马铃薯栽培种与野生种叶肉细胞融合及体细胞杂种鉴定

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

以马铃薯栽培种Solanumtuberosum‘中薯二号’的无性系 3# 、 8# (2n =4 8)和二倍体野生种Solanumchacoense (2n =2 4 )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 10 0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 ,5 2 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NTCP 9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。

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 effects on the protoplast fusion. The results showed that no much difference existed in the fusion efficiency between the two methods, but higher cell plating frequency and a stronger plant differentiation ability of callus were observed in electrofusion. Among 100 plants differentiated in earlier stage, 97 showed somatic hybrids by RAPD marker detection. Further ploidy level analysis with flow cytometry demonstrated 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 NTCP-9, The results 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.