Studies On The Transformation Of Chitinase And GFP Genes Into Rapeseed Mediated By Agrobaeterium Tumefaciens

As one of the most important commercial crops all over the world, rapeseed is thought to be the most valuable target crop to study. Nowadays,31% of the world's commercially grown rapeseeds are GM ones. As this crop is easily infected by diseases such as Sclerotinia during its growing period, which causes severe reduction of product, studies of rapeseed's genetical modification mostly concentrate on the introduction of foreign genes into rapeseed to enhance its resistance to diseases.Agrobacterium-mediated transformation which is generally accepted to be the most efficient genetic transferring method in rapeseed gene modification, has been used in this study. Through the exploration of medium components optimizing and comparison between lower hypocotyl explant and petiole explant, an optimized transformation system for rapeseed was obtained. Then the target gene chitinase and the reporter gene gfp were successfully transferred into four Brassica napus lines.This study shows, the petiole explants which had been germinating for 4 days can be dipped in the Agrobacterium suspension with OD 0.05 (650nm) immediately without pre-cultivation. Then, the explants should be transferred into callus culture medium after cocultivating for 2 days. This is a more simple and efficient method than the one using lower hypocotyl explant. The most suitable mediums used at different stages of transformation and concentration of kanamycin used to screening transformants was 50mg/L. The total transformation rate reaches 20.3%.