The Role Of Arabidopsis Transcription Factors LFY2 And MYB2 In The Regulatory Network Of Anther Development

The development of flowers and anthers is closely related to the reproduction of higher plants and human life,for which the development of anthers has become one of the current research hotspots.Previous study suggested that the absence of LFY2 and MYB2 would affect the degradation of tapetum and pollen fertility.This research further studied the molecular mechanism of transcription factors LFY2 and MYB2 regulating flower and anther development in Arabidopsis,which laid a foundation for the research of angiosperms flower development.Using Arabidopsis as experimental material,the expression characteristics of LFY2 and MYB2 were analyzed by GUS staining and q PCR.Prokaryotic expression vectors of LFY2 and MYB2 were constructed and recombinant proteins were obtained after purification by affinity chromatography.The interaction between the two transcription factors and the known related genes was verified by EMSA in vitro.In addition,the role of transcription factors LFY2 and MYB2 in anther development regulation network was explored through the summary and analysis of existing literatures and databases.The research results are as follows:(1)LFY2 gene expresses in many tissue sites during the growth stage of Arabidopsis thaliana,and appears to be highly expressed in flowers near the stage of tapetum degradation.MYB2 also expresses in flowers near the stage of tapetum degradation,indicating that these two transcription factors may be involved in the regulation of anther development and affect pollen fertility in addition to participating in physiological processes of other tissues.(2)It was proved by EMSA that LFY2 does not directly regulate CEP1.Based on the analysis of existing literature and databases,it is predicted that there are binding sites of transcription factors AG,AP1,AP3,MYB33,MYB65,DYT1 and MYB2 related to anther development on the promoter of LFY2,indicating that LFY2 should be a member of the regulatory network of Arabidopsis flower and anther development.It was also verified that MYB2 could directly bind to the promoter sequence of CEP1,LFY2 and ?VPE in vitro.The MYB2 binding sites can be concluded as double-stranded fragments composed of 5'-CNGTTR-3' and 5'-YAACNG-3' complementary sequences and 5'-AAACCA-3' and 5'-TGGTTT-3' complementary sequences.It is speculated that MYB2 can regulate the expression of these three genes by directly combining the promoters of these three genes.(3)Combined with the results of previous studies and the results of this study,the regulatory network of flower development and anther development was linked by transcription factors AP1,LFY2 and MYB2.The regulatory network of flower and anther development was perfected.LFY2 and MYB2 were important components in the anther development regulation network.