Regulation Mechanism Of Histone Deacetylase Subunit SAP30C In Rice Heading

Based on the significant conclusion of the association analysis between OsSAP30C which is encoding a deacetylase subunit gene and heading dates,we have construct the gene editing Crisp-cas9 vector which is transformed into wild-type NIP(Nipponia).In comparison with heading dates of mutant lines and wild type under the short-day and long-day condition,the homozygous mutants showed delayed heading date obviously under SD condition.In this research,we study the photoperiod pathway and molecular mechanism of controlling rice headings.Solutions presented in this research can be summarized as follows:1.As a subunit of histone deacetylase complex,OsSAP30C is involved in regulating circadian of rice flowering under SD condition.Mutants showed the delayed heading by 14 days compared with wild type.2.OsSAP30C is an epigenetic protein located in the nucleus by subcellular localization strategy.3.Transgenic plants of GUS fused with OsSAP30C promoter displayed strong activity in vascular bundles mainly in rice leaf veins.4.It was found that transcripts level of Hd3a have decreased observably under SD circumstance whereas other heading-related genes of deficient plants showed inalteration by qPCR detection.5.Based on the rhythmic expression detection,it was found that the peak value of Hdl in ossap30c(ZT=16h)under SD condition moved forward compared with the wild type with resulting in abnormal circadian.6.Through the detection of total histone H3 and H4 acetylation,no significant changes appeared in these histone modifications under SD environment,suggesting that deacetylase defects may affect other type of amino acid modifications which promotes genes concerning to heading stage and yield to participate in histone modification.7.Through the assays of histone deactylation enzamy activity of SAP30C and SAP 18 which are purified from prokaryotic expression,we obtained that SAP30C can play a histone deacetylated function in vitro strongly,but the activity of SAP30C weakened by mingling with SAP18.8.Yeast point-to-point double hybrid experiments,BIFC verifications,CoIP and LCI tests showed that OsSAP30C,as a component of the deacetylase complex can interact with SAP 18,HDA9,HDA19,SNL2,HAD and SAP30B directly or indirectly.9.It was proved that SAP30C is deposited on upstream sequence of Hdl but not Ehdl flanking regions by ChIP-qPCR inspection in ossap30c and wild type.