Identification of a Pm4 allele conferring powdery mildew resistance in wheat breeding line GR18-1

Powdery mildew caused by Blumeria graminis f. sp. tritici (Bgt) is a serious fungal wheat disease of wheat worldwide. Host resistance is considered to be the most environmental-friendly and efficient approach against this disease. Wheat breeding line GR18-1 showed resistance to powdery mildew at both seedling and adult stages for several years. Genetic analysis indicated that a single dominant gene, tentatively designated as PmGR-18, conferred powdery mildew resistance in GR18-1. Bulked segregant analysis (BSA) and marker analysis showed that PmGR-18 was located in the Pm4 interval on chromosome arm 2AL and flanked by the markers Xwgrc763 and Xwgrc872, respectively, with genetic distances of 0.5 and 1.0 cM corresponding to a physical interval of 1.13 Mb based on the Chinese Spring reference genome sequence v2.1. Using homology-based cloning and Sanger sequencing, we found that the sequence of PmGR-18 was totally consistent with that of Pm4d. qRT-PCR analysis showed that the expression levels of two splicing variants Pm4d_V1 and Pm4d_V2 in GR18-1 were significantly upregulated after inoculating with Bgt isolate E09, and the level of Pm4d_V2 was lower significantly than that of Pm4d_V1 at most of the time points, suggesting a different resistance pattern may be involved in genotype. To facilitate the transfer of PmGR-18 in marker assisted selection (MAS) breeding, the flanked markers Xwgrc763, Xwgrc872 and the functional marker JS717/JS718 were tested and confirmed to enable the tracking of PmGR-18 when it transferred into those susceptible cultivars.