结球甘蓝(Brassica oleracea var. capitata)和青花菜(Brassica oleracea var. italica)小孢子胚再生植株频率低是目前影响游离小孢子培养技术有效应用的关键问题之一, 研究其小孢子胚植株再生频率的影响因素, 提高胚再生植株频率, 对促进游离小孢子培养技术在甘蓝类蔬菜育种中更好地应用具有重要意义。该文以结球甘蓝中甘11和青花菜TI-111等基因型为试材, 对影响游离小孢子胚再生成植株的固体培养基类型、琼脂浓度、胚的类型及胚在液体培养基中的滞留时间等因素进行了研究。结果表明: 游离小孢子培养25天的子叶胚在琼脂浓度为1%–1.25%的B5培养基上植株再生频率最高。进一步通过8个不同基因型对上述实验结果进行了验证, 结果显示, 游离小孢子培养25天的子叶胚在1%琼脂浓度的B5培养基上植株再生频率达77.8%–97.2%。
The application of microspore culture still produces low frequency of plant regeneration from microspore-derived embryos in many cabbage and broccoli genotypes. To improve the frequency of plant regeneration, we studied the effect of medium, agar concentration, embryoid type and days of culture of microspores on plant regeneration in hybrid F1 generations of a cabbage, ‘Zhonggan 11’, and a broccoli, ‘TI-111’. B5 medium with 1%–1.25% agar was optimal for germination and regeneration of embryos, and the frequency of plant regeneration from cotyledonary embryos was significantly higher than that from other embryos, especially cotyledonary embryos from the 25-day-cultured microspores. The frequency of plant regeneration for cotyledonary embryos from 25-day-cultured microspores in 8 cultivars of cabbage and broccoli transferred to B5 medium with 1% agar ranged from 77.8% to 97.2%.