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Doubled-haploid production in chickpea (Cicer arietinum L.): role of stress treatments

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An Erratum to this article was published on 01 April 2010

Abstract

This is the first report on the production of double-haploid chickpea embryos and regenerated plants through anther culture using Canadian cultivar CDC Xena (kabuli) and Australian cultivar Sonali (desi). Maximum anther induction rates were 69% for Sonali and 63% for CDC Xena. Under optimal conditions, embryo formation occurred within 15–20 days of culture initiation with 2.3 embryos produced per anther for CDC Xena and 2.0 embryos per anther for Sonali. For anther induction, the following stress treatments were used: (1) flower clusters were treated at 4°C for 4 days, (2) anthers were subjected to electric shock treatment of three exponentially decaying pulses of 50–400 V with 25 μF capacitance and 25 Ω resistance, (3) anthers were centrifuged at 168–1,509g for 2–15 min, and finally (4) anthers were cultured for 4 days in high-osmotic pressure (563 mmol) liquid medium. Anthers were then transferred to a solid embryo development medium and, 15–20 days later, embryo development was observed concomitant with a small amount of callus growth of 0.1–3 mm. Anther-derived embryos were regenerated on plant regeneration medium. Electroporation treatment of anthers enhanced root formation, which is often a major hurdle in legume regeneration protocols. Cytological studies using DAPI staining showed a wide range of ploidy levels from haploid to tetraploid in 10–30-day-old calli. Flow cytometric analysis of calli, embryos and regenerated plants showed haploid profiles and/or spontaneous doubling of the chromosomes during early regeneration stages.

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Abbreviations

CDC:

Crop Development Centre

CLIMA:

Centre for Legumes in Mediterranean Agriculture

DAFWA:

Department of Agriculture and Food in Western Australia

ABA:

Abscisic acid

BAP:

6-Benzyl aminopurine

DAPI:

4, 6-Diamidino-2-phenylindole

2, 4-D:

2, 4-Dichlorophenoxy acetic acid

GA3 :

Gibberellic acid

2iP:

6-γ, γ Dimethylallyl-amino purine

IAA:

Indole acetic acid

NAA:

1-Naphthalene acetic acid

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Acknowledgments

We would like to thank Dr. Alison Ferrie (PBI/NRCC) for use of the flow cytometer. We are grateful to Tim Dament for his technical support. We also appreciate Dr. Tajinder Grewal’s and Lasantha Ubeyseana’s help with the statistical analyses. This work was supported by Saskatchewan Agricultural Development Fund Project No. 20050723.

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Authors and Affiliations

  1. Crop Development Centre, University of Saskatchewan, 51 Campus Drive, Saskatoon, SK, S7N 5A8, Canada

    Ravinder Kaur Grewal, Monika Lulsdorf, Albert Vandenberg & Thomas D. Warkentin

  2. Centre for Legumes in Mediterranean Agriculture, The University of Western Australia, 35 Stirling Hwy, Crawley, WA, 6009, Australia

    Janine Croser

  3. Institut National de la Recherche Agronomique, B.P. 86510, 21065, Dijon Cedex, France

    Sergio Ochatt

Authors
  1. Ravinder Kaur Grewal
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  2. Monika Lulsdorf
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  3. Janine Croser
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  4. Sergio Ochatt
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  5. Albert Vandenberg
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  6. Thomas D. Warkentin
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Corresponding author

Correspondence to Ravinder Kaur Grewal.

Additional information

Communicated by M. Jordan.

An erratum to this article can be found at http://dx.doi.org/10.1007/s00299-010-0844-6

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Grewal, R.K., Lulsdorf, M., Croser, J. et al. Doubled-haploid production in chickpea (Cicer arietinum L.): role of stress treatments. Plant Cell Rep 28, 1289–1299 (2009). https://doi.org/10.1007/s00299-009-0731-1

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  • DOI: https://doi.org/10.1007/s00299-009-0731-1

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