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In vitro plant regeneration through direct organogenesis in Populus deltoides clone G48 from petiole explants

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Abstract

A rapid and efficient protocol is developed for in vitro plantlet regeneration of Populus deltoides clone G48 using petiole explants. The highest frequency of shoot regeneration (74.75%) from petiole was obtained on MS medium supplemented with 0.50 mg/l BAP and 0.20 mg/l IAA. The regenerated shoots started turning brown and necrotic after 10–15 days in culture. To overcome the browning problem, the explants along with the developing shoot buds were transferred to modified MS medium containing 0.50 mg/l BAP, 0.20 mg/l IAA, 15 mg/l AdS, 0.1% PVP, 100 mg/l casein hydrolysate, 50 mg/l L-glutamine, 250 mg/l (NH4)2SO4 and 0.5% agar. Shoot multiplication and elongation took place on the same medium. Indole-3-acetic acid at 0.10 mg/l was most effective for root regeneration. Using the current protocol, it took 2 months to regenerate plantlets. The in vitro regenerated plantlets were successfully acclimatized and established in greenhouse conditions. This regeneration system using petiole explants provides a foundation for Agrobacterium-mediated genetic transformation of P. deltoides clone G48 for incorporation of various silviculturally important traits.

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Abbreviations

AdS:

Adenine sulphate

BAP:

6-Benzylaminopurine

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

Kn:

Kinetin

MS:

Murashige and Skoog’s medium

NAA:

Naphthaleneacetic acid

PVP:

Polyvinylpyrrolidone

TDZ:

Thidiazuron

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Acknowledgements

The authors are thankful to Professor and Head, Department of Biotechnology for providing all the facilities to carry out the research work. The assistance in providing P. deltoides clone G48 plant material by the Head, Department of Tree Improvement and Genetic Resources is highly acknowledged.

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  1. Ajay Kumar Thakur

    Present address: Directorate of Rapeseed-Mustard Research, Bharatpur, 321 303, Rajasthan, India

Authors and Affiliations

  1. Department of Biotechnology, Dr. Yashwant Singh Parmar University of Horticulture & Forestry, Solan, H.P., 173 230, India

    Ajay Kumar Thakur, A. Saraswat & D. K. Srivastava

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  1. Ajay Kumar Thakur
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  2. A. Saraswat
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  3. D. K. Srivastava
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Correspondence to Ajay Kumar Thakur.

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Thakur, A.K., Saraswat, A. & Srivastava, D.K. In vitro plant regeneration through direct organogenesis in Populus deltoides clone G48 from petiole explants. J. Plant Biochem. Biotechnol. 21, 23–29 (2012). https://doi.org/10.1007/s13562-011-0067-0

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  • DOI: https://doi.org/10.1007/s13562-011-0067-0

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