马铃薯栽培种与野生种叶肉细胞融合及体细胞杂种鉴定

 以马铃薯栽培种Solanum tuberosum ‘中薯二号’的无性系3 # 、8 # (2n = 48) 和二倍体野生种Solanum chacoense (2n = 24) 的无菌苗为原生质体来源, 比较了PEG (聚乙二醇) 融合和电融合两种方式马铃薯原生质体融合效果的影响。结果显示, 两种融合方式对细胞的融合率没有显著差异, 但电融合的细胞植板效率和愈伤组织分化能力均显著高于PEG融合法。在早期挑选的生长旺盛的愈伤组织再生的100个株系中, 经RAPD 标记检测有97 个系为体细胞杂种。杂种植株经流式细胞仪倍性分析表明, 52.6 %为六倍体, 14.4 %为八倍体, 14.4 %为非整倍体, 18.6 %为混倍体。叶绿体SSR 引物NTCP29 用于细胞质重组检测, 结果显示, 体细胞杂种中叶绿体具有偏亲现象, 大多数杂种只含有单一亲本的叶绿体类型, 只有16.5 %株系为叶绿体重组的杂种植株。

Mesophyll Protoplast Fusion of Solanum tuberosum and Solanum chacoense and Their Somatic Hybrid Analysis

The protoplasts were isolated from the leaves of Solanum tuberosum clones 3 # and 8 # (2n =48) , which were derived from cv. ‘Zhongshu 2’, and dipliod species S . chacoense (2n = 24) . The PEG and electric fusion methods were employed for looking into their effect s on the protoplast fusion. The results showed that no much difference existed in the fusion efficiency between the two methods , but higher cell platingf requency and a stronger plant differentiation ability of callus were observed in elect rofusion. Among 100 plants differentiated in earlier stage , 97 showed somatic hybrids by RAPD marker detection. Further ploidy level analysis with flow cytometry demonst rated that 52.6 % were hexaploids , 14.4 %octoploids , 14.4 % amphiploids and 18.6 % mixoploids. Cytoplasm recombination of the somatic hybrids were investigated using a specific chloroplast SSR primer NTCP29 , The result s indicated that most of the hybrids contain the chloroplast from one of the fusion parents. Only 16.5 % of the hybrids combined the two parental chloroplasts.