Jafar Mammadov1, Xiaochun Sun2, Yanxin Gao3, Cherie Ochsenfeld4, Erica Bakker5, Ruihua Ren6, Jonathan Flora7, Xiujuan Wang8, Siva Kumpatla9, David Meyer10, Steve Thompson11. 1. Dow AgroSciences, 9330 Zionsville Road, Indianapolis, IN, 46268, USA. jamammadov@dow.com. 2. Dow AgroSciences, 9330 Zionsville Road, Indianapolis, IN, 46268, USA. sun1@dow.com. 3. Dow AgroSciences, 9330 Zionsville Road, Indianapolis, IN, 46268, USA. ygao3@dow.com. 4. Dow AgroSciences, 9330 Zionsville Road, Indianapolis, IN, 46268, USA. caochsenfeld@dow.com. 5. Dow AgroSciences, 9330 Zionsville Road, Indianapolis, IN, 46268, USA. bakker2@dow.com. 6. Dow AgroSciences, 9330 Zionsville Road, Indianapolis, IN, 46268, USA. rrren@dow.com. 7. Dow AgroSciences, 9330 Zionsville Road, Indianapolis, IN, 46268, USA. jpflora@dow.com. 8. Dow AgroSciences, 9330 Zionsville Road, Indianapolis, IN, 46268, USA. xwang10@dow.com. 9. Dow AgroSciences, 9330 Zionsville Road, Indianapolis, IN, 46268, USA. spkumpatla@dow.com. 10. Dow AgroSciences, 9330 Zionsville Road, Indianapolis, IN, 46268, USA. dhmeyer@dow.com. 11. Dow AgroSciences, 9330 Zionsville Road, Indianapolis, IN, 46268, USA. sathompson@dow.com.
Abstract
BACKGROUND: Gray Leaf Spot (GLS causal agents Cercospora zeae-maydis and Cercospora zeina) is one of the most important foliar diseases of maize in all areas where the crop is being cultivated. Although in the USA the situation with GLS severity is not as critical as in sub-Saharan Africa or Brazil, the evidence of climate change, increasing corn monoculture as well as the narrow genetic base of North American resistant germplasm can turn the disease into a serious threat to US corn production. The development of GLS resistant cultivars is one way to control the disease. In this study we combined the high QTL detection power of genetic linkage mapping with the high resolution power of genome-wide association study (GWAS) to precisely dissect QTL controlling GLS resistance and identify closely linked molecular markers for robust marker-assisted selection and trait introgression. RESULTS: Using genetic linkage analysis with a small bi-parental mapping population, we identified four GLS resistance QTL on chromosomes 1, 6, 7, and 8, which were validated by GWAS. GWAS enabled us to dramatically increase the resolution within the confidence intervals of the above-mentioned QTL. Particularly, GWAS revealed that QTLGLSchr8, detected by genetic linkage mapping as a locus with major effect, was likely represented by two QTL with smaller effects. Conducted in parallel, GWAS of days-to-silking demonstrated the co-localization of flowering time QTL with GLS resistance QTL on chromosome 7 indicating that either QTLGLSchr7 is a flowering time QTL or it is a GLS resistance QTL that co-segregates with the latter. As a result, this genetic linkage - GWAS hybrid mapping system enabled us to identify one novel GLS resistance QTL (QTLGLSchr8a) and confirm with more refined positions four more previously mapped QTL (QTLGLSchr1, QTLGLSchr6, QTLGLSchr7, and QTLGLSchr8b). Through the novel Single Donor vs. Elite Panel method we were able to identify within QTL confidence intervals SNP markers that would be suitable for marker-assisted selection of gray leaf spot resistant genotypes containing the above-mentioned GLS resistance QTL. CONCLUSION: The application of a genetic linkage - GWAS hybrid mapping system enabled us to dramatically increase the resolution within the confidence interval of GLS resistance QTL by-passing labor- and time-intensive fine mapping. This method appears to have a great potential to accelerate the pace of QTL mapping projects. It is universal and can be used in the QTL mapping projects in any crops.
BACKGROUND: Gray Leaf Spot (GLS causal agents Cercospora zeae-maydis and Cercospora zeina) is one of the most important foliar diseases of maize in all areas where the crop is being cultivated. Although in the USA the situation with GLS severity is not as critical as in sub-Saharan Africa or Brazil, the evidence of climate change, increasing corn monoculture as well as the narrow genetic base of North American resistant germplasm can turn the disease into a serious threat to US corn production. The development of GLS resistant cultivars is one way to control the disease. In this study we combined the high QTL detection power of genetic linkage mapping with the high resolution power of genome-wide association study (GWAS) to precisely dissect QTL controlling GLS resistance and identify closely linked molecular markers for robust marker-assisted selection and trait introgression. RESULTS: Using genetic linkage analysis with a small bi-parental mapping population, we identified four GLS resistance QTL on chromosomes 1, 6, 7, and 8, which were validated by GWAS. GWAS enabled us to dramatically increase the resolution within the confidence intervals of the above-mentioned QTL. Particularly, GWAS revealed that QTLGLSchr8, detected by genetic linkage mapping as a locus with major effect, was likely represented by two QTL with smaller effects. Conducted in parallel, GWAS of days-to-silking demonstrated the co-localization of flowering time QTL with GLS resistance QTL on chromosome 7 indicating that either QTLGLSchr7 is a flowering time QTL or it is a GLS resistance QTL that co-segregates with the latter. As a result, this genetic linkage - GWAS hybrid mapping system enabled us to identify one novel GLS resistance QTL (QTLGLSchr8a) and confirm with more refined positions four more previously mapped QTL (QTLGLSchr1, QTLGLSchr6, QTLGLSchr7, and QTLGLSchr8b). Through the novel Single Donor vs. Elite Panel method we were able to identify within QTL confidence intervals SNP markers that would be suitable for marker-assisted selection of gray leaf spot resistant genotypes containing the above-mentioned GLS resistance QTL. CONCLUSION: The application of a genetic linkage - GWAS hybrid mapping system enabled us to dramatically increase the resolution within the confidence interval of GLS resistance QTL by-passing labor- and time-intensive fine mapping. This method appears to have a great potential to accelerate the pace of QTL mapping projects. It is universal and can be used in the QTL mapping projects in any crops.
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