| The treatment of additional CaCl2under proper dose and timing not only promoted the female gametophyte, but also increased the number of archegonia of Adiantum reniforme var. sinense. The Ca2+concentration in cellular was higher after the treatment and sex organs formation and mainly distributed along the cell membrane, which suggested that the exist of foreign calcium promoted the sex organ formation. In this research, some filial gametophytes could produce sporophytes in agamic reproduction way along the gametophyte in different places at the same time and formed sporophyte in sexual way meristem area after additional CaCl2treated. All of the agamic reproduction belonged to misogamy.At gene level, MSAP were used to analyze the DNA methylation level in this study. Results showed that about26-30%cytosine of CCGG sequences in A. reniforme var. sinense genome was methylated.1)in the different stages of gametophyte without sex organs, with sex organs and with mass sex organs, the average of methylation was increased with sex organs formation and number increasing. DNA hypermethylation and demethylation happened mostly in mass sex organs stage. Many fragments were found to be the homology gene with WRKY and WAK in rice.2)Changes of DNA methylation during somatic embryogenesis were detected:in gametophyte cell(19.2%), in embryonal connective tissue (17.9%), in embryo (20.1%), in young sporophyte (19.5%). Hypermethlation happened in embryonal connective tissue. Results in this study showed that DNA methylation was connected with sex organ formation and somatic embryo development closely and directly. |
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