Biochemistry Department, Rothamsted Experimental Station, AFRC Institute of Arable Crops Research, AL5 2JQ, Harpenden, Herts., UK.
Theor Appl Genet. 1988 Aug;76(2):260-6. doi: 10.1007/BF00257854.
Conditions are described for large scale electrofusion of mesophyll protoplasts of dihaploid S. tuberosum with those of diploid S. brevidens. Overall fusion frequencies of 20%-30% were achieved, and following fusion, large numbers of protoplast-derived calli were obtained. Putative somatic hybrid plants were selected from the regenerated shoots by examining their morphological characteristics. Twenty-one somatic hybrids were confirmed by isoenzyme analysis and six somatic hybrids were further confirmed by Southern hybridization. Tetraploid hybrids were obtained, but cytogenetic studies indicated that more of the regenerated hybrids were hexaploid than had previously been found following chemical fusion of the same partners. Some advantages of electrofusion over chemical fusion are discussed.
描述了用二倍体 S. brevidens 的叶肉原生质体大规模电融合的条件。实现了 20%-30%的总体融合频率,并且融合后获得了大量的原生质体衍生愈伤组织。通过检查再生芽的形态特征,从再生芽中选择了拟体细胞杂种植物。通过同工酶分析确认了 21 个体细胞杂种,通过 Southern 杂交进一步确认了 6 个体细胞杂种。获得了四倍体杂种,但细胞遗传学研究表明,与用相同亲本进行化学融合相比,再生杂种中六倍体的比例更高。讨论了电融合相对于化学融合的一些优势。