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Initiation and development of microspore embryogenesis and plant regeneration of Brassica nigra

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Abstract

Brassica nigra is generally regarded as a recalcitrant species for microspore culture among Brassica crops. Conditions for reliable induction of microspore embryogenesis of B. nigra were studied in this context. Flower bud length and microspore developmental stage were correlated with further embryogenesis. The optimal bud size range was 2.0–2.5 mm for the highest proportion of totipotent, late uninucleate microspore and the highest frequency of microspore embryogenesis. Treatment of a short heat shock by incubating the microspore culture at 32°C for 24 h was suitable for the microspore survival, sustained cell divisions, and further induced embryogenesis. Subsequently, the use of NLN medium with the addition of 13% sucrose and 0.1% activated charcoal (AC) provided the optimal conditions for the development of microspore-derived embryos (MDEs). The early cotyledonary (EC) stage embryos cultured on MS medium fortified with 4.6 μM zeatin (ZT) and 0.12 μM indole-3-acetic acid (IAA) resulted in the most efficient rates of plantlet regeneration. The ploidy levels of regenerated plants of B. nigra were determined by flow cytometry, revealing that 50.6% were diploid. The results enable the advancement of breeding programs and genetic studies in B. nigra.

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Acknowledgments

This work was financially supported by the Natural Science Foundation of China (31201629), Zhejiang Province for Key Agriculture Development Projects (2012C12903), Science and Technology Department of Zhejiang Province, and Zhejiang Academy of Agricultural Sciences.

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Authors and Affiliations

  1. Institute of Vegetables, Zhejiang Academy of Agricultural Sciences, Hangzhou, 310021, China

    Honghui Gu, Xiaoguang Sheng, Zhenqing Zhao, Huifang Yu & Jiansheng Wang

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  1. Honghui Gu
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  2. Xiaoguang Sheng
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  3. Zhenqing Zhao
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  4. Huifang Yu
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  5. Jiansheng Wang
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Correspondence to Honghui Gu.

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Editor: John W. Forster

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Gu, H., Sheng, X., Zhao, Z. et al. Initiation and development of microspore embryogenesis and plant regeneration of Brassica nigra . In Vitro Cell.Dev.Biol.-Plant 50, 534–540 (2014). https://doi.org/10.1007/s11627-014-9612-6

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  • DOI: https://doi.org/10.1007/s11627-014-9612-6

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