原生质体非对称融合获得花椰菜与Ogu CMS甘蓝型油菜种间杂种

甘蓝型油菜Ogu CMS系幼叶原生质体经剂量分别为0.025 0,0.075 0,0.315 0,0.468 0和0.960 0 J/cm2的紫外线辐照后,与花椰菜下胚轴原生质体通过PEG法诱导融合,其中0.025 0~0.468 0 J/cm2获得再生植株68株,0.960 0 J/cm2辐射剂量下没有获得再生株。通过根尖染色体计数、流式细胞仪倍性鉴定、同工酶酶谱分析、RAPD及Ogu CMS线粒体基因orf138 STS(Sequence Tagged Site)标记引物扩增等方法对再生株进行了鉴定。结果表明,其中32株为杂种植株。同时也发现,紫外线辐照剂量的增加对供体染色体的丢失程度影响不大。

Interspecific Somatic Hybrids Between Brassica oleracea var botrytis and Ogura Type CMS Brassica napus Obtained from Asymmetric Protoplast Fusion

Asymmetric protoplast fusion beween hypocotyl protoplasts of Brassica oleracea var.botrytis and leaf protoplasts of Ogura type CMS Brassica napus was performed by PEG method.Leaf protoplasts from donor plant were treated with UV light at a dose of 0.025 0,0.075 0,0.315 0,0.468 0 and 0.960 0 J/cm~2 respectively.68 plants regenerated in the dose range from 0.025 0 to 0.468 0 J/cm~2,and none at the dose of 0.960 0 J/cm~2.Results of chromosome counts,ploidy analysis by flow cytometry,isozyme analysis,RAPD and specific amplification(STS primers) of Ogura-CMS-associated mitochondrial gene orf138 revealed that 32 somatic hybrid plants were obtained.It was showed in our experiment that the degree of elimination of UV-irradiated donor chromosomes did not depend on the irradiation dose.