The Study Of Medicago Truncatula Plant Defensin Gene (MsDef1) Genetic Transformation Of Sugarcane

Sugarcane(Saccharum officinarum L.) is a very important sugar and economic crops, and also a very important energy crops. But damage by disease has greatly reduced the sugar content, output and economic benefits. Sugarcane smut, caused by Ustilago scitaminea Syd, is one of the severe diseases in sugarcane. Traditional chemical prevention affect weakly on Sugarcane smut. Breeding antivirus sugarcane varieties through genetic engineering will provide an effective way to prevent this disease.MsDefl is one kind of plant defensins divided from Medicago truncatula. It Shows very effective antifungal activity in vitro. We hope to get some broad spectrum resistance plants against fungal through genetic engineering.The main results are as follows:1. The plant expression vector pMsDN containing the native MsDefl gene and bar gene was constructed. In addition, another vector pMsDS containing synthetic MsDefl gene and bar gene was constructed.2. Two expression vectors were introduced into sugarcane by Agrobatorinm mediated transformation.34 PCR positive plants containing native MsDefl and 33 PCR positive plants containing synthetic MsDefl were obtained. The results showed that the foreign genes had been probably integrated into sugarcane genome.3. Four native MsDef1 and six synthetic MsDefl PCR positive plants were selected to detect by RT-PCR.3 native MsDefl and 4 synthetic MsDef1 transgenic plants were positive. The results showed that the foreign genes expressed on the transcription level.4. Five synthetic MsDefl PCR positive plants were selected to detect by Southern Blot. It had been showed hybridization bands in transgenic plants No.3 and 4.