, Volume 195, Issue 3, pp 467–475 | Cite as

Efficient doubled haploid production in microspore culture of loose-curd cauliflower (Brassica oleracea var. botrytis)

  • Honghui Gu
  • Zhengqing Zhao
  • Xiaoguang Sheng
  • Huifang Yu
  • Jiansheng Wang


We present an improved protocol for highly efficient production of doubled haploid loose-curd cauliflower plants (Brassica oleracea var. botrytis) via microspore culture. Our experiment explored factors such as donor plant treatment, flower bud pretreatment, embryo germination medium, and ploidy characterization of regenerated plants. Our technique efficiently produced embryos from both tight- and loose-curd donor plants, although the embryo yields were genotype dependent. We achieved a germination rate of around 30 % by employing a hormone combination of zeatin, indole-3-acetic acid, and 6-benzylaminopurine pretreatment culture. We also used 1–4 days of cold pretreatment of the flower buds, which were submerged into NLN-13 medium, to induce microspore embryogenesis. Analysis using an FCM Ploidy Analyzer showed that more than 50 % of regenerated plants were spontaneously doubled haploids, more than 25 % were tetraploids, and fewer than 7 % were haploid. Visual examination of plants in the field revealed that they had distinct phenotypic characteristics relating to their ploidy level. The efficient production of double haploids using our improved microspore culture technique is a promising approach that can be applied in loose-curd cauliflower breeding programmes and genetic research.


Loose-curd cauliflower (Brassica oleracea var. botrytisMicrospore culture Doubled haploid Spontaneous doubling Cold pretreatment 





Doubled haploid(s)


Petal length/Anther length

NLN medium

Nitsch and Nitsch medium

MS medium

Murashige and Skoog medium


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Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Honghui Gu
    • 1
  • Zhengqing Zhao
    • 1
  • Xiaoguang Sheng
    • 1
  • Huifang Yu
    • 1
  • Jiansheng Wang
    • 1
  1. 1.Institute of VegetablesZhejiang Academy of Agricultural SciencesHangzhouChina

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