Study On Agrobacterium- Mediated Transformation Of Brassica Napus With Binary Insect Resistant Genes

The production of oil rape (Cruciferae B+assica species) is often affected by pest damage .The wide use of chemical pesticides inevitably brings about serious safety and pollution problems. With the development of biotechnology, selection for pest-resistant introducing transgenic plants by introducing foreign pest resistant genes into the crops has been and more used by crop breeders.In this project the petioles from the sterile seedlings of Brassica Napus (Jin4.H165) were used as experimental matenals. Factors influencing B. Napus transformation mediated by A. tumefaciens have been investiguted.The results showed that petioles from 6-7/d old sterile seedlings had the highest differentiation frequency and were appropriate for transformation. When the petioles were infected by A.tumefaciens for 5 min, the efficiency of transformation was the highest and the calli produced the lest browning. Carbenicillin 500mg/L was used for excluding A. tumefaciens and 15mg/L kanamycin was used for screening transgenic calli. An efficient and stable B. Napus transgenic system was established.Integration of the insect-resistant genes into the genome of transgenic plants was verified by PCR amplification. All the transformants were normal in morphology. The bioassay of transgenic plant is now under way.