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Microspore culture protocol for Indonesian Brassica oleracea

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Abstract

A microspore culture protocol for Brassica oleracea of Indonesian origin (cv. ‘Kemeh’) has been successfully established. A high number of embryos formed with high microspore density i.e. 15 × 104 cells/ml. Embryo formation was improved by using flower buds (4.5–4.6 mm in length) as explants, a temperature treatment at 30.5°C for 48 h and then transfer to 25°C continuously until embryos formed. A total of 295 embryos were obtained from 189 buds, 30% of which were abnormal (i.e. with an abnormal cotyledon or lacking hypocotyls). All normal embryos that grew and survived, 165 in total, were successfully transferred to soil and grew well in plastic bags (15 cm in diameter) containing a mixture of burned-rice husk and organic manure (1:1, v/v).

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Acknowledgments

Budi Winarto would like to express his gratitude to Dr. Jan B. M. Custers and his assistant Tjitske Rikson for their invaluable guidance and encouragement during his training on Brassica microspore culture in conjunction with transfer of haploid technology to Indonesia at Plant Research International, Wageningen, The Netherlands. Dr. Winarto is also thankful to the Agency of The Dutch Ministry of Economic Affairs and International Agricultural Centre (IAC), Wageningen, The Netherlands for financial support.

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Authors and Affiliations

  1. Indonesian Center of Horticultural Research and Development, Jl. Ragunan No 29A, Pasar Minggu, Jakarta Selatan, Indonesia

    Budi Winarto

  2. Faculty of Agriculture and Graduate School, Kagawa University, Miki-cho, Kagawa, 761-0795, Japan

    Jaime A. Teixeira da Silva

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  1. Budi Winarto
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  2. Jaime A. Teixeira da Silva
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Correspondence to Budi Winarto or Jaime A. Teixeira da Silva.

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Winarto, B., Teixeira da Silva, J.A. Microspore culture protocol for Indonesian Brassica oleracea . Plant Cell Tiss Organ Cult 107, 305–315 (2011). https://doi.org/10.1007/s11240-011-9981-z

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