紫羊茅高频植株再生系统的建立与GUS基因的转化

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,3～5 mg/L 2,4-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15 d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.

Establishment of high frequency plantlet regeneration system of Festuca rubra and GUS gene transformation

Mature seeds of Festuca rubra were used for explants. Effects of 2,4-D, organic material, and illumination, on callus induction, growth state and shoot regeneration were studied. Results indicated that a 2,4-D concentration of 3-5 mg/L was optimal for callus induction and callus subculture. The frequency of callus induction on the medium supplemented with 3-5 mg/L of 2,4-D ranged from 46.6%-57.9% with the callus keeping an embryogenic state during subculture. The rate of plant regeneration from callus which were subcultured more than two years was approximately 99% in the medium when supplemented with 3-5 mg/L (2,4-D) + 500 mg/L proline + 130 mg/L L-Asparginem, and used in conjunction with 15 day's illumination every one to two months during callus darkness subculture. By using the callus which was subcultured more than two years, transient expression of the GUS gene was observed on the callus after particle bombardment.