羽衣甘蓝小孢子胚胎发生观察及再生植株倍性鉴定

In vitro Microspore Embryogenesis and Chromosome Doubling of Kale

以5个不同基因型羽衣甘蓝品种为试材进行游离小孢子培养,研究小孢子发育时期与花蕾形态的关系、基因型对小孢子出胚率的影响、胚成苗及再生植株加倍和倍性鉴定。结果表明:(1)适宜于羽衣甘蓝游离小孢子培养的花蕾形态指标为:蕾长3.5～4.5mm、花瓣/花药为0.85～1.10。(2)在5个供试品种中,‘4105’出胚率最高,每蕾2.45个,其次是‘7341’,每蕾2.18个,‘7340’和‘7348’小孢子出胚情况较差,分别为每蕾1.36和0.51个,而‘7342’没有出胚。(3)采用150mg.L-1秋水仙碱浸泡幼苗根部24h,结合流式细胞仪快速检测植株倍性,发现二倍体总加倍率为78.4%,大大提高了小孢子再生植株的利用效率。

Five different genotypes of kale （Brassica oleracea L.var.acephala） were used to study the relationship of microspore developmental stage with bud size,effects of genotype on embryogenesis,embryos transferred to plantlets,chromosome doubling and ploidy identification.The results showed that：（1）The bud size suitable for using as index in microspore culture was 3.5～4.5 mm length and the ratio of anther to petal was 0.85～1.10.（2）In 5 genotypes,the embryogenesis efficiency of ‘4105’ was the best,that was 2.45 embryos per bud,then ‘7341’ with 2.18 embryos per bud;the efficiency of ‘7340’ and ‘7348’ was lower,that was 1.36 and 0.51 embryos per bud,respectively;‘7342’ had no embryos produced.（3）The microspore-derived seedlings were treated with 150 mg·L-1 colchicine for 24 h,then used to identify the ploidy level with flow cytometry.We found that the frequency of total doubling diploids was 78.4%,which was better for utilization in production.