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采用埃及海岛棉辐射突变系226、188、197和埃及棉吉扎69的下胚轴为外植体,接种子改良的 MS 培养基(MS 无机盐+B_5有机元素)上,补加不同外源激素诱导愈伤组织和分化,然后经过悬浮培养,获得大量胚状体。经过重复实验,226、197能够产生大量胚状体,胚状体的发生频率分别为90%和70%左右,发生指数3.0和2.0。最适诱导培养基为改良 MS+2,4-D 0.05mg/L+KT 0.1mg/L,最适继代次数3～5次,其中需要在 IAA 1mg/L+ZT 0.1mg/L 培养基中继代2次。分化培养基为 MS+KT 1mg/L+NAA 4mg/L。然后选择淡黄或灰白松散的愈伤组织接种到悬浮培养基MS+KT 0.05mg/L 中,培养15～20d,即产生大量胚状体和胚性细胞团。在不加激素的培养基上,胚状体萌发成苗生根。对不同状态愈伤组织的过氧化物酶和酯酶同工酶进行分析表明,其酶活性和谱带差异较明显。
Using the hypocotyls of Aegilops taurus mutants 226,188,197 and Giza 69 in Egypt as explants, the MS medium supplemented with different exogenous The hormone induces callus and differentiation, and then undergoes suspension culture to obtain a large amount of embryoid body. After repeated experiments, 226,197 can produce a large number of embryoid bodies, embryoid body frequency of occurrence of 90% and 70%, the occurrence of 3.0 and 2.0 index. The optimum induction medium was MS + 2,4-D 0.05mg / L + KT 0.1mg / L, and the optimal number of subcultures was 3 ~ 5 times. The optimum medium was IAA 1mg / L + ZT 0.1mg / Relay on behalf of 2 times. The differentiation medium was MS + KT 1 mg / L + NAA 4 mg / L. Then choose light yellow or gray loose callus inoculated into suspension medium MS + KT 0.05mg / L, cultured 15 ~ 20d, that is, a large number of embryoid bodies and embryogenic cell mass. In hormone-free medium, embryoid body germinates into rooted plants. The analysis of peroxidase and esterase isozymes in different state callus showed that the enzyme activity and band difference were obvious.