Genetic Research On Disease Resistance To Sclerotinia Sclerotiorum In Brassica Napus

Sclerotinia stem rot caused by S. sderotiorum is a kind of stubborn disease of rapeseed, which results in serious yield loss. The scientists in the world have made a wide spread and profound research on the disease, but the development is not satisfying. Based on the resource of our lab, explorating technique of studying on Sderotinia stem rot, investigating the resistant resource, and cloning the gene resistant to S. sderotiorum is our main aim.This research includes three aspects, verifing transgenic breeds with technique of PCR, southern blotting and antibiotic screening, testing the resistance to S. Sderotiorum of 12 breeds with detached leaves inoculation and ethane diacid toxin at the seedling stage and mapping QTLs relating to resistance to S. sderotiorum in a DH population (derived from cross combination of Tapidorxningyou 7, and including 202 lines). The research results are as follows:1) Transgenic materials testifyingPCR results of transgenic descendant lines with AP24-Glu gene indicated: all the transgenic plants were positive to NPTII primer amplification and negative to AP₂₄, 35S, 35S-AP₂₄ primer amplification; Southern blotting results indicated: all plants were negative to NPTII, AP₂₄ and 35S probe hybridization.PCR results of transgenic descendant lines with Chi-Oxo gene indicated: all the transgenic plants were positive to the NPTII primer amplification; Southern blotting results indicated: there is one plant was blotting result indicated: there only one line was positive to NPTII probe hybridization and all the lines were negative to AP₂₄ and 35S probe hybridization.Seeds of the the two transgenic descendant line were sensitive to kanamydn.2) Resistance to ethanedioic acid and S. sderotiorum analysisThe results indicated that the difference of the two resistances among breeds is significant. Tapidor, Hui 5200, Zhongyou 821 and 6-203 show strong resistant to ethanedioic acid, and Xiangyou 15, Ningyou 7, Ning RS-1 and Huashuang 3 are strong sensitive to ethanedioic acid, Tapidor and 6-203 show better on resistance to Sderotiniasclerotiorum, and Westar is prone to sensitive to Sclerotinla sclerotiorum. Variant breeds for resistance to S. sclerotiorum and ethanedioic acid were different. 3) QTLs mapping in TN populationBased on the genetic map construct, 19 QTLs were detected in all with WinQTLCart 2.0, and covered 9 linkages, accounted for 3.53%-11.24% of the total phenotypic variation respectively. Among which, the peak LOD of rsO4j is O.OlcM away from the nearest marker, the peak LOD of rsO6g is 0.009 cM away from the left and 0.914cM from the right nearest marker, the peak LOD of rsO4h is 0.01 cM away from the left and 1.04 cM from the right nearest marker, the peak LOD of rsO7c is only 0.007cM away from the left marker...