Fuction Analysis Of Promoter Of PDF Gene In Vitis Vinifera And Expression Regulation Of Transcription Factor ERF For PDF Gene

Various biotic and abiotic stresses were the important factors that affected crop growth, development and yield. The interactions between transcription factor and promoter play a very important role during the expression and regulation of stress-related genes. The regulation relationship investigation for the transcription factor and stress-related genes has important theoretical and practical significance in terms of improving the resistance of plants. In this paper, systematic studies were performed about the interaction between the cis-acting element of grape plant defensins gene (PDF) promoter and ethylene response factor (ERF). The following results were obtained.1. Amplified by PCR, the VvPDF2 and VvPDF1-2 gene promoter sequences of European grape (Vitis vinifera) were obtained with the length 1819bp and 1586bp respectively. The cis-Acting elements of the promoter sequences were analyzed by use of online procedures PLACE and PlantCARE. The results indicated that several cis-acting elements involved in plant hormone response and response to biotic and abiotic stress in the VvPDF1-2 and VvPDF2 gene promoter, such as the ERE element, TC-rich element, HSE components and W-box elements.2. The expression vectors were transfect into the Arabidopsis protoplasts using PEG-Ca2 +-mediated method and the reporter gene expression were detected by fluorescence intensity of LUC in the transient expression system. Then we analyzed the activity of the promoters.3.Analysis the interaction between the VvERF3b and VvERF2-1 transcription factors and VvPDF1-2 gene promoter were performed by Arabidopsis protoplast transient expression system.To further study the interaction between the VvERF2-1 and VvERF3b transcription factors and the VvPDF1-2 gene promoter, we constructed the VvERF3b and VvERF2-1 transcription factor expression vectors. Double plasmids were transfect into Arabidopsis protoplast transient expression and relative LUC expression was detected. The results showed that, compared with the control, VvERF2-1 and VvERF3b transcription factors could work with VvPDF1-2 gene promoter. VvERF2-1 transcription factor, which enhanced the VvPDF1-2 gene transcription activity, could activate the transcription of the 5 'deletion fragments of VvPDF1-2 promoter. VvERF3b transcription factors, which inhibited the activity of the VvPDF1-2 gene promoter, could inhibit the transcription of the 5 'deletion fragments of VvPDF1-2 gene promoter. In summary, this study obtained two VvPDF gene promoter sequences by PCR and constructed the related expression vectors of the 5 'deletion of the promoter. Using Arabidopsis protoplast transient expression system to express reporter genes and testing report gene activity to analysis the promoter activity. We analyzed the relationship of the VvERF2-1, VvERF3b promoters and VvPDF1-2 transcription factor. The experimental results not only establishes the foundation for the further studies about the mechanism of VvPDF1-2 gene promoter and the VvERF transcription factor, but also provides a theoretical basis for studying the molecular mechanism of resistance of grape.