Zhang Li, Zhao Hong, Liu Fan
Beijing Vegetable Research Center, Beijing Academy of Agriculture and Forestry Sciences, Beijing.
Fen Zi Xi Bao Sheng Wu Xue Bao. 2008 Aug;41(4):265-74.
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.
通过聚乙二醇(PEG)介导不同剂量紫外线辐射的原生质体融合,获得了甘蓝型花椰菜与黑芥之间的不对称种间体细胞杂种。黑芥基因型为St.461,对甘蓝生产中常见的黑腐病、黑胫病和根肿病具有抗性。通过形态观察、流式细胞术(FCM)测定相对DNA含量、染色体计数、DNA分子标记等多种方法对再生植株进行了分析。形态观察表明,再生植株形态从中间类型到花椰菜类型各不相同。20%的再生植株DNA含量低于供体和受体之和。23%的再生植株染色体数目少于双亲之和。DNA分子标记分析(SRAP)表明,杂种中受体亲本的遗传信息或多或少完整保留,而供体基因组的特异性扩增条带在20%至97.77%的杂种中丢失。此外,对22株再生植株进行了黑腐病抗性测试,其中17株对野油菜黄单胞菌野油菜致病变种表现出良好抗性。这初步证明了通过紫外线介导的不对称体细胞杂交,将野生黑芥的外源病原菌抗性基因转移到甘蓝型花椰菜中。总之,紫外线辐射诱导了花椰菜与黑芥之间不对称杂种的产生。大多数杂种中发生了染色体消除或供体染色体的有限导入,然而,消除程度与紫外线剂量无关。