The Establishment Of Wheat Genetic Transformation System Mediated By A. Tumfaciens And The Studies On Transgene

Immature embryos from 24 different wheat genotypes were cultured, comparing the efficiency of callus induction and regeneration among them and the factors influencing induction and regeneration of callus were studied. The materials with higher induction and regeneration were transformed with A .tumefaciens N105 containing GUS gene, studying the factors influencing the efficiency of transformation mediated by A. tumefaciens. Abased on that ,Embryo-derived callus from shannong995604 were transformed with A.tumefaciens LBA4404 containing NptIIand CpTI. The results were as follows:1, Shannong995604 and F281-10 with higher induction and regeneration frequency were obtained from 24 different wheat genotypes with good agronomic traits, the study on the factors influencing the induction and differentiation of wheat callus showed: keeping higher 2,4-D concentration in subculture media,or alternating subculture was beneficial to maintain the higher regeneration frequency of wheat callus. Good callus with higher regeneration capacity can be obtained when scultem of embryos after pollination 15 days was cultured.2, Shannong995604 were transformed with A.tumefaciens N105 containing GUS gene,The factors influencing the efficiency of transformation were studied. It showed: The higher efficiency was observed when A.tumefaciens cell density was adjusted to OD6oo=0.6 and the inoculation duration was 1 hour; The efficiency can be increased by adding AS to the media of pre-culture and co-culture, The transgenic system mediated by A.tumefaciens was established, and the efficiency of transformation in this system is 2.86%.3, Using the system of transformation,shannong995604 were transformed with A.tumefaciens LBA4404 containing NptIIand CpTI. As a result,three transgenic wheat plants were obtained, and they were proved that foreignDNA have been intergrated into their wheat genome by using PCR amplification and Southern blotting analysis of the three transgenic wheat plants. The three transgenic wheats' agronomic traits were not different from the same genotype without being transformed.