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Doubled haploid production in rocket (Eruca sativa Mill.) through isolated microspore culture

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Abstract

Haploid induction in rocket (Eruca sativa Mill.), a novel and increasingly important vegetable, was studied in microspore culture. A procedure based on a high sucrose NLN medium and heat shock treatment resulted in nuclear divisions and embryo induction. The effect of genotype both among seed lots and among single plants was a major factor influencing embryo formation. The addition of activated charcoal was essential for obtaining reproducible results, 0.2 mg l−1 being superior to 1.0 mg l−1. A 24 h heat shock treatment at 32°C doubled the embryogenic response compared to a 48 h treatment. Embryo conversion was only efficient (23%) for embryos that had been cultured on medium with activated charcoal and subcultured on solid B5 medium; pretreatment of embryos with ABA or desiccation for 1–3 weeks inhibited embryo conversion. Analysis of ploidy level revealed that the majority (65.6%) of 489 regenerated plantlets tested were diploid. Breeding programs and genetic studies of rocket are likely to benefit substantially from the established method.

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Abbreviations

ABA:

Abscisic acid

AC:

Activated charcoal

DAPI:

4′,6-Diamidino-2-phenylindole

MTT:

3-4,5 Dimethylthiazol-2-yl-2,5 diphenyl-tetrazol bromide

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Acknowledgements

The authors thank Dr. JBM Custers (Plant Research International, Wageningen, NL) for his generous help with initial stages of protocol establishment and critically reading the manuscript. This work was supported by Slovenian Research project P4-0077.

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Authors and Affiliations

  1. Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1000, Ljubljana, Slovenia

    Lucija Leskovšek, Marijana Jakše & Borut Bohanec

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  1. Lucija Leskovšek
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  2. Marijana Jakše
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  3. Borut Bohanec
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Correspondence to Borut Bohanec.

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Leskovšek, L., Jakše, M. & Bohanec, B. Doubled haploid production in rocket (Eruca sativa Mill.) through isolated microspore culture. Plant Cell Tiss Organ Cult 93, 181–189 (2008). https://doi.org/10.1007/s11240-008-9359-z

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  • DOI: https://doi.org/10.1007/s11240-008-9359-z

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