Transformation Of Petris Vittata L. Phytochelatins Synthases Gene, PvPCS1 In Arabidopsis Thaliana

Along with the human activities development and industry technologydevelopment, the content of heavy metal in soil becomes higher and higher. Heavymetal pollution has increasingly threatened human health and life quality, and hasbecome a serious society and environment issue. Arsenic usually exists in theformation of compounds. The content of arsenic in nature is luxuriant. As people allknow, the toxicity of arsenic is quite strong. It can result in malformation, cancer andmutation effect. U.S Environmental Protection Agency (EPA) has classified arsenicthe A-Carcinogen. In recent years, a number of arsenic leaked into soil because ofhuman activities and industry development. Nowadays, Arsenic contaminationbecomes one of the public concerns.The phytoremediation technology has been developed in recent years. Becauseof the lower costs, better effects and the character of enviromnental amity, thistechnology gradually becomes the promising tool for eliminating the heavy metalpollution. Especially the discovery of Pteris vittata L., the first plant identified as anarsenic hyperaccumultor, makes the utilization of phytoremediation technology afeasible approach to handle the arsenic-contaminated soil. Pteris vittata L. almostbecomes the modle plant in the investigation of arsenic-hyperaccumulating plants.Numerous scholars analyze its characteristics from physical, chemical, biologicalaspects, and hope to provide some theoretical and scientific basis during theapphcation of the plant for remediating arsenic-contaminated soils.However, due to Pteris vittata L. is a fern which has a little biomass, and growsvery slowly. With the development of molecular biology and genetic engineering, it isvery possible to construct a new transgenic plant which bears the characteristics ofhyperaccumulating arsenic, fast growing and having more biomass at the same time.Thus, the transgenic plant can be used in the phytoremediation more effectively.In another aspect, the discovery, cloning and transformation of thehyperaccumulating critical genes, such as phytochelatin synthase gene, makes itpossible to construct very efficient transgenic heavy-metal hyperaccumulating plants. In this study, the phytochelatins synthase gene in P. vittata, PvPCS1, was clonedand fused with the promoter Ca MV 35S to construct the overexpression vectorpMON530-PvPCS1. Subsequently, it was transformted into wild-type Arabidopsis.After dried, the T₀ seeds was screened on the PNS media with 50μg/mL kanamycinand got 72 resistent plants. Then the plants were identified by PCR, and 69 positiveplants were got. Final, RT-PCR experiment was performed to investigate theexpression of PvPCS1 and the result showed that the gene, PvPCS1, was expressed inthe transgenic plant at a high level. All the result of experiments proved that PvPCS1was successfully transformed into Arabidopis thaliana.The significance of this study is to construct a transgenic plant that express thegene, PvPCS1, in Arabidopsis. Then we can investigate the function of PvPCS1 inhyperaccumulating arsenic. Up to now, we have got the TI seeds. The further study isto screen the homozyouse plants, so that we can use it to investigate the sensitive andthe resistive ability of the transgenic plants. The ultimately purpose of this research isto analysis the function of PvPCS1 in high plants and to construct a more efficientplant that can be used in the phytoremediation of arsenic-contaminated soil. Thisstudy was the first of this research and established a stabile foundation for the furtherexperiment.