Xu Z Q, Jia J F
Provincial Key Laboratory of Biotechnology, Northwest University, Xi'an.
Sheng Wu Gong Cheng Xue Bao. 2000 Mar;16(2):173-8.
Somatic hybrid plants between alfalfa and sainfoin were regenerated by protoplast fusion and culture. DNA samples of the hybrid plants, hydroxyproline-resistant sainfoin plants, alfalfa cell line transformed with Agrobacterium tumefaciens 702 were isolated with a new and simple method. The hybridity was identified by random amplified polymorphic DNAs and Southern hybridization. Significant differences can be seen in the sequences amplified, which are specific for each parent/primer combination under the amplification conditions used. In 20 random oligonucleotide primers used, six could amplified more DNA fragments and had better polymorphisms. The results suggested that besides containing nuclear substances of two parents, the hybrid genome was inclined to eliminate sainfoin chromosome with DNA reconstruction. However, the somatic genome also could produce the sainfoin-specified DNA fragments which further confirmed by Southern hybridization. The hybrids were asymmetric and had certain regeneration ability just because the intervention of sainfoin DNA.
通过原生质体融合和培养再生出了苜蓿和红豆草之间的体细胞杂种植物。采用一种新的简单方法分离了杂种植物、抗羟脯氨酸红豆草植物、用根癌农杆菌702转化的苜蓿细胞系的DNA样本。通过随机扩增多态性DNA和Southern杂交鉴定杂种性。在所使用的扩增条件下,扩增出的序列存在显著差异,这些差异对每个亲本/引物组合都是特异的。在使用的20种随机寡核苷酸引物中,有6种能扩增出更多的DNA片段且具有更好的多态性。结果表明,杂种基因组除含有双亲的核物质外,还倾向于通过DNA重建消除红豆草染色体。然而,体细胞基因组也能产生红豆草特异的DNA片段,这一点通过Southern杂交得到了进一步证实。杂种是不对称的,并且由于红豆草DNA的介入而具有一定的再生能力。