Analysis Of Early Stage Expression Mode Of Brassica Napus Response To Sclerotinia Sclerotiorum

Sclerotinia stem rot is the most devastating disease of Brassica napus in China,Up to now, a entire-immuned cultivar was not found. For breeders, it is very impormant to understand the interaction of Brassica napus and Sclerotinia sclerotinia in molecular level. We analyzed the resistance genes involved in the interaction process post-innoculated 0.5 to 28.5 hours by Suppression subtractive hybridization and reverse northern blot. The main results are as follows:1.Took leaves post-innoculated 0.5-28.5 hours as positive material,leaves un-inoculated as negative material.same amount of sample in different period were blended and exacted mRNA,constructed a forward library which contained 1920 clones and a backward library which contained 1008 clones.The lenth of the inserted fragment are about 300 bp -900bps, most of them are 500-600bps.The resistant fragment are accumulated about 1000 times.2.Total RNA was extrated from Leaves post-innoculated 0.5,4.5,8.5 hours and synthesized cDNA as probes, conducted reverse northern blot. 50 unrepeated resistance-related ESTs were indentified. According to the putative function analysis, the 50 ESTs from Brassica napus were related to the primary metabolism, energy metabolism, secondary metabolism, disease defence, cell structure, transcription, protein synthesis, and signal transduction. The transcription products involved in the oxgen burst and jasnonade pathway and related metabolism pathway. Fnuctions of 18 ESTs were still unknown. We deduced that Brassica napus confine Sclerotinia Sclerotinia mainly through jasmonade pathway in early stage. We made a primary analysis of expression mode and interaction of some genes in different period post-inoculated.3.To confirmed the realibility of the reverse northern blot and analyzed expression mode of different kinds of genes,we conducted northern blot in different period with probes synthesized from 3 clones which classified into 3 different kinds.The results showed the reverse northern blot were a reliable method and different kinds of genes had different expression mode.