Institute of Genetics, Biological Research Center, Hungarian Academy of Sciences, 6701 Szeged, Hungary.
Proc Natl Acad Sci U S A. 1987 Dec;84(23):8434-8. doi: 10.1073/pnas.84.23.8434.
Transfer of methotrexate and 5-methyltryptophan resistance from carrot (Daucus carota) to tobacco (Nicotiana tabacum) was achieved by fusion between leaf mesophyll protoplasts of tobacco and irradiated cell culture protoplasts of carrot. Some of the regenerated somatic hybrids exhibited normal tobacco morphology with coexpression and independent segregation of the transferred resistance markers. Chromosomal instability resulted in aneuploid somatic hybrids with significantly lower chromosome number than predicted by simple addition of parental chromosome number. The methotrexate resistance phenotype was correlated with the expression of carrot-specific dihydrofolate reductase as judged by isozyme and immunological characteristics of the enzyme. The genomic construct of these somatic hybrids made the transmission of the resistance character into the next sexual generation possible.
通过将烟草的叶肉原生质体与经辐照的胡萝卜细胞培养原生质体融合,实现了氨甲喋呤和 5-甲基色氨酸抗性从胡萝卜(Daucus carota)向烟草(Nicotiana tabacum)的转移。一些再生的体细胞杂种表现出正常的烟草形态,同时表达和独立分离转移的抗性标记。染色体不稳定性导致非整倍体体细胞杂种的染色体数目明显低于双亲染色体数目的简单加和。氨甲喋呤抗性表型与胡萝卜特异性二氢叶酸还原酶的表达相关,可通过同工酶和酶的免疫学特性来判断。这些体细胞杂种的基因组构建使抗性特征传递到下一个有性世代成为可能。