Genetic Transformation Of Chorispora Bungeana Cold-Induced Genes In Sugarcane

It has known that sugarcane (Saccharum officinarum L.) is one of the most important sugar crops in China and throughout the world. And Guangxi province has the largest number of raw materials of sugarcane. However, due to the frequence of cold stress in recent years, it has lead to large reduction of sugarcane production and sugar content, resulting in huge economic losses. As sugarcane, one of the vegetatively propagated crops, is a complex polyploid, such as aneuploid, highly heterozygous, and other characteristics. So it is probably very difficult to manage the cold stress using conventional breeding strategies. While application of transgenic technique could accelerate the breeding of cold tolerant sugarcane varieties.In this study, cold tolerance gene CbCOR15a from Chorispora bungeana was used to construct the dual expression vector pCambia3300-CbCOR15a which carried the selection marker bar gene and CaMV35S promoter. Then, the constructed vector was applied to transform sugarcane embryonic calli by Agrobacterium tumerfaciens-mediated method.The main results were listed as following:1The results show that different hormone combinations affected the differentiation of sugarcane callus, and the combination use of2.0mg/L KT and0.2mg/L NAA differentiation medium has the highest differentiation rate of89%.2The statistical data indicated that the rooting medium containing2.0mg/L NAA and70g/L sucrose was most suitable for rooting.3The most proper herbicide PPT selection concentrations were0.75mg/L and 0.5mg/L for callus and seedlings, respectively.4The results suggested that the best antimicrobial concentration is500mg/L cefotaxime sodium (Cef) for3days callus after gene transferred, and300mg/L for the seedlings after differentiation,200mg/L for the seedlings after rooting.5Nine transgenic plants were obtained and four plants were confirmed by using the PCR method. It has primarily shown that the COR gene has been transferred into the sugarcane.The present experiment has established the genetic transformation system of sugarcane and successfully obtained the transgenic plants.