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Take-all is a serious disease found in wheat across the world. Haynaldia villosa is considered to be resistant to take-all at a high level. TH3 was an amphiploid (2n=42, AABBVV) between Triticum durum and Haynaldia villosa with significant resistance to take-all fungus isolated from China. In greenhouse experiment, the derivatives of the hybrid between wheat and TH3 showed better resistance to take-all than that of the wheat control. One of the derivatives named HW918-5 was selected for further analysis. Cytological and genomic in situ hybridization (GISH) analysis indicated that a monotelosome originated from H. villosa existed in the genome of the offspring of the line HW918-5. The monotelosome with promising resistant gene for take-all was located on the 3V chromosome of H. villosa in the further PCR-based molecular analysis.