Li Haonan, Wang Zhe, Zhou Weina, Li Jinyan, Zhao Zifan, Zheng Chao, Liu Zhiyong, Zhang Yun
College of Horticulture, Shenyang Agricultural University, Shenhe District, 120 Dongling Road, Shenyang, China.
Protoplasma. 2025 Jul;262(4):801-813. doi: 10.1007/s00709-025-02032-6. Epub 2025 Jan 22.
Microspore culture is an efficient and rapid method that produces doubled haploid (DH) lines for hybrid breeding in crops and vegetables. However, the low frequency of microspore embryogenesis and spontaneous diploidization in Chinese kale still require improvement. In the present work, an efficient microspore culture protocol was constructed and used for DH producing in Chinese kale breeding. The results showed that kinetin (KT) and methylene blue (MB) improved microspore embryogenesis and embryo development. The spontaneous diploidization rates were below 14.48%. Therefore, artificial doubling was used for haploids; the doubled haploid rate was significantly increased, and the highest doubling rate reached 33.75% with pendimethalin. 98 DH lines were obtained in this study. This study also identified the horticultural characters of the DH lines, which showed significant differences between them. All DH lines were self-incompatible. The hybrid crosses were performed using five DH lines selected for the Chinese kale breeding. This study was helpful to accelerate the application of microspore culture and provide DH lines for Chinese kale hybrid breeding.
小孢子培养是一种高效快速的方法，可用于培育作物和蔬菜杂交育种所需的双单倍体（DH）系。然而，芥蓝小孢子胚胎发生频率低和自发二倍体化现象仍有待改善。在本研究中，构建了一种高效的小孢子培养方案，并用于芥蓝育种中双单倍体的产生。结果表明，激动素（KT）和亚甲基蓝（MB）可促进小孢子胚胎发生和胚胎发育。自发二倍体化率低于14.48%。因此，采用人工加倍处理单倍体；双单倍体率显著提高，使用二甲戊灵时最高加倍率达到33.75%。本研究获得了98个DH系。本研究还鉴定了DH系的园艺性状，各系之间表现出显著差异。所有DH系均表现为自交不亲和。利用筛选出的5个DH系进行了芥蓝杂交育种。本研究有助于加速小孢子培养的应用，并为芥蓝杂交育种提供DH系。
