Flanders Research Institute for Agriculture, Fisheries and Food (ILVO) - Plant Sciences Unit, Melle, Belgium.
Department Plants and Crops, Ghent University, Ghent, Belgium.
Sci Rep. 2021 Feb 25;11(1):4553. doi: 10.1038/s41598-021-83970-y.
Celeriac F hybrid seed production is currently complicated due to the instability of cytoplasmic male sterile lines. To develop alternative alloplasmic CMS lines, an asymmetric protoplast fusion and hybrid screening methodology was established. Celeriac suspension cells protoplasts were used as the acceptor and carrot, coriander and white celery mesophyll protoplasts as the donor for protoplast fusion experiments. Acceptor cytoplasmic inheritance was inhibited by iodoacetamide treatment and donor nuclear genome inheritance was prevented by UV exposure. Protoplasts were selectively stained and fused using electroporation and polyethylene glycol, and candidate hybrid shoots were obtained. One chloroplast and three mitochondrial markers that could distinguish acceptor and donors organelles were used to characterize over 600 plants obtained after fusion events, without identifying any cybrid. In order to increase the testing efficiency a high number of micro plantlets were pooled and hence the presence of the carrot specific Atp1 marker in one of the pooled samples was detected. We demonstrated that fusion took place between celeriac and a carrot indicating that the creation of viable hybrids is possible although at a very low frequency. These findings open the path for new cytoplasmic hybridization and the isolation of novel CMS lines of celeriac.
芜菁甘蓝 F1 杂种种子的生产目前较为复杂，因为细胞质雄性不育系不稳定。为了开发替代的异质细胞质 CMS 系，建立了不对称原生质体融合和杂种筛选方法。芜菁甘蓝悬浮细胞原生质体作为受体，胡萝卜、芫荽和白芹叶肉原生质体作为供体进行原生质体融合实验。碘乙酰胺处理抑制受体细胞质遗传，紫外线照射防止供体核基因组遗传。使用电穿孔和聚乙二醇对原生质体进行选择性染色和融合，并获得候选杂种芽。使用能够区分受体和供体细胞器的一个叶绿体和三个线粒体标记物，对融合事件后获得的 600 多株植物进行了特征分析，未鉴定出任何胞质杂种。为了提高测试效率，将大量微型幼苗进行了混合，因此在其中一个混合样本中检测到了胡萝卜特异性的 Atp1 标记物。我们证明了芜菁甘蓝和胡萝卜之间发生了融合，表明尽管融合频率非常低，但仍有可能产生可育杂种。这些发现为新的细胞质杂交和芜菁甘蓝新型 CMS 系的分离开辟了道路。
