Subcellular Localization And Expression Analysis Of BASL Protein Family

Cell polarity is the basis of cell diversity and the cornerstone of organism morphogenesis and function.The polarity produced by plants at the cell and tissue level is usually related to the cell polarity localization of the protein complex.In the study of Arabidopsis stomatal development,BASL protein was found to exhibit dynamic polar localization at the subcellular level and plays a role in regulating asymmetric cell division.However,studies have shown that the subcellular polar localization of BASL is not limited to stomatal lineages,which indtates BASL may also be involved in regulation of other plant tissue development.In order to analyze the subcellular localization and molecular function of the BASL protein from an evolutionary perspective,we chose to explore it in Physcomitrella patens and rice.This study includes four parts,and the following results have been achieved:(1)The homologous genes of the BSAL protein in rice are Os02g0582100 and Os04g0467100;in P.patens are Pp1s25 and Pp1s37.We constructed the CRISPR/Cas9 knockout vectors and the promoter reporting system vectors of P.patens and rice.The sequencing results show that all vectors were constructed accurately.(2)The CRISPR/Cas9 knockout mutants of rice were obtained by Agrobacterium-mediated transformation of japonica rice.After hybridization and screening,we identified 111 transgenic rice plants,and a total of 8 stable T3 homozygous transgenic lines were obtained.No obvious phenotype was observed in the vegetative stage.(3)Through PEG-mediated protoplast transformation,the CRISPR/Cas9 knockout mutants and promoter reporter strains of P.patens were obtained.We observed that the BASL homologous have very clear localization signal in the axillary tissue.(4)To obtain sufficient embryogenic callus and transgenic seedlings,we optimized the cotton transgenic system,which will help to explore the function of BASL protein in higher dicotyledonous cotton plants.