秋水仙碱对甘蓝型油菜离体小孢子胚胎发生的影响

研究了秋水仙碱不同浓度和处理时间对甘蓝型油菜23个基因型离体小孢子胚胎发生的影响.3个基因型的小孢子被10、50和100mg/L秋水仙碱处理24h或48h,胚产量是2.55～14.75胚/蕾,10～50mg/L处理72h则是0.94～2.43胚/蕾.这表明处理72h对小孢子胚发生有抑制作用.用200、400、500和800mg/L处理2个基因型小孢子16～48h,胚产量为0.6～1.33胚/蕾,未处理对照是6.25和9.36胚/蕾.可见200～800mg/L浓度对胚再生有不同程度的阻碍效应.结果还证明,小孢子对秋水仙碱的反应与其基因型有关.当用10、20、50和100mg/L处理48h时,22B5-6和903-3小孢子的胚产量为37.09～69.47胚/蕾,而F1-29、W592和SF10-12是0.28～1.45胚/蕾,相互之间差异很大.秋水仙碱处理小孢子的目的是使其再生植株的染色体高频率加倍,因此应根据胚产量和染色体加倍率来确定秋水仙碱浓度和处理时间.本试验中,采用10～50mg/L处理48h或者用100mg/L处理24h,约80%基因型的小孢子胚产量在5胚/蕾以上,约70%基因型的再生植株加倍率达60%以上,可有效地用于油菜遗传和育种研究等领域.

Effects of Colchicine on in Vitro Microspore Embryo genesis in Brassica napus

The effects of colchicine on in vitro microspore embryogenesis of Brassica napus were investigat- ed using 23 genotypes. 2.55 - 14.75 embryos/bud were regenerated from microspores of 3 genotypes by colchicine treatment at the concentrations of 10, 50 and 100 mg/L with 24 h or 48 h treatment but only 0.94 2.43 with 10 - 50mg/L for 72 h, indicating that the treatment of 72h has suppressive influence to microspore embryogenesis. The doses of 200 - 800mg/L colchicine with 16 -48h reduced 0.6 - 1.33 embryos/bud in 2 genotypes and 6.25 -9.36 in the untreated controls. Futhermore, the respone of microspore embryogenesis to colchicine is correlative with genotypes used. When appling 10, 20, 50 and 100mg/L colchicine and 48h treatment, lines 22B5-6 and 903-3 produced 37.09 - 69.47 embryos/bud, however 0.28 - 1.45 from genotypes F1- 29, W592 and SF10-12. The purpose of colchicine treatment microspore is dihaploid of microspore chromosome. Therefore, the colchicine dose and treatment duration used not only could produce a lot of embryos but also form higher rate of dihaploid plant. In present expriment using 10 - 50mg/L with 48h or 100mg/L with 24h obtained over 5 embryos/bud in about 80% of genotypes with more than 60% dihaploid plants. This suggested that our method can be used efficiently to the field of genetics and breeding in rapeseed etc.