Study On Biological Effect Of Ion Beam Irradiation On Arabidopsis Thaliana

Since the late 80 s of last century,Institute of Plasma Physics Chinese Academy of Sciences,Mr Yu put forward the biological effect of ion beam,many domestic and international agencies have studied the biological effects and mechanism of ion implantation by different plants and microorganisms.The researchers have found many mutagenic effects,including epigenetic traits,biochemical indicators,cell morphology and molecular effects,to create greater economic and social benefits.In recent years,heavy ion beam has become a new type of radiation induced with its unique physical characteristics,and the application of heavy ions in clinical medicine to mutation breeding,also yielded the results.With the rapid development of high-throughput sequencing technology,a new method-RNA-seq technology is widely used in life science research.It can be very effective qualitative and quantitative analysis of transcriptome,it is widely used in gene expression regulation research.Through RNA-seq,the gene expression of the material can be analyzed in detail,and the differential expression between different materials can be studied.In this paper,using N+ low energy ion beam and 12C6+ heavy ion beam to irradiate the Colombian type(Col)Arabidopsis seeds and the Xindao 18 rice,the phenotypic statistical analysis of seedlings cultured after irradiation to screen material further experiment.Through phenotypic statistical analysis,it is found that the phenotype of N+ irradiation is relatively chaotic,and the results of 12C6+ irradiation is relatively stable,therefore,12C6+ ions are selected for further study.The seeds of Arabidopsis thaliana were irradiated by 12C6+ heavy ion beam with initial energy of 80 MeV/U and irradiation dose of 50 Gy,75Gy,100 Gy,150Gy and 200 Gy respectively.By 10 days of culture,all the materials to be sequenced according to phenotypic analysis classified as growth inhibitory YZ(Z75,Z100 S,Z150,Z200)and non-growth inhibitory CJ(Z50,Z100H)transcriptome sequencing and analysis.Using a Illumina Hiseq4000 platform,6 accept radiation type group and 1 wild type control group were sequenced,three biological samples were set up each group,total of 21 samples,each sample average output 2.12 GB data.By comparing the data of the sequencing data with the reference genome,the gene expression level of each group was obtained,and the expression of the gene was calculated by using the expression level.The expression of 39 transcripts in YZ group was changed,These transcripts are functionally annotated and found to function in a variety of biological pathways and gene families,such as photosynthesis,resistance to stress and transcription factors;In the CJ group,a total of 54 transcripts were changed,and the results of their functional notes were involved in the synthesis of stress and transcription factors and metabolic enzymes in addition to various biological pathways and gene families.The expression of differentially expressed genes was verified by q-PCR technique,and the differentially expressed genes(upregulated or down-regulated)were verified.The results of the selected genes were consistent with the sequencing results.Finally,by analyzing the partial phenotype and differential expression of 100 S and 100 H,it was found that the genes with significant changes in the expression of 100 S and 100 H were metabolized and responded to external stimuli,suggesting that the phenotypic differences were metabolized Changes caused by its specific formation mechanism remains to be further studied.The current ion beam irradiation research in the field of biology mainly includes two points,one is the application of ion beam irradiation,and the other is to explore the mechanisms of ion beam mutagenesis.Its main application mutation breeding and ion beam mediated transgene,etc.,and it has been widely recognized by many researchers;it's hard for researchers to study on ion beam mutagenesis,and it has attracted the attention of many researchers in global world,but has not yet given reasonable explanation.At present,the mechanism of ion beam mutagenesis is mainly from phenotype,cell level and physiological and biochemical level to carry out the work,relatively little research on the molecular level.In this paper,the irradiation of the plant material was carried out by ion beam irradiation.The phenotype and gene expression of Arabidopsis thaliana were changed by phenotypic screening and transcriptome sequencing.The sequencing data were analyzed by bioinformatics.The differential expression of related genes and the use of molecular biology methods to verify,to understand the ion beam mutagenesis mechanism has theoretical significance.The work in this paper,which is of guiding significance in the next experimental work,and puts forward a research direction and possibility for further study of ion beam mutagenesis mechanism.