Transformation And Expression Of UGPase And Anti-sense 4CL Genes In Tobacco

What impacts on cellulose and lignin synthesis in transgenic tobacco with two genes coming from Amorpha fruticosa were studied, one is UGPase gene that coding UDP-glucose pyrophosphorylase, the other is 4CL gene that coding 4-coumarate:CoA ligase. This study provides theory and technique can support for cultivating lower content of lignin, higher content of cellulose plant, and raveling out the problem of pollution which disposing lignin causes during paper pulping by genie engineering.This study made use of UGPase and 4CL genes firstly to insert plasmid pBI121 with PCR amplification and Restriction endonuclease, to construct plant express vector pBU, pBa4CL and vector with two genes pBU-a4CL. Three vectors were transferred into Agrobacterium tumefaciens LBA4404.Based on that, mediated by Agrobacterium tumefaciens, the UGPase gene, anti-sense 4CL gene and fused gene synthesized those two genes, was transformed into tobacco separately. Results of PCR and Southern blot determination indicated that they are all integrated into the genome successfully. The holocellulose and klason lignin content of transgenic tobaccos growing 5 months were measuredrthe transformed tobaccos with UGPase genes showed that enhancing the expression of UGPase gene could improve the content of cellulose by 2.4%-6.3%, but which did not affect the content of lignin; the transformed tobaccos with wti-4CL genes showed that down-regulating the expression of 4CL gene could reduce the concent of lignin by 29.6%～41.2%, but which did not affect the content of cellulose; the transformed tobaccos with UGPase and anti-4CL genes showed that the content of holocellulose increased by 3.4%～6.8%, and their content of klason lignin reduced by 28.1%～42.8% at the same moment.In addition, this study compares the thickness of cell wall and the color of stem between transgenic or not tobacco. Results show that, the cell wall thickness of transgenic tobacco with UGPase gene becomes thicker; the stem color of transgenic tobacco with anti-4CL gene displays brown strips. Which illuminates the change of cellulose and lignin content can affect tissue and cell in transgenic tobacco, that result provides method for differentiating transgenic...