Transformation of Bar Gene and Regeneration ofPhosphinothricin-tolerant Plants in Maize

At present,the works of weeding are tedious,time consuming and labour intensive in maize(Zea mays L.) planting by popular artificial manner.However,it is an effective approach to overcome the problem by breeding herbicide-resistant varieties.In order to obtain phosphinothricin-tolerant transgenic elite maize materials,in this study,the bar gene was introduced into embryonic calli derived from immature embryos of maize elite inbred line 18-599hong via microprojectile bombardment.Bombarded calli were selected on selection medium containing 6,10,15 mg·L-1 Bialaphos(Phosphinothricin,PPT).After that,the generated young plants from resistant calli were first exercised in laboratory,then,the live plants were transplanted to field.At last,75 plantlets were regenerated and grown,and 8 of 75 were harvested.The results of PCR(polymerase chain reaction)and Southern blotting analysis demonstrated that bar gene has been integrated into the genome of transgenic plants.PCR-positive progeny plants were artificially daubed with herbicide,and the damage of each plant was investigated after 10 days.The results showed that the non-transgenic plants were dead,while,the presence of the bar gene provided resistance to the herbicide.