Trauma Embryo of Genetic Transformation System with Agrobacterium Mediated Method
ZHANG Li-jun;LIU Long-long;ZHANG Jian-zhen;CUI Lin;Crop germplasm resources research institute,Shanxi Academy of Agricultural Sciences;Key Laboratory of Crop Gene Resources and Germplasm Enhancement on Loess Plateau,Ministry of Agriculture,P. R. China;Research Institute of Applied Biology,Shanxi University;
[Objective]The purpose was to optimize agrobacterium-mediated plant germination seed transformation method,increase conversion rate,breed GM varieties with specific functions,and improve viability and adaptability of crops. [Method]This paper consulted agrobacterium-mediated plant germination seed transformation method and selected germinating seeds of oat,maize and sorghum as test materials.Germinating seeds of oat,maize and sorghum were infected and transformed by agrobacterium strain,which included plasmid P3301 ubiA c.Transformation conditions were optimized. Genetic transformation systems of oat,maize and sorghum were studied. Concentration of bacterium,seed germinating time,ultrasonic power,and influence of concentration of acetosyringone and glufosinate on the above three crops were compared and analyzed. [Result]The experiment showed that when 0. 6 OD600≥0. 4,transformation achieved the best effect; Ultrasonic power was 900 W at the first time and treatment time was 10 minutes; Ultrasonic power was 200 W at the second time and transformation achieved the best effect when treatment time was 4 minutes; Acetosyringone could affect germination of oat,sorghum and maize significantly. It could increase transformation rate of maize and lower that of sorghum,but for oat,its effect was not obvious; in terms of tolerance to glufosinate,the difference was not significant among transformed plants of oat,maize and sorghum. Compared with non-transformed plants,seed germination of oat,maize and sorghum were restrained to some degree under the stress of 0. 8 mg / L herbicide solution. With 1 mg / L herbicide solution to select seedlings of T0 and T1transformed plants,the result showed survival rates of transformed plants is higher than those of control group.[Conclusion ]After herbicide selection and PCR test on T0 and T1seedlings,it was proved preliminarily that transformed plants were got.