摘要
选用兼具抗黑腐病、黑胫病、根肿病的野生种质黑芥(B.nigra)作为供体,具有良好原生质体培养能力的花椰菜(B.oleracea var.botrytis L.)基因型"0307"作为受体,供体叶肉原生质体经不同剂量UV照射处理后,通过PEG介导,与不经任何处理的受体下胚轴原生质体融合,培养获得再生植株。形态学观察显示再生植株的表型多样,呈偏向受体花椰菜类型及中间类型分布;SRAP分子标记特征表明,杂种中受体的遗传信息较为完整,而来自供体的特异扩增带丢失量约为20.0%-97.8%,且不同杂种中存在热点丢失序列;约23.0%的再生植株其染色体数目小于供、受体染色体数之和;采用流式细胞仪对再生株的细胞DNA含量进行分析,结果显示,20%的植株其DNA含量小于供、受体DNA含量总和。对22棵再生植株进行了黑腐病人工接种抗性鉴定,17棵再生植株表现良好抗性,初步证明通过非对称体细胞杂交技术已将外源抗性基因转入花椰菜中。实验表明UV辐射处理,导致再生植株中供体遗传信息的部分丢失或者染色体在不同程度上的消减,获得了花椰菜与黑芥的非对称体细胞杂种,但UV的剂量效应不明显。
Asymmetric interspecific somatic hybrids between Brassica oleracea var. botrytis and B. nigra were produced by PEG-induced fusion of protoplasts radiated by different doses of UV. B. nigra, genotype St.461, has resistance to black rot, black leg and clubroot diseases which are popular in cabbage production. The regenerated plants were analyzed by several means including morphology observation, relative DNA content measurement by flow cytometry （FCM）, chromosome counting, DNA molecular marker. Morphology observation indicated that the regenerated plants varied in morphology from intermediate type to cauliflower type. DNA contents of 20% regenerated plants were less than the sum of donor and recipient. Number of chromosomes in 23% of the regenerated plants were less than the sum of both parents. DNA molecular marker analysis （SRAP） demonstrated that the genetic information from the recipient parent was retained more or less complete and intact in the hybrids, while specific amplification bands of donor genome were lost in the hybrids from 20% to 97.77% . Furthermore, black-rot resistance test was performed in twenty-two regenerated plants, and seventeen of which showed good resistance against Xamthomonas campestris pv campestris. That gave the elementary proof of transferring of alien pathogen resistant genes from wild B. nigra to B. oleracea via UV mediated asymmetrical somatic hybridization. In conclusion, UV irradiation induced production of asymmetric hybrids between cauliflower and black mustard. Chromosome elimination or a limited introduction of donor chromosomes occurred in most of the hybrids, however, the degree of elimination was independent on UV doses.
出处
《分子细胞生物学报》
CSCD
北大核心
2008年第4期265-274,共10页
Journal of Molecular Cell Biology
基金
国家自然科学基金(30771206)
北京市自然科学基金(5073045)
北京市科技创新能力建设项目资助
关键词
花椰菜
黑芥
非对称体细胞杂交
UV辐射
SRAP分析
Cauliflower. Brassica nigra. Asymmetric somatic hybridization. UV irradiation. SRAP
作者简介
通讯联系人：电话：010-51503016，E-mail：liufan@nercv．com
