大麦不同基因型游离小孢子的直接培养再生及培养体系的优化

Plant Regeneration and Optimization of Culture System in Direct Culture of Isolated Microspores from Different Genotypes of Barley

以微搅拌法建立了小孢子直接游离的预处理和培养程序。在大田生长的４个对培养反应不同的大麦基因型上，以新鲜幼穗游离小孢子进行直接培养，均成功地诱导了胚状体并获得再生绿色植株。小孢子的发育进程说明，直接游离的小泡子在预处理过程中的发育要慢于在花药中预处理的小孢子，而且其培养效率也较低。直接游离小孢子的培养密度以０．８～１．０×１０５／ｍｌ较理想，至少应不低于６×１０４／ｍｌ．８％－１０％的糖浓度可明显提高小孢子分裂频率和胚状体诱导频率。实验结果也表明两种培养基ＦＨＧ和ＭＮ６无明显差异，均适宜于直接游离的小孢子培养。

Using four barley genotypes responses in anther culture, embryo - like structures (ELS) and green which were grown in field and showed different plants were successfully obtained through direct culture of the microspores freshly - isolated from the spikes by microblending. The developmental course showed that during pretreatment, freshly - isolated microspores developed more slowly than the microspores pretreated in anthers, and the culture efficiency was also inferior to the latter. The microspore density of 0.8- 1.0x 105 per ml was optimal and 0.6x 105 per ml was minimal for direct culture. lt was found that 8%- 10% maltose concentration increased microspore division and ELS development. The results indicated that the media of FHG and MN6 showed no significant difference in culture response and were all suitable for direct culture. The significance of direct culture of microspores in theory and application was discussed.