Establishment Of A New Type Of The Whole Seed Of Wheat Genetic Transformation System And A Preliminary Study On The Anti Bacterial Peptide Rev4

Wheat is one of the most important food crops in the world.Recently,there were many researches about genetic breeding.The main methods include biolistic bombardment,Agrobacterium tumefaciens,pollen-tube pathway,ultrasonic wave,ion implantation,laser microbeams puncture,PEG(Polyethylene glycol)and electroporation.The application of biolistic bombardment and Agrobacterium tumefaciens are the most wide.Agrobacterium is a natural genetic transformation system which has many advantages,including relatively simple Integration model,good genetic stability,easy operation,and large fragment transfer.So it has been one of the main methods of the research to the transgenic wheat.During the process of the transformation of agro bacterium-mediated plant genetic,the conversion receptors which are chosen usually have the capacity of meristem,such as immature embryos,Shoot tips,Floral organs,mature embryos callus and so on.Although the immature embryo of the wheat has strong regenerative capacity,we can't get it for many reasons such as the seasons.Besides,it is difficult for mature embryo callus to regenerate which control the application of the plant genetic transformation.Our experiment provided a way of making the seed of wheat overall infected by the imbibition curve of seed.This way can establish quick and effective system of wheat genetic transformation.Seed imbibition was one of the steps of the germination.So our experiment made use of this step to infect the seed and the process of sharp absorption would be beneficial to the agrobacterium adsorption and the transferred foreign gene fragment.First establish system of wheat genetic transformation,we used the imbibition curve of seed to know the best Infection time was 15 h.According to the chemical analysis of the Gus,The results show the best infection time and infection bacterial concentration was 12 h and A600=0.5,explored the Kanamycin to thelethal concentration was 35mg/L.Then we transformed the wheat seed with agrobacterium rhizogenes.After the transformation,the wheat has hairy root,developed root system,more lateral roots.In addition,there are more elongated and more closely of apical cell elongation zone.Finally,we used agrobacterium tumefaciens that has the gene of antimicrobial peptides Rev4 to infect the wheat.Then byextracting the RNA of the infected root of wheat and then RT-PCR,identificated of the antimicrobial peptide gene expression in wheat roots.Moreover,transgenic Receptor was ripe whole wheat seed.So it was convenient to get.We needn't to induce the explants to the callus.There were no more problems about the difficulty of plant regeneration and it was quick and effective.This new system of transformation solved many problems in the traditional wheat genetic transformation method such as plant regeneration difficulties,complicated experimental process,and long experimental period.This is not only magnificent to the research of wheat genetic engineering,but also to the practice of hereditary breeding.And it also supplies technical support to other plant genetic transformations.