Construction Of Degradation System SMAX1-likes-Venus Induced By Strigolactone In Arabidopsis Thaliana

Strigolactones(SLs)is a new class of novel plant hormones associated with plant branching.Although great progress in biosynthesis and signal transduction were made in recent years,a little was known in the temporal-spatial localization and monitoring for SL.In this study,the SMAX1-likes genes,homologous gene of rice D53,an inhibitor for SL signaling,were found in Arabidopsis thaliana according to the method of bioinformatics.The SMAX1-like gene were cloned and fused with VENUS,and transformed into Arabidopsis thaliana,which will lay the foundation for the temporal-spatial localization and monitoring of SL in Arabidopsis thaliana.1.The SMAXL6,SMAXL7 and SMAXL8 genes homologous gene of OsD53 in Arabidopsis thaliana were cloned and SMAXL6-VENUS,SMAXL7-VENUS,SMAXL8-VENUS fusion expression vector were constructed.2.SMAXL7 showed highly similar spatial structure with D53,and SMAXL7 was a possible transcriptional inhibiting factor of SL signaling pathway in Arabidopsis thaliana.SMAXL7,D1-SMAXL7,N-SMAXL7,D2-SMAXL7 and VENUS fusion expression vectors were constructed since SMAXL7 contains four functional domains M,D1,N and D2.3.SMAXL7-VENUES was well expressed in nucleus through fluorescence expression analysis in transient transformation protoplasts of Arabidopsis thaliana.In order to obtain stable Arabidopsis thaliana transgenic lines,M-SMAXL7-VENUS,D1-SMAXL7-VENUS,N-SMAXL7-VENUS,D2-SMAXL7-VENUS,SMAXL7-VENUS were transferred into Agrobacterium and transformed into the wild-type(Col-0)Arabidopsis thaliana,and then all stable transgenic lines were screened.4.M-SMAXL7,D1-SMAXL7,N-SMAXL7-VENUS,D2-SMAXL7 fusion protein were obtained by in vitro expression in prokaryotic expression system,which laid the foundation for the further study of SLs.