N~+Ion Beam Implantion In Glycyrrhiza And Analysis Of Differential Gene Expression

Since the80’s of the last century Ion beam biotechnology had been advanced, it has a fast development in just a few decades and becomes one of the major mutagenesis methods in mutation breeding and the acquisition of excellent characters of crops, plants and microorganism in our country. Ion beam biotechnology has obtained a widely of achievements. Comparing to other physical and chemical mutagenesis methods, ion beam implantation technology has some advantages just like the low injury, the high mutation frequency and the high mutation frequency. This technology is advantageous to screen and gain mutants.Glycyrrhiza is a kind of high medicinal value plant because of its metabolite, glycyrrhizic acid, andother compounds have very high value in medical use. Secondly, Glycyrrhiza is one kind of the major forage grass in pastoral areas providing materials of mixed forage grass for livestocks and also with the positive effects of windbreak and sand fixation. In the recent years, related researchs of Glycyrrhiza are increasing. The application of ion beam mutation breeding in Glycyrrhiza have been reported, mainly concentrated in ion implantation of Glycyrrhiza contemporary stress resistance, enzyme activity and gene polymorphism analysis. But for ion implantation of Glycyrrhiza contemporary polymorphisms of the gene expression studies have not been reported.This paper studied that the different doses of N+ion implantation in Glycyrrhiza effect on the germination of Glycyrrhiza and analysis of differential gene expression of radical of the plants by N+ion implantation.Main conclusions are as follows:1. Using6different doses of N+ion implantation to handle Glycyrrhiza seeds. The doses are including2.6×1016ions/cm2,3.9×1016ions/cm2,5.2×1016ions/cm2,6.5×1016ions/cm2,7.8×1016ions/cm2,9.1×1016ions/cm2.By determining the germination rate, germinability and germination index we obtain the "saddle-shaped" curve of these three major germination datas. All of six doses treatment groups are superior to control group in germination rate, geminabiity and germination index. This outcome confirms the promoting effects for Glycyrrhiza seeds germination within the scope of these doses of N+ion implantation. The treatment of2.6×1016ions/cm2dose reflects the highest germination ability and the treatment of7.8×1016ions/cm2is the second crest value in the curve of germination rate.2. The N+ion implantation of Glycyrrhiza seeds induced differences in gene expression of contemporary roots were analyzed by cDNA-AFLP molecular marker technology. Obtained N+ion implantation Glycyrrhiza radicle and without N+ion implantation of Glycyrrhiza radicle cDNA by reverse transcription technique. The use of Pstl and Msel two kinds of restriction endonuc lease enzyme digestion, design of joints and pre amplification primers and the corresponding9pairs of selective amplification primers for the specific fragment amplification and sequencing. After the analysis,9pairs of selective amplification primers with81primers amplified results, there were bands of1576, the difference in band number286, accounting for about20%of the total bands, the polymorphic fragments were cloned and sequenced, A total of92sample length and purification fragments are basically identical with reliability, by using BLAST analysis the homology comparison of sequencing results, of which53retrieved sequencing sequence homology with high known function of the protein sequences of DNA or mRNA sequences.