Aflp Analysis Of Three Early Maturing Rice Variation Lines

The Marize genome DNA was transferred into Rice Yujing 6 by Ion Beam mediated technology. We obtained a large number of variation materials of the contemporary transformation. Through many years cutivating and scerrning, we obtained a crop of variation lines of Rice. Three early maturing variation lines which had been stable heritabled for four years were selected as research materials. The maturity of three variation was much earlier than control Rice, and the length was lower than control Rice olso.AFLP marker technique was used to study three early stable heritable maturing variation lines, 64 pairs of selective primers were used to analysised the polymorphism of genome DNA of variation lines and controls. The result showed that the amplified bands between the variation lines and the negative control (Yujing-6) had significant differences. The similarity ratio of amplified bands in the variation lines (A, B and C) and the negative control with the positive control A (i.e. maize) were 19.9%,20.1%,20.0% and 18.8% each, it indicated that compared with maize the similarity of genome DNA in variation lines was higher than that of negative control. There were 966 DNA bands in variation line A and totol number of 90 polymorphic bands in which there were 55 new bands, 28 absent bands, and 7 target bands, reached the polymorphic ratio of 9.3%. There were 963 DNA bands in variation line B and totol number of 81 polymorphic bands in which there were 51new bands, 22 absent bands, and 8 target bands, reached the polymorphic ratio of 8.4%. There were 974 DNA bands in variation line C and totol number of 98 polymorphic bands in which there were 59 new bands, 31 absent bands, and 8 target bands, reached the polymorphic ratio of 10.1%. It revealed that the genome DNA of variation lines had changed after mediated transferred Maize DNA by Ion Beam. Some new bands of AFLP were cloned and sequened, then made these sequences retrieved in NCBI Genome database. From the results of the retrieval, we can retrieve highly matching rice sequences in genome database besides A-P6M4-N. The results showed that these new bands came from receptor rice, and the digestion sites of the rice genome DNA might have been changed by ion beam mediated. Two matching sequences of A-P6M4-N came from 6 and 11 chromosomes, it showed that the chromosome translocation had happened in receptor rice. The DNA chains might be fractured by ion beam injection, then resulted variation on the process of DNA recombinant.The target bands from 4 primers were cloned and sequenced. Through analyzing the similarity of the target bands by DNAman, the target bands from 3 primers had low similarity contrast with control marize, reached the ratio of 30% to 45% besides the primer P6M8. Though Blast retrieval, we recovered that the target bands of P6M8 might be the homologous sequences of rice and marize. And other target bands could retrieve the highly similiarity sequences in the genomo database, but no marize DNA sequences retrieved. This revealed that the false positive phenomenon existed in target bands from AFLP amplification. It can not decide the target bands came from marize DNA, but might be the result of ion beam mediated.