Obtainment Of Transgenic Wheat With The Pinellia Pedatisecta Agglutinin (PPA) Gene By Agrobactrium Tumefaciens

The wheat is an important food crop in the world.The wheat aphid is one of theimportant pests.The way which it harms the wheat is that they can feed our wheat directlyor spread the virus.Importantly,The wheat aphids multiply fast, when the wheatheading,filling.Therefore,The wheat aphids threat the production of our wheat seriously.Inorder to combat the wheat aphids,we always use a large number of pesticides on the wheatfields.These pesticides can not only cause a serious zoology problem, but also increase theviability of aphids.So to get a new aphid resistance of wheat varieties has become theprimary goal of the wheat production.Now the resistance gene of the wheat aphid isGalanthus nivalis agglutinin (GNA) gene.While this experiment choses the Pinelliapedatisecta agglutinin (PPA) gene.In this experiment,I obtain the transgenic wheat withthe PPA gene by Agrobactrium tumefaciens.At the same time,I estabilished a more maturegenetic transformation system in the wheat.In this experiment,I used three kinds of wheat varieties,including Shi4185,Yangmai12,Bobwhite.I studied some factors,including different varieties, different hormones,ect.Inthe experiments,I studied the transienting expression of the GUS gene in the differentAgrobactrium tumefaciens strains,different promoters,and different varieties of wheatcallus.And then,I got a series of positive plants,after a varities of screening, differentiation,cultivating seedling. The results showed that the wheat immature embryo callus were more suitable than thewheat mature embryos callus as a transformation of the receptor material.Shi4185andBobwhite were better than Yangmai12in transient expression of the gus gene.Theconcentration of Agrobacterium can affect the results.I can effectively control the growthof Agrobacterium,when the OD600between0.6-0.8.It’s30mins,the process Agrobacteriuminfecting the wheat callus.In the antibanterial medium,the density of Timentin is500mg/L.The time of antibanterial is30mins.Compared with the UBI promoter,the35Spromoter is better in firing the gus expression.During the four different Agrobacteriumstrains,EHA105(35S-PPA-GUS) is the best in the transient expression of the GUSgene.The density of Hygromycin is50mg/L,in the screeing medium.the wheat callusdifferentiation medium contains KT2.0mg/L,NAA0.1mg/L,Timentin250mg/L.