Department of Molecular Biology, University of Aarhus, C. F. Møllers Allé 130, DK-8000, Aarhus, Denmark.
Theor Appl Genet. 1996 Mar;92(3-4):403-10. doi: 10.1007/BF00223686.
Fourteen somatic hybrids generated by electrofusion of mesophyll protoplasts from a non-flowering dihaploid S. tuberosum clone, DHAK-11, and a male-sterile dihaploid clone S. tuberosum, DHAK-33, were grown in the greenhouse and subjected to morphological assessments and tests for fertility and resistance to the white potato cyst nematode Globodera pallida pathotypes Pa2 and Pa3. The ploidy level of the hybrids ranged from 38 to 63 chromosomes. All hybrids developed flowers with violet petals except for one, hy-56, that possessed red petals. The colour of the tuber skin was purple in all hybrids except in hy-56 where the tuber skin was red. All of the hybrids were female fertile and generated viable seeds. Near-tetraploid hybrids produced the highest number of seeds per fruit and these seeds had a normal size. Hybrids with 58 or more chromosomes produced smaller seeds and less seeds per fruit. The germination frequency of the seeds was not influenced by the chromosome number of the hybrids. Pollen viability was determined and the male fertility of three hybrids was tested. Pollination with these three hybrids gave rise to fruit development, but only one produced viable seeds. The hybrids were tested for resistance to G. pallida pathotypes Pa2 and Pa3. A high level of resistance to Pa3, inherited from one parental clone, DHAK-11, and a high level of resistance to Pa2, inherited from the other parental clone, DHAK-33, was combined in four hybrids. These results demonstrate, that protoplast fusion is an efficient method for restoring the fertility of somatic hybrids generated from sterile parent clones, and is a powerful procedure for the complementation of multigenetic disease resistance traits in potato breeding lines.
十四株体细胞杂种通过电融合由非开花二倍体 S. tuberosum 克隆 DHAK-11 的叶肉原生质体和雄性不育二倍体克隆 S. tuberosum DHAK-33 产生,在温室中生长,并进行形态评估以及对活力和对白色马铃薯胞囊线虫 Globodera pallida 致病型 Pa2 和 Pa3 的抗性测试。杂种的倍性水平从 38 到 63 条染色体不等。除了一个具有红色花瓣的杂种 hy-56 外,所有杂种都发育出紫罗兰色花瓣的花。除了杂种 hy-56 外,所有杂种的块茎皮都是紫色的,而杂种 hy-56 的块茎皮是红色的。所有杂种都是雌性可育的,并产生有活力的种子。近四倍体杂种每果产生的种子数量最多,并且这些种子大小正常。具有 58 条或更多染色体的杂种产生的种子较小,每果种子较少。种子的发芽频率不受杂种染色体数的影响。测定花粉活力并测试三个杂种的雄性育性。用这三个杂种授粉导致果实发育,但只有一个产生了有活力的种子。杂种对 G. pallida 致病型 Pa2 和 Pa3 进行了抗性测试。对致病型 Pa3 的高水平抗性,来自一个亲本克隆 DHAK-11,对致病型 Pa2 的高水平抗性,来自另一个亲本克隆 DHAK-33,在四个杂种中结合在一起。这些结果表明,原生质体融合是一种有效的方法,可恢复来自不育亲本克隆的体细胞杂种的活力,并且是在马铃薯育种群中补充多基因疾病抗性性状的有力程序。