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Isolated microspore culture of chickpea (Cicer arietinum L.): induction of androgenesis and cytological analysis of early haploid divisions

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Abstract

Routine production of haploid plants has not been reported for any legume, despite the major role these species play in sustainable farming systems and human nutrition. It is within this context that we report a protocol for the induction of haploid development in chickpea (Cicer arietinum L.) using isolated microspore culture. The cultivars “Rupali”, “Narayen”, and “Kimberley Large” were identified as responsive to isolated microspore culture. Flower bud length and microspore developmental stage were correlated for these cultivars. Depending on the cultivar, buds 2.85–3.5 mm in length contained uninucleate microspores. Microspores from donor plants grown in winter and spring were more responsive than those grown in summer. A cold treatment (4°C) of between 24 and 48 h enhanced microspore response in winter- and spring-grown material but was not effective in summer-grown material. A medium developed by the authors was effective for microspore induction and early-stage embryo development. The addition of hormones to this medium was promotive of microspore induction in winter- and spring-grown material, but not in summer-grown material. The initial haploid division predominantly occurred via symmetrical division of the vegetative nucleus. Further research is under way to convert pro-embryos into plants.

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Acknowledgment

The authors are indebted to Ms Kylie Edwards for technical expertise and Dr Sergio Ochatt for his ongoing support, advice and collaboration. Thanks to Ms Padmaja Raman (The University of Western Australia) for statistical support and Dr Christine Davies (Tweakediting) for manuscript editing. The authors wish to acknowledge and thank their funding bodies, including the Grains Research and Development Corporation (GRDC), Canberra, Australia, Australian Research Council (ARC), Canberra, Australia, Council of Grain Grower Organisations Pty Ltd. (COGGO), South Perth, Australia and the Department of Agriculture and Food Western Australia (DAFWA), South Perth, Australia. This research was supported under Australian Research Council’s Industry Linkage Projects funding scheme (project number LP 056211).

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Authors and Affiliations

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

    Janine S. Croser & Kayley M. Usher

  2. Crop Development Centre, The University of Saskatchewan, 51 Campus Dr, Saskatoon, Canada

    Monika M. Lulsdorf & Ravinder K. Grewal

  3. Institute of Agriculture, Faculty of Natural and Agricultural Sciences, The University of Western Australia, 35 Stirling Hwy, Crawley, WA, Australia, 6009

    Kadambot H. M. Siddique

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  1. Janine S. Croser
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  2. Monika M. Lulsdorf
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  3. Ravinder K. Grewal
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  4. Kayley M. Usher
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  5. Kadambot H. M. Siddique
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Correspondence to Janine S. Croser.

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Editor: Y. Takahata

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Croser, J.S., Lulsdorf, M.M., Grewal, R.K. et al. Isolated microspore culture of chickpea (Cicer arietinum L.): induction of androgenesis and cytological analysis of early haploid divisions. In Vitro Cell.Dev.Biol.-Plant 47, 357–368 (2011). https://doi.org/10.1007/s11627-011-9346-7

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