Application Of Ion Beam To Mutation And Transferring On Arabidopsis

Through the study on Arabidopsis(Landersberg erecta wild type) callus inducing and plant differentiation system, it was found that the callus inducing medium was B5+ 5mg/l 2,4-D+ 0.5mg/l KT and differentiation medium was MS+5mg/L KT +0. 1mg/1 IAA. The system was efficient. At the same time, we set up the liquid culture system that produced seedlings.The system of ion beam implantation to Arabidopsis callus and seedlings were set up on the base of the study of protection from the hazard of Arabidopsis calluses and seedlings in process of ion beam implantation. The protection effect of DMSO and vitrification from hazard was compared. It was found that vitrification could efficiently protect callus and seedlings from hazard. After optimize the implantation parameters, we set up the experiment system implanting calluses and seedlings.During the study on the mutation effects of ion beam on Arabidopsis seeds, it was found that the sensitivity degree of different ecotype seeds was different. Plant survival rate could be affected by different ions' energy and ions' type. Arabidopsis seeds were more sensitive than that of wheat, rice, soybean etc. With the study on the relationship between seeds survival rate and implantation dose, we found the suitable mutation dose and transformation dose. Using RAPD, we studied the variation aroused by ion beam implantation in Ml and M2 generations. 2.6,13 and 26 x1015 N+/cm2 were used in the experiment. Variation frequency increased with theincrease of dose. They were 13.9%, 14.9% and 28.7%. Compared with other same type mutation methods, ion beam had the higher variation frequency. Some polymorphism bands of Ml could be found in M2.We transformed GUS gene mediated by ion beam into Arabidopsis seeds, GUS gene temporary expression was not obtained. But when seeds were bourgeoned 24-48h and then implanted by ion beam and dealt with transformation, we obtained 10% temporary expression.With the precondition that invitro tissue could survive after implantation, the study on transferring GUS gene into Arabidopsis callus was done. And we got the transformants. Transformation efficiency was up to 2%.Through the study on ion beam assistanted agrobacterium transformation Arabidopsis seedlings, in this experiment system, group that used agrobacterium transformation seedlings directly had no transformants. But in groups that ion beam assitanted agrobacterium transformation Arabidopsis seedlings, we got transformants and transformation efficiency was 3%.