Department of Genetics, Wageningen Agricultural University, Dreijenlaan 2, NL-6703 HA, Wageningen, The Netherlands.
Theor Appl Genet. 1991 Dec;83(2):225-32. doi: 10.1007/BF00226255.
This paper describes the analysis of the elimination of potato DNA from potato-tomato somatic cell hybrids. The hybrids were obtained by fusion of protoplasts of a cytoplasmic albino tomato genotype with leaf mesophyll protoplasts of a potato genotype carrying the β-glucuronidase (GUS) gene of Escherichia coli. The potato protoplasts were either isolated from unirradiated plants or from plants irradiated with 50 or 500 Gy of γ-rays. Green calli were selected as putative fusion products. The hybridity of these calli was confirmed by isoenzyme analysis. All of the green calli tested contained a potato-specific chloroplast DNA restriction fragment, and most of the calli analysed were positive for β-glucuronidase activity. In 72 of the hybrid calli we determined the percentage of potato nuclear DNA using species-specific probes. All of the tested green calli contained a considerable amount of potato genomic DNA, irrespective of the dose of irradiation of the potato parent. The limited degree of potato DNA elimination and the absence of true cybrids are discussed.
本文描述了从马铃薯-番茄体细胞杂种中消除马铃薯 DNA 的分析。杂种是通过融合具有大肠杆菌β-葡聚糖酶(GUS)基因的马铃薯基因型的叶肉原生质体与细胞质白化番茄基因型的原生质体而获得的。马铃薯原生质体要么从未辐照的植物中分离得到,要么从辐照 50 或 500 戈瑞γ射线的植物中分离得到。绿色愈伤组织被选为可能的融合产物。通过同工酶分析确认这些愈伤组织的杂种性。所有测试的绿色愈伤组织均含有马铃薯特异性叶绿体 DNA 限制片段,并且大多数分析的愈伤组织均对β-葡聚糖酶活性呈阳性。在 72 个杂交愈伤组织中,我们使用种特异性探针确定了马铃薯核 DNA 的百分比。无论马铃薯亲本的辐照剂量如何,所有测试的绿色愈伤组织均含有相当数量的马铃薯基因组 DNA。讨论了马铃薯 DNA 消除的有限程度和不存在真正的胞质杂种的情况。