The Analysis Of Differential Display Of Responsive Genes In Brassica Napus For Sclerotinia

The study based on the significant difference between Zhong R888(Brassice napus) and Zhongyou821 (Brassice napus) on the Sclertiorum tolerance to establish BC₅ near isogenic line (zhongR888×zhongyou821) with zhongyou821 as the donor parent. Then the Sclertiorum tolerance difference between the BC₅ (zhongR888 xzhongyou821 )and zhongyou821 was analyzed by cDNA-AFLP technique. The main resμLts are as follows:We use the MINI-PROTRAN3 (?) gel electrophoresis of BIO-RED Company with Small PAGE (10×8.5cm) to optimize cDNA-AFLP with silver staining which make it units the advantages of low cost and less pollution and accelerate the application of cDNA-AFLP technique to popμLar biochemistry laboratory.we get some special TDFs (Transcripts-derived Fragments), and Information of defense relative-fragments were got by bioinformatics analysis, then two of them were especially focused on .The first TDF was 40-2 which was a leucine-rich repeat family protein. Furthermore, it was more transcripted in zhongyou821 than in BC₅ identified by semi-quantified PCR which was based on the proper control of protein extension factor-2, and northern blotting also show the same resμLt.The second TDF 18-2 was extended to about 1260bp which include a ORF by theanalysis of ORFfinder. And the similarity of the fragments between in the BC₅(zhongR888×821) and in zhongyou821 was 89.9% and the enzyme site mapdemonstrated that MseI enzyme (used in cDNA-AFLP) site was disappeared inBC₅888×821 but not in zhongyou821. Northern blotting indicate that only BC₅(zhongR888 xzhongyou821 )have positive blot signal. Recombine the fragment with expression vector and induced the GST fusion protein. And the unknown protein's function shouLd be elucidated in further research.