DLO-Centre for Plant Breeding and Reproduction Research (CPRODLO), P.O. Box 16, NL-6700, AA Wageningen, The Netherlands.
Theor Appl Genet. 1996 Mar;92(3-4):316-25. doi: 10.1007/BF00223674.
Results are reported on the transfer of single, specific chromosomes carrying kanamycin resistance (Kan(R)) and β-glucuronidase (GUS) traits from a transformed donor line of potato (Solanum tuberosum) to a recipient line of the tomato species Lycopersicon peruvianum through microprotoplast fusion. Polyethylene glycol-induced mass fusion between donor potato microprotoplasts containing one or a few chromosomes and normal recipient diploid L. peruvianum protoplasts gave several Kan(R) calli. A high frequency of plants regenerated from Kan(R) calli expressed both Kan(R) and GUS, and contained one or two copies of npt-II and a single copy of gus. Genomic in situ hybridization showed that several microprotoplast hybrid plants had one single potato donor chromosome carrying npt-II and gus genes and the complete chromosome complement of the recipient L. peruvianum (monosomic additions). Several monosomic-addition hybrid plants could be regenerated within the short time of 3 months and they were phenotypically normal, resembling the recipient line. These results suggest that the transfer of single chromosomes is tolerated better than is the transfer of the whole donor genome. The unique advantages of microprotoplast fusion are discussed: these include the direct production of monosomic addition lines for the transfer and introgression of economically important traits in sexually-incongruent species, the construction of chromosome-specific DNA libaries, high-resolution physical mapping and the identification of alien chromosome domains related to gene expression.
本文报道了通过微原生质体融合,将携带卡那霉素抗性(Kan(R))和β-葡萄糖醛酸酶(GUS)特性的单个特定染色体从马铃薯(Solanum tuberosum)的转化供体系转移到番茄种 Lycopersicon peruvianum 的受体系。聚乙二醇诱导的含一个或几个染色体的供体马铃薯微原生质体与正常的受体二倍体 L. peruvianum 原生质体之间的大规模融合产生了几个 Kan(R)愈伤组织。从 Kan(R)愈伤组织再生的植物中,有很高的频率同时表达 Kan(R)和 GUS,并含有一个或两个拷贝的 npt-II 和一个拷贝的 gus。基因组原位杂交表明,一些微原生质体杂种植物有一条携带 npt-II 和 gus 基因的单个马铃薯供体染色体和受体 L. peruvianum 的完整染色体组(单体附加)。在短短的 3 个月内,可以再生出几个单体附加杂种植物,它们表型正常,类似于受体系。这些结果表明,单个染色体的转移比整个供体基因组的转移更能被容忍。还讨论了微原生质体融合的独特优势:这些优势包括在性不亲和物种中转移和导入经济上重要性状的单体附加系的直接产生,染色体特异性 DNA 文库的构建,高分辨率物理图谱的构建以及与基因表达相关的外来染色体区域的鉴定。