Functional Research Of Cinnamyl Alcohol Dehydrogenase In Lignin Of Bambusa Multiplex (Lour.)

Bambusa multiplex (Lour.)Raeuschel is a Bambusa plant belonged to Bambusoideae of Gramineae. It’s all know that China is the center of the world’s Bamboo resources, the area of bamboo plants is one third of the total area of world bamboo plants. The Bambusa multiplex (Lour.)Raeuschel, which has the characteristics of fast growth and high cellulose content and so on, the most widely distributed cluster bamboo in China. The Bambusa multiplex (Lour.)Raeuschel is a good papermaking raw material, with high economic value. However, the content of Lignin in bamboo plants is too lager to make paper. We should remove the Lignin in the process of paper industry using bamboo. In the remove process, the Chemicals is large used, which not only increases the cost of paper making, but also pollutes the environment seriously. So, it is very important significance to use molecular biology and genetic engineering to cultivate new varieties of bamboo with low Lignin content under the premise of no affect the growth of bamboo.The cinnamyl alcohol dehydrogenase (CAD), which affects the last process of synthesis reaction of monolignol, is one of the key enzymes in pathway of lignin biosynthesis, and is important in the regulation of lignin biosynthesis. In our research, the fusion expression vector and over-expression vector of CAD gene were constructed. And through particle bombardment onion epidermal cells to identify the position of CAD gene in cell; In addition, we measured the Lignin and cellulose content in the transgenic plants. In the research of the function of CAD gene, we got the following result:1) Identified the length of CAD gens in Bambusa multiplex (Lour.)Raeuschel by PCR amplification, the fusion expression vector named p1302-GFP-CAD and the over-expression vector named CPB-CAD were constructed, respectively.2) Through particle bombardment, turned the fusion expression vector p1302-CAD into the epidermis cell of onion. Under fluorescence microscope, we found that the CAD gene was in the epidermis cell of onion, it implying that the CAD protein has a signal in nucleus.3) The over-expression vector CPB-CAD was transferred into rice by agrobacterium-mediated method. We identified two positive transgenic plants by PCR detection.4) The content of Lignin and Cellulose of transgenic plants and wild plants were measured. The results show that compared with the wild type, lignin content of positive transgenic plants had largely enhanced, average increased50.0%; Cellulose content also corresponding reduced, a drop of about2.6%on average。...