Institute of Plant Physiology, Biological Research Center, HAS, P.O. Box 521, H-6701, Szeged, Hungary.
Plant Cell Rep. 1993 May;12(7-8):399-402. doi: 10.1007/BF00234700.
An early identification of fusion products was based on the presumed vigorous growth of hybrid calluses after fusion between Solanum brevidens and S. tuberosum leaf protoplasts. The S. brevidens protoplasts were unable to form multicellular colonies under the applied culture conditions. Three size groups of calluses were separated and analyzed at two different early phases of culture period. "Squash blot" hybridization with a S. brevidens specific repetitive DNA probe showed that the group of the largest calluses consisted of putative somatic hybrids with a frequency of 80-100% in three independent experiments. Furthermore, approximately 80-95% of the middle sized calluses and 33-90% of the smallest ones were shown to be hybrid. The unexpectedly high percentage of fusion products, even in the case of the smallest calluses, may result from the suppression of the development of parental potato colonies in cultures with mixed cell population. Till this time 120 independent colonies selected as putative hybrids have been regenerated into plants. All of them exhibited hybrid phenotype, and their hybrid origin was proved by cytological and restriction fragment length polymorphism analyses.
早期融合产物的鉴定是基于杂种愈伤组织在 Solanum brevidens 和 S. tuberosum 叶原生质体融合后的旺盛生长。在应用的培养条件下,S. brevidens 原生质体无法形成多细胞集落。在培养期的两个不同早期阶段,分离并分析了三组大小不同的愈伤组织。用 S. brevidens 特异性重复 DNA 探针的“压片”杂交显示,最大的愈伤组织组由假定的体细胞杂种组成,在三个独立的实验中频率为 80-100%。此外,大约 80-95%的中等大小的愈伤组织和约 33-90%的最小的愈伤组织是杂种。即使是最小的愈伤组织,融合产物的比例也出乎意料地高,这可能是由于混合细胞群体培养中双亲马铃薯集落的发育受到抑制。到目前为止,已经从 120 个独立的假定杂种菌落中再生出 120 株植物。它们都表现出杂种表型,通过细胞学和限制性片段长度多态性分析证明了它们的杂种起源。