| Arabidopsis TRANSPORTIN 1(TRN1)belongs to Importin ? family,whose main function is to mediate nuclear and cytoplasmic transport of related proteins as receptors.If the nuclear-cytoplasmic transport can not proceed smoothly,the target protein can not function normally,thus affecting the normal life activities of cells and even the whole plant.Based on the results of previous studies in our laboratory,the comparative analysis of homozygous mutant trn1 and wild type(Col)plants in apical meristem-related marker gene expression,plant hormone response,and quantitative analysis of genes interacting with TRN1 were carried out to explore the function of TRN1 in plant growth and development.The stem apical meristem of trn1 homozygous mutant developed slowly and irregularly,and the root apical meristem developed defectively.In order to explore its molecular mechanism,we introduced WUS or CLV3 gene promoter-driven GUS gene(pWUS: GUS and pCLV3: GUS),QC-specific marker gene promoter-driven GUS gene(QC46: GUS and QC184: GUS)into trn1 mutant by hybridization respectively,and DR5: GUS,cell cycle marker gene pCYCB: GUS,PLT1,PLT2 and SHR gene promoter-driven pPLT1: CFP and pPLT2: CFP,as well as fusion genes such as pPLT1: PLT1: YFP,pPLT2: PLT2: YFP and pSHR: SHR: eGFP,which play important roles in the regulation of root meristem cells.By detecting the expression of these genes,it was found that all these genes in homozygous mutants of trn1 were affected to a certain extent,regardless of the location or quantity of expression.In addition,we explored the response of homozygous mutant trn1 and WT to cytokinin and auxin,and analyzed the expression levels of related genes in their signaling pathways.The results showed that the sensitivity of homozygous mutant trn1 to cytokinin increased,but the sensitivity to auxin decreased,and found that the expression levels of genes related to hormone response signaling pathway changed.We quantitatively analyzed the results of yeast hybridization and found that the expression of related genes in homozygous mutants of trn1 had changed.Based on this,we further analyzed the function of interacting genes.Through this study,we found that TRN1 affects the development of plant apical meristem by affecting the expression of WUS,CLV3,PLT1,PLT2,SHR and CYCB genes during plant growth and development,and TRN1 affects plant response to cytokinin by regulating the expression of ARRs response factor in cytokinin signaling pathway.The expression of related genes affects plant response to auxin.In addition,TRN1 can interact with a variety of proteins,ultimately affecting a variety of biological processes.Therefore,we believe that TRN1,as a nuclear transport protein in Arabidopsis thaliana,participates in many biological processes by affecting the transport of a series of substrate proteins,and creates conditions for us to search for TRN1 substrates in the future. |
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