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

In the processes of growth and development, plants often encounters biotic and abiotic stresses, which can retard growth and productivity of plants. Studies have shown that while the plants sense stresses, many resilience-related genes will be affected by the transcription factors, resist to external stress and protect the plant. It is thus clear that resilience-related genes play an important role in plant defense responses, so the understanding of molecular regulation mechanism of resilience-related genes is very necessary. The gene of pathogenesis-related protein (PRs) is one of resilience-related genes, in this study, the promoter of VvPR3gene was cloned from grape, and then analysis of cis-acting element and the interaction relationship between cis-acting element and ethylene response factor(ERF) wre carried out. The main results are as follows:1. the VvPR3gene promoter of European grape (Vitis vinifera) were obtained with the length1775bp by PCR, the analysis indicated this sequence contains kinds of cis-acting element, some of these answer various plant hormones, such as ERE, CGTCA-motif, W-box, TCA-element, MYBGAHV and P-box et al; and others response to biotic and abiotic stress, such as MBS, CCAAT-box and ARE et al.2.5’deletion of VvPR3gene promoter were cloned to construct four expression vectors that contain different length. These four reorganized expression vector were transferred into A. thaliana protoplast cells by PEG-Ca2+,and then detected the expression level of Luciferase reporter gene. The result showed that with the increase of the5’deletion fragment in length, the expression amount of reporter gene showed downtrend. That meaned there were cis-acting elements which can restrain the downstream gene expression in every5’deletion fragment.3. The VvERF2and VvERF3b transcription factors that has been build in laboratory and four deletion fragment expression vector were transferred into A. thaliana protoplast cells by PEG-Ca2+, and then detected the expression level of Luciferase reporter gene. The results shown that VvERF2transcription factor, which enhanced the VvPR3gene promoter5’ deletion fragments activity, could activate the transcription of the VvPR3gene promoter.; VvERF3b transcription factors, which inhibited the activity of the VvPR3gene promoter5’ deletion fragments, could inhibit the transcription of the VvPR3gene promoter.In summary, in this preliminary study we made clear the type and characteristics of cis-acting element in VvPR3gene promoter, and the interaction relationship between cis-acting element and ethylene response factor(ERF). The results laid the foundation for the further research on molecular mechanism of the regulation of gene expression, and provide a theoretical basis for the breeding of high-resistant plants.