The Study Of Comparison And Establishment Of Wheat Genetic Transformation System Mediated By Agrobacterium

Explant of immature embryos from 4 genotypes wheat and mature embryos from 12 genotypes of wheat were cultured for transformation by tissue culture mediated with Agrobacterium LBA4404 and C58C1 containing hygomycin and GUS gene, in order to research the factors influencing wheat transformation by Agrobacterium,establishing and optimizing the transformation system. At time, mature embryos cutted directly and stalk point were cultured for transformation by non-tissue culture. Studying the factors of the system medited by Agrobacterium-mediated, comparing the efficiency between tissue culture and non-tissue culture, the plant regenerations system for transformation was established and optimized. The results were follows:1, The wheat immature embryo callus were acceptor in tissue,through adjusting the concentration and compatibility of hormone in the MS , N6, SD2.The paper acquired the SK medium: SD2+KT 1mg/L +6BA 0.5 mg/L +ZT 3 mg/L;The wheat genotypes and the states of suitable starting material were shown to be important in the competence for transferation by explant of immature embryos.2 , The study improved the method of acquiring mature embryo by isolating whole embryo into power embryo with scalpel, then revulsiond calli in inducing medium. The method iproved avoided to sprout from mature embryo completely and increasd calli frequencies. Abased on that, the study established the plant transformation system with mature embryos. The mature embryo were cultured on induction medium with 2, 4—D 4 mg/L and AS. Infectione and co-cultivation was pormed with AS. It showed that induction frequencies were not influenced wheat genotypes and regeneration frequencies were affected wheat genotypes.3 , The paper studied the factors influencing the efficiency of transformation with mature embryos cutted directly by non-tissue culture and obtained: wheat was immersed at 37℃ and continuouing 3-4 hours,selected with Hygomycin 100 mg/L, adjusted Agrobacterium OD650=0.6. It showed that induction frequencies were not influenced wheat genotypes .4, The paper studied the factors Agrobacterium influencing the efficiency of transformation with stalk point by non-tissue culture and obtained: stalk point were co-infectioned with Agrobacterium in 20 min. regeneration frequencies were not affected wheat genotypes.5 , Generation resistance plant were identified by GUS and PCR amplifying ,the result showed the alien genes were integrated into plant cells,transgenic wheat plant were obtained by Mature embryo , mature embryos cutted and stalk point.