| Sugarcan.e(Saccharum officinarum L.) is an important sugar and economic crops, and else an important energy crops. But damage by disease and insect 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.MtDef4 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.This study constructe two vectors screened by bar gene, the native MtDef4/KDEL gene’s plant expression vector pADN and the synthetic MtDef4/KDEL gene’s plant expression vector pADS. Then we introduced them into agrobacterium tumefaciens EHA105 through freezing and thawing method, and utilized agrobacterium-mediated transformation to transform sugarcane’s embryogenic callus.92 and 47 PCR positive plants are obtained respectively. Then we analyzed parts of PCR positive plants using RT-PCR.5 of 6 plants, choosed from 92 PCR positive plants, and 4 of 8 plants, choosed from 47 PCR positive plants, are positive in RT-PCR. These results primary showed that both genes have been integrated into the sugarcane genome.This study was also carried out pathogen isolation of the collected 24 Sugarcane smut Samples of 24 Cane-growing areas, from eight cities of Hainan Province. According to morphological characteristics of colonies, combination of optical microscope observation and identification of molecular biology, we could basically determine that the pathogens isolated are Ustilago scitaminea Syd. All of these studies laid the foundation to the later related experiments, such as sugarcane breeding resistant to smut, resistance mechanism,and so on. |
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