秋水仙素对草莓离体叶片再生和多倍体诱导的影响

Effects of colchicine on regeneration and polyploid induction from leaf in vitro of strawberry(Fragaria×ananassa)

以草莓(Fragaria×ananassa Duch.)栽培品种‘雪蜜’(2n=8X=56)的离体叶片为外植体,研究了不同浓度秋水仙素对愈伤组织诱导率、不定芽再生率以及多倍体植株诱导的影响,并采用流式细胞仪对多倍体植株的倍性进行鉴定。结果显示,用质量体积分数0.1%、0.3%、0.5%和0.7%的秋水仙素浸泡2、4和6 d,草莓离体叶片均能诱导出愈伤组织和不定芽,但随秋水仙素浓度的提高和处理时间的延长,愈伤组织诱导率和不定芽再生率均显著下降。用不同浓度秋水仙素处理均能产生多倍体植株,倍性为9X、10X、11X、12X、14X和16X;随秋水仙素浓度的提高,多倍体诱导率呈现先上升后下降的变化趋势。用质量体积分数0.3%秋水仙素浸泡处理4 d是最佳的草莓离体叶片诱导方法,不定芽再生率达到40.5%,多倍体诱导率为100.0%,并且诱导产生出16X的植株。

Taking leaf in vitro of strawberry （ Fragaria x ananassa ＇ Xuemi＇ , 2n = 8X = 56） as explant, effects of different concentrations （0.1%, 0.3 %, 0.5 % and 0.7 % ） of colchicine on callus induction rate and regeneration rate of adventitious bud as well as polyploid induction from leaf in vitro were studied, and the ploidy determination of polyploid was carried by means of flow cytometer. The results show that callus and adventitious bud can be induced from leaf in vitro treated with 0.1%, 0.3%, 0.5% and 0.7% colchicine for 2, 4 and 6 d, and with colchicine concentration rising and treatment time prolonging, the callus induction rate and regeneration rate of adventitious bud decrease obviously. The polyploid can be induced by using different concentrations of colchicine, and chromosome ploidy of regeneration plantlets are 9X, 10X, 11X, 12X, 14X and 16X. Induction rate of polyploid appears the trend of first increasing then decreasing with colchicine concentration rising. The best method of inducing polyploid of strawberry is that leaf in vitro is treated by using 0. 3% colchicine for 4 d. Using this method, the regeneration rate of adventitious bud and induction rate of polyploid reach 40. 5% and 100.0%, respectively, and 16X plantlets can be obtained.