Cloning And Analyzing The Disease Resistance Gene Of Powdery Mildew In Wheat (Triticum Aestivum L.)

Powdery mildew caused by Blumeria graminis f.sp.tritici is one of the most serious diseases the wheat production all over the world. Breeding resistant cultivars has been proved the most effective way to control this disease. Near iosogenic line (NIL) Mardlar/7×Bainong3217 (Pm2) was developed by Key Laboratory of Crop Germplasm and Biotechnology, Ministry of Agriculture, Institute of Crop Germplasm Resources. Pm2 is effective against 15 biotype of Blumeria graminis in China. Cloning of Pm2 gene and the related genes for Powdery mildew resistance is significance for understanding disease-resistance mechanism and disease resistance breeding. In this study, Gene expression analysis by cDNA-AFLP in wheat after powdery mildew inoculation revealed the resistance related gene. The full-length cDNA sequence of wheat was obtained via silicon cloning and RT-PCR technique analysis whether resistance disease, and get resistance related gene.283 TDFs were obtained with cDNA-AFLP technique in Near iosogenic line (NIL) Mardlar/7×Bainong3217. The material of this expriment select NIL resistance Mardler/7×Bainong 3217 (Pm2) and Bainong 3217,and then designed the treatments : Mardler/x7Bainong3217 inoculation, Mardler/7x Bainong 3217 water treat (CK ) , Bainong 3217 DCINA treated, Bainong 3217 inoculation and Bainong 3217 water treat (CK) , cDNA-AFLP differential display the collect leaves respectively after 0,1, 3,6, 12,24 hours inoculation,Using 553 primer combinations select 46 primer and then select out 283 differential display fragments. Detection the resistance of thesis wheats with the left leaves. The result of resistance detection indicate that Mardler/7×Bainong3217 is immunity, Bainong 3217 treated by DCINA is partialy resistance, Bainong 3217 is suspective.A gene with ORF was got via silicon cloning and RT-PCR technique. There are 80 amplified fragments by Second PCR, select 54 amplified fragments sequenced, there are 4 fragments related to resistance through bioinfomatic analysis. One full-length cDNA sequence of wheat lipid transfer protein (LTP) gene which has...