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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,并对游离小孢子直接培养在理论和应用上的意义进行了讨论
The microspore direct free pretreatment and culture program was established by micro-mixing method. In the four barley genotypes with different response to culture, all the cultivars were directly cultured with fresh spikelet-free microspores, and embryoid bodies were successfully induced and regenerated green plants were obtained. The development of microspores showed that the direct free vesicles developed more slowly during pretreatment than the microspores pretreated in anther, and their culture efficiency was also lower. Direct free microspore culture density of 0.8 ~ 1.0 × 105 / ml is more desirable, at least not less than 6 × 104 / ml. 8% -10% sugar concentration can significantly improve the microsporocyte division frequency and embryoid body induction frequency. The experimental results also showed that there was no significant difference between the two medium FHG and MN6, which is suitable for the direct free microspore culture, and the significance of direct microspore culture in theory and application was discussed