Differential Display Of Badila Infected Sugarcane Mosaic Virus

Differential display (DD) is a effective method to clone unknow expressed gene. Since its advent, it was widely applied. In order to find out how Sugarcane mosaic disease happened, expressive difference of gene about Badila which was virus free by way of tissue culture and Badila infected SCMV was studied by DD.The experimental materials used to DD must have the similar genetic background. In order to detect the virus of the experimental materials, a pair of primer combination was designed according to the 3'end sequence of the sugarcane mosaic virus (SCMV). The detected result suggested that the Badila which was virus free by way of tissue culture did not have any PCR band and the Badila infected SCMV had a PCR band. Consequencely, two kinds of experimental materials had the similar genetic bankground except for the difference of whether it had the SCMV or not, and were suitable for DD anylysis.Total RNA used to DD must have the high quality. A method system of extracting total RNA was established. The quality of RNA obtained by the method system was higher and fit for differential display (DD) anylysis, but one of the two methods had its own advantages and shortcomings.In addition, a set of flouresent DD operating method was founded. 9 differentially expressed cDNA fragments were obtained by this set ofoperating method and selected 23 primer combinations. The size of 9 fragments was between 250bp and 350bp. These fragments were grouped into two types:4 differentially expressed cDNA fragments which exist or not in the two compared experimental materials ; 5 differentially expressed cDNA fragments which had the intensity difference between the two compared experimental materials. 5 fragments were obtained from Badila which was virus free, and 4 fragments were obtained from Badila infected SCMV.The 9 fragments were amplified by agarose gel again and had the same size as the fragments in the fluoresent DD Gel.