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An efficient protocol for shoot regeneration and genetic transformation of pigeonpea [Cajanus cajan (L.) Millsp.] using leaf explants

  • Genetic Transformation and Hybridization
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Abstract

A protocol for efficient plant regeneration from leaf explants of pigeonpea [Cajanus cajan (L.) Millsp.] was developed for the production of transgenic plants. Leaf explants from 4- to 5-day-old in vitro raised seedlings were most efficient in producing multiple adventitious shoots in 90% of the explants on shoot induction medium [Murashige and Skoog (MS) medium +5.0 μM benzyladenine +5.0 μM kinetin]. Shoot buds originated from the petiolar cut end of the explants and elongated rapidly on medium containing 0.58 μM gibberellic acid. Over 80% of the elongated shoots rooted well on MS medium containing 11.42 μM indole-3-acetic acid and were transplanted with 100% success. The procedure reported here is very simple, efficient and reproducible, and is applicable across diverse genotypes of pigeonpea. The usefulness of this system for further studies on the genetic transformation of pigeonpea has been demonstrated in biolistics-mediated gene transfer by using nptII and uidA as marker genes, where 50% of the selected plants showed gene integration and expression.

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Fig. 1A–K.
Fig. 2.
Fig. 3.
Fig. 4A–D.

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Abbreviations

BA :

N6-Benzyladenine

GA 3 :

Gibberellic acid

IAA :

Indole-3-acetic acid

IBA :

Indole-3-butyric acid

2-iP :

N6-[2-Isopentenyl]adenine

MS :

Murashige and Skoog medium

RIM :

Root induction medium

RT-PCR :

Reverse transcriptase polymerase chain reaction

SEM :

Shoot elongation medium

SIM :

Shoot induction medium

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Acknowledgements

We thank C.J. Mohan Reddy and D. Pandary for their excellent technical help. The financial support from DGIC, Belgium, and Andhra Pradesh-Netherlands Biotechnology Project is gratefully acknowledged.

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Authors and Affiliations

  1. Genetic Transformation Laboratory, International Crops Research Institute for the Semi-Arid Tropics, Patancheru, 502 324, Andhra Pradesh, India

    S. Dayal, M. Lavanya & K. K. Sharma

  2. Department of Botany, Osmania University, 500 006, Hyderabad, India

    S. Dayal & P. Devi

Authors
  1. S. Dayal
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  2. M. Lavanya
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  3. P. Devi
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  4. K. K. Sharma
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Correspondence to K. K. Sharma.

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Communicated by P.P. Kumar

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Dayal, S., Lavanya, M., Devi, P. et al. An efficient protocol for shoot regeneration and genetic transformation of pigeonpea [Cajanus cajan (L.) Millsp.] using leaf explants. Plant Cell Rep 21, 1072–1079 (2003). https://doi.org/10.1007/s00299-003-0620-y

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  • DOI: https://doi.org/10.1007/s00299-003-0620-y

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