Dna Methylation Level Study Of Common Wheat Under The Gametocidal Chromosome Monosomic Addition

Common wheat Chinese Spring(CS), CS-gametocidal chromosome 2C monosomic addition(CS-2C), CS-gametocidal chromosome 3C monosomic addition(CS-3C), CS-gametocidal chromosome 3CSAT monosomic addition(CS-3CSAT), common wheat Norin 26(NL26) and NL26-gametocidal chromosome 3C monosomic addition(NL26-3C) were used as materials in this research. To analyze different gametocidal chromosome monosomic additions under the same genetic background of wheat and same gametocidal chromosome monosomic additions under the differen genetic background of wheat, MSAP method was used to identify the levels of DNA methylation of anthers of those wheat lines above, moreover, homology exploration of differential methylation fragments caused by gametocidal chromosomes was studied to lay the foundation for relvealing the relation between DNA methylation modification and the molecular function mechanism of gametocidal chromosome. The main results obtained by this study as follows:1.MSAP analysis of different gametocidal chromosome monosomic addition under genetical background of same wheat genotype.Methylation levels were identified by MSAP technology, and the results showed that levels of DNA methylation of CS-3C, CS-3CSAT, and CS-2C were 48.51%, 46.46% and 45.33% respectively, all of which were higher than that of CS(40.35%). Analysis of methylation variation showed that both CG and CHG sites were hypermethylated, and the hypermethylated frequencies of CS-3C were 13.09% and 3.33% resctively; CS-3CSAT were 11.31% and 4.59% respectively; CS-2C were 11.17% and 4.55% respectively; moreover, levels of hypermethylation of CG sites were higher than CHG sites. The overall methylation level of NL26-3C(36.85%) was a little higher than NL26(35.61%) using MSAP technology.2.MSAP analysis of same gametocidal chromosome monosomic addition under genetical background of different wheat genotype.Analysis of 3C monomer addition system in the background of CS and NL26. Level of hypermethylation of gametocidal chromosome 3C added to CS was 20.22%, while NL26 of that was3.48%, which suggested the extent of variation of gametocidal chromosome 3C added to CS was higher than that of NL26. Analysis of methylation variation showed that hypermethylated frequency of CG sites of NL26 with chromosome 3C was 9.12%, CHG sites(4.44%)of that was lower than CG sites, which was identical with the result of CS with gametocidal chromosome 3C.3.Sequence analysis of DNA methylation of differential sites of gametocidal chromosome monosomic addition.Partial differential fragments were recovered and sequenced.Blastn results showed that differential sequences were homology to MITE(Miniature inverted-repeat transpsable elements) transposon of Aegilops, Gypsy TREP retrotransposon, Sukkula-like transposon, SNF2 P gene, HMW glutenin protein gene sites and 3B chromosome of common wheat, mRNA predicted protein of barley and genomic chromosome of rice.