Cloning And Functional Analysis Of BpbZIP1 Gene From Birch

bZIP transcription factor(basic-region leucine zipper)is a basic leucine zipper transcription factor.It is one of the largest transcription families.It has many functions to regulate and participate in many important abiotic stress responses.The anti-stress function of these bZIP transcription factors has also been verified a lot.Identification of the BpbZIPl gene of Betula platyphylla Suk resistant to abiotic stress provides data and materials for studying the molecular basis of resistance formation and the genetic improvement of resistance.In this study,birch seedlings were used as materials,and genomic information was used to clone the BpbZIP1 gene of birch.Phylogenetic analysis of resistance-related genes using bioinformatics analysis.It is grouped with Arabidopsis resistance-related gene proteins.The abiotic stress response analysis of the BpbZIPl gene,the experimental results are as follows:The quantitative expression of BpbZIPl gene and several resistance-related SOD and POD genes under salt and drought stress in birch was analyzed by real-time quantitative fluorescence method.The expression pattern of BpbZIPl gene is similar to that of SOD and POD genes.It is induced by NaCl treatment,and the expression change is not obvious under mannitol treatment.The BpbZIP1-PBI121-GFP fusion expression vector was transformed into onion epidermal cells for subcellular localization using a gene gun-mediated transient transformation method.The results showed that the BpbZIP1 gene was localized on the nucleus.The BpbZIP1 gene overexpression vector was transformed into Arabidopsis thaliana by Agrobacterium-mediated transformation.Histochemical staining(NBT,DAB,Evans Blue)and physiological and biochemical indicators(SOD/POD activity)were performed after the transgenic material was treated with salt and drought stress.analysis.Under the condition of NaCl stress,the transgenic lines had stronger ability to scavenge active oxygen than the wild-type control,and the activities of SOD and POD increased;under the condition of mannitol stress,the scavenging ability of active oxygen was not significantly different from that of the control,SOD activity increased,and POD activity increased.less.The BpbZIPl gene can improve the salt tolerance of Arabidopsis transiently transformed strains by eliminating reactive oxygen species.Drought resistance needs to be verified in the homologous birch transgene.A rapid gene function analysis was performed by a transient transformation method.The BpbZIPl gene overexpression vector was transiently transformed into white birch,and histochemical resistance(NBT,DAB,Evans Blue)and physiological indicators(SOD/POD activity)were used to determine the drought and salt tolerance functions.Under the condition of NaCl stress,the transgenic lines had stronger ability to scavenge active oxygen than the wild-type control,and the activities of SOD and POD increased.Under the condition of mannitol stress,the scavenging ability of active oxygen was not significantly different from that of the control.less.The results showed that the BpbZIP1 gene could improve the salt tolerance of the birch transiently transformed lines by eliminating active oxygen,and the drought resistance was not obvious.The yeast one-hybrid was used to preliminary identify the binding of transcription factors to the ABRE/G-box element.BpbZIP1 is able to specifically bind to the ABRE/G-box element and specifically to the promoter fragment containing the ABRE/G-box element.Identification of the birch BpbZIP1 gene and its downstream gene-binding elements with salt tolerance capacity provided basic data for further analysis of the salt-tolerant molecular regulation mechanism of birch BpbZIP1 gene.