The Effect Of DNA Methylation On The Strength Of Gametocidal Chromosome Action

DNA methylation and gametocidal chromosomes are somehow linked,but the mechanism of action needs further exploration.In this study,the strong gametocidal chromosome and mild gametocidal chromosome chromosome were the main research material,and the real-time PCR,chromosome cytology identification and methylation sequencing analysis were used to further study the influence of DNA methylation on gametocidal strength,the main results are as follows:1.Preparation of the gametocidal hybrid material and its cytological identificationThe intense chromosome-plus（CS-3C3C）and the mild chromosome(CS-3C^SAT3C^SAT)were used as parents,and the hybrid materials CS-3C3C^SATand CS-3C^SAT3C were obtained.Using the somatic chromosome production technique,the number of somatic chromosomes in the four materials was stable at 44,and five samples were observed in the hybrid material,ensuring the accuracy of the experimental material.2.Analysis of promoter methylation levels and gene expression changes in specific genesBisulfite sequencing analysis of material for different gametoidal additional lines,And compared with the gene expression changes,The results found an inverse relationship between the expression levels of CS-3C3C^SATand CS-3C^SAT3C in 1D,2D,6A,and 7B and total DNA methylation in leaves,This indicates that the expression of related genes is regulated by DNA methylation;Very low expression of CS×3C and CS×3C^SATin 6A,Ta ORF3,Into an inverse relationship with total methylation,Higher expression of the hybrid CS-3C3C^SATand CS-3C^SAT3C than their parents,This is inversely related to the methylation changes at 2D and Ta ORF3,Indicating that changes in DNA methylation affect gene expression,Thus leading to the weakening of the gametocidal action strength.3.Effect of methylation modifiers on apical chromosome aberrations in gametocidal wheatTreat wheat root tip with different concentrations of methylation modifier and set the control group（distilled water treatment）,many types of chromosome aberration phenomenon can be observed under ordinary light microscope and fluorescence microscope,and the aberration rate of different gametocidal attachment materials are obvious difference,which shows that DNA methylation plays a certain role in maintaining the chromosome stability of gametocidal chromosome attachment material somatic cells.4.Effect of methylation modifier treatment on DNA methylation in wheatBisulfite sequencing analysis of apical DNA before and after treatment with methylation modifiers.The results showed that both CS and CS-3C3C tips in 6A,Ta ORF2,and Ta ORF3 had lower CG methylation levels compared to controls,whereas the MTFMS treated group increased at different methylation levels compared to the5-aza C treated group.This suggests that DNA methylation levels in apical cells can regulate the expression of specific genes,and subsequently affect chromosomal stability.Chromocytological observation shows that methylation modifiers can cause chromosome aberrations;through the bisulfite sequencing analysis,methylation promoter can increase DNA methylation level,but decrease DNA methylation level,gene expression and DNA methylation level and gene expression,indicating that DNA methylation can affect the strength of gamete chromosome by regulating the expression of gene.