Synthetic Of PRX Gene In Vitis Vinifera And Functional Analysis Of The Gene Using Arabidopsis Thaliana

In recent years, due to the continued deterioration of the global environment, plants had to suffer from a lot of abiotic stresses, such as soil drought, salinity, extremely weather. These abiotic stresses generated serious adverse effects on plants. The improvement of fruit trees for abiotic stress tolerance is particularly important because the growth of fruit trees is a long-term process, often taking more than 10 years. However, traditional fruit trees breeding method has many shortcomings. It is time consuming process and affected by many factors with the improvement of fruit trees. With the development of genetic engineering technology, it has became possible to use the genetic engineering technology to solve some of the traditional problems in the process of fruit trees breeding.Peroxiredoxins (Prxs) are a class of peroxidase present in all organisms, and they play major role in plants’growth. Some studies suggest that Prxs have a strong antioxidant capacity. Prxs can remove active oxygen and free radicals in plants and also have protective effect on photosynthesis. Grape is the popular fruit tree. Due to its highly economic value and nutritional value, grape has been cultivated widely in the world. In this experiment, we have optimized the gene of peroxiredoxin in grape successfully by synthetic method and studied its function. The results show that peroxiredoxin of grape significantly improved salinity resistance of plants. It has important application value for grape or other fruit trees breeding, meanwhile, it has provided theoretical foundation in research of resistance and character improvement of fruit.In this paper, we chemically synthesized the peroxiredoxin gene in Vitis vinifera based on the amino acid sequences released on the NCBI. We optimize the nuclear sequence by balancing the radio of G+C/A+T, deleting six or more consecutive AT sequence and more then five GC sequence, removing reverse repeats sequences and sequences to form stem-loop structure and transcription termination signal, and increasing the free energy of nuclear at the 5’domain, decreasing the free energy of nuclear at the 5’domain. The synthesized gene, named VvPRXI, was transferred into wild Arabidopsis thaliana by agrobacterium floral dip method.We treated the wild type and transgenic type with different concentrations of NaCl salinity stress on seeds, seedlings and plants and detected several resistance physiological indicators under 150 mM NaCl salinity stress. Comparison the seed germination rate percentage between the transgenic type and wild-type, we found that the inhibition of high salinity stress on wild-type is significantly stronger than transgenic plants. Under high salinity stress, growth of the transgenic plants was better than wild-type plants. Moreover, the changes of chlorophyll, proline and malondialdehyde contents intuitional explain the transgenic type plants have stronger tolerance on high salinity. All these results showed that, under high salinity stress, the VvPRXI over-expressing transgenic plants exhibited more resistance to salinity stress than wild-type plants.