Inorganic Pyrophosphatase Gene Genetic Transformation Of Sugarcane

Sugarcane(Saccharum officinarum L.), which is not only the most importment sugar crops, but also an important energy crops in china. The main goal of sugarcane breeding is to increase the sugar content in sugarcane. Enhance the ability for sucrose synthesis in sugarcane is an efficient method to increase the sugar content in sugarcane.The sucrose synthesis of sugarcane is taken place in the mesophyll cells. During the processing of sucrose synthesis, a large amount of PPi would be produced. The accumulation of PPi in mesophyll cells largely can inhibit the sucrose synthesis. The PPi can be cleaved by PPase. Transfer the PPase gene into sugarcane and make it express in mesophyll cells can decrease the accumulation of PPi and therefore increase the sucrose synthesis. Ultimately it is possible to increase the sugar content in sugarcane.According to the sequence of inorganic pyrophosphatase gene in Saccharomyces cerevisiae, the PPase gene was amplified by using PCR technique from Saccharomyces cerevisiae. A plant expression vector, pCBIPR, in which, the PPase gene was driven by the mesophyll cell specificity expression promoter rbcS, was constructed. Then the pCBIPR was introduced into the super-poison Agrobacterium tumerfaciens EHA105 through the"Freezing and thawing"method and used for transformed sugarcene embryonic calli by Agrobacterium tumerfaciens-mediated method. The transformed shoots were screened under the selective pressure of PPT and 56 resistant shoots were obtained. Two transgenic plants were confirmed by using the method of PCR from 9 resistant shoots. It primary showed that the PPase gene was integrated into the sugarcane genome.