots was confirmed through histological studies.

Key words

Callus Camellia sinensis 2,4-D Leaf explant Shoot regeneration 



2,4-dichlorophenoxyacetic acid




indole-3-acetic acid


indole-3-butyric acid


Murashige and Skoog (1962) medium


naphthalene acetic acid


thidiazuron (N-phenyl-N-1,2,3-thiadiazol-5-yl urea)


plant growth regulator


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Copyright information

© Springer 2005

Authors and Affiliations

  • Indra Sandal
    • 1
  • Ajay Kumar
    • 1
  • Amita Bhattacharya
    • 1
  • Madhu Sharma
    • 1
  • Adarsh Shanker
    • 1
  • Paramvir Singh Ahuja
    • 1
  1. 1.Division of BiotechnologyInstitute of Himalayan Bioresource TechnologyPalampurIndia

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