花椰菜与黑芥种间体细胞杂种的获得和鉴定  

Development and Identification of Interspecific Somatic Hybrids Between Cauliflower and Black Mustard

作　　者：张丽[1] 赵泓[2] 陈斌[2] 刘凡[2] 

机构地区：[1]首都师范大学生命科学学院,北京100037 [2]北京市农林科学院蔬菜研究中心,北京100097

出　　处：《植物学通报》2008年第2期176-184,共9页Chinese Bulletin of Botany

基　　金：北京市自然科学基金(No.5073045);北京市自然科学基金重大项目(No.5050001);北京市科技创新能力建设项目

摘　　要：利用非对称体细胞杂交技术,获得芸薹属花椰菜(Brassicaoleraceavar.botrytis)与黑芥(B.nigra)的种间杂种,实现了野生种质抗病基因向甘蓝类蔬菜作物的渗透。以具有良好再生能力的花椰菜下胚轴原生质体作为融合受体,具有抗黑腐、黑胫和根肿病优良性状的黑芥叶肉原生质体作为融合供体,用不同强度的UV射线处理后,利用PEG方法诱导供、受体原生质体融合。培养后获得170棵再生植株,选取来自40个不同愈伤组织的40棵单株进行形态学观察及RAPD和SRAP分子标记检测,结果表明其中30棵为体细胞杂种。染色体计数显示,约23%杂种植株的染色体数目小于供、受体染色体数之和。用流式细胞仪测定DNA含量显示,杂种植株DNA含量是受体的2-4倍,20%杂种植株DNA含量小于供、受体之和。Interspecific somatic hybrids of black mustard and cauliflower were produced by asymmetric somatic hybridization to introduce disease-resistance genes of wild species into Brassica crops. Hypocotyl protoplasts of Brassica oleracea var. botrytis, genotype 0307, possess high regeneration capacity. B. nigra, genotype St.461, shows resistance to black rot, black leg and clubroot diseases. Hypocotyl protoplasts of 0307 were fused with UV-irradiated mesophyll protoplasts of B. nigra by the PEG method. Approximately 170 regenerated plants were obtained, and 30 of 40 plants were verified as somatic hybrids by morphological observation, and with RAPD and sequence-related amplified polymorphism （SRAP） molecular markers. Chromosome analysis showed 23% of the hybrid plants with fewer than 34 chromosomes, the sum of donor and recipient parents. Flow cytometry revealed most of the hybrid plants with DNA content two to four times greater than that of the fusion recipient but 20% with a content less than that of the sum of donor and recipient parents.

关 键 词：非对称体细胞杂交 黑芥 花椰菜 杂种鉴定 植株再生 

分 类 号：S635[农业科学—蔬菜学] Q943[农业科学—园艺学]