Cloning Of AtPSK Genes And Expression Of AtPSK2 Gene In Roselle Cell

Plant cell culture techniques possess great potential application for the production of Chinese medicines, phytomedicines and the other natural compounds. It can solve many problems once and for all in resource availability and environmental protection, which arise in the production of these substance with traditional methods, including chemical synthesis, extraction from wild plant material and field cultivation. However, duo to the slower growth, longer growth cycle and easier contamination of plant cells, the industrialization process of plant cell culture techniques was very slow. Therefore, in order to obtain a transgenic Roselle cell line that can initiate cell division and proliferation at low cell density, a efficient genetic transformation system of Roselle cell was established at first, and then AtPSK2, the phytosulfokine gene from Arabidopsis thaliana genome, that encodes precursor of phytosulfokine was introduced into Roselle cell by the above genetic transformation system and expressed functionally. The main results of the research are summarized as follows.1. By using Agrobacterium tumefaciens LBA4404 and EHA105, harboring pBI121 plasmid, as donor strains and by studying systematically on the basic conditions of genetic transformation, two efficient genetic transformation systems were initiatively established. When both cotyledon or hypocotyls of sterile seedlings and callus were used as recipient material, the transgenic callus expressed stably GUS activity and transgenic cell line expressed stably NPT II activity were gained, however, the transformation rate of cotyledon or hypocotyls was much higher than that of callus, the former was 29.4% and the latter only 4%. As a inducer, acetosyringone can greatly raise the transformation rate when callus was used as recipient material. In addition, Agrobacterium tumefaciens LBA4404 strain was more suitable for the transformation of Roselle.2. According to the sequence information in Arabidopsis genome database, two pair of specific primers were designed and synthesized for the...