以花椰菜-黑芥体细胞杂种自交及回交后代为材料,结合形态学特征,分子标记和荧光原位杂交(FISH)等群体分析技术,在16份高代回交及自交材料中筛选获得了花椰菜类型渐渗系材料12份,偏花椰菜类型异附加系材料4份。形态学调查及FISH分析表明:12份渐渗系材料除茎、叶等营养器官特征与花椰菜近似外,已具有较正常的花球形成,染色体数为18条,均来源于花椰菜;植株花粉母细胞减数分裂行为基本正常。SSR和AFLP分子标记检测表明,渐渗系材料除扩增到数量不等的黑芥多态性位点外,还扩增到埃塞俄比亚芥特异的多态性位点和少量新位点,另外发现渐渗系材料有丢失亲本花椰菜多态性位点的现象。FISH分析4份异附加系材料的染色体组成,结果显示:PFCN29BC2S1-4为9条花椰菜染色体附加8条黑芥染色体;PFCN15-2S1BC5-6-2和PFCN29BC4-37-7为16条花椰菜染色体分别附加7条和14条黑芥染色体;PFCN29BC4-38-5为花椰菜染色体附加了5-7条黑芥染色体。
In this research, we analyzed the asymmetrical somatic hybrids of cauliflower (Brassica oleracea var. botrytis, CC, 2n = 18) and black mustard (B. nigra, BB, 2n = 16) and their selfed or backcrossed progenies by integrating morphological, cytological and molecular information. Among 16 advanced selfing and backcrossing descendants in analysis, 12 introgression lines displaying the cauliflower-like morphology were identified with chromosome number 18. Simple sequence repeat (SSR) and amplified fragment length polymorphism (AFLP) analysis showed that those introgression lines preserved some DNA banding patterns from B. nigra, B. carinata (BBCC, 2n = 34) and some of new bands, and all of these introgression lines showed an absence of some cauliflower-specific sites. 4 plants (PFCN29BC2S1-4, PFCN15-2S1BC5-6-2, PFCN29BC4-37-7 and PFCN29BC4-38-5) were confirmed to have 9, !6, 16 and 18 cauliflower chromosomes added with 8, 7, 14 and 5 - 7 B. nigra-derived chromosomes respectively by fluorescence in situ hybridization (FISH) .