Study Of DNA Methylation In Gametophyte Development And Sexual Organs Differentiation Of Adiantum Reniforme Var Sinense

Nowadays,the study of epigenetic involves not only animals also plants, one of the common mechanism of epigenetic studies is DNA methylation,mainly researching on the whole genome level of DNA methylation research and specific gene methylation and use of methylation. The whole genome DNA methylation study widely uses the method of Methylation-Sensitive Amplification Polymorphism analysis(MSAP), this method can accurately estimate the whole genome degree of methylation from a certain level, and according to the gene sequencing results finds useful target genes for subsequent function verification response. MSAP technique mainly use two kinds of methyalation sensitive isoschizomers to cut genomic DNA and through selective amplification PCR reaction to analyze genome-wide DNA methylation level.Adiantum reniforme var. Sinense has high ornamental value in leaf and plant shapes, which is a kind of rare and precious plants of the ferns. Due to its high ornamental value, now its application in the field of family and flower markets is deepening. More and more studies are for Adiantum reniforme var. Sinense. But due to starting late and Adiantum reniforme var. Sinense is generally located in the place such as mountain rock, so research on it is from the beginning of the introduction to study the rapid propagation system.And the research on molecular is just started.This study selects Adiantum reniforme var. Sinense asexual and sexual reproduction samples of each three periods to do the MSAP analysis. The MSAP technical analysis results shows that the Adiantum reniforme var. Sinense asexual reproduction and sexual reproduction have frequent phenomenon of DNA methylation, and with the differentiation of sexual organs and meristem, methylation level increased obviously, and with the formation of sporophyte differentiation, methylation level began to reduce, but the rate is still high; Research also illustrates that as the growth and development of Adiantum reniforme var. Sinense, the obvious phenomenon of excess methylation and demethylation is found and in different period it showes certain specificity.From a certain extent, this shows that the degree of DNA methylation differences have relevance with Adiantum reniforme var. Sinense growth and development in different periods;After sequencing, two kinds of different genes, namely S2 P family proteins and WRKY transcription factors, are found. CpG island projections for these two genes find that S2 P has not CpG island, but the GC content reaches 55%; WRKY transcription factors exist in the two CpG island and GC content is up to 80%.And their different expression quantity in six different development period by real-time quantitative calculation illustrates that S2P’s expression quantity form is highest in sporophyte differentiation period of asexual reproduction.And S2 P sequence is cutting from selective amplification in sporophyte differentiation period of asexual reproduction,which speculates that S2 P methylation may play an important role in asexual spores differentiation; WRKY transcription factor has high expression guantity in the formation of sexual reproduction sporophyte and asexual reproduction of meristem differentiation period. WRKY sequence is cutting from selective amplification in sexual sporophyte, which speculates that WRKY methylation may play an important role in the formation of sexual sporophyte.