Transformation And Screening Of Herbicide Resistant Bar Gene In Maize

Gtaet progress had been in transgenic maize and molecular breeding.But,building a quik,high efficient and cheap transformation method is anxious to be solved by investigator. A non-tissue culture transformation technique at original site that was mediated by agrobacterium was innovated in Center of Agri-Biotechnology, Jilin Academy of Agriculture Sciences.PTF102 vector with Bar gene was introduced into maize by this technique in our experiment,transforments were obtained,some of them were analyzed,the result proved that the foreign gene was inherited in maize plant, The average transformation rate was 10%. The influence factors of the transformation method were studied, optimum transformation time is 26～28 hours afer pollination,the optimise OD₆₀₀ of agrobacterium is 1.0,AS(100μmol/L) provided an increase in the transformation efficiency from a baseline 0.56% to 7.52%.In present paper ,three screening method of herbicide were compared in early generation of transgenic maize,spraying seedings is the best.92 percent of herbicide-resistant plants recomfirmed positive by detection of CR, PCR and southern blotting validations.In present paper we developed a quickly batch screening technique by using the herbicide- resistant selection marker of transgenic maize.It might be about 70% to distinguish low expressive transgenic lines and to eliminate the pseudo-transgenic lines inT₂ and T₃ generation, which used to be as transgene positive. Obviusly we have strong suggestions that this approach for screening technique should be more efficient and time-saving and also should come true by selections in the plots and plates in transgenic breeding program.