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以紫羊茅的成熟种子为外植体材料,研究了2,4-D、有机物及光照对愈伤组织诱导发生、生长状态及绿苗分化能力的影响。结果表明,3~5mg/L2,4-D为愈伤组织诱导和继代培养基的适宜浓度,诱导率为46.6%~57.9%,并使愈伤组织保持胚性状态。在继代培养基中添加脯氨酸和天门冬酰氨,并且每培养1~2个月给予15d左右的散射光培养,使培养了2年多的愈伤组织仍保持99%以上的绿苗分化率。利用基因枪将GUS基因转入继代2年多的愈伤组织,通过组织染色法检测到了高频率的GUS基因瞬时表达。
The effects of 2,4-D, organic matter and light on the induction of callus, the growth status and the ability of green shoots to differentiate were studied using the mature seeds of the red fescue as explants. The results showed that 3 ~ 5 mg / L 2,4-D was the suitable concentration of callus induction and subculture medium, the induction rate was 46.6% ~ 57.9%, and the callus maintained embryogenic status. Proline and asparagine were added to the subculture medium, and cultured for about 15 days with 1 ~ 2 months of culture, the callus cultured for more than two years still maintained more than 99% of the green seedlings Differentiation rate. The gene gun was used to transfer GUS gene into the callus for more than two years. The transient expression of GUS gene was detected by tissue staining.