苜蓿红豆草属间体细胞杂种的分子生物学鉴定

通过原生质体融合和培养获得苜蓿红豆草属间体细胞杂种植株。采用一种简便方法从杂种组织再生植株叶片、红豆草羟脯氨酸抗性系再生植株叶片和苜蓿根癌农杆菌702转化系愈伤组织提取DNA用于RAPD和Southern杂交分析。随机引物扩增结果显示两种亲本的RAPD多态具有明显差异。在所用20种随机引物中,6种产生较多的DNA片段。杂种组织具有两种亲本特有的DNA片段,但倾向于排除红豆草亲本的染色体,表明该杂种为非对称杂种,两种亲本染色体之间可能发生了重组。由于红豆草DNA的介入,杂种组织表现出较强的分化能力。分别利用RAPD扩增得到的OPA141000bp红豆草羟脯氨酸抗性系特异产物和OPA141600bp苜蓿根癌农杆菌702转化系特异产物为探针进行Southern分子杂交,证明杂种组织同时具有这两种DNA片段的同源序列。

Molecular Identification of Intergeneric Somatic Hybrid Plants between Alfalfa and Sainfoin

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.