Transformation Of The Glyphosate Oxidoreductase Gene To Rape (Brassica Napus L.)

To establish a simple,practical transformation technique in rape(Brassica napus L.), the pollen-tube pathway transformation was used in this research.The full-length sequence of glyphosate oxidoreductase gene(GOX) was amplified via PCR from the transgenic glyphosate resistant rape.The rape varieties "Qushi 00S-76","Taikong 3-19","Fan 4" and "Fan 5" were used as recipients,and the plant expression vector pPZP221 containing GOX gene was used as exogenous gene donor for the pollen-tube pathway transgenic research.The plasmid DNA solution was applied to without cut and cut 1/3 of stigma at 20,24 and 30 h after hand-pollination,and PCR results of To generation plants showed that the positive rates were 1.1%,3.2%and 1.6%,respectively.The transformation rate of cutting 1/3 of stigma was 3.0%,while the transformation rate of without cutting stigma was 1.0%.This research also studied the resistance of T₀ generation plants,there are 0.7%and 1.2%T₀ generation plants appeared high resistance when the seedlings were selected by 160mg/L gentamicin and 0.02%glyphosate.The pollen-mediated transformation approach is a simple,easy and widely used technology.It avoids the tissues culture procedures used in Agrobacterium infection and particle bombardment,etc.