Cloning Of SpsLAS In Satix Psammophila,Tissue Specific Expression And Expression Vector Construction

Branching development,playing an important role in plant architecture,is the vital tree trait which would make effects on plant spatial patterning,photosynthetic efficiency,biomass,carbon sequestration efficiency,afforestation and landscape.In this research,LAS(LATERAL SUPRESSOR)gene was cloned succssfully by homologous cloning method from Salix psammophila.Bioinformatics analysis,tissue specific expression and construction of overexpression vector were finished.The main results are as follows:1.The coding sequence of SpsLAS gene was obtained by homologous cloning method.Amino acid conserved sequence analysis showed that the protein has typical characteristics of GRAS protein family and belongs to the LS subfamily.2.Phylogenetic analysis of homologous protei n showed that Salix psammophila has high homology with LAS protein of other plant species,especially is closely related to Salix purpurea and Populus trichocarpa in woody plants.3.Tissue-specific expression results of the SpsLAS gene showed that the gene’s highest expression is detected from the bud,and then decreased respectively from the root,stem 3,leaf axils,mature leaves,stem 1,phloem,xylem and stem 2.4.To make the LR recombination reactions between enzyme-digesed products and pMDC32 overexpression vector,then transform into the kanamycin-contained LB medium,screening and sequencing results showed that the overexpression vector of SpsLAS was successfully constructed.The above results showed that the SpsLAS gene is doned successfully and SpsLAS function may have an important relationship with branch controlling.This research laid the foundation for further analysis of LAS gene in woody plants and exploration of GRAS gene family in woody plants.