[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.